Sixty-one percent of the confiscated products contained a SC and 31% contained both XLR-11 and CID

Posted on July 13, 2023 by hemp2022

Future studies should examine the impact that within-individual changes in psychiatric problems have on substance use in the current context. Our study shows that when adolescent boys experience an increase in conduct disorder problems, they subsequently experience an increase in the quantity and frequency of substance use, while an increase in alcohol use can also subsequently result in increased anxiety problems in adolescence. Reducing fluctuations in conduct disorder problems and substance use at sensitive developmental turning points such as early and late adolescence may have lasting effects in preventing psychiatric and substance use problems by young adulthood. Synthetic cannabinoids are a class of drugs that are becoming increasingly popular throughout the United States and Europe. Also known as “K2,” “spice,” spike,” or “legal marijuana,” SC are causing intoxication requiring emergency department visits in epidemic and unparalleled numbers.1 Patients present with a wide array of symptoms, ranging from nausea and vomiting to confusion, agitation, short-term memory loss, cognitive impairment, psychosis, seizures, arrhythmias, strokes and even death.SC have often been associated with sympathomimetic effects such as mydriasis, hypertension and tachycardia.We present a case series of patients with SC intoxication who presented atypically with central nervous system and cardiovascular depression over a five-month period; in addition, we present an analysis of blood, urine and SC samples using mass spectrometry. Intoxication with SC products should be considered for patients with undifferentiated psychomotor depression and bradycardia in addition to the excitatory effects previously described.Samples were extracted with organic solvent and concentrated to isolate any drugs present on the plant material. Briefly, 5 mg aliquots of an unknown plant material, or 100 μL of submitted blood/urine,vertical grow rack systems were transferred to screwtop centrifuge tubes. Two mL of ethyl acetate were added and the samples were thoroughly mixed. Samples were extracted for 10 minutes on a nutating mixer at 24 revolutions per minute. The solvent was transferred to clean test tubes and the extracts were evaporated to dryness under nitrogen at 45°C.

Samples were reconstituted in 50 μL methanol and 50 μL 0.1% formic acid in water and transferred to conical autosampler vials for analysis by liquid chromatography time-of-flight mass spectrometry. Similarly, samples were reconstituted in 50 μL ethyl acetate for GC/MS confirmation analysis. Biological samples underwent a 20-minute room temperature hydrolysis period prior to liquid-liquid extraction. Hundreds of distinct SC compounds have been identified.SCs are responsible for a rapidly growing number of presentations to EDs throughout the U.S. in the past several years.SC use causes intense highs and has become popularized due to accessibility, affordability and limited detectability in common drug screens.Intoxications often present in clusters due to local distribution of a single product and great variability in the herbal mixtures.In 2011, SCs were the second most commonly used drug in the 10th grade and the third most common in eighth grade following marijuana and inhalants.Despite the federal ban on SCs that year, there was no decline in frequency of use in high school students the following year. However, use declined in each of the next three years.Users of SCs vary greatly in both demographics and motivation, but are typically males aged 13-59, most with polydrug use and are found in larger, urban populations.SCs are known to interact with the cannabinoid receptors, CB1 and CB2 , leading to changes in levels of multiple neurotransmitters including acetylcholine, dopamine, noradrenaline, glutamine and GABA.Genetic polymorphisms in enzymes responsible for metabolism of SCs can lead to increased blood levels of the parent compound and prolonged duration of action, and therefore a potential increased risk of adverse events.In addition, many SC metabolites retain biological activity.Combination of these metabolites with accumulation of the parent drug creates complex pharmacodynamics, especially when the multitude of other compounds typically found within herbal mixtures is considered. SCs have been reported to exhibit a wide array of effects. CNS effects include psychosis, anxiety, agitation, irritability, memory changes, sedation, confusion and hallucinations,in addition to lowering the seizure threshold in susceptible individuals.Reported cardiovascular effects include tachycardia, chest pain, dysrhythmias, myocardial ischemia and cerebrovascular accident caused by embolisms due to cardiac arrhythmias or reversible cerebral vasoconstriction syndrome.By the end of 2012, JWH-018 was not detected in samples, and XLR-11 became the most common SC detected,as exhibited in our sample analysis.

In our case series, CID and alkyl SC derivatives, such as INACA compounds and XLR-11,were the most commonly detected with no opiates, imidazoline receptor agonists, benzodiazepines or other sedative-hypnotics detected that might explain the atypical presentations.Seventy-five percent of blood samples and 77% of urine samples tested positive for SC. Unlike their predecessors, novel SC appear to be associated with significant CNS depression and bradycardia. The compounds detected in our case series tended to be full agonists at the cannabinoid receptor and are more potent than Δ9-THC.The lack of other CNS and cardiovascular depressants suggests that the clinical findings are due to the combination of these compounds and not coingestants or adulterants. It is important to note that many substances detected in the plant samples were not detected in the blood or urine samples. Some examples include 5-Fluoro-NNEI 2’-naphthyl isomer, 5-fluoropentylindole, NM-2201 and NPB-22. There are multiple explanations for these findings. The patient may have used SC products that were not included in our plant samples and therefore would not be associated with the urine and blood samples. It is also possible that the metabolites of the compound were not in the database or that the level was below the LC TOF detection limits. Furthermore, the metabolite may have been metabolized to a common XLR metabolite that was detected, or the drug had already been eliminated from the body. More than one-quarter of a million women in the United States are currently living with HIV , and many women living with HIV fare poorly on the HIV Care Continuum . In 2015, only 50% of WLHIV were retained in care and 48% achieved HIV viral suppression . Despite the broad availability of effective antiretroviral medications, WLHIV also experience high rates of morbidity and mortality compared to the general population . Trauma is increasingly recognized as a near-universal experience among WLHIV and as a key contributor to HIV acquisition, morbidity, and mortality. Defined as “an event, series of events, or set of circumstances that is experienced by an individual as physically or emotionally harmful or threatening and that has lasting adverse effects” , trauma can include childhood and/or adult physical, sexual, or emotional abuse or neglect,vertical grow racks cost as well as sociostructural violence such as racism, sexism, homophobia, transphobia, xenophobia, or living in a community where violence is common. People living with HIV experience disproportionately high rates of trauma , including rates of childhood sexual abuse that are more than twice the rates among the general population .

Trauma exposure in PLHIV is associated with nonAIDS related deaths , and is predictive of experiencing later violence . It is also closely associated with mental health disorders including depression, PTSD, and anxiety , as well as with increased HIV-risk behavior, including substance use disorders . HIV diagnosis is itself often highly traumatic . Among PLHIV, trauma and substance use often function syndemically, as “epidemics interacting synergistically and contributing, as a result of their interaction, to excess burden of disease in a population” . The syndemic of violence/trauma, substance use, and HIV has been identified as one of the main drivers of HIV infection and of poor health outcomes among women living with HIV . Research has consistently shown high rates of substance use among people living with HIV, and rates that are higher than among the general population . Substance use has also been shown to have a negative impact on HIV treatment adherence and virologic suppression .The link between trauma and health outcomes has led to calls for increased attention to trauma in health care by advocates and government leaders, including the U.S. Preventive Services Health Task Force, the Institute of Medicine, and the Agency for Healthcare Research and Quality . While an emerging literature describes interventions to address trauma and PTSD among PLHIV , no prospective study has evaluated the impact of a comprehensive model of trauma-informed health care delivery on health outcomes. To address this gap, we initiated implementation of a model of traumainformed health care in one clinic serving WLHIV in the San Francisco Bay Area. As part of this effort, we are conducting a broad evaluation of the impact of TIHC on patient health outcomes. Here we report results of baseline data analyses, examining the association of trauma with physical, behavioral, and social health indicators, with particular attention to quality of life and undetectable viral load. We then consider how the results of the investigation serve to inform efforts within health care settings to improve outcomes. Women were recruited from the waiting room during regular clinic hours on two half-days each week. Researchers approached patients in the waiting room, briefly explained the purpose of the study and, if a patient was interested, met with her in a private room. At that time, the researchers reviewed consent documents, explained the study procedures including data abstraction from the electronic health record , and answered any questions. Individuals were eligible to participate if they self-identified as cisgender or transgender women who were 18 years of age or older, were currently receiving primary HIV care at the clinic, and were English-speaking and cognitively able to complete the interview. If the patient was eligible and willing, she signed a general consent form and an EHR data abstraction consent form. Following consent, the researcher conducted the interview by reading each question aloud and marking responses in a survey booklet. At the end of the interview, the participant received a $25 gift card inappreciation of her time. Most interviews took 30-45 minutes to complete. After the interview, researchers abstracted relevant data from the participant’s EHR. Anxiety symptoms were assessed using the Generalized Anxiety Disorder scale , a 7-item self-report scale to measure symptom severity. A score of 10 or above indicates at least moderate anxiety. Substance use was measured in three ways. Alcohol use was assessed using the short Alcohol Use Disorders Identification Test , a 3-item screen to identify individuals who may be hazardous drinkers or who have alcohol use disorders . The instrument provides a raw numerical score ; an indicator of binge drinking; and a diagnostic of AUD . Drug use was measured using one question from the Alcohol, Smoking and Substance Involvement Screening Test asking about substance use in the past three months. We dichotomized this into any non-prescribed drug use in the past 3 months, and a similar variable of “hard” drug use that excludes marijuana. Drug abuse was measured using the Drug Abuse Screening Test 10 , which yields a score range of 0-10. A score of 3 or greater indicates at least a moderate level of drug abuse. Quality of Life was measured using the five-item WHO-Five, developed crossculturally by the World Health Organization . The instrument measures self-reported quality of life over the past two weeks in the areas of mood, physical vitality, and interest in life. A score below 13 indicates poor quality of life . Mental well-being was measured using the seven-item Short WarwickEdinburgh Mental Well being Scale, which focuses on emotions and mental functioning , and yields a score of 7-35. Undetectable viral load. For participants who consented to having data abstracted from their electronic health record , we abstracted HIV viral load and CD4 counts. Data were abstracted only if the person had had a test within the past year, and the most recent test result was used. Viral load was dichotomized as detectable or undetectable . For the analysis, we focused on undetectable viral load as the outcome of interest. Patient-Provider relationship was measured using the Engagement with Health Care Provider scale , a 13-item instrument in which clients rate their interactions with their providers on a scale of 1 “always” to 4 “never”. Responses are summed to get a total score of 1-52, with lower scores indicated greater engagement. Appointment Adherence. We abstracted EHR data to examine appointment adherence as a proxy for engagement in care.

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CDT did not significantly predict performance on any neurocognitive measure

Posted on July 12, 2023 by hemp2022

The direct biomarkers include ethyl glucuronide , ethyl sulfate , and phosphatidylethanol. Past research has shown that several indirect alcohol use biomarkers are correlated with cognitive performance in individuals with alcohol use disorders: AST,ALT,MCV,and GGT.Thus, alcohol use biomarkers may not only be used to screen for alcohol problems or abstinence,but also have a specific role in screening for cognitive impairment in individuals with alcohol use disorders. These biomarkers may offer more than simply getting a history of the amount and frequency of recent alcohol use. Though several indirect biomarkers have been explored with respect to cognitive performance, the newer direct biomarkers, such as EtG and EtS, have not received any attention in the literature. Although the direct biomarkers are minor metabolites of alcohol,these biomarkers, such as EtG, can also be found in the brain.Whether any of the direct biomarkers are associated with cognitive performance in individuals with alcohol use disorders remains an open question. In an effort to add to this scarce literature on the association of alcohol use biomarkers and cognitive performance, we conducted a secondary analysis of baseline data from a recently completed pharmacological pilot clinical trial among veterans with alcohol dependence and post traumatic stress disorder . This study included the measures, at baseline, of indirect and direct alcohol use biomarkers and neurocognitive measures, which allowed us to explore the relationship between biomarkers and cognitive performance. Because this study was conducted in alcohol-dependent veterans with comorbid PTSD, we were also able to explore the unique relationship between alcohol use biomarkers and cognitive performance in a group having particularly poor clinical outcomes.To the best of our knowledge,vertical led grow lights the relationship between alcohol use biomarkers and cognitive performance specifically in veterans with alcohol dependence and PTSD has not been previously explored.

In this sample of veterans with alcohol dependence and PTSD, we hypothesized that the indirect biomarkers would predict baseline cognitive performance. On the basis of the evidence that they can be found in the brain, we also hypothesized that the direct biomarkers would predict baseline cognitive performance. Demographic data, such as age, sex, race, ethnicity, years of education, marital status, and occupational status, were collected. Psychiatric diagnoses and concurrent medication use were captured by a review of each participant’s electronic medical record at the SFVAMC. Substance use disorder diagnoses were assessed using the Substance Use Disorders module of the Structured Clinical Interview for DSM-IV-TR, Research Version, Patient Edition .The level of substance use for the past 90 days was assessed using the Timeline Follow back Method.PTSD was diagnosed by the Clinician-Administered PTSD Scale.The level of depression was assessed using the 21-item self-report Beck Depression Inventory.Blood samples were obtained for CDT , GGT, MCV, AST, and ALT levels. Urine samples were obtained for EtG and EtS levels. Standard operating procedures were followed by the Clinical and Translational Science Institute at the SFVAMC to obtain these samples. Levels of GGT, MCV, AST, and ALT were analyzed locally at the SFVAMC Department of Laboratory Medicine. CDT sample was shipped and analyzed at the Clinical Neurobiology Laboratory in the Institute of Psychiatry at the Medical University of South Carolina. EtG and EtS samples were shipped and analyzed at the Department of Laboratory Medicine at the Yale University School of Medicine. The Trail Making Test part A was used to assess psychomotor speed and simple visual attention and part B was used to assess task switching and cognitive flexibility; the raw scores were converted to T scores.The Hopkins Verbal Learning Test—Revised was used to assess verbal memory.We used the %retention score for this analysis, where the raw score was converted to a T score; the assessment of retention is relatively free of effortful memory search and retrieval.The Balloon Analogue Risk Task was used to assess risk taking; we used the primary score of “adjusted average number of pumps on unexploded balloons.” The Delay Discounting Task was used to assess impulsivity; we used the Kln score, defined as the log-transformed DD after applying the hyperbolic function. All analyses were conducted using IBM SPSS Statistics, version 20 .

