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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Anandamide synthesis can be elicited in vitro by a variety of agents that elevate intracellular Ca2+ levels

Posted on June 29, 2023 by hemp2022

Although separate enzymes cata lyze the syntheses of anandamide and NAPE, the two events are likely to occur simultaneously because Ca2+- stimulated anandamide production is often accompanied by de novo formationof NAPE .For example, the Ca2+ ionophore ionomycin stimulates [ 3 H]anandamide formation in cultures of rat striatal and cortical neurons labeled by incubation with [ 3 H]ethanolamine . In the same neurons, Ca2+-dependent [3 H]ana ndamide production may be elicited by the glutamate receptor agonist, kainate, by the K+ channel blocker 4- aminopyridine, and by membrane-depolarizing con centrations of K+ ions . Depolarizationof neural cells was also shown to evoke Ca2+-dependent anandamide release in vivo . Along with Ca2+ entry, activation of certain G protein-coupled receptors can also initiate anandamide generation. Administration of the dopamine D2-recep tor agonist quinpirole causes a profound stimulation of anandamide synthesis in the rat basal ganglia, which is prevented by the D2 antagonist raclopride . Importantly, cocaine elicits a similar response , suggesting a role for anandamide in the actions of these psychostimulant drugs. The ability of the anandamide transport inhibitor AM404 to reduce D2 agonist induced hyperactivity, discussed below,vertical farming market further sup ports this possibility .The biological elimination of anandamide proceeds through two successive steps of high-affinity transport into cells, followed by intracellular degradation . Brain neurons and astrocytes in culture internalize anandamide through a process that fulfills all key requirements of a carrier-mediated transport. Plots of the initial rates of [3 H]anandamide internalization in rat brain neurons and astrocytes in culture yield apparent Michaelis constants that are consistent with a saturable process and are comparable to the KM values of brain amine or amino-acid transporters .

Moreover, neurons and astrocytes in culture internalize [3 H]anandamide, along with a select group of structurally related compounds, in a stereoselective manner . Even further, [3 H]ana ndamide internalization can be inhibited by drugs that have no effect on the uptake of non-cannabinoid lipid mediators such as fatty acids and eicosanoids. Nevertheless, anandamide is internalized in a Na+- and energy-inde pendent manner , a feature that differentiates this lipid mediator from most conventional neurotransmitters.The prototype of this class of drugs, the arachido nate derivative AM404 , has provided important information on the properties of anandamide transport, not only aiding the in vitro characterization of this process, but also helping to reveal its possible functions in animals. Importantly, the partial cannabimetic profile exhibited by this agent in vivo suggests that anandamide transport might provide a useful tar get in disease conditions in which the endocannabinoid system is hypofunctional . Evidence indicates that one such condition could be opiate withdrawal, which is markedly reduced in rodents by administering AM404 . These theories have been hindered by the fact that the putative transport system responsible for anandamide internalization is still uncharacterized at the molecular level. In fact, the presence of such a system has been recently questioned, based on the observation that [ 3 H]anandamide uptake in certain cell lines is saturable at longer , but not at shorter incu bationtimes . This finding has been interpreted to suggest that fatty-acid amide hydrolase —a key enzyme of intracellular anandamide degradation, described in a subsequent section—may be responsible for the saturation of uptake noted at longer incubation times . However, the result may also be explained on purely technical grounds, as the high concentration of serum albumin used in the experiments of Glaser and collaborators was previously shown to prevent [3 H]anandamide internalization .

