Cases missing outcome data were typically dropped from analyses

While the women in the qualitative study explicitly described these experiences related to drug and not alcohol use, it is plausible that they apply to alcohol as well. Thus, policies that require informing women that their substance use may have already harmed their fetus, such as MWS policies, could lead women to avoid prenatal care. Policies that mandate reporting to CPS, that define alcohol use during pregnancy as child abuse/neglect, or that allow civil commitment for alcohol use during pregnancy could also lead women to avoid prenatal care. A positive association between prenatal care utilization and birth outcomes has been documented , and if pregnant women who drink alcohol avoid prenatal care, prenatal care providers miss opportunities to provide other health promoting interventions that 1) support women to reduce or stop drinking, 2) provide other important components of prenatal care, such as monitoring for pre-eclampsia, and 3) link them to other supportive services. Punitive policies that lead pregnant women who drink to avoid prenatal care could thus increase the chances of adverse birth outcomes. In addition, policy contexts that allow criminal justice prosecutions or require CPS reporting could also influence effectiveness of alcohol-related interventions such as screening and brief interventions, which are widely recommended for pregnant women, including at-risk drinkers . Screening in an environment where being reported to CPS is a possible outcome from disclosing substance use may make women less likely to disclose use to providers and thus less likely to get support and services to help them reduce their use . To date, however, there has been no comprehensive research examining whether and how either supportive or punitive state-level policies targeting alcohol use during pregnancy are associated with birth outcomes and prenatal care utilization. Research on this topic is crucial because 1) policies continue to be debated and enacted in individual states , 2) the federal government is now incorporating them in federal legislation , and 3) some of these laws are being challenged in state court .

In addition, findings from research examining the effects of policies targeting alcohol use during pregnancy can help inform how state policymakers respond to opioid and cannabis use during pregnancy,vertical agriculture farming which are timely given the opioid crisis and legalization of both recreational and medical cannabis in several states. This study combines state- and individual-level data to examine associations between state-level policies targeting alcohol use during pregnancy and birth outcomes across 50 states over 42 years. We hypothesize that each supportive policy will be associated with decreased negative birth outcomes and each punitive policy will be associated with increased negative birth outcomes. We also hypothesize that each individual punitive policy will be associated with decreased prenatal care utilization, while prohibitions against criminal prosecution will be associated with increased prenatal care. We do not expect to see associations between mandatory warning signs and prenatal care or between priority treatment and prenatal care because we do not foresee them contributing to an environment of trust or mistrust between women and providers. If there are associations, we expect associations with increased prenatal care because they may lead women to be more motivated to seek information from providers or more able to get treatment and thus have more support to engage in prenatal care. Primary outcomes were low birthweight and premature birth . Secondary outcomes were any prenatal care utilization, late prenatal care utilization , inadequate prenatal care ), and an APGAR score ≥ 7. All outcome data came from birth certificates. We also took steps to address changes in data collection over time. For example, prior to 1980, NCHS did not impute continuous gestational age when the last menstrual period day was unavailable. After 1980, NCHS began imputing gestational age when the last menstrual period day was unavailable. We applied this imputation method to1972-1980 data to be able to have more complete data to construct the adequacy of prenatal care variable . Analyses of APGAR scores were for the years 1978-2013 because APGAR scores were not reported on birth certificates prior.The main exposure variables were time-varying state-level indicators regarding whether states had particular policies in the month and year of conception. These policies were: Mandatory Warning signs, Priority Treatment for Pregnant Women, Priority Treatment for Pregnant Women and Women with Children, Reporting Requirements for Data and Treatment Purposes, Prohibitions on Criminal Prosecution, Civil Commitment, Reporting Requirements for Child Protective Services Purposes, and Child Abuse/Child Neglect. These policies have been detailed elsewhere and are briefly described in Table 1.