All continuous variables were checked for normality , and nonparametric tests were used when appropriate. All continuous variables were also checked for extreme values; values with a z-score > 3.29 or < −3.29 were adjusted to the next highest value. Where adjusted results differed from the original data, the adjusted results are presented. Because most values were undetectable at <100 ng/mL, EtG was dichotomized into <100 ng/mL vs. >100 ng/mL. Because most values were undetectable at <50 ng/mL, EtS was dichotomized into <50 vs. >50 ng/mL. Two multiple regression models were estimated and tested for each neurocognitive measure . The first model included the alcohol use biomarker alone as the predictor. The second model included the alcohol use biomarker along with the following 3 additional predictors: Beck Depression Inventory , Clinician-Administered PTSD Scale , and receiving medications . As mood symptoms,PTSD symptoms,and medications can affect cognitive performance, we included these 3 additional predictors in the second model to determine if they would make a significant contribution. Because this was an exploratory secondary analysis, we did not control for type I error; pvalues < 0.05 were considered statistically significant. Assumptions in each regression model were checked by assessing several parameters such as Durbin–Watson statistic , collinearity , standardized residuals , Cook’s distance , linearity/homoscedasticity , and normality of residuals . All of these assumptions in each multiple regression model for each neurocognitive measure were met. Finally, previous evidence shows that alcohol intake itself can affect cognitive performance.We explored whether the number of drinks significantly correlated with any of the neurocognitive measures. Table I presents baseline demographic and clinical data. Table II presents baseline substance use, alcohol use biomarker, and neurocognitive data. Tables III and IV present the multiple regression analyses between alcohol use biomarker data and neurocognitive data. Table III presents the results with the first model that included the alcohol use biomarker alone as the predictor; Table IV presents the results with the second model that included the alcohol use biomarker along with the 3 additional predictors .In both models, GGT significantly predicted performance on the HVLT-R %Retention; the Beck Depression Inventory and the Clinician-Administered PTSD Scale also significantly contributed to the second model along with GGT. In only the first model, GGT significantly predicted performance on the TMT-A.

GGT did not significantly predict performance on the BART, DD, TMT-B, and in the second model on the TMT-A. In only the first model, MCV predicting performance on the BART approached significance. It did not significantly predict performance on any other neurocognitive measure.In both models, AST significantly predicted performance on the HVLT-R %Retention. In only the first model, AST predicting performance on the TMT-A approached significance. AST did not significantly predict performance on the BART, DD, TMT-B, and in the second model on the TMT-A. In the first model, ALT predicting performance on the TMT-A approached significance. It did not significantly predict performance on any other neurocognitive measure. EtG and EtS did not significantly predict performance on any neurocognitive measure. The number of drinks did not significantly correlate with any of the neurocognitive measures . These results were nonsignificant for the number of drinks in the past 4 to 90 days. Also, because GGT and AST were the only two measures to predict performance on the HVLT-R %Retention, we assessed whether these were correlated; GGT and AST were correlated in this analysis . Baseline alcohol use biomarker and neurocognitive data from a pilot clinical trial among veterans with alcohol dependence and PTSD were analyzed in this secondary analysis. GGT and AST significantly predicted performance on the HVLT-R %Retention; the Beck Depression Inventory and the Clinician-Administered PTSD Scale also significantly contributed to predicting performance on the HVLT-R %Retention along with GGT. GGT alone,vertical cannabis grow without any other predictors, significantly predicted performance on TMT-A. Without any other predictors, AST and ALT alone predicting performance on the TMT-A approached significance. Without any other predictors, MCV alone predicting performance on the BART approached significance. Thus, the initial hypotheses were partially supported. The indirect biomarkers may predict neurocognitive performance for several reasons such as by serving as a surrogate marker for heavy alcohol use, thereby representing alcohol’s potential for direct neurotoxicity; by serving as a marker of hepatic dysfunction for transaminases, thereby representing hepatic effects on brain function; and by having a direct neurotoxic effect of their own. The finding that GGT and AST predicted performance on some neurocognitive measures is consistent with that of previous research.For example, increases in GGT may increase the transport of amino acids into the brain across the blood–brain barrier, which may alter cognitive performance.GGT has also been associated with gray matter decline and brain shrinkage,which may affect cognitive performance. GGT is known to be a marker of oxidative stress and has been found to be elevated in patients with Alzheimer’s disease,which highlights a potential association of GGT with cognitive performance. Cognitive changes because of poor liver function may be due to the liver failing to catabolize circulating neurotoxins,and GGT and AST may help identify patients who show a change in visual attention and verbal memory performance. ALT significantly predicted performance on the TMT-A, but the limitations of the sample might have contributed to ALT not fully achieving significance. Approaching significance, the MCV predicting BART performance is interesting. Though MCV may appear to be unrelated to cognition, some studies have shown that erythrocyte volume may influence cognition,and that MCV can predict delirium after surgery.MCV has also been associated with gray matter decline and ventricular enlargement.One possibility is that the increased erythrocyte volume, which is found in alcohol dependence and during times of stress,may lead to erythrocytes having difficulty passing through narrow brain capillaries and subsequently affecting cognitive performance.CDT not predicting performance on any neurocognitive measure is consistent with previous reports.It is important to note that other studies in individuals with alcohol use disorders have similarly shown no association of indirect biomarkers with any neurocognitive measure.One plausible explanation for this is that because the direct biomarkers are minor metabolites of alcohol,30 the concentrations of these biomarkers in the brain may not have been sufficient to affect the neural pathways underlying cognitive performance.

Another plausible explanation may be that the direct biomarkers represent alcohol use for a much briefer time than the indirect markers, which represents anywhere from several weeks to several months; therefore, the indirect biomarkers represent more chronic measures of heavy drinking and more likely represent the direct toxic effects of alcohol on brain function. This analysis suggests that in addiction settings, some of the indirect alcohol use biomarkers serve as an indicator of a subset of patients who are at high risk for cognitive impairment. Alcohol use biomarkers cannot replace a comprehensive neurocognitive evaluation for assessing cognitive impairment. Rather, in settings where a comprehensive neurocognitive evaluation is not feasible, alcohol use biomarkers might be the next best tool that clinicians could potentially use to identify veterans with alcohol dependence and PTSD who are likely to show cognitive impairment. Cost and practicality of ordering alcohol use biomarkers would be some hurdles for a clinician to implement these biomarkers in routine clinical practice. For example, in our own San Francisco Veterans Affairs clinical setting, the two indirect biomarkers in this analysis that predicted cognitive performance can more easily be ordered through our computerized medical record system, compared to the direct biomarkers that require special ordering and processing. Thus, in addition to the scientific relationship between alcohol use biomarkers and cognitive performance, clinicians must consider cost and practicality of ordering alcohol use biomarkers when implementing these biomarkers in routine clinical practice. This analysis has several strengths. First, seven alcohol use biomarkers were analyzed. Second, three additional predictors were integrated into the second regression model and yet still found significance with a few biomarkers. Third, a naturalistic sample of veterans was analyzed, which can help generalize these findings to veterans with alcohol dependence and comorbid PTSD. Finally, this is the first known analysis to explore the relationship between alcohol use biomarkers and cognitive performance in veterans with both alcohol dependence and PTSD. Inevitably, this analysis also has limitations. First, the study was not specifically designed to assess the aims of this post hoc analysis. As a result, the number of exploratory analyses conducted likely produced some type I errors. Second, because the sample size was small, this may have been the reason for only obtaining approaching significance level findings for some biomarkers.

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Participants were given a thirty-dollar drugstore gift-card upon completion of the interview

Posted on July 11, 2023 by hemp2022

Although baseline sleep patterns did not predict conversion to psychosis, our findings demonstrate that disturbed sleep is strongly related to increased severity of CHR symptoms over time. This association held in half the sleep characteristics when explored independently. Adjusting for depression attenuated the association between sleep and symptoms considerably. Furthermore, while effect sizes were similar bidirectionally, baseline sleep was a significant predictor of CHR symptoms two months later but not vice versa. These findings advance the current literature in several important ways. First, our findings correspond with previous CHR studies in which sleep difficulties at baseline did not predict conversion.This may suggest that sleep operates as an indicator of conversion only in conjunction with several other important markers. This hypothesis is supported by a predictive risk model of conversion in which “sleep disturbance” was one of the six factors that, jointly, yielded a markedly high positive predictive value. Sleep disturbances may have predictive value when they occur close to time of conversion, since sleep alterations are early indicators of psychosis relapse.Poor sleep may also predict conversion among specific groups, such as those with prolonged sleep disturbance or those who only recently developed sleep problems. The role of disturbed sleep as a potential catalyst for conversion in our study remains speculative because 30% of converters completed only one sleep assessment and 27% transitioned to psychosis 10 months post-baseline, at which point sleep was no longer tracked longitudinally. Future studies would benefit from looking at sleep metrics tracked longitudinally in large samples followed through the time of conversion. Second, our findings reveal long-term and robust correlations between a wide range of sleep disturbances and symptoms across all four CHR domains. Although poor sleep did not predict conversion,plant racks its relation with symptom exacerbation is clinically important considering that even non-converting CHR youth often have poor functional outcome and persistent symptoms.

Our current findings using self-reports are supported by electrophysiological studies relating REM latency and REM density to positive symptoms in psychotic disorders, reduced delta EEG activity and reduced REM latency to negative symptoms in schizophrenia, and reduced sleep spindle activity to both positive and negative symptoms,although these findings vary.In further concordance with our findings on disorganized and general symptoms, individuals with non-affective psychoses and comorbid sleep disorders had greater cognitive disorganization, depression, anxiety and tension/stress levels than those without the latter morbidity.Likewise, EEG delta activity in early-course non-affective psychoses were inversely related to disorganization syndrome.Although associations between sleep parameters and clinical symptoms vary largely across studies, individuals, and stages of illness, sleep characteristics of psychosis-afflicted individuals unequivocally differ from those who are unaffected, and these differences are present prior to illness onset. Third, symptoms of depression showed a strong attenuating effect, such that relationship strengths between sleep and all CHR symptom domains were reduced by roughly half or more. Disturbed sleep is a symptom of and risk factor for depression,and unipolar depressive disorders are common comorbidities among CHR youth. In 744 CHR individuals, >40% met DSM-IV criteria for current depression and almost 20% for past depression.Similarly, depression mediated the association between sleep and suspiciousness in CHR youth, and consistently partially mediated the association between sleep and psychotic experiences.The attenuating role of depression in the current study served to strengthen existing cross-sectional evidence and demonstrate its validity over time. Lastly, we aimed to address the directionality of the relationship between sleep and CHR symptoms. Between baseline and 2-month follow-up, total sleep score was a significant predictor of total, positive, negative, and general CHR symptoms. While general symptoms were the only domain with a statistically significant bidirectional association with sleep, bidirectional effect sizes, as indicated by regression coefficients , were all nearly equal.

These findings suggest that sleep is a driving force in symptom exacerbation, and that promoting healthy sleep may be a useful target for the maintenance of CHR symptoms. Evidence has been accumulating in support of cognitive behavioral therapy for insomnia, which not only reduced insomnia but also improved symptoms of paranoia and hallucinations in a large sample of university students and in a small CHR group.Adjacent research has found that sleep quality is malleable and thus can be improved even in clinical populations.These findings offer promising evidence for advancement in clinical staging models and future sleep-related therapies for both CHR and overt psychosis, such as an upcoming trial by Waite et al.Another budding direction of research involves computational advances in causal discovery analysis, offering innovative approaches to addressing causality in the context of observational data.Such methods require careful consideration of assumptions and properties underlying the data at hand,but could be utilized in future analyses of these and other prospective longitudinal datasets involving CHR and other clinical populations. Limitations of this study include the use of a self-report to assess sleep. The PSQI and the RU-SATED have high validity and reliability and subjective sleep problems have been the primary focus of sleep treatments. Such assessments, however, are fundamentally different from electrophysiological sleep characteristics that may be differentially associated with conversion and symptom levels. Therefore, the use of both self-report and electrophysiological sleep measures over time may inform clinical risk models and constitute targets for intervention. Other notable limitations include the significant dropout rates, the absence of ongoing sleep assessments succeeding the 8-month follow-up, and the relatively small sample size of our converting group . Furthermore, we acknowledge that pre-registering our study hypotheses would have strengthened our findings.The 1.6 to 3.5 million homeless youth in the United States may be staying in shelters or temporary housing , or living on the street – those between the ages of 18-24 years – who are experiencing homelessness are challenging to quantify due to the transiency of homelessness and the lack of consistent definitions of homelessness within the scientific literature . In San Francisco, approximately one in five homeless individuals is a TAY . Young people who experience homelessness have tumultuous lives and must continuously prioritize basic needs within a context of limited resources .