Consistent with this interpretation, recent studies have provided additional evidence for the existence of an anandamide transport system independent of FAAH . In particular, one of these studies has shown that cultures of cortical neurons isolated from the brain of FAAH null mice internalize anandamide as efficiently as do neurons that express normal levels of the enzyme. The same study also demonstrated that the transport inhibitor AM404 is equally effective at reducing anandamide internalization in neurons of FAAH-null and wild-type mice. These results indicate that FAAH does not pro vide the driving force for anandamide uptake or serve as a target for AM404. Invivo experiments further support this conclusion, showing that AM404 not only enhances the actions of exogenous anandamide in FAAH-null mice, but acts more effectively in this mutant strain than it does in control animals. This implies that AM404 is not in fact a FAAH inhibitor, as it has been proposed , but a FAAH substrate. In support of this idea, it was found that membranes prepared from the brains of normal mice rapidly hydrolyze AM404, whereas those prepared from mice that lack FAAH are unable to carry out this reaction.The fact that FAAH is not directly involved in anandamide internalization raises the question of what mechanism provides the driving force for this process. One possibility is that an intracellular protein may sequester anandamide at the membrane, driving its internalization and facilitating its movement to the mitochondria and the endoplasmic reticulum, where FAAH is primarily localized . If selective for anandamide, such a protein might participate in the transport process as well as serve as a target for transport inhibitors. This hypothetical model is consistent with fatty acid transport into cells, which is also thought to require the cooperation of membrane transporters and intracellular fatty-acid binding proteins .AM404 increases endogenous anandamide levels in brain tissue and peripheral blood of rats and mice . This effect is accompanied by a series of behavioral responses that, though blocked by the CB1 antagonist rimonabant , are clearly distinguishable from those of direct cannabinoid agonists. For example, administra tion of AM404 into the cerebral ventricles of rats decreases exploratory activity without producing cata lepsy and analgesia, two hallmarks of direct CB1 receptor activation.

Inaddition, AM404 reduces two characteristic effects caused by activationof D2 family receptors: the yawning response elicited in mice by low doses of the D1/D2-receptor agonist apomorphine; and the stimu lationof locomotor activity evoked inrats by the D2- receptor agonist quinpirole . These effects are observed at doses of AM404 that selectively target anandamide transport and produce only mild hypokinesia when the drug is administered alone . The results of this study, which have been confirmed in several subsequent reports , demarcate the pharmacological profile of AM404 from those of direct-acting cannabinoid drugs. This distinction may result from the ability of AM404 to enhance anandamide signaling in an activity-dependent manner, causing anandamide to accumulate in discrete regions of the brain and only when appropriate stimuli initiate its release. Pharmacological activation of D2 receptors may represent one such stimulus, suggesting that blockade of anandamide transport might offer an innovative strategy to correct abnormalities associated with dysfunction in dopaminergic transmission. Initial tests of this hypothesis have shown that systemic administration of AM404 normalizes movement in spontaneously hypertensive rats , an inbred line in which hyperactivity and attention deficits have been linked to a defective regulation of mesocorticolimbic dopamine pathways .FAAH was first identified as an amide hydrolase activity present in rat liver tissue, which catalyzes the hydrolysis of the fatty-acid ethanolamides palmitoy lethanolamide and oleoylethanolamide . That anandamide serves as a substrate for this activity was first suggested onthe basis of biochemical evidence and later demonstrated by molecular cloning,vertical farming pros and cons heterologous expression and generation of FAAH null mice by homologous recombination . FAAH belongs to a group of enzymes known as ‘amidase signature family’ and catalyzes the hydrolysis not only of anandamide and other fatty-acid ethanola mides, but also of primary amides such as oleamide and even of fatty-acid esters such as 2-AG . Elegant site-directed mutagenesis and X-ray diffraction studies have demonstrated that this unusually broad substrate preference is due to a novel catalytic mechanism involving the amino-acid residue lysine 142. This residue may act as a general acid catalyst, favoring the protonation and consequent detachment of reaction products from the enzyme’s active site . Three serine residues that are conserved in all amidase signature enzymes also may be essential for enzymatic activity: serine 241 may serve as the enzyme’s catalytic nucleophile, while serine 217 and 218 may modulate catalysis through an as-yet-unidentified mechanism . Electron microscopy experiments in the rat and mouse brain have shown that FAAH is predominantly, if not exclusively localized to intracellular membrane com partments, particularly to the endoplasmic reticulum and the mitochondria . Although FAAH appears to be the predominant route of anandamide hydrolysis in the brain, other enzymes are likely to participate in the breakdown of this endocannabinoid in peripheral tissues. An acid amide hydrolase activity catalytically distinct from FAAH has been characterized in human mega karyoblastic cells and shown to be highly expressed in the rat thymus, lungs and intestine .The search for small-molecule inhibitors of intra cellular FAAH activity has led to the emergence of several potent and selective agents, which include substituted sulfonyl fluorides , alpha-keto-oxazolopyridines an d carbamic acid esters .