The first policies, Reporting Requirements for CPS and Child Abuse/Child Neglect, went into effect in Massachusetts in 1974. Next, Washington DC adopted Mandatory Warning Signs in 1985 and Kansas adopted Reporting Requirements for Data and Treatment Purposes in 1986. In 1989, California established Priority Treatment for Pregnant Women, and both Florida and Washington established Priority Treatment for Pregnant Women and Women with Children. Kentucky, Missouri, and Virginia put Prohibitions on Criminal Prosecution into effect in 1992. South Dakota and Wisconsin established Civil Commitment in 1998. All policies were still in effect in at least four states in 2013. Each policy indicator variable is dichotomous, coded as 0 if it was not in effect for that state in the month/year of conception and 1 if it was in effect for the month/year of conception. Linking the policy indicators to the month and year of conception improves the accuracy of exposure timing .Models controlled for both individual-level maternal characteristics and for state level characteristics and policies in effect during the pregnancy. Individual-level maternal characteristics included maternal age, race, marital status, education, nativity, and parity. If data for individual-level controls were missing, we created a missing category to include all available data. Version of birth certificate was also included as an indicator variable. State-level controls included state- and year- specific poverty, unemployment, per capita cigarette consumption, and per capita total ethanol consumption, as well as indicators for whether government control of wine sales and government control of spirit sales were in effect for that state in that year. Data for state-level controls came from secondary sources, including the U.S. Census, the U.S. Centers for Disease Control and Prevention, APIS, the National Highway Traffic Safety Administration, National Beverage Control Association, and published research . State-level per capita cigarette consumption and per capital alcohol consumption were included because these variables could not be controlled at the individual-level due to lack of data documented on birth certificates in the earlier years and concerns with the quality of these data in the later years . Multivariable logistic regression was used for all outcomes. Regression models included all policy indicators simultaneously, fixed effects for state and year, state specific cubic time trends, and adjusted for both individual and state-level control variables. Regression models also accounted for clustering of standard errors according to mother’s state of residence.

Taking the most conservative approach, analyses included year fixed effects and birth certificate version indicator variables to account for changes in Vital Statistics data gathering over time as well as other relevant events in those states and years. State-specific cubic time trends were added to address possible concerns with endogeneity. All analyses were performed in Stata v14.2. This is the first study to comprehensively assess whether state-level policies targeting alcohol use in pregnancy are related to adverse birth outcomes, outcomes that indicate measurable harms due to alcohol use during pregnancy. We find that most policies targeting alcohol use during pregnancy – MWS, CACN, CC, PCP, RRDATA, and PTPREG – appear associated with increased adverse birth outcomes, possibly due to some of these policies leading women to avoid prenatal care. In addition, it appears that generally applicable alcohol policies – specifically retail control of wine sales and any other policies that lead to decreased population-level consumption – are associated with improved birth outcomes. Although the magnitudes of effects are generally small,cost of vertical farming they are still meaningful in such a large population. Overall, these findings do support our hypotheses that policies punishing alcohol use during pregnancy are associated with increased adverse birth outcomes and may lead to avoidance of prenatal care. They do not, however, support our hypothesis that the more supportive policies – including Mandatory Warning Signs – are associated with decreased adverse birth outcomes. They also are inconsistent with our expectation that supportive policies would be unlikely to be associated with prenatal care utilization. With a few exceptions , scholars have consistently distinguished policies targeting substance use during pregnancy as either supportive or punitive; our study findings do not support this distinction. Rather, our findings suggest that state level policies targeting alcohol use during pregnancy at best do not improve birth outcomes and, at worst, lead to increases in adverse birth outcomes and lead women to avoid prenatal care. This pattern of findings is not completely surprising for three key reasons. First, qualitative research has found that information that leads women to worry that their substance use has already irreversibly harmed their fetus leads women to avoid prenatal care . Similarly, our findings suggest that rather than providing women with information that helps them change their behavior and engage with health care services that may support such behavior change, MWS may operate by scaring women and leading women to avoid such help. Second, this same previous qualitative research has found that policies related to CPS and child removal lead women to avoid prenatal care. Our findings related to CACN policies are consistent with this previous research, and extend prior findings by indicating that this avoidance of prenatal care may be linked to worse birth outcomes. This is crucial, as ongoing research on alcohol outcomes has found some associations between states with CACN and less alcohol use during pregnancy . The current analyses show that even though defining alcohol use during pregnancy as child abuse/neglect is associated with decreases in self-reports of binge and heavy alcohol during pregnancy, this does not translate to better birth outcomes. Third, and perhaps most vitally, previous research indicates that policy making related to alcohol use during pregnancy appears more related to policy making in the area of reproductive rights than to policy making that reduces public health harms from alcohol use in the population overall .