Thirty-four percent of homeless youth in San Francisco reported trading drugs to help meet basic needs such as shelter . Instability and risky behaviors inevitably place youth in situations that may cause physical or emotional harm, including exposure to social and structural violence such as racial discrimination, sexism, homophobia or living in a community where violence occurs frequently . Housing instability and the accompanying exposure to external stressors and violence have an impact on mental health and patterns of substance use. Several studies indicate the presence of psychiatric disorder in more than 48.4% of homeless youth . Homeless youth also experience higher rates of poor mental health symptoms and higher consumption of alcohol and drugs compared to their housed peers . These youth may use substances as a coping mechanism for mental health symptoms and for the daily challenges experienced while homeless ; Stress and trauma are common risk factors for substance use . TAY are also at a greater risk of experiencing violence, physical injuries, and psychological consequences over peers who are housed . Past histories of abuse, transphobia, dangerous living situations, limited financial and emotional resources, engagement in substance use and high-risk sexual activity,multi-tiered growing and irregular patterns of sleep and eating all contribute to poor mental health and substance use in youth . Evidence of the impact of trauma and substance use on health outcomes, specifically with TAY who are experiencing homelessness, is growing. This paper investigates relationships between past traumatic experiences and current substance use and mental health symptoms. We conducted a cross-sectional study of 100 homeless TAY in San Francisco, California, in close collaboration with a local community-based organization that serves this population. Each year, the CBO delivers housing, employment, and education services for 2,500- 3,000 youth aged 12-24 years who are experiencing homelessness or at risk of becoming homeless. Clients include individuals who are actively living on the street, in temporary housing or shelters, living in single-room occupancy housing, living with friends, in foster care, or otherwise unstably housed. The organization offers a broad range of programs, which includes referral centers, education and employment training programs, temporary emergency shelter, residential programs, medical care, behavioral health, and case management services. Recruitment occurred at multiple service sites of the CBO, including drop in-centers, medical clinics, agency community meetings, CBO events, transitional housing sites, and at the CBO’s housing site that exclusively serves clients with an HIV diagnosis. Study flyers were posted at CBO sites, announcements were made at site meetings, and CBO staff members referred clients to approach research staff for inclusion in the study. TAY clients who were interested in participating contacted study personnel onsite or via a secure Google Voice telephone number to schedule an appointment with one of the trained research assistants. At the meeting, potential participants were screened for eligibility. Individuals were eligible if they were between 18 and 24 years old and were recipients of services provided by the CBO. Eligible individuals then proceeded through an informed consent process. Following the informed consent process, the research assistant conducted a one-on-one survey interview with the participant, reading each question aloud and marking responses in a Computer-Assisted Survey Information Collection system via an iPad tablet. The CBO specifically recommended that the research assistants, rather than the clients, navigate the CASIC system due to client differences in reading and concentration levels that could affect their ability to complete the questionnaire by themselves. Each interview was conducted in a private room and lasted 45-120 minutes.

Exposure to traumatic events prior to the age of 18 was measured with the Adverse Childhood Experiences instrument , which has been used extensively including among young adults . The number of reported ACEs was dichotomized into less than 4, or 4 or greater. Scores of 4 or greater indicate a greater level of household dysfunction, abuse, and neglect and have been shown to be associated with morbidity and poor school performance in TAY . PTSD symptoms were measured with the PTSD Checklist for DSM-5 , which has demonstrated internal consistency, reliability, and validity . The PCL-5 can be used for screening and for monitoring change in symptom intensity. A total score was calculated , with a score of 33 or above indicating potential PTSD and the need for a more thorough psychiatric evaluation . The CES-D was used to determine symptoms of depression. The CESD has shown good reliability and validity for assessment of depression in various populations including adolescents . A total score was calculated , with a score of 16 or above indicating risk for clinical depression . Anxiety symptoms were assessed using the GAD-7 which has been shown to have strong validity and reliability including among adolescents . A total score of 10 or above on a scale of 0-21 indicates at least moderate anxiety . To evaluate substance use, participants first answered the NIDA Quick Screen V1.0 , which identifies participants who have reported alcohol binge drinking in the past year , and report past year use of tobacco products, illicit drugs, or prescription drugs for non-medical reasons. Responses were dichotomized . Participants who reported use of illegal drugs or prescription drugs for non-medical reasons in the past year then completed the NIDA-Modified ASSIST V2.0 . The original ASSIST was developed by the World Health Organization and has demonstrated strong reliability and validity . In the NIDA-Modified ASSIST, participants are asked about past three-month’s use of cannabis, cocaine, prescription stimulants, methamphetamine, inhalants, sedatives, hallucinogens, street opioids, and prescription opioids. If a substance used was not listed, participants could specify it in the “other substances” option . For each substance reported, a Single Substance Involvement Score was calculated. A mean SSIS between 0-3 indicates low-risk of harmful consequences for the user; a score of 4-27 indicates moderate-risk; a score greater than 27 indicates high-risk. Moderate-risk level scores indicate that the participant may be misusing substances but may not currently meet diagnostic criteria for a substance use disorder. Study participants ranged from 18-24 years of age; the majority were male , and 52% identified as lesbian, gay, bisexual, transgender, or queer . More than two-thirds were persons of color. At the time of the survey, 23% of participants were living with HIV, 50% were experiencing literal homelessness, and almost one-third had been previously incarcerated for more than three days. Over three-quarters of participants reported 4 or more ACEs, 80% reached the diagnostic threshold for PTSD, 74% for depression, and 51% for at least moderate anxiety.

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Clustering analysis also revealed two subgroups within schizophrenia

Posted on July 10, 2023 by hemp2022

Ascorbic acid, a well-known antioxidant, reduces Cu cations to Cu0 only when the cations are bound to organic substrates such as DNA in the presence of oxygen in the dark or via autocatalysis on Cu metal seeds in the absence of stabilizing organic ligands . As an example of synthetic control, pHdependent conformation of histidine-rich peptides has led to larger nanocrystals of Cu0 at pH 7–10 than at pH 4–6 . Plants produce ascorbic acid for many functions and rhizospheres often contain the breakdown products of ascorbic acid, which facilitates electron transfer during mineral weathering . Plants produce more ascorbic acid when grown in soils contaminated with heavy metals including copper . Fungi, which proliferate over plants and bacteria in metal-contaminated soils, can stabilize excess copper by extracellular cation binding or oxalate precipitation , but mechanisms probably also require enzymes, thiol-rich proteins and peptides, and antioxidants . The formation of electron-dense Cu granules within hyphae of arbuscular mycorrhizal fungi isolated from Cu- and As-contaminated soil suggests that fungi also can produce nanoparticulate copper. Some copper reduction possibly occurred in response to the European heat wave of the summer of 2003 . Elevated expression of heat shock protein HSP90 and metallothionein genes has been observed in hyphae of an arbuscular mycorrhizal fungus in the presence of 2 × 10–5 M CuSO4 in the laboratory . This suggests that a single driving force can trigger a biological defense mechanism that has multiple purposes. Thus, reduction of toxic cations to native elements may increase as rhizosphere biota fight metal stress and stresses imposed by elevated temperatures expected from global warming.Despite schizophrenia being a debilitating disorder affecting 1% of the population, there are no extant biomarkers to aid the clinician in identifying this disorder. Studies predict the genetic risk to be up to 80%,but despite strenuous research efforts the genes and polymorphisms found to be associated with schizophrenia account for very little of the genetic risk. Environmental risk such as urbancity,migrant status,childhood maltreatment,prenatal infections,cannabis use and maternal vitamin D deficiency also contribute to schizophrenia susceptibility. However, not all individuals exposed to environmental risk develop schizophrenia.This observation suggests that interaction between susceptibility genes and environmental factors may better account for schizophrenia. DNA methylation has been identified as a key mechanism for environmental regulation of gene expression.DNA methylation is an epigenetic modification that is essential for normal human development via regulation of gene function.

DNA methylation results in the addition of a methyl group on the cytosine of CpG dinucleotides,vertical growing systems which can then be inherited through cell division. These cytosine modifications can affect gene expression by altering the binding of transcription factors to promoter regions or changing mRNA processing. DNA methylation studies of the brain and peripheral tissue have previously been reported for schizophrenia. However, to our knowledge, no study has published results from an Illumina Infinium HumanMethylation450 Beadchip in the brain tissue of patients with schizophrenia. Studies to date have typically been performed in peripheral tissues and have been limited to the analysis of CpG islands in the promoter regions. A recent DNA methylation study analysed 27 578 CpG sites in peripheral blood cells from 18 patients with schizophrenia and 15 normal controls.This study revealed 603 CpG sites that had significantly different DNA methylation levels between schizophrenia and controls. Among these genes were HTR1E, COMTD1 and SLC6A3, which have previously been found to be associated with schizophrenia. An epigenetic study of monozygotic twins discordant for schizophrenia identified a number of loci differentially methylated in peripheral blood.Selected gene promoters have also been analysed for differential DNA methylation in the brain tissue from small numbers of patients with schizophrenia. Some of these genes include RELN,COMT,SOX1016 and HTR2A.An earlier study of 12 000 CpG islands in the frontal cortex of 35 schizophrenia and 35 controls revealed differential DNA methylation in genes associated with glutamatergic and GABAergic pathways.Apart from the present study, the only extant study using a 450 000 genome-wide methylation array was performed in leukocytes from patients with schizophrenia.DNA methylation analysis of schizophrenia has been more widely performed in peripheral tissue, because it can be readily obtained from living patients. The epigenetic profile differs in the brain compared with the peripheral tissue; however, some regions may have common patterns,which would make these regions ideal as potential biomarkers for schizophrenia. Some of the genes found to be differentially methylated in peripheral tissue of schizophrenia patients include HTR1A,HTR2A, BDNF,GRM2,GRM5and COMT.Brain tissue from the Human Brain and Spinal Fluid Resource Centre, CA, USA, was obtained in order to examine tissue involved in the etiology of schizophrenia. We analysed this tissue in a genome-wide methylation study of schizophrenia. We report significant differences in methylation status in brain tissue from schizophrenia patients compared with that from controls. In addition, unsupervised clustering analysis revealed two distinct groups corresponding to schizophrenia and controls.

Results of future epigenetic studies hold great promise of a schizophrenia biomarker and treatment, as epigenetic processes can be reversed.In order to assess differences in methylation between groups, the original n = 385 167 β-values were converted to M-values via the logit transformation as recommended by Du et al.Differentially methylated probes were detected using the limma package.The limma procedure uses linear models to assess differential methylation, whereby information is shared across probes.A major benefit of the limma procedure is that it allows the inclusion of covariates or other factors in the specification of the linear model. As such, we were able to adjust for age and PMI in the detection of differentially methylated probes by including age and autolysis covariates in the specification of the design matrix. Although most studies have found that methylation status is unaffected by PMI, we decided to adjust for PMI as a confounder.Probes were considered to be differentially methylated if the resulting adjusted P-value was o0.05. The Benjamini–Hochberg method was used to adjust the P-values and ensure that the false discovery rate was o0.05. The corresponding gene list was derived from the gene annotations associated with the probes.For cluster analysis, the top 3000 most variable probes were selected . A recursively partitioned mixture model was used to cluster the β-scores. RPMM is a model-based unsupervised clustering algorithm developed for measurements that lie between 0 and 1. This algorithm was implemented using the RPMM Bioconductor package.The implementation of RPMM was identical to Hinoue et al.who used a fuzzy clustering algorithm for initialisation and level-weighted version of Bayesian Information Criterion as a split criterion. In order to adjust for age and PMI, a series of linear models were fitted to the M-values using the function lmFit in the limma package. Coefficients for age and PMI, along with an intercept were estimated for each probe. Owing to this model specification, the residuals of the linear model represent the methylation values adjusted for the effect of age and PMI. The residuals were then back-transformed and clustered using the RPMM method implemented for the unadjusted probes. To allow visualisation of the distance between samples and to further reinforce the RPMM clustering, multidimensional scaling with a Euclidian distance metric was performed on both the adjusted M-values and the adjusted β-values. The first two coordinates, along with the RPMM clusters, are visualised in Figures 1a and b.