The latter were identified during structure– activity relationship studies aimed at determining whether esters of carbamic acid such as the insecticide carbaryl inhibit FAAH activity. It was found that, although carbaryl is ineffective in this regard, variations in its template result in significant inhibitory potencies. Further structural optimizations yielded a group of highly potent inhibitors, a representative example of which is provided by the compound URB597 . Kinetic and dialysis experiments indicate that URB597 interacts non-competitively with FAAH, which is suggestive of anirreversible or slowly reversible association with the enzyme. Importantly, URB597 has no notable effect on CB1 or CB2 binding, anandamide transport, or rat brain monoglyceride lipase , a cytosolic serine hydrolase that catalyzes the hydrolysis of the second endocannabinoid, 2-arachido noylglycerol. Following administration to rats in vivo, URB597 produces profound, dose-dependent inhibition of brain FAAH activity. After injection of a maximal dose of compound , FAAH inhibitionis rapid , persistent and associated with a 3-fold increase in brain anandamide levels. Furthermore, the inhibitor intensifies and pro longs the effects produced by exogenous anandamide, yet it elicits no overt cannabinoid-like actions when administered alone; for example, it does not cause hypothermia, hot-plate analgesia, or hyperphagia .Although URB597 does not display a typical canna binoid profile in live animals, it exerts several pharma cological effects that might be therapeutically relevant. One such effect, the ability to reduce anxiety-like beha viorsinrats, was demonstrated in two distinct experimental models: the elevated ‘zero maze’ test, and the isolation-induced ultrasonic emission test . The ‘zero maze’ consists of an elevated annular platform with two open and two closed quad rants and is based on the conflict between an animal’s instinct to explore its environment and its fear of open spaces where it may be attacked by predators . Benzodiazepines and other clinically used anxiolytic drugs increase the proportion of time spent in, and the number of entries made into, the open com partments. In a similar fashion, URB597 elicits anxio lytic-like responses at a dose that corresponds to those required to inhibit brain FAAH activity. Moreover, these effects are prevented by the CB1-selective antagonist rimonabant. Analogous results were obtained in the ultrasonic vocalization emission test, which measures the number of stress-induced vocalizations emitted by rat pups removed from their nest . If confirmed in further behavioral models, these findings would suggest that inhibition of intracellular FAAH activity might offer  an innovative target for the treatment of anxiety , which is also a feature of marijuana withdrawal .2-AG was identified as a second endocannabinoid substance in 1995 . The multiple roles of this lipid compound in cell metabolism and its high levels in brain tissue— about 200-fold higher than those of anandamide—suggest that much of cellular 2-AG may be involved in housekeeping functions. The diversity of roles played by this compound also complicates our efforts to establish biochemical route involved in its physiological formation. Nevertheless, one pathway has emerged as the most likely candidate . This pathway starts with the phospholipase-mediated generation of 1, 2- diacylglycerol . This serves as a substrate for two enzymes: DAG kinase, which catalyzes DAG phosphorylationto phosphatidic acid; and DAG lipase , which hydrolyzes DAG to monoacylglycerol . Pharmacological inhibition of phospholipase C and DGL prevent the Ca2+-dependent accumulation of 2-AG in rat cortical neurons, which suggests a key role of this pathway in2-AG generation . However, additional routes of 2-AG synthesis also may exist, including phospho lipase A1 , hormone-sensitive lipase or a lipid phosphatase . In neurons and glia, 2-AG synthesis may be initiated by a rise incytosolic Ca2+ levels. For example, incultures of rat cortical neurons, the Ca2+ ionophore ionomycin and the glutamate receptor agonist N-methyl d-aspartate stimulate 2-AG productionina Ca2+-dependent manner .