This means that the problem of alcohol use during pregnancy likely has not benefited from the same public health policy development process used to address public health harms from alcohol use in the general population. The current results show that reduced population-level alcohol consumption and government control over wine retail sales are associated with improved birth outcomes, which is in line with previous studies; therefore policymakers and public health professionals who wish to improve birth outcomes through state-level policies targeting substance use should look to the broader alcohol policy field for lessons and approaches, rather than continuing with the types of policies currently in effect. We do note that some of the patterns of findings are more difficult to understand. For example, the policy that mandates priority treatment for pregnant women was related to lower odds of inadequate PCU, but higher odds of low birthweight, premature birth, and late PCU. These mixed findings could be because the policy indicator does not capture actual treatment availability. States prioritizing treatment for pregnant women might have fewer treatment slots than states without such laws, meaning our finding could be just an indication of lack of treatment availability; future research should examine this. Laws giving pregnant women priority could also prevent women from getting treatment prior to becoming pregnant, especially in states with limited treatment availability. Similarly, laws giving pregnant women priority might prevent other people – including partners of women who become pregnant – from getting treatment, leading to adverse birth outcomes due to harms from others’ drinking. Our findings are inconsistent with the only other published study that examined associations between MWS and adverse birth outcomes across both states and time. In that study, MWS were associated with decreased odds of very low birthweight and very preterm birth . This discrepancy could be because the previous study 1) only examined MWS without accounting for other policies; 2) used data only for the years 1989-2006; 3) examined different outcomes; 4) only used a subset of states; 5) did not link policy data to individual outcomes based on the month of conception; 6) controlled for state-level policies alcohol and tobacco policies and not actual per capita consumption; and/or 7) controlled for individual-level alcohol use data from birth certificates, which are of poor quality and which could be more likely to be assessed and documented in cases of adverse birth outcomes.

Measurements of PPI can be a great additional tool in this multivariate approach

Mice that model the 22q11 deletion syndrome were genetically engineered and displayed PPI impairments . Because redoxdysregulation and allelic variants of the genes coding for the rate-limiting glutathione synthesizing enzyme glutamate-cysteineligase modifier have also been implicated in BD, mice with this GCLM knocked out were tested and exhibited PPI deficits . In a genome-wide association study, a relationship between the NCAN gene and BD was observed. Ncan KO mice were subsequently tested and displayed decreased PPI levels, together with a range of other mania-like behaviors , which were normalized after lithium treatment . In another recent study, treatment with valproate reversed PPI deficits observed in transgenic mice over expressing corticotropin-releasing factor, which is implicated in the etiology of BD . Finally, PPI deficits observed in BD have been recapitulated in other animal models that exhibit mania-like behavior such as the Na+, K+-ATPase α3 mutant mice or ClockΔ19 mutant mice , but not in other animal models such as the N-ethyl-N-nitrosourea -generated mutant mouse strain . Furthermore, Black Swiss mice have often been reported as a model for mania yet they exhibited better PPI compared to C57BL/6 mice . Given the observed PPI deficits across numerous disorders, investigating the exploratory and/or other relevant behaviors of these genetic mutants will prove useful for BD research in the future. As previously mentioned, DAT KO mice have also been suggested as a model for mania and their sensorimotor gating capacities have also been assessed. Significant PPI deficits were observed in these mice,commercial vertical farming systems which were attenuated by pretreatment with the DA D2 antagonist raclopride and the antipsychotics clozapine and quetiapine .

DAT KD mice, that have approximately 10% expression of the DAT gene compared to WT mice, initially appeared to have normal PPI levels . More recently however, we have observed PPI deficits in these DAT KD mice . Interestingly, the COMT Val allele, which contributes to slower DA clearance in the prefrontal cortex, is associated with reduced PPI in Caucasian healthy humans . Therefore, a link between reduced DA clearance and PPI deficits has been established. Together, these pharmacological and genetic studies can be helpful in the screening of valid animal models for BD and development of novel therapeutics , although the PPI paradigm by itself is not selective for assessing BD mania treatment efficacy. Furthermore, it is clear that the circuitry underlying PPI is diverse and complex , yet it is unclear whether deficits in models described here are a result of alterations in different circuitry, or whether they exert an effect that converges on one particular circuit. Comparing these manipulations for this convergence would be useful in order to establish whether novel treatments could be elucidated from this work. Importantly, the strength of high-quality translational research is the capacity to assess a battery of different behaviors relevant to the disorder across species .In the case of PPI and BD, mixed results are observed across both models and patients.Neurocognitive capabilities closely correlate with a patients’ functional outcome . Developing treatments targeted at the neurocognitive deficits of patients should therefore enhance their quality of life. Impaired cognition as a quantifiable symptom of BD should be included in the multivariate assessment of screening animal models for BD. Working memory, vigilance, inhibitory control, decision-making under risk, and processing speed can all be affected in BD patients . Some of the cognitive deficits that can be quantified in both humans and rodents include impaired attentional performance, disinhibition, and impaired risk-based decision-making , but further studies in other cognitive domains are required. Translational paradigms assessing such behavior in a similar fashion across species can help bridge the gap between cognitive difficulties in BD and their assessment in animal models for BD.Decision-making performance in a clinical setting is most often measured using a task called the Iowa gambling task .