The results of the clustering indicate that the methylation profiles in those with schizophrenia are a heterogeneous group. There were some profiles that were consistently deemed distinct from the controls, whereas there were others that were not found to be significantly dissimilar. Twelve samples in particular tended to exhibit the former trait. When comparing these two potential subgroups of those with schizophrenia,vertical grow system we can see that the two subgroups exhibit no obvious difference in characteristics . Thus, there is potential for methylation arrays to be used to detect differences within these two potential subgroups. Differential methylation analysis between the two schizophrenia subgroups indicated that there were 73 222 probes that were differentially methylated . Of those probes, 6681 were promoter-associated and 2006 were both promoter-associated and located at a CpG island. After adjusting for age and PM1, 56 001 probes were found to be differentially methylated , 4779 being promoter-associated and 1238 both promoter-associated and located at a CpG island. The abundance of differentially methylated probes suggests significant groupings within the schizophrenia methylation profile. By contrast, a history of completed suicide or the presence of another psychiatric disorder revealed no significant differences in methylation.Differential DNA methylation in schizophrenia has been reported in several studies to date, although most of these studies involve the use of non-functional tissues such as blood. In this study, we analysed DNA methylation status in brain tissue, the primary tissue of pathology in schizophrenia, employing a genome-wide methylation array with very extensive coverage of the potential methylation sites in the human genome. After adjusting for age and PMI, 4641 probes corresponding to 2929 unique genes were found to be differentially methylated. When we compared the differentially methylated gene list with past studies using peripheral leukocyte samples, we found a high concordance rate, particularly for genes previously found to be associated with schizophrenia. Of the 589 genes Nishioka et al.11 found to be differentially methylated in peripheral blood cells from patients with schizophrenia, we were able to replicate 99 of these in the brain tissue. This shows promise for the use of non-invasive tissue such as blood or saliva to be used as a future diagnostic indicator of schizophrenia. We are aware of only one other study that used the 450 Illumina array in schizophrenia, although peripheral leukocytes were analysed rather than the brain tissue.That study identified 10 747 differential DNA methylation sites in medication free subjects.One of the genes they identified was RAI1, which has altered DNA methylation in the present study as well as an earlier schizophrenia study using the brain tissue.Other genes found to be differentially methylated in both the leukocyte study and the present brain tissue study includes HDAC4, GFRA2 and GDNF. The leukocyte study did not replicate COMTD1 and HTR1A that were found to be differentially validated from a previousstudy in the peripheral tissue.However, we report that these genes are differentially methylated in the brain. Although we were able to validate many of the previously identified CpG sites, experimental validation using an alternative method, such as pyrosequencing, would also confirm our results. Functional significance of genes found to be differentially methylated should also be tested by gene expression.

Unsupervised clustering of the top 3000 most variable probes revealed two distinct groups after adjusting for age and PMI. Cluster 1 comprised 88% patients with schizophrenia and 12% controls, whereas cluster 2 comprised 27% patients with schizophrenia and 73% controls. To our knowledge, this is the first report of DNA methylation profiling that is able to significantly differentiate between those with schizophrenia and control subjects. Although Nishioka et al.was able to identify site specific DNA methylation changes in patients with schizophrenia, they were unable to discriminate between controls and schizophrenia patients using unsupervised clustering.11 DNA methylation patterns differ in brain cells compared with peripheral tissues such as blood,and this may explain the lack of separation reported by Nishioka et al.Although some genes may have the same epigenetic profiles in peripheral and brain tissue, a more comprehensive list of tissue-specific genes may be required to differentiate controls from those with schizophrenia. Another reason may be the analysis of fewer CpG sites in the previous study. This is potentially important, as a DNA methylation signature across the whole genome is required to identify the most important differentially methylated probes. The results of our clustering analysis will need to be confirmed in an independent brain tissue cohort.It is possible that these two subgroups have specific symptomatology that warrants further investigation in a sample set with a comprehensive clinical history. After adjusting for age and PMI, DTNBP1, COMT and DRD2 were found to be differentially methylated between the two schizophrenia subgroups. Interestingly, these are genes that we have previously found to be associated with schizophrenia.A recent study has found significant DNA methylation changes in the early stages of development and suggest that aberrant DNA methylation during the transition from the fetal to the postnatal period of development could be critical for the pathogenesis of schizophrenia.

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Stress management strategies are needed to preserve well-being during this abrupt isolation period

Posted on July 7, 2023 by hemp2022

Drawing on this approach, we understand experience as meaningful sensory perception in temporal context and within particular cultural, social, and interpersonal settings and subjectivity as the more or less enduring structure of experience. With respect to mental illness, this approach invites anthropological recognition of struggle as a fundamental human process that comes to light in the context of lived experience . Second is the ongoing anthropological concern with adolescence as a stage in the life course at which identity is consolidated and people approach full cultural membership but which is also fraught with challenges to well-being that anthropology can contribute to understanding in a way relevant to mental health policy and practice . The contemporary anthropological approach to childhood is strongly influenced by child standpoint theory that aims at an account of society from where children are socially positioned and in which they are not passive social “others” but agentive participants in social life, hence co-constructors of social knowledge and by extension of knowledge generated by research2 . In particular, anthropologists have taken up the idea that “children have agency and manifest social competency” . Guided by these concerns, we will focus specifically on self-cutting among a group of adolescents who have been psychiatric inpatients; by attending to experience and subjectivity articulated in the youth’s own voices, we will come to understand self-cutting as a crisis of agency enacted on the terrain of their own bodies. There is scant literature on how young people conceive and understand mental health ,vertical farm indoor let alone experiential accounts of adolescent mental illness from the standpoint of the child .

In addressing the experience of cutting among a clinically defined and diagnosed group of youth, our stance is not to fall prey to accepting a false dichotomy between ethnographic and clinical sensibilities; that a young person is following a regimen of psychotropic medication is as much an ethnographic as a clinical fact, and that a young person lives in a fragmented family environment may have clinical as well as ethnographic implications. Self-cutting can be understood as a troubling symptomatic behavior or as a creative struggle for agency and may exhibit elements of both pathological obsession and ritual transformation, but in either case it is an enactment of a vexed relation between body and world.This discussion is based on SWYEPT, our study of youth in New Mexico who were inpatients in the state’s flagship Children’s Psychiatric Hospital at the University of New Mexico . New Mexico is a state whose total population according to the 2010 United States Census was 2,059,179. In 2010, according to the US Census Bureau’s categories, by race the largest population proportions were designated “white” and American Indian/Alaska Native , with 23 federally recognized Indian tribes in the state comprising various groups of Pueblos, Navajos, and Apaches; other racial categories were minimally represented. By ethnicity, Hispanics or Latinos accounted for the largest single block , while among non-Latinos the largest blocks identified themselves as generically white or American Indian . New Mexico is one of the poorest states in the nation. According to the US Statistical Abstract, as of 2008 the median household income was $43, 508 or 44th among the 50 states, and the proportion of persons living below the poverty level was 17.1% or 5th in rank among the states. New Mexico ranks as one of two states within the United States hardest hit by child poverty, with the rate of 30% in New Mexico . Relatedly, home foreclosures have also been inordinately high. Along with poverty comes a serious drug problem, with parts of the state severely afflicted by heroin and methamphetamine use, and the presence of violent gangs, with one antigang website listing 178 in the Albuquerque area. The SWYEPT study examines cultural meaning, social interaction, and individual experience among adolescent patients and their families, with the long-term goal of producing knowledge of broad use to those concerned with the treatment of adolescents suffering from mental illness in the context of significant cultural differences.

The aspects of this knowledge include: types of problem, illness, or psychiatric disorder experienced by afflicted adolescents; trajectories of adolescent patients from the community into treatment and back into the community; patient experience of therapeutic process and family response to that process; alternative and complementary resources brought into play by families on behalf of patients; difference between the experience of afflicted adolescents and that of counterparts who have not been diagnosed or treated for emotional disturbance. Notably for present purposes, ours was not explicitly a study of self-cutting or self-harm, but cutting emerged within the ethnographic interviews as a theme deserving of the particular attention we devote to it here. We recruited participants for the study with the assistance of three clinicians at Children’s Psychiatric Hospital who referred to us patients aged 12–18 they judged as not so severely cognitively disabled or developmentally impaired as to be unable to participate in interviews and not so emotionally fragile or clinically vulnerable that their participation would be unduly stressful. We obtained informed consent from youth and their parent or primary guardian based on these referrals, recognizing the ethical responsibility of respecting the vulnerabilities of individual patients and the need for continued rapport in the relationship between therapists and families, as well as the importance of our respect as researchers for the clinical expertise of the referring therapists. All participants entered the project as inpatients at CPH. Assisted by a team of graduate student ethnographers and clinically trained diagnostic interviewers, we conducted ethnographic interviews covering life history and experience with illness and treatment with the young people and their primary parent/guardian three times at approximately six months’ intervals.During this period, we also conducted the child version of the Structured Clinical Interview for DSMIV , a clinician-administered research diagnostic interview , the Adolescent Health Survey , and the Youth Self Report and Child Behavior Check List for children and their parent/guardian respectively.

Although initial interviews occasionally took place in the hospital, it was rare for a participant still to be there at the time of the second and third ethnographic interviews. Yet it was not always the case that they were at home, since it was not uncommon for them to be placed instead at another treatment facility of in-treatment foster care. This often led us far afield from the hospital in Albuquerque, such that our ethnography ranged across the entire state of New Mexico and occasionally beyond. In this respect, our work was not strictly speaking a clinical ethnography in the sense of ethnography primarily situated in a clinical context that focuses on the institutional cultural milieu and interactions among patients and staff . Our focus was instead on the experience and subjectivity of the troubled youth along their trajectory back to their families, back again to the hospital, to other institutions, or to treatment foster care. Whenever possible we conducted interviews in participants’ homes both for their convenience and so that interviewers could conduct ethnographic observation of the domestic environment and neighborhood setting. Our primary ethnographic locus was thus the family rather than either the clinic or the community, following the methodological premise that families are the principal formative inter subjective locus for adolescents and for the mentally ill, no less for mentally ill adolescents . Given these caveats, our work could be described as clinical ethnography in a different sense, insofar as it synthesizes clinical and ethnographic sensibilities and approaches . This means not only a balanced attention to diagnostic profile and life experience,vertical farm supplier but recognition that narrative data generated by diagnostic and ethnographic interviews can be complementary by identifying different kinds of experientially relevant events and themes . The participants constituted an ethnically diverse group including New Mexican Hispanics and Latinos of Mexican descent, Anglo-Americans, and Native Americans. While an ethnically diverse group of youths whose economic and residential conditions vary, the life situations of most are shaped by features of structural violence . Of the 47 adolescents who participated in the research, 57% reported having cut or harmed themselves at some time, comparable to 61% among adolescents hospitalized for psychiatric problems in a previous study by DiClemente, Ponton, and Harley in another North American location.

This rate can be understood against the background of a reported rate of 1–4% of self-injurious behavior in the general population , while the rate among adolescents has been placed by various researchers as ranging between 1 and 39% . Let us now take a closer look at cutting among several of these young people in order to get a sense of how they talk about it and what it means to them, its place in the overall configuration of their experience, and the similarities and differences among them that might allow us to characterize cutting as a crisis of agency.SWYEPT participants represent a wide range of diagnostic profiles from a clinical standpoint and a diversity of life experiences from an ethnographic standpoint , but our purpose here is to present a series of vignettes that summarize the range of experiences and utterances centered around the phenomenon of cutting. Lacking space to present full case studies, we briefly examine how they describe their own cutting behavior and what that behavior means in the context of their troubled lives and in the constitution of their subjectivity. We have selected these instances and interview excerpts based on the young person’s relative ability and/or willingness to elaborate on how cutting has been a part of their lives. Each profile includes the biographical and ethnographic context of the young person’s experience, their diagnostic and functional status, medication history, their own experiential commentary, and a brief interpretative commentary. We first met Maria, a Hispanic 17-year-old female, when she was living in a ranch style home in a lower-income neighborhood that was bustling with the activities of her extended family and infant son. During the course of the study, she later lived in an apartment with her son, boyfriend and mother, moving again two years later into a very small apartment with her toddler. Maria was the youngest of three daughters to a single mother with multiple boyfriends and father figures. She bore the physical and emotional marks of a major arm injury in mid-childhood from a car accident in which her extremely drunk mother was driving, as well as enduring severe and catastrophic stressors related to abuse, neglect, and sustained psychosocial instability in her early life. While Maria is close to her older sisters, they were not a significant source of personal or financial support and held an antipsychiatric opinion about how Maria did not need medication. She relayed that she had to “grow up fast” since her mother had severe problems with alcohol. Indeed, her mother’s alcohol abuse severely affected their relationship. Maria was sexually assaulted at the age of 11 by her mother’s ex-boyfriend, which disturbed her greatly; she marks this as a time of pain and confusion. She grew up believing her adoptive father was her biological father, but when Maria was 15, her father went to fight in Iraq. While in Iraq, he wrote her two letters saying that she was not his biological daughter, and he was getting remarried and could not support her financially or otherwise. She said she was devastated by this, by the fact that he would “cut me out of his life.” Maria responded by cutting on herself. In this instance, we have a patent metaphoric and literal alignment of psycholinguistic and palpablebodily expression. These events have created major emotional and psychiatric challenges for Maria. When Maria began the study, she had been admitted for a suicide attempt and ongoing postpartum depression. Prior to being admitted, she had an eating disorder, along with self-harming and abusing cannabis for two years. Her SCID diagnosis shows mood disorder due to a general medical condition, postpartum major depressive disorder, brief psychotic disorder related to postpartum depression, separation anxiety disorder, PTSD, alcohol abuse, cannabis dependence, and an eating disorder.For Maria, cutting was an intended if fraught means of communication in the face of the emotional pain of abandonment. This was not the first time she cut; her practice began at age 11 following the sexual assault by her mother’s boyfriend.