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A parallel mixed-effects, logistic regression model was used for the dichotomous outcome

Posted on June 28, 2023 by hemp2022

Standardized fidelity ratings were made on 50% of randomly selected CBT sessions. These showed acceptable to excellent fidelity on all domains.The study employed a Yang-style tai chi delivered weekly for one hour following each CBT session. Each session started with a 10-minute warm-up, stretching and review of tai chi principles followed by 30 minutes of tai chi exercises, including five animal forms, a walking meditation, and a partnered activity known as push hands. Each class ended with a 10-minute cool down and a 5-minute closing that included a review of the material presented. A tai chi instructor with 18 years of experience trained two APAIT exercise program staff to lead the tai chi exercises. The staff underwent training for 1 hour weekly for 3 months before they began to lead tai chi sessions in the study. The trainer also attended four tai chi study sessions with each of the study instructors during the trial to monitor staff members’ instruction and provide feedback and adjustments as needed. Between November 2015 and April 2016, participants were recruited by a research assistant who distributed flyers at venues serving PLWH, including APAIT, gave presentations at nearby health agencies, and approached potential participants at health centers. A flow diagram shows the number of individuals approached, screened, and recruited, as well as losses to follow-up. Randomization was conducted by research staff who used consecutively numbered, sealed envelopes containing assignment information using a computer-generated set of random numbers to select permutated blocks of six. Within each block, equal numbers were assigned to each of the three groups. Participant follow-up concluded in July, 2016. Participants were compensated for their time via gift cards. Participants in the CBT/TC/TXT arm could be compensated up to $280,indoor vertical farm those in the SG arm could be compensated up to $200, while subjects in the AO arm could be compensated up to $120. Compensation included $10 for attending each CBT, TC or SG session.

Demographic and health-related data included date of birth, gender, race/ethnicity, education, marital status, housing arrangement, employment status, number of years living with HIV, most recent CD4+ T lymphocyte count and HIV-1 RNA [detectable or undetectable ] and number of non-HIV chronic medical conditions. Participants also completed mental and physical health  measures . Substance use data included most often used substance and the total number of substances used. Substances included both drugs and alcohol. Substance use measures included the WHO ASSIST-Version 3 and the Timeline Follow back . The TLFB was used to determine the number of days in the past 30 days of a) using a preferred substance; b) using any substance; c) using any drugs; and d) heavy drinking . Pain data included number of years of chronic pain, and medications used to treat pain. Pain measures included two items from the abbreviated Brief Pain Inventory , i.e., average self-rated pain intensity and percent pain relief in the past 24 hours from pain treatments , the Pain Self-Efficacy Questionnaire, and the modified Roland-Morris Disability Questionnaire. Data were collected via self-administered paper and pencil surveys. Physical performance was assessed with the Short Physical Performance Battery . We calculated a total SBBP score for each participant and determined the percentage of participants with low physical performance versus high physical performance. In person assessments were conducted at APAIT at baseline, and at 8- and 12- weeks after the baseline assessment. All measures were collected at baseline. In addition, the substance use measures based on the TLFB, the pain measures including the BPI, PSEQ and RMDQ and the physical performance measure, the SPPB were collected again at 8- and 12-weeks. Feasibility was assessed by a) success of recruitment and randomization, b) retention and treatment engagement rates, and c) feedback about the study from participants. Bivariate analyses were used to a) compare groups at baseline to assess the success of randomization, b) compare treatment retention and engagement between groups, and c) determine associations between baseline variables, treatment adherence and engagement. Measures of treatment efficacy included reductions in substance use, pain, and pain-related disability, as well as improvements in physical performance. Changes in outcome variables between baseline and both 8 weeks and 12 weeks are reported as means, standard deviations and medians for each group. Because the data were not normally distributed, we used the non-parametric, Wilcoxon signed rank test in each group to test if the within-individual change in the group was statistically significantly different from zero change.