The IGT is based on repeatedly making choices between four different card decks in order to maximize gains in the long-term . Unknown to the test subject, two options will deliver high monetary gains but with occasionally large losses , while the other two options pay smaller amounts of money but also incur smaller penalties . Initially, healthy individuals will sample all decks but as the task progresses they preferentially choose the advantageous options significantly more as they deliver the highest gains in the long-term. Patients with BD however, choose more disadvantageous or risky choices and learn slower . Hence, BD patients make poorer decisions under risk in a laboratory-based task. In order to study biological mechanisms behind decision-making in more detail, several different rodent versions of the IGT have been developed . One procedure uses an operant-based chamber in which the rodent chooses from four different light cues . Each option is associated with a specific reward/punishment schedule similar to the one used in human IGT. One technique uses such an IGT but trains the rodents on the reinforcement schedules over many sessions with decision-making assessed after contingency acquisition. Using this procedure, the effects of pharmacological manipulations of DA, serotonin, and norepinephrine as well as lesions in the orbitofrontal cortex and amygdala on decision-making for learned contingencies could be studied. When this multi-plesessions IGT paradigm was adopted for use in mice, it was discovered that the DAT KD mouse model for BD tended to exhibit an increased risk-preference compared to WT mice after 25 training sessions . Furthermore, secondary measures indicated elevated levels of motivation and motor impulsivity in these mice. Using the same IGT procedure, the effects of selective DAT inhibition was also studied in mice. GBR12909 and modafinil both increased measures of motor impulsivity and motivation significantly, but affected risk-preference only subtly . In contrast, the mixed DAT/NET inhibitor amphetamine induced a more conservative strategy in both mice and rats , resulting in a risk-averse preference without affecting motor impulsivity. Hence, in this between-sessions IGT, amphetamine reduced risk preference in rodents , while selective DAT inhibition only subtly increased risk preference. Importantly however, the human IGT requires subjects to learn to select the advantageous options over a single session. BD patients learn more slowly in this single task. Thus, a similar time-course of this decision-making process in a rodent IGT may more accurately reflect human decision-making. When tested within a single session, it was possible for rats to learn to select the advantageous options over time in a single 60 min session consistent with healthy humans.

This single-session IGT test was successfully adopted for use in mice, resulting in a single-session IGT more analogous to the human IGT . Using this procedure, it was established that both GBR12909- treated and DAT KD mouse models of BD exhibited poor learning of reward contingencies over a single session consistent with what was observed in manic BD patients performing the human IGT. Additionally, DAT inhibition also increased motivation and motor impulsivity in mice consistent with earlier observations . Perhaps most striking was that using post-hoc choice analyses, it was discovered that decision-making deficits of the BD mania patients and both mouse models were driven by increased sensitivity to rewards,horticulture vertical farming specifically a preference to shift to high reward options. Such findings in patients are consistent with previous reports for mania patients and differ by state of BD, with depressed patients being more sensitive to punishment . Combined, these data indicate that selectively reducing DAT functioning induces a behavioral profile that resembles the impaired decision-making and hedonia behaviors observed in BD mania . These data also highlight that species-specific versions of the IGT provide another useful translational tool that can be used to investigate the biological underpinnings of poor risk-based decision-making in patients with BD . Another typical neuropsychological test that is often used to quantify attention and inhibitory deficits – another core aspect of cognitive dysfunction in BD – is the continuous performance test . Different versions of the CPT exist, but all include both target and non-target stimuli with which attentional processes such as vigilance and response inhibition can be quantified. When patients with BD are tested in the CPT, several studies have demonstrated impaired attentional performance and inhibitory processing deficits compared to healthy subjects . Because vigilance deficits present in BD can be measured this way, vigilance may be another potential target to assess across species . Using the same operant chambers as utilized for the rodent IGT, a 5-choice -CPT was developed for mice , and is also being used in rats . Hence, several measures of vigilance and response inhibition processes can successfully be measured with the 5C-CPT in both mice and rats wherein improved performance was observed in healthy mice and rats , and PCP impaired performance in rats . Subsequently, the 5C-CPT was reverse-translated for use in humans with recent reports describing the first translational assessment of vigilance wherein patients with schizophrenia exhibit impaired performance in the 5C-CPT that is also fMRI compatible . Hence, the human task is clinically sensitive while also demonstrating translational predictive validity. In support of the translational predictive validity of the 5C-CPT, both mice and healthy humans displayed impaired attentional functioning after 36 hours of sleep deprivation . Using the mouse 5C-CPT, it was discovered that DAT KD mice exhibited inhibitory deficits and poor vigilance similar to that of patients with BD mania performing the human 5C-CPT . This translational paradigm could therefore prove useful when assessing attention in rodent models for BD and comparing it with that of patients. These studies are also of importance for future BD research given that sleep deprivation can induce mania in BD patients. While some have therefore suggested that sleep deprivation induces mania-like behavior in rodents, e.g., hyperactivity, increased aggression, hypersexuality, and stereotypy , this premise is somewhat flawed since healthy humans are not ‘manic’ as a result of sleep deprivation. Rather, because sleep deprivation can precipitate manic states in patients , sleep deprivation could be combined with genetic susceptibility models related to BD and tested in the 5C-CPT in an attempt to more closely relate to clinical findings.