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The fact that someone has used a drug in the past month does not mean they use the drug frequently

Posted on July 6, 2023 by hemp2022

For a decade there have been claims that methamphetamine, originally a West Coast biker drug, is spreading east, but if so, the diffusion has been fairly slow, and there is no evidence that use is significantly increasing. As seen in Figure 2, cigarette users tend to be daily users, but most recent users of cocaine and the hallucinogens did so only once or twice in the past month. About a third of marijuana users use almost every day; surprisingly, only half as many alcohol users do so. The prevalence of drug use provides a distorted picture of the actual health and safety harms posed by psychoactive drugs. There are several reasons for this. First, because drug use is only probabilistically related to drug harm, most harms to self and others are attributable to heavy users, and for most substances , a minority of users account for a majority of the quantity consumed. Second, school- and household-based surveys under represent hard core users, who are more likely to be truants, school dropouts, homeless, or institutionalized. Another major category of drug-related harm is the transmission of HIV. Injection drug use accounts for about a third of all AIDS cases in the US , and non-injection use is associated with an elevated risk of unsafe sexual practices. But there is no reliable way to attribute some fixed percentage of AIDS cases to cocaine use, to marijuana use, and so on. In keeping with our general analytical framework, recall that total harm is the product of prevalence x quantity x average harm. Thus, marijuana – by far the most prevalent illicit – accounts for a sizeable share of all three harm indicators. But relative to their much lower prevalence, it is clear that heroin, cocaine, and methamphetamine are disproportionately harmful substances. A 2007 Lancet article by David Nutt and colleagues offers what is to date the most sophisticated attempt to rate psychoactive substances by their “intrinsic” health and behavioral harms. In decreasing order of harmfulness,vertical farming rack the worst five drugs were heroin, cocaine, the barbituates, street methadone, and alcohol.

Tobacco was ninth, cannabis eleventh, LSD fourteenth, and MDMA eighteenth. If one were starting a society from scratch, it is unclear where one might paint a bright line separating licit from illicit substances, but it is difficult to see why alcohol and tobacco would be more accessible than cannabis or MDMA. But of course, we are not starting a society from scratch. The data in Table 2 are misleading if one wants to rank the intrinsic psychopharmacological harms of drugs – marijuana looming too large – but they are valid indicators if one wants to know the contributions of different substances to social harm under the current regime and with current patterns of use. These kinds of indicators tells us little about how harmful each substance might be in a regime of regulated legal access. Average harm per use might approximate the rating levels in Nutt et al. , though heroin use would become a lot safer per dose. Total harm might differ, if for example legal LSD were to become massively popular. But this might be a short term effect; societies seem to learn from experience and scale back on drugs that are obviously dysfunctional . The FY2008 national drug control budget allocates 36 percent of drug funding for interdiction and source-country controls, 28 percent for domestic drug law enforcement, 23 percent for treatment, and 12 percent for prevention . We can only offer a whirlwind tour of the empirical literature on these interventions. Readers can find comprehensive assessments in recent monographs by Boyum and Reuter and the National Academy of Sciences . The drug policy literature is enormous and yet remarkably thin, in that rigorous program evaluations are rare. There are some valuable cost-effectiveness analyses , but these are limited by the available descriptive data and some daunting problems of causal inference , and many of the available evaluations were conducted by program developers, raising concerns about intellectual and financial conflicts of interest.In a classic analysis, Reuter and colleagues explained why we should not expect big impacts of efforts to thwart drug production and trafficking. First, it is not possible to completely “seal the borders” against relatively small packages of chemicals that will be sold at very high prices; there are too many possible smuggling routes and tactics, and dealers are very adaptive. Second, the price structure of illicit markets is such that bulk drug products in source countries are “dirt cheap” compared to the high retail street prices in the US.

At the source, the economic value is low by US standards but high by local standards. But most of the markup in US prices occurs in the last few links of the distribution chain, within US borders. For example, Caulkins and Reuter note that the wholesale price of cocaine or heroin in a source country is only about 1 percent of its US retail street price. For example $1500 of cocaine in Colombia may be worth $15,000 at the US border, and $110,000 in the US retail street market. Thus even very large seizures in other countries are unlikely to have big effects on local prices. In recent years, defense analysts have used time-series data to argue that interdiction and source-country campaigns actually do have a significant impact on street prices and US demand. But these analyses have been debunked by a National Academy panel , arguing that the apparent correlations are spurious and amplified by selective focus on certain source-country interventions that happened to precede short-term price drops. It does not follow, however, that we could eliminate these programs entirely without a detectable effect. Most analysts believe that interdiction risks do raise prices; it is just that there are probably steeply declining marginal returns to such efforts. Presumably, these programs serve other US political, diplomatic, and economic goals beyond drug policy, laudable or otherwise. But we could probably cut back significantly on these efforts without seeing an increase in US drug consumption. What has this massive social experiment bought us? Early in the growth period, around 1992, one could argue that it was correlated with a considerable drop in drug use relative to the late 1970s . But this period of optimism was short-lived. By 1996, about half of the gains were gone, and levels of use have remained fairly stable since then, even as the drug prison population continued to rise. In fact, illicit drug prices have plummeted during a period when massive law enforcement sought vigorously to make drugs more expensive . This is troubling, because prices do matter; contrary to widespread belief, even addicts have been shown to be sensitive to drug prices . From the perspective of prevalence and quantity reduction, falling prices are a serious problem. But conceivably, falling prices may be beneficial from the perspective of harm reduction, because addicts might be expected to conduct fewer income-generating crimes to feed their habit. This is another illustration of the need to confront hard trade offs in thinking about drug policies.

The harshness of US marijuana enforcement has long received considerable criticism, and indeed it is difficult to defend . But Caulkins and Sevigny warn against exaggerated concerns about unlucky marijuana smokers rotting away in a prison cell. Although 38 percent of state and federal incarcerations for drug offenses involved simple possession, “for only 2 percent of imprisoned drug-law violators was there no reason whatsoever to suspect possible involvement in distribution…depending on how strict a definition one preferred, one might argue that anywhere from 5,380 to 41,047 people were in prison in the United States solely for their drug use.” On the other hand, many who avoid time in prison do spend time in jail – as much as a third of arrestees in a study of three counties in Maryland . More troubling is the disproportionate imprisonment of African American men. African Americans accounted for about half of all drug incarcerations . A major factor is the differential severity of mandatory minimum sentences for crack vs. powder cocaine. Under these laws, a dealer would have to sell 500 grams of powder cocaine but only 5 grams of crack cocaine to receive the same five-year sentence. Since crack is more likely to be sold in African American communities, this has greatly widened the racial gap in sentencing. Even putting aside the questionable pharmacological and moral aspects of this differential policy, there is no evidence whatsoever for its effectiveness in controlling crime. Caulkins and colleagues show that conventional sentencing is significantly more cost effective. Although the crack mandatory sentences were trimmed somewhat in 2007,commercial indoor vertical farming and the Supreme Court recently acted to restore some judicial discretion in these cases . Whether these changes will translate into a closing of the large racial differential remains to be seen. The optimal level of drug law enforcement is surely well above zero, but just as surely, well below current levels . Caulkins and Reuter argue that we could reduce the drug prisoner population by half without harmful consequences; they note that this would still leave us with system “a lot tougher than the Reagan administration ever was.” Kleiman suggests tactics for getting more mileage out of less punishment through the use of small, quick sanctions, strategically deployed. In 2005, there were about 1.8 million people in substance abuse treatment in the US, about 40 percent for alcohol, 17 percent for the opiates, 14 percent for cocaine, and 16 percent for marijuana . There are certainly many thousands of people who need treatment and are not receiving it. Whether expanding the available treatment capacity would bring them in is an open question. We should be wary of assuming that a purely “public health” approach to drugs can work; the police and courts play a crucial role in bringing people into treatment – increasingly so with the expansion of drug courts and initiatives like California’s Proposition 36, the 2001 law which permits treatment in lieu of incarceration for those convicted for the first or second time for nonviolent drug possession . For most primary drugs of abuse, criminal justice referrals are a major basis for treatment: in 2005, 57 percent of marijuana treatment, 49 percent of methamphetamine, and 27 percent of smoked cocaine. But 36 percent of clients in alcohol treatment were referred by the criminal justice system, so legal status may not be the crucial lever. In a sophisticated cost-effectiveness analysis, Rydell and Everingham estimate that the U.S. could reduce cocaine consumption by 1 percent by investing $34 million in additional treatment funds, considerably cheaper than achieving the same outcome with domestic drug law enforcement , interdiction , or source country controls .

But because treatment effects are usually estimated using pre-post change scores that are vulnerable to two potential biases . First, the posttreatment reduction could reflect a simple “regression to the mean” in which an unusually extreme period of binge use would be followed by a return to the user’s more typical levels, even in the absence of treatment. Second, treatment pre- and posttests are vulnerable to selection biases because clients who enter and remain in treatment until post-treatment measurement are a non-random and perhaps very unrepresentative sample of all users. Regression artifacts would inflate treatment estimates; selection biases could either inflate or deflate the estimates. We believe that the full weight of the evidence makes it clear that treatment is both effective and cost-effective, but until these problems are better addressed, we cannot be sure that the benefits of expanded treatment would be as large as Rydell and Everingham implied. Even its most passionate advocates recognize that treatment’s benefits are often fleeting. About three quarters of heroin clients and half of cocaine clients have had one or more prior treatment episodes . Forty to sixty percent of all clients will eventually relapse, though relapse rates are at least as high for hypertension and asthma treatment . Importantly, Rydell and Everingham recognized that treatment can provide considerable health and public safety benefits even if it only reduces drug use while the client is enrolled. Held up to a standard of pure prevalence reduction , treatment is unimpressive. But by the standards of quantity reduction and harm reduction, treatment looks pretty good.

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Seven percent of women admitted to treatment centers in 2009 abused MA at time of admission

Posted on July 5, 2023 by hemp2022

A recruiter sits across from the participant in a quiet screening room to ensure privacy during the consenting process. Typically, only one recruiter is present with one participant, unless particular expertise is needed from additional staff. In this case, and with the participant’s permission, another recruiter may be brought in to assist and offer additional information. The consent process involves conveying study information to a prospective participation and usually requires approximately 30-minutes, but the time can vary depending on the complexity of the study. During the informed consent process, the recruiter gives the prospective participant a copy of the “Experimental Subjects Bill of Rights” to review, which is required by California state law when enrolling in biomedical research that involves medical experimentation. The recruiter explains the HNRP’s privacy practices and data confidentiality procedures, including that participants’ identifying information is stored securely and separately from information collected for the study and only specific research staff have access to that information. Next, the recruiter reviews each paragraph of the IRB approved study consent document with the prospective participant. At the end of each consent section, the recruiter asks if the prospective participant has any questions or concerns. Across studies hosted by the HNRP, the consent documents contain language required by the IRB, standard HNRP practices,indoor vertical farming system and study specific information resulting in documents that range from 4 to 17 pages. Lastly, participant’s knowledge of the study is tested with a validated assessment tool , which is used to evaluate understanding of the study purpose and procedures, including possible risks and benefits.

Two raters independently coded participants’ responses to questions 1, and 2, which were openended prompts where participants could write as much or as little as they wanted. Each rater established their own code book organized by emerging patterns and themes. After initial coding of the responses, the raters and senior author met to review the themes generated. The majority of codes and resulting themes were initially agreed-upon by both raters with discrepant themes discussed with the senior author until reaching inter-rater agreement. Theme labels were determined by group consensus.In research studies, it is the responsibility of the researcher to ensure that the consent process is implemented correctly. While decisional capacity questionnaires are common, particularly in populations with higher risk of impaired decisional capacity, it is not common practice to solicit feedback from participants with regard to the informed consent process. Research participants at the HNRP, a research center with multiple years of working with PWH and dedicated research staff with experience taking potential research participants through the consent process, found the consenting process, on average, to be extremely informative and extremely consistent with what they did during the study. This indicates that the traditional informed consent process involving a face-to-face discussion with a prospective research participant combined with a written consent document can be successful. Additionally, several different themes were endorsed as “most important” and the most common answer for what was “least important” was that everything was important. This would indicate that although some participants do not find all portions of informed consent to be useful, some participants do; including the goals of the study, procedure, risks, and information about protection of confidentiality. This may also suggest the consenting process should be tailored to individual participants, and participants should be able to choose how much additional information they are provided beyond the required information. We did not observe differences in response to any of the questions by HIV status. Unfortunately, there is little research examining what PWH value most from research study participation, the informed consent process in general, or in comparison to HIV-negative persons to compare with our findings.