To assess the preliminary efficacy of the intervention, we conducted an intention-to-treat analysis-all individuals randomized in the study were included in the analyses whether they received treatment or not using baseline, 8-, and 12-week follow-up data for each dependent variable. Linear, mixed-effects models were used to evaluate the five continuously scaled outcomes , with treatment group as a between-individuals factor , time as a within-individuals factor , and a group-by-time interaction to examine differences in the magnitude of change between groups.Poisson regression models were used for the four count outcomes . From the fitted model, to estimate treatment effects, we evaluated three between-group pairwise comparisons of change over time. Poisson regression models estimate within-group change as a count ratio , and the treatment effect estimate is the ratio of two CRs. The treatment effect estimates from the linear regression models are between-group differences in the magnitude of the change from baseline . The logistic regression model estimates within-group change as an odds ratio ,and the treatment effect estimate is the ratio of two ORs. Since change at 12 weeks was greater than the change at 8 weeks for most outcomes, we treated time as a continuous variable: actual number of days elapsed from baseline to the date of the follow-up, divided by 30 . These models were adjusted for baseline number of different types of substances used because 12-week follow-up participation rates varied significantly by baseline values of this variable Most used substances included alcohol, cannabis and stimulants, and the mean number of substances used by each participant was 2.69 . ASSIST scores indicated that most participants had moderate or high risk substance use. On average, in the prior 30 days, participants reported using their preferred substance, any substance, and drugs on more than half of the days, as well as 3.2 days of heavy drinking . Participants reported having chronic pain an average of 10.6 years. The mean pain intensity score was 6.9, and percent pain relief provided by any treatment in the past 24 hours was 50.7%. The mean score on the Short Physical Performance Battery was 8.0 and the percentage of those with low physical performance on the SPPB was 48%.

From baseline to 12 weeks, the CBT/TC/TXT group had statistically significant reductions in all four substance use outcomes: days of using preferred substance in the past 30 days , days of using any substance in past 30 days , days of using drugs in past 30 days , and days of heavy drinking in past 30 days . The support group also showed a statistically significant reduction in the number of days of preferred substance use in the past 30 days,vertical farming equipment from baseline to 12 weeks .Statistically significant improvements were observed in both physical performance outcomes in the CBT/TC/TXT group from baseline to 8 weeks and from baseline to 12 weeks . The median change in % with low physical performance was 0 but the signed rank sums are statistically different at each time point. The SG showed a statistically significant improvement in SPPB score from baseline to 12 weeks .Significantly greater reductions were observed in the CBT/TC/TXT group, relative to the AO group in three of the four substance use outcomes . As compared to the AO group, the CBT/TC/TXT group had a 2% greater relative reduction per month in the number of days of using their preferred substance, a4% greater relative reduction per month in the number of days of using drugs in the past 30 days, and a 19% greater per month reduction in number of days of heavy drinking in the past 30 days. We observed similar findings when comparing the CBT/TC/TXT group to the SG . Sixteen CBT/TC/TXT participants provided feedback about the study. Overall feedback was very favorable; two-thirds wished that the eight-week program and the one hour sessions lasted longer, while the remainder thought the individual program components and session length were adequate. Most rated the CBT group and tai chi sessions as extremely useful and enjoyed working with the therapist/instructor . About 70% of the CBT/TC/TXT participants indicated that they would definitely attend this type of group therapy if it were offered by APAIT with no compensation and not as part of a study. Suggestions for additional topics to address included stigma, HIV and aging, and more education about various medical conditions. Two participants suggested including a male facilitator living with HIV and one suggested having the groups available in Spanish. Related to the text messaging component, 69% reported it was extremely important in helping them to make or maintain a change. Fifty-six percent would have liked to receive the messages for longer than 8 weeks, and 50% thought two motivational texts a week was not enough. This study demonstrated that a multi-component behavioral intervention addressing substance use, pain, and physical performance in older PLWH with comorbid substance use and pain disorder conducted in partnership with a community-based agency is feasible, acceptable and has preliminary efficacy.