Since patients with bipolar depression can actually benefit from sleep deprivation , the same procedure may also be used to validate predictive treatment models of BD depression because if in a ‘depressed’ state, mice should recover from sleep deprivation. Thus, the 5CCPT provides an additional instrument to the behavioral tool set with which behaviors relevant to BD can be screened and associations between environmental manipulations and genetic susceptibility for BD can be studied . Selective inhibition of the DAT recreates several behavioral profiles of BD mania . Perhaps most importantly, risk-preference in the mouse IGT correlated modestly with specific exploration in the mouse BPM . In human studies, a relationship was observed between specific exploration in the BPM and poor performance in the frontal-mediated Wisconsin card-sorting test . In other words, the higher the specific exploration, the more likely subjects were to exhibit impaired cognitive functioning. It is therefore feasible that developing treatments that would attenuate the pervasive specific exploration profile of BD patients seen in mania and euthymic phases may also positively affect a number of cognitive functions. To date, no such attenuation has been observed in published studies in which AMPT or VPA was administered to the DAT mice in the mouse BPM . Instead, both treatments increased levels of specific exploration even further, reflective perhaps of a deleterious effect on cognition. AMPT treatment can indeed impair cognitive functioning as was previously assessed by reward processing and probabilistic learning in human patients with MDD and bulimia nervosa . The mood stabilizer VPA, just as lithium, has also been associated with cognitive deficits in healthy subjects and BD patients . These data highlight the difficulty in studying the predictive pharmacological validity of animal models for BD without a ‘gold standard’ treatment that is efficacious for all symptoms and underscore the need to develop novel treatments that benefit cognition in the already negatively affected BD population. The current review has focused on efforts made to determine what neural mechanisms might contribute to abnormal behaviors that are relevant to BD, particularly mania. This review has focused on translational tests of behavior that are sensitive to deficits in BD patients and in which an equivalent task exists in rodents . Significant progress has been made, but a great deal of work is still required in this field. Reduced DAT functioning appears key to recreating many aspects of behavioral abnormalities seen in BD mania patients . This idea stemmed in part from observations that a DAT polymorphism reduces DAT expression and lower DAT levels are observed in drug-free patients with BD, although these genetic observations in humans are inconsistent. Future studies should determine if patients with genetic polymorphisms exhibit reduced DAT levels and abnormal behaviors in these translational paradigms. Furthermore, other aspects of cognitive functioning rarely tested in animal models of BD such as working memory have not been tested in the DAT model animal.

We tested one- to five-group quadratic trajectory models to find a best-fitting model

Results showed deterioration of white matter integrity in youth who drank heavily compared with age- and sex-matched controls. Moreover, the slope of this reduction over time corresponded with days of drinking since the study entry.Within-subject analyses contrasted developmental trajectories of youth before and after they initiated heavy drinking. These analyses suggested that drinking onset was associated with, and appeared to precede, disrupted white matter integrity. This disruption was greater in younger adolescents than in older adolescents, and was most pronounced in the genu and body of the corpus callosum.It is possible that these brain structure changes may occur concomitantly with modifications in certain neurotransmitter and hormone secretion systems, which markedly influence the refinement of certain brain areas and neural circuits.Along with altered development and maturation of gray and white matter, studies have reported neurocognitive consequences of underage drinking, such as impairments in attention, verbal learning, visuospatial processing, and memory.Neurocognitive deficits linked to moderate to heavy drinking during this critical developmental period may lead to direct and indirect changes in neuromaturational course, with effects that may extend into adulthood. Squeglia et al. examined neurocognitive function in adolescents who