One review examining barriers to participation in HIV drug trials found societal discrimination and distrust of researchers, among other things , were barriers to participation . Based on this review, as well as historical and ongoing stigma toward PWH, confidentiality may be something PWH highly value in research studies. However, in this study, we did not observe HIV serostatus differences in proportion of participants reporting confidentiality to be “most important.” The majority of PWH participants who filled out the consent questionnaire have been in multiple studies at the center and, on average, have completed more study visits than HIV-negative participants. The center is also actively involved within the community. Therefore, participants prior experience with the center may have had a higher level of trust going into the informed consent process, which may have influenced their responses. Additionally, confidentiality is likely valued by many individuals regardless of HIV status, which also may be why we did not observe any difference by HIV serostatus. While this study adds valuable insights into informed consent literature, there are limitations. First, the center in which this study took place is an established community research facility with highly trained staff experienced both working with this study population and consenting participants. Studies without these resources, trained staff, or established study enrollment practices may have difficulty with establishing the practices described in this study. Second, participants in the study were already willing to come in for the research study, so their responses may not accurately reflect what the general population values from the informed consent process or what aspects of the informed consent process may persuade individuals that are not willing to participate in research. Moreover, we were unable to examine responses by other variables of interest due to sample size restrictions. Future studies should aim to assess participants’ preferences with respect to information presented during the informed consent process and how those preferences differ at the individual level. Lastly, not all participants provided responses to every question, particularly the open-ended response questions, which may be due to participants not having feedback to provide or found the open-ended response questions to be burdensome.

Asking a more specific question or interviewing participants about what changes they would suggest for improving the informed consent process may have elicited more responses.This study demonstrates that implementing the informed consent process with trained staff can be successful. Participants reported the experience as both informative and believed what they were told during the consent process was consistent with what they experienced during the study. This is in line with systematic reviews that have found that the most effective way to improve understanding of the informed consent was to have a one-on-one discussion with study participants . This would suggest that IRBs and researchers should be invested in the training of those who implement the informed consent process as well as monitor how the informed consent is presented to research participants. Furthermore, our recommendation to researchers working with PWH is to view the informed consent process as an opportunity to build trust, educate and show a true appreciation for the participants’ time, which will hopefully encourage continued participation in research.There are opportunities to continue to improve the informed consent process, and there have been recent updates to the “Common Rule”, which will influence the presentation of the informed consent moving forward . Often in practice the informed consent falls short of what it aims to accomplish . For example, a recent meta-analysis found that participants’ understanding of portions of the informed consent ranged from 52–76% . Even more discouraging is that Tam et al. found that the proportion of participants who understood the informed consent process has not improved in the past 30years. In hopes of improving informed consent to make it more engaging and understandable, two studies compared a simplified and concise informed consent with a traditional consent form. Both studies reported that participants found the shorter informed consent more engaging, and one study reported that comprehension was equivalent to the standard consent form whereas the other study found improved understanding with the shorter consent form . Another study found that implementing a fact sheet and engaging in a question and answer feedback session improved open-ended questions to assess understanding of the informed consent . Additionally,indoor vertical farming kits in a study that compared ways to assess understanding of the informed consent, found that commonly used forced choice or self-report questionnaires may overestimate the level of understanding of the informed consent as recognition of information does not ensure comprehension of information . Therefore, free-response questions may be a better measure of comprehension indicating that IRBs that review decisional capacity questionnaires and researchers who are trying to improve the informed consent process must be mindful of how understanding of the informed consent is assessed.

As research moves more toward digital studies and trials where the in-person interaction is not feasible, as we are currently facing with the COVID-19 crisis, it will be important to design the consent process that can build upon elements of existing successful consent models. Standards of practice and design features for digital consent, also known as eConsent, are in development. Some groups have begun to create open-source and customizable tools for low-risk, mobile-mediated research , which has been used in patient populations . Digital studies and eConsents are advantageous as they allow for use of multimedia methods , which can be standardized and reviewed by an IRB to ensure that participants are receiving the necessary information. Studies that have examined multimedia methods have shown that they successfully relay information to participants, with some studies reporting that use of video or PowerPoint is related to an increase in engagement and comprehension . Additionally, as more participants want their study results returned to them, digital consent could allow participants to tailor the consent to their personal preferences and select which data they would like access to. However, there are additional considerations when designing eConsent . For example, in a focus group study of patients underrepresented in research, participants overall found eConsent easy to use and interesting; however, minority and rural participants raised concerns about accessibility, trust, and confidentiality . We anticipate our findings demonstrating an engaging informed consent process can be adapted for digital deployment for populations that value privacy and confidentiality such as PWH.Methamphetamine abuse is a significant problem in the United States, particularly in the West and Midwest. Worldwide, amphetamines are second only to cannabis as the most widely abused drugs, with a prevalence of 14 to 57 million, or 0.3 to 1.3%, of all 15-64 year olds. In 2010, an estimated 353,000 people age 12 or older in the United States reported using MA, with 105,000 estimated new users. Women account for a substantial subset of MA users; data from treatment centers in 2003 showed 45% of patients treated for amphetamine abuse were women, increasing to 46% in 2009.Moreover, MA abuse among pregnant women is a persistent problem. The Infant Development, Environment and Lifestyle study found approximately 6% of women reported drug use during pregnancy. Further, the prevalence of MA abuse in pregnant women admitted to federally funded treatment centers in the U.S. rose to 24% in 2006. Similarly, international data from 2004 demonstrates that amphetamines were used by 23% of substance-abusing mothers. The effects of prenatal MA exposure on childhood outcome are not well characterized. MRI data has shown that MA exposure is associated with reductions in striatal and caudate volume, which may be associated with cognitive deficits. Volumetric assessments of MRIs in exposed children have also demonstrated smaller subcortical volumes including the putamen, globus pallidus, and hippocampus, potentially impacting attention and memory. These findings are consistent with behavioral issues described in a small cohort of children exposed to methamphetamine. In this small non-randomized sample, prenatal MA exposure is associated with deficits in executive function and spatial performance, aggressive behavior and problems with peers, as well as delays in math and language. Relatively little is known about the effects of MA during early infancy. In a study of cocaine and MA exposed newborns, there was an increased incidence of intraventricular hemorrhage and white matter densities observed on cranial ultrasound. In another study, prenatal amphetamine exposure has been associated with increased drowsiness in exposed infants in the first few months of life that resolved by twelve months of age. However, these previous findings were retrospective and utilized a small sample size. The Infant Development, Environment and Lifestyle study is a prospective longitudinal investigation of neurobehavioral outcome related to MA exposure in utero.

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BD is a major psychiatric disorder affecting approximately 2% of people worldwide

Posted on July 4, 2023 by hemp2022

Emotional dysregulation and impaired stress response, other important features of bipolar disorder , may be caused by disturbances in corticotropin metabolism and dysfunction in the hypothalamic–pituitary–adrenal axis . Interestingly, a recent rat model study demonstrated significant long-term loss, shrinkage of cell soma size, and axonal degeneration of corticotropin-releasing factor-positive neurons in the amygdala following neonatal hypoxia–ischaemia . These changes were associated with increased locomotor activity and exploratory behaviour , behavioural abnormalities that are also observed in patients with bipolar disorder . Moreover, increased anxiety has been reported following perinatal asphyxia and is associated with dopamine-innervated neurocircuitries in the amygdala, among other structures . Dopaminergic pathways are particularly vulnerable to perinatal asphyxia , and are also involved in the pathophysiology of psychotic disorders . Taken together, it seems biologically plausible that perinatal asphyxia is associated with long-term alterations in the structure of the amygdala, as our results suggest. Such alterations may be functionally associated with the distinct behavioural abnormalities observed in bipolar disorder. Instead of confirming our initial hypothesis that the associations between perinatal asphyxia/severe OCs would be stronger in patients with psychotic than non-psychotic bipolar disorder, the results indicate different patterns of associations in psychotic versus non-psychotic bipolar disorder. Within a psychosis continuum, psychotic bipolar disorder would be considered to be closer than non-psychotic bipolar disorder to schizophrenia. In schizophrenia, smaller hippocampal volumes have been associated with preand perinatal trauma . Surprisingly, we found no associations between severe OCs or perinatal asphyxia and smaller hippocampal volume in patients with psychotic bipolar disorder,agriculture vertical farming but we did find such associations in patients with non-psychotic bipolar disorder.

The biological validity of this association is, nevertheless, supported by the literature. First, animal models have demonstrated the pyramidal neurons within the hippocampus to be sensitive to prenatal hypoxia . Second, in the human neonate, hippocampal neurocircuitries are reported to be particularly vulnerable to hypoxia , and, third, healthy adolescents who have suffered perinatal asphyxia exhibit reduced hippocampal volumes . Based on these findings, one could expect to find associations between perinatal hypoxia or severe OCs and hippocampal volume in the healthy controls as well, as we did in a previous study of schizophrenia patients and healthy controls . The hippocampus is, however, one of the brain regions in which neurogenesis occurs , and the number of hippocampal neurons , as well as the hippocampal volume as measured by MRI , might increase in response to different training tasks. On the other hand, hippocampal volume may be reduced in alcohol dependence and heavy cannabis use , as well as in several mental disorders including schizophrenia , unipolar depression and bipolar disorder , although we did not confirm the latter in this study. As such, a variety of factors may confound and interfere with putative associations between pre- and perinatal trauma and adult hippocampal volume. The number of possible known and unknown confounders not accounted for constitute one limitation in the current study. Although we did control for current lithium use, which is known to affect hippocampal and amygdala volumes in bipolar disorder, we did not have reliable data on cumulative medication use. Another possible limitation to the generalizability of the current study is the inclusion of patients across mood states, i.e. depression, mania/hypomania, and euthymia, since it has been suggested that amygdala volume may fluctuate across mood states . Third, the subject groups were relatively small when the bipolar disorder group was split into psychotic and non-psychotic subgroups, which may have caused type II errors within the statistical analyses. Finally, by studying severe OCs, and comparing subjects with them versus all other subjects , we placed high demands on the strength of the relationship, and may have missed possible associations between brain structure and less severe OCs.

Strengths of the current study include the use of unbiased birth registry data, thorough clinical characterization of participating subjects, and the use of one MRI scanner with no upgrades during the study period. In summary, we report perinatal asphyxia to be related to smaller amygdala volume in patients with bipolar disorder. This suggests a neurodevelopmental component in the brain morphology of bipolar disorder. The different associations between preand perinatal complications and brain morphology observed in patients with psychotic and non-psychotic bipolar disorder, as well as their possible functional consequences, warrant further investigation.Treatments for bipolar disorder have been very limited and no novel therapeutics specifically for BD have been developed in a long time. Treatments that target the neural circuitry specifically affected in BD patients are urgently required in order to ameliorate the symptoms of this disorder.It is characterized by episodes of mania and depression, with periods of absence of symptoms that meet diagnostic criteria for mania or depression in-between . The symptomatology of BD is heavily dependent on the state in which patients present. Mania includes long periods of euphoria, reduced sleep, hypersexuality, extreme irritability, racing thoughts, aggression, hedonic behavior, and hyperactivity, which is a conspicuous feature during acute manic states . Indeed, the latest edition of the Diagnostic and Statistical Manual of Mental Disorders states that changes in activity and energy are required for diagnosis of a manic episode in addition to an ‘increase in goal-directed activity’ or ‘psychomotor agitation’ . Symptoms of depression are largely the opposite and consist of long periods of dysphoria, reduced libido, increased sleep, feeling tired or ‘slowed down’, anhedonia, and greater risk of suicide. Bipolar depression is differentiated from unipolar depression or major depressive disorder diagnostically by the requirement of at least one manic or hypomanic episode for BD diagnosis . In addition to mood and behavioral changes, patients with BD also suffer from a variety of neurocognitive deficits in domains such as executive functioning, vigilance, impulsivity, and decision-making .

Although BD has typically been described as a mood disorder, these cognitive impairments are irrefutably important since they correlate closely with a patient’s functional outcome . Hence, by understanding and improving cognitive function, the patient’s quality of life will likely improve. Untreated or poorly maintained BD can be devastating to the quality of life of patients and their families. Treatments that are commonly used include the mood stabilizer lithium, anticonvulsants such as valproate and lamotrigine, and several antipsychotics . None of these treatments completely stabilize behavior nor improve cognitive functioning. In fact, current treatments, particularly lithium, can worsen cognitive functioning and ingenuity . Hence, increased creativity and individuality felt by many people with BD are eroded by these treatments, limiting adherence compliance . Instead of using generic ‘band aid’ treatments for subduing behavior, developing targeted treatments for problematic symptoms experienced by BD patients would likely generate better treatment outcomes and adherence. Indeed, none of the currently approved treatments were developed with BD as a target, but were in fact discovered serendipitously . Together with the symptoms suffered by patients, these poor treatment options contribute to an increased suicide mortality rate ; recent estimates indicate that one in three BD patients attempt suicide . Novel therapies based on the underlying mechanisms of abnormal behavior in BD patients are urgently required to improve quality of patient care. Both in vitro and in vivo preclinical studies play essential roles in the discovery of novel targets to develop useful therapeutic outcomes. Modeling the abnormal behaviors present in people with BD across different species remains a challenging task . The complexity of the abnormal behaviors in patients both within and between bipolar states makes modeling BD extremely difficult. Even targeting a single behavioral abnormality can be challenging; for example,china vertical farming racing thoughts or hopelessness is measured using questionnaires in humans, which would be impossible to mimic in rodent studies. The limitations of mimicking psychiatric illnesses in non-human mammals has resulted in the frequent simplistic use of increased motor activity as a primary measure of animal models of BD mania . Besides activity, other relevant BD behaviors modeled in animals include aggression, hypersexuality, hedonia-like behavior, inattention, and risk-taking or decision-making . The use of a comprehensive sequence of tests assessing different BD-like behaviors – instead of focusing on one aspect of the illness – can aid the identification of global mechanisms underlying symptoms as well as treatments . Combining this approach with the reverse-translational behavioral tasks that characterize behaviors from patients to rodents and vice versa would likely yield more relevant information . The majority of approaches used in the field still tend to focus on the assessment of relative basic behaviors in rodents. The observations of simple behaviors in rodents have limited validity with regards to modeling human behaviors often interpreted from self-report measures . More elaborate tasks provide a high-quality method to quantify behaviors exhibited by BD patients in a laboratory setting and subsequently assess such behaviors in rodents. Therefore in this review, we will describe models of BD that specifically utilize translationally relevant tasks. While the use of basic tests with little specific relevance to abnormal patient behavior can be useful as a first pass in the screening of rodent models for BD, our focus will be placed on tasks that exist in both humans and animals from which direct comparisons of behavior performance can be made. Several lines of research have begun to utilize this methodology, resulting in promising behavioral paradigms in humans and rodents. Indeed, this translational approach is being used in other disorders as well, such as HIV and methamphetamine dependence , schizophrenia , and other substance use disorders . Here, the quantification of abnormal behavior in BD and attempts to recreate these behaviors in etiologically relevant models will be discussed. This review provides an invited summary of research presented as a Keynote Lecture at the International Behavioral Neuroscience Society in 2014, highlighting our laboratory’s approach toward delineating the neural mechanisms underlying BD. Mechanisms underlying the complex umbrella of symptoms in BD are as yet unresolved, complicating targeted treatment development. Nevertheless, several mechanisms have been elucidated that likely mediate both states of mania and depression in BD. The catecholaminergic-cholinergic balance hypothesis of BD states that functional levels of catecholamines are increased during manic states, whereas increased cholinergic functioning is more relevant to depression .