Importantly, our enrolled population used multiple substances, had ASSIST scores indicating moderate to high risk substance use, long term chronic pain and high rates of low physical performance at baseline. Feasibility and acceptability indicators showed moderate levels of participant enrollment , excellent 12-week assessment completion , acceptable to excellent CBT treatment fidelity ratings, and high attendance at CBT and tai chi sessions . Our qualitative data highlight overall positive program feedback and provide suggestions for changes to study procedures to enhance study efficacy. Anecdotally, we also learned from study staff about some of the barriers to attending the group based sessions including illness, drug use, lack of stable housing, and transportation issues. We also learned that most participants did not complete the homework assigned to them as part of the CBT component. We hypothesized that the intervention would lead to reductions in substance use and pain outcomes and improve physical performance. Looking first at within-group changes, statistically significant improvements from baseline to 12 weeks were observed in the CBT/TC/TXT group for all four substance use outcomes, one pain outcome and both physical performance measures. In the SG, from baseline to 12 weeks, statistically significant improvements were seen for one substance use and one pain and one physical performance outcome. In the AO group, no statistically significant within group improvements were observed from baseline to 12 weeks. Looking next at between-group changes, we observed several statistically significant between-group changes, most importantly in days of heavy drinking and in the SPPB score. The substance use change scores were modest except for days of heavy drinking in which, compared to the AO group, the CBT/TC/TXT group and SG had large relative reductions . We also observed a significant between group treatment effect with respect to perceived relief obtained from pain treatments over the past 24 hours suggesting that the intervention may improve quality of pain management. This is important given that both the mean and median self rated pain intensity among participants at the time of enrollment was moderately severe . We did not, however, observe any meaningful treatment-related reductions in pain intensity or perceived disability due to pain. Possible explanations for these latter findings include that most participants’ pain was of a neuropathic origin, which has not been shown to respond to psychological interventions when used as stand-alone therapy . In addition, our intervention merged elements of pain and substance use behavioral treatments such that the dose of pain coping skills training, behavioral activation, and cognitive restructuring may have been insufficient. Also as participants did not complete the homework exercises, which serve to reinforce the use of the techniques and suggests that adoption of the behavioral strategies to manage pain both during and after the intervention period was limited. With respect to the physical performance outcomes, SPPB score improved in the CBT/TC/TXT group relative to both the AO and SG of a magnitude indicative of meaningful improvement .

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Multi-Tier Mobile Grow System Widely Used in Vertical Farming Business

Posted on June 27, 2023 by hemp2022

A widely used multi-tier mobile grow system in vertical farming is the vertical racking system. This system allows for efficient space utilization and easy mobility within a vertical farm. Here are some key features and benefits of this system:

  1. Vertical Space Utilization: The vertical racking system consists of multiple tiers or shelves that can be stacked vertically, maximizing the use of vertical space in a controlled environment. This enables growers to grow a larger number of plants in a smaller footprint.
  2. Easy Mobility: The mobile feature of the system allows the racks or shelves to be moved easily within the facility. This flexibility enables better utilization of space, simplifies maintenance and harvesting processes, and provides easier access for workers.
  3. Customizable Configurations: The racks or shelves can be designed and configured to accommodate different sizes of plant containers, such as trays, pots, or grow bags. This flexibility allows for the cultivation of a variety of crops and facilitates easy management of different growth stages.
  4. Adjustable Lighting and Environmental Control: The vertical racking system can be equipped with adjustable LED grow lights, climate control systems, and irrigation systems. This enables precise control over lighting, temperature, humidity, and nutrient delivery, optimizing plant growth conditions.
  5. Improved Workflow and Efficiency: With a mobile system, growers can easily reposition the racks or shelves according to their workflow needs. This reduces labor requirements, streamlines cultivation processes, and improves overall operational efficiency.
  6. Scalability and Expansion: The modular design of the vertical racking system allows for scalability and expansion as the business grows. Additional tiers or units can be added to increase growing capacity and meet growing market demands.
  7. Better Pest and Disease Management: The controlled environment provided by the vertical racking system helps minimize the risk of pest infestations and diseases. It allows for better monitoring and implementation of preventive measures, reducing the need for chemical pesticides and promoting healthier plant growth.
  8. Easy Maintenance and Cleaning: The mobile nature of the system makes it easier to clean and maintain the growing area. It allows for convenient access to all plants and equipment, facilitating routine maintenance tasks such as pruning, plant inspections, and system cleaning.

Vertical racking systems have gained popularity in vertical farming due to their versatility, efficient space utilization, and improved workflow. They are suitable for a wide range of crops, including leafy greens, herbs, microgreens, and even some fruiting plants. However, it’s important to consider specific business requirements, facility constraints, and crop characteristics when selecting and implementing a multi-tier mobile grow system in a vertical farming business.

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