Among the catecholamines, dysfunctional dopamine neurotransmission is recognized to be a central factor in the pathophysiology of BD . The DA transporter is the primary reuptake mechanism of extracellular DA by which homeostasis is maintained . Polymorphisms in the gene encoding for DAT have been associated with BD , although these results have not been replicated in every genome wide association study . SNPs can reduce cell surface migration of DAT , down-regulating its functional expression. Subsequently, reduced striatal DAT levels have been observed in drug-free depressed and euthymic BD patients and in the postmortem tissue of BD patients . On the other hand, higher DAT binding has also been observed in the striatum of medicated depressed and unmedicated euthymic BD patients. Another symptom associated with BD is an abnormal circadian rhythm . For instance, altered rhythms in physiological parameters such as body temperature, plasma cortisol, and melatonin have been observed in patients with depression and BD , who also often experience altered sleep-wake cycles. Altered circadian rhythms can influence the release, synthesis, and levels of neurotransmitters such as DA . Taken together, these mechanistic findings provide targets which can be used to aid the development of animal models as well as treatments for BD.Previously, we described how increased motor activity and exploratory behavior described in patients with BD could be characterized using a multivariate translational approach called the human behavioral pattern monitor . In brief, the human BPM consists of a room that is novel to the test subjects and contains furniture and several items placed around the room to encourage exploration . Subjects are asked to wait in the room with no specification of the duration, unaware of simultaneously being tested on several ambulatory parameters. By measuring exploratory behavior of patients in this novel environment, a unique pattern of activity and exploration of manic and euthymic BD patients was identified. This abnormal exploratory pattern exhibited by BD patients included hyperactivity, increased specific exploration, and more linear patterns of movement, which importantly differed from control subjects , adult attention deficit hyperactivity subjects , and patients with schizophrenia . Because the human BPM was developed using a “reverse-translational” approach based on the BPM originally designed for testing rats or mice , it has been possible to examine mechanisms that underlie this specific pattern of exploration using the same manner of assessment of locomotor exploration in rodents. In brief, the BPM for rodents consists of a Plexiglas arena containing both floor and wall holes .

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Conditioned gaping in rats appears to be a selective index of conditioned nausea

Posted on July 3, 2023 by hemp2022

However, the data use agreement is designed to maximize transparency in the science, by requiring those who use the data resource to reference a versioned release of the data so that other investigators can repeat, expand, and challenge the results—particularly important in this era of renewed focus on scientific rigor and reproducibility. New standards for responsible use of large shared databases are emerging and are likely to guide editorial practices in the future, but the benefits of such databases for increasing transparency, enabling rapid replication, and generally accelerating scientific progress should outweigh the challenges. Finally, the ABCD Study may set new records for the number and disciplinary reach of scientists engaged by a single study. Over 200 investigators are actively engaged to date, and as the study evolves and the data emerge, a large number of additional scientists are expected to participate in the evolving design, data collection, analysis and interpretation, and many novel uses of the data. Furthermore, the availability of the data may draw scientists from wide-ranging disciplinary back grounds to the cause of understanding development of the human brain and finding innovative approaches for preventing adverse health and mental health outcomes and thus improving life trajectories.Chemotherapy-induced nausea and vomiting can be classified into three categories: acute onset, occurring within 24 h of the initial chemotherapy administration; delayed onset, occurring 24 h to several days after the initial treatment; and anticipatory nausea and vomiting . Anticipatory nausea develops in response to chemotherapy treatments,vertical cannabis farming in which cytotoxic drugs are delivered in the presence of a novel context . Developing in approximately 30% of patients by their fourth treatment , AN has traditionally been understood in terms of classical conditioning.

After one or more treatment sessions, a conditional association develops between the distinctive contextual cues of the treatment environment and the unconditioned stimulus of chemotherapy treatment that results in the unconditioned response of post-treatment illness experienced by the patient. Subsequent exposure to the treatment environment results in the patient experiencing a conditioned response of nausea and/ or vomiting before initiation of chemotherapy treatment. Once it develops, AN has been reported to be especially refractive to anti-emetic treatment . The evaluation of potential treatments for AN would be accelerated by the establishment of a reliable rodent model of nausea. Although rats are incapable of vomiting, they display characteristic gaping reactions when exposed to a flavoured solution previously paired with lithium induced nausea. In fact, this gaping reaction in the rat requires the same orofacial musculature as that required for vomiting in other species . Only drugs that produce emesis in species capable of vomiting produce conditioned gaping in rats, although many non emetic drugs produce conditioned taste avoidance . Furthermore, anti-emetic drugs interfere with the establishment of conditioned gaping reactions elicited by a nausea-paired flavor, presumably by interfering with the nausea .Not only are flavor cues capable of eliciting conditioned gaping reactions when paired with lithium chloride – induced nausea in rats, but recently Limebeer et al. have demonstrated that re-exposure to LiCl-paired contextual cues also elicit conditioned gaping reactions in rats. This paradigm more closely resembles that reported to produce AN in chemotherapy patients. Rats were injected with LiCl or saline before placement in a vanilla-odor laced chamber with lights and texture different than their home cage on each of four trials, separated by 72 h. To equate both groups for experience with illness, the rats in group unpaired were injected with LiCl and those in group paired were injected with saline 24 h after each conditioning trial but were then simply returned to their home cage. When the rats were returned to the conditioning context, 72 h after the final conditioning trial, rats in group paired showed the conditioned gaping reaction, as a measure of AN. Although classical anti-emetic treatments such as the 5- hydroxytryptamine-3 antagonist, Ondansetron , effectively reduce unconditioned nausea and vomiting, they are ineffective in the alleviation of conditioned nausea once it develops in humans . Indeed, OND also did not suppress the conditioned gaping reactions displayed during re-exposure to the LiCl-paired context .

Further more, using the emetic species, Suncus murinus as an animal model for AN, pre-treatment with a dose of OND that was shown to alleviate acute vomiting , did not reduce the display of conditioned retching reactions during re-exposure to a nausea-paired context . Thus, although OND has proven effective in the reduction of acute post-treatment nausea and vomiting, it does not appear to relieve conditioned nausea when it does develop. The endocannabinoid system has been implicated in control of nausea and vomiting . The psychoactive component in marijuana— delta-9-tetrahydrocannabinol —has been shown to interfere with the expression of vomiting in shrews and ferrets and conditioned gaping reactions elicited by a lithium-paired flavor in rats . The Δ9 -THC-induced suppression of conditioned nausea could be reversed by a CB1 receptor antagonist/reverse agonist , implicating the CB1 receptor in this effect . Limebeer et al. demonstrated that, unlike OND, Δ9 -THC also interfered with the expression of conditioned gaping elicited by the LiCl paired contextual cues in rats. This finding was consistent with the demonstration that Δ9 -THC, unlike OND, also suppressed the expression of conditioned retching in shrews when returned to a LiCl-paired context . The primary non-psychoactive compound found in marijuana, cannabidiol , has also been shown to suppress nausea and vomiting. In shrews, vomiting elicited by LiCl is suppressed by low doses of CBD, while higher doses were found to facilitate vomiting, rather than reducing its expression . Additionally, CBD reduced conditioned retching in shrews elicited by a lithium-paired context . In rats, a dose of 5 mg/kg CBD interfered with the establishment of conditioned gaping elicited by a LiCl paired flavor, as well as its expression . Because CBD has not been shown to bind with known CB receptors, this suppression of nausea and vomiting does not appear to be linked to activity of the CB1 or CB2 receptors . These results suggest that the primary nonpsychoac tive compound found in marijuana may be a useful treatment for conditioned nausea.

Arachidonylethanolamide or anandamide is an endogenous agonist for cannabinoid receptors which is rapidly degraded by the fatty acid amide hydrolase that is distributed throughout the brain and periphery . The action of AEA can be prolonged by inhibiting its degradation, through the use of URB597 , an FAAH enzyme inhibitor, that can increase basal levels of AEA in the rat brain . URB administration has been shown to reduce the establishment of conditioned gaping elicited by LiCl-paired saccharin solution in rats . Thus, prolonging the action of AEA with the FAAH inhibitor URB has been shown to suppress the establishment of conditioned nausea in rats. The experiments described below evaluated the potential of the non-psychoactive component of marijuana, CBD, and the FAAH inhibitor, URB, to interfere with conditioned gaping elicited by a LiCl-paired chamber in rats. On each of four conditioning trials,vertical farm company rats were injected with LiCl-paired immediately before placement in a distinctive context laced with the odor of vanilla extract. After the conditioning trials, the rats were injected with CBD or URB before placement in the distinctive CS context. Additionally, experiment 2 evaluated the potential of the CB1 antagonist/ inverse agonist, SR141716A , to reverse the suppression of conditioned gaping produced by URB. Finally, in experiment 3, the ability of URB to interfere with the establishment of conditioned gaping was also assessed. In each experiment, to ensure that the suppression of conditioned gaping was not merely an artefact of drug-induced suppression of general activity, the number of seconds that the rats remained immobile was recorded as a measure of activity. The doses of CBD were selected on the basis of our previous work demonstrating that these lower doses suppressed lithium induced vomiting in the shrew, but higher doses potentiated vomiting. Furthermore, a dose of 5 mg/kg of CBD interfered with the establishment and the expression of conditioned gaping elicited by a lithium paired flavour . The doses of URB were chosen based upon results showing that in vivo FAAH activity is blocked with a half-maximal inhibitory dose of 0.15 mg/kg ip with concurrent increase in brain AEA . Additionally, a dose of 0.3 mg/kg has been shown to attenuate the establishment of conditioned gaping elicited by a flavored stimulus .The subjects were male Sprague-Dawley rats and Long-Evans rats .

The change in strain across experiments coincided with a change in laboratory locations. There were no strain differences in baseline measures. The animals were group-housed in Plexiglas cages in the colony room at an ambient temperature of 21°C with a 12/12 light dark schedule and were maintained on an ad libitum schedule of food and water. All procedures adhered to the guidelines of the Canadian Council of Animal Care and were approved by the Animal Care Committee of University of Guelph.The distinctive context utilized for conditioning varied the dimensions of location, visual, tactile and olfactory cues from the home cage environment. The room was dark with two 50-Watt red lights on either side of the conditioning chamber. The conditioning chamber was made of opaque Plexiglas sides with an opaque lid. The chamber was placed on a table with a clear Plexiglas top. A mirror beneath the chamber on a 45° angle facilitated viewing of the ventral surface of the rat. Four plastic containers were permanently attached to holes on each side of the chamber in which a cotton dental roll saturated with vanilla flavour extract was placed to create the olfactory cue in the chamber. The cotton roll was inaccessible to the rat, with a newly saturated cotton roll used for each rat placed in the context.The rats received four conditioning trials, during which the contextual chamber was paired with 127 mg/kg LiCl. On each conditioning trial, each rat was injected with LiCl and immediately placed in the distinctive context for a 30-min period. This procedure was followed for a total of four conditioning trials, with 72 h between each trial.When re-introduced to a context previously paired with LiCl, rats display conditioned gaping ,a measure of conditioned nausea. The expression of this gaping reaction is not reduced by pre-treatment with a classic anti-emetic agent, OND, as has been reported by human chemotherapy patients; however, the psychoactive component in marijuana, Δ9 -THC, did suppress conditioned gaping elicited by a LiCl-paired chamber . In agreement with the gaping data with rats, Parker et al. reported that shrews display a conditioned retching reaction when re-introduced to a chamber previously paired with LiCl and this conditioned retching reaction was sup pressed by pre-treatment with Δ9 -THC or CBD, but not OND. The present findings provide additional support for the potential of cannabinoid treatments to suppress AN on the basis of results with the rat gaping model. Experiment 1 demonstrated that low doses of CBD that suppressed LiCl-induced vomiting and conditioned retching in the shrew as well as the establishment and the expression of gaping elicited by a LiCl-paired flavor in the rat , also suppressed the expression of gaping elicited by LiCl-paired context in the rat. A low dose of 5 mg/kg CBD has also been reported to serve as an effective anti-inflammatory agent , as well as a neuroprotective antioxidant . As CBD is a non-psychoactive component in marijuana, the ability of CBD to suppress conditioned gaping is promising for its use in the suppression of AN. The effective anti-nausea range of CBD appears to be narrow because at the highest dose tested , CBD did not suppress the expression of gaping elicited by a LiCl-paired context. Our previous work with the Suncus murinus revealed that although doses of 1–10 mg/kg CBD reversed LiCl-induced vomiting, higher doses potentiated LiCl-induced vomiting. It is possible, that CBD also produces a biphasic effect on the expression of conditioned gaping with lower doses reducing the expression of conditioned gaping, but higher doses enhancing the expression of gaping elicited by a LiCl-paired context. Experiments 2 and 3 demonstrated that the FAAH inhibitor, URB, also suppressed the display of AN as measured by gaping in rats. Presumably, prolonging the action of endogenous AEA produced an anti-nausea effect when rats were re-exposed to the LiCl-paired context. The effect of URB on the expression of conditioned gaping was dose-dependent, with 0.3 mg/kg serving as the effective dose.

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Electron micrographs were taken at 30,000– 50,000× magnifications with a Hitachi 7100 electron microscope

Posted on June 30, 2023 by hemp2022

Further high-resolution immunostaining and electron microscopic analysis in the human hippocampal formation and neocortex have narrowed down the presence of CB1 receptors to GABAergic boutons and also to glutamatergic axon terminals . Together, these findings contribute to the hypothesis that 2-AG may be a synaptic messenger in the human nervous system. However, despite their potential therapeutic significance and their prominent mRNA expression levels in the human hippocampus , the precise location of two key enzymes, DGL-α and MGL, known to regulate 2-AG signaling at chemical synapses in rodents have not yet been investigated in detail in the human brain. The aim of our study was therefore to uncover the precise molecular organization of the 2-AG signaling pathway at excitatory synapses in the human hippocampus by using novel antibodies with confirmed target specificity for DGL-α and MGL, as well as light and high-resolution electron microscopy. Control hippocampi were kindly provided by the Lenhossék Human Brain Program, Semmelweis University, Budapest. Control subjects died suddenly from causes not directly involving any brain disease, and none of them had a history of any neurological disorders. The control subjects were processed for autopsy in the Department of Forensic Medicine of the Semmelweis University Medical School, Budapest, and the brains were removed 2–5 h after death. Informed consent was obtained for the use of brain tissue and for access to medical records for research purposes. Tissue was obtained and used in a manner compliant with the Declaration of Helsinki. All procedures were approved by the Regional and Institutional Committee of Science and Research Ethics of Scientific Council of Health . After postmortem removal, the hippocampal tissue was immediately dissected into 3- to 4- mm-thick blocks, and immersed in a fixative containing 4% paraformaldehyde, 0.1% glutaraldehyde, and 0.2% picric acid in phosphate buffer . The blocks were first rinsed for 6 h at room temperature in the fixative, which was replaced every hour with a fresh solution.

The blocks were then post fixed overnight in the same fixative solution,vertical farming startup but without glutaraldehyde. In the case of one control brain, both the internal carotid and vertebral arteries were cannulated 4 h after death, and the brain was perfused with physiological saline followed by a fixative solution containing 4% paraformaldehyde and 0.2% picric acid in PB . The hippocampus was removed after perfusion, and was cut into 3- to 4-mm-thick blocks, and was post fixed in the same fixative solution overnight. Subsequently, 60-µm-thick coronal sections were prepared from the blocks with a Leica VTS-1000 Vibratome for immunohistochemistry.Immunostaining of the slices was performed as described previously . Briefly, after washing in PB , sections were cryoprotected in 10% sucrose and in 30% sucrose in PB for 15 min and overnight, respectively, then freeze thawed four times in an aluminium-foil boat over liquid nitrogen to enhance penetration of the antibodies without destroying the ultrastructure. Residual sucrose was washed from the tissue in PB and then endogenous peroxidase activity was blocked for 10 min by treatment with 1% H2O2 dissolved in PB. Subsequently, all washing steps and antibody dilutions were carried out in Tris-buffered saline . After extensive washing in TBS , sections were first blocked with 5% normal goat serum for 45 min, washed in PB for 15 min, and then incubated with the primary antibody of interest for 48 h. The following polyclonal, affinity-purified primary antibodies were used in the present study: rabbit anti-DGL-α ; rabbit anti-MGL ; and rabbit anti-MGL . The specificity of the DGL-α-INT antibody was confirmed by the lack of immunostaining in hippocampal sections derived from DGL-α knockout mice . The specificity of the two MGL antibodies was supported by immunostaining in HEK293 cells transiently expressing a V5 epitope-tagged MGL; by the lack of immunostaining in neurons pre incubated with 5 µg/ml of the corresponding immunizing protein; and by the identical staining pattern in hippocampal sections at the electron microscopic level with the two antibodies raised against independent epitopes of the MGL protein .

After primary antibody incubation, human and mouse hippocampal sections were washed extensively in TBS , then treated first with biotinylated goat anti-rabbit IgG for 2 h, washed again three times in TBS, and then incubated with avidin-biotinylated horseradish peroxidase complex for 1.5 h. This step was followed again by washing in TBS and in Tris buffer , and finally the immunoperoxidase reaction was developed using 3,3- diaminobenzidine as chromogen and 0.01% H2O2 dissolved in TB. After the development of immunostaining, sections were washed in PB, treated with 1% OsO4 in 0.1 M PB for 20 min, dehydrated in ascending series of ethanol and acetonitrile, and embedded in Durcupan . During dehydration, sections were also treated with 1% uranyl acetate in 70% ethanol for 20 min. After overnight incubation in Durcupan, the sections were mounted onto glass slides and coverslips were sealed by polymerization of Durcupan at 56 °C for 48 h. Light microscopic analysis of immunostaining was carried out with a Nikon Eclipse 80i upright microscope , and light micrographs were taken with a DS-Fi1 digital camera . For electron microscopic investigations, selected immunoreactive profiles and regions were photographed and re-embedded for further ultrathin sectioning. Series of consecutive ultrathin sections were collected on Formvar-coated single-slot grids and counter stained with lead citrate for 2 min.For immunofluorescence double staining, after freeze-thawing and intense washing, the sections were first blocked with 5% normal donkey serum for 45 min, and then incubated with mouse anti-NeuN and either with rabbit anti DGL-α , or rabbit anti-MGL , or rabbit anti-MGL primary antibodies for 48 h. Afterward, the sections were washed again in TBS three times for 15 min each, then incubated with secondary antibodies Alexa 594- conjugated goat anti-mouse IgG and DyLight 488- conjugated donkey antirabbit IgG for 2 h.

Excess secondary antibody was washed out three times in TBS, and three times in 0.1 M PB for 15 min each. Finally, the sections were mounted in Vectashield onto glass slides, and the coverslips were sealed with nail polish. Image acquisition was performed with an inverted Nikon Eclipse Ti-E microscope equipped with an A1R confocal system . Images of double labeling were obtained of a single focal plane by a 4× objective in sequential scanning mode using a four channel PMT detector. For the adjustment of digital light and electron micrographs, Adobe Photoshop CS2 was used. In all imaging processes, adjustments were done in the whole frame and no part of an image was modified separately in any way. To reveal the site of synthesis of the endocannabinoid 2-AG in the human hippocampus by determining the localization of its predominant synthesizing enzyme DGL-α , we first sought to identify an antibody with unequivocal specificity for this transmembrane serine hydrolase. Therefore, DGL-α- immunostaining was performed and compared in hippocampal sections derived from wild type or DGL-α knockout mice . Using an affinity-purified antibody raised against a large intracellular loop on the C-terminus of DGL-α , immunoperoxidase reaction revealed at low magnification that the general dense distribution of DGL-α-immunostaining followed the topographic arrangement of glutamatergic pathways in the wild-type hippocampus . In contrast, the immunoreactive material was almost fully absent in the DGL-α knockout hippocampus confirming the specificity of the “DGL-α INT” antibody . At higher magnification, the differences in staining intensity between the somatic and dendritic layers were even more pronounced . While nuclei and cell bodies in the principal cell layers were largely devoid of DGL-α-immunore activity, an intense punctate staining pattern was observed throughout the neuropil in those layers, which contain a high density of excitatory synapses in the hippocampus . This was in accordance with the observations we have reported earlier using this antibody in the hippocampus and in other regions . On the other hand, this punctate labeling was largely missing in DGL-α knockout hippocampi . Therefore, in the next set of experiments,vertical urban farming we incubated hippocampal sections derived from human subjects together with hippocampal sections derived from wild-type C57BL/6 mice using the “DGL-α INT” antibody. At low magnification, immunofluorescence staining for DGL-α was unevenly distributed throughout the human hippocampal formation . This pattern followed the laminar organization of the hippocampus and was found to be largely similar in mice . At higher magnification, the highest density of DGL-α- immunoperoxidase reactivity was observed in the strata oriens and radiatum of the cornu ammonis subfields, and in the inner molecular layer of the dentate gyrus , whereas somewhat weaker, but still significant density of DGL-α-immunoreactivity was found in the strata pyramidale and lacunosum-moleculare of the cornu ammonis and in the outer two-third of the stratum moleculare . Somata of pyramidal cells and dentate gyrus granule cells contained only very low amount of DGL-α-immunolabeling. At even higher magnification, the punctate staining pattern also showed striking similarities with the pattern observed in wild-type mice . This widespread granular pattern of DGL-α-immunoreactivity was visible throughout the hippocampal formation, but its distribution varied with regard to given subcellular profiles. For example, in the stratum radiatum of the CA1 subfield, DGL-α-positive granules were distributed along the main trunk of the apical dendrites of pyramidal cells, whereas the trunk itself was devoid of immuno staining . Similarly, apical and possibly oblique dendrites of granule cells also appeared to be outlined on their surface by dense DGL-α-immunolabeling.

To reveal the precise subcellular position of DGL-α in principal cells of the human hippocampus, we first tested the specificity of the “DGL-α INT” antibody at the ultrastructural level . Hippocampal sections from mice with different genotypes were processed together within the same incubation wells to ensure identical treatment throughout the imunostaining procedure. Further high resolution electron microscopic analysis in samples taken from the stratum radiatum of the CA1 subfield of wild-type hippocampus revealed that DGL-α-immunoreactivity was predominantly concentrated in dendritic spine heads receiving asymmetric, putative excitatory synapses, in accordance with previous findings . Altogether, at least ~24% of dendritic spine heads were unequivocally positive for DGL-α immunoreactivity in our wild-type random samples ; this ratio should be treated as a conservative estimation restricted by epitope accessibility. In contrast, under identical staining condition, only two out of 201 spine heads contained weak immuno peroxidase reaction end product in sections taken from the DGL-α knockout mouse , indicating the low level of background in this immuno staining experiment. To determine whether in the human hippocampus the same subcellular domain, namely the postsynaptic spine head, corresponds to the punctate staining pattern observed at the light microscopic level, hippocampal sections from human subjects with DGL-α- immunostaining were also processed for further electron microscopic analysis. Two regions were selected for detailed investigations, the stratum radiatum of the CA1 region and the inner third of the stratum moleculare of the dentate gyrus . In both regions, the DAB end product of the immunoperoxidase staining procedure, representing the subcellular position of DGL-α, was concentrated in dendritic spine heads protruding from DGL-α-immunonegative dendritic shafts. Because the majority of hippocampal GABAergic interneurons, including for example basket cells are aspiny, therefore the widespread occurrence of DGL-α in this characteristic subcellular compartment also reveals that principal cells express this enzyme in the human hippocampus. Notably, the DAB precipitate was consistently present within the spine heads through consecutive ultrathin sections. In contrast to this high concentration of DGL-α in dendritic spines, intensity of DGL-α-immunoreactivity did not reach the detection threshold in other subcellular profiles like excitatory and inhibitory axon terminals, or glial processes in the human hippocampus . Taken together, these data ultimately confirm previous findings that DGL-α accumulates postsynaptically in dendritic spines of principal cells in the mouse hippocampus and suggest that this 2-AG-synthesizing enzyme has a conserved function in the regulation of retrograde endocannabinoid signaling based on its entirely similar postsynaptic localization at excitatory synapses in the mouse and human hippocampus. If the enzyme responsible for 2-AG biogenesis is postsynaptically located , whereas its receptor is presynaptically positioned , then the next important question is where the 2-AG signal is terminated at excitatory synapses in the human hippocampus. Because MGL knockout mice have not yet become available to use as specificity controls, we employed two independent antibodies recognizing different epitopes of the MGL protein to characterize the regional distribution and subcellular localization of 2-AG’s principal hydrolyzing enzyme, MGL in the human hippocampal formation.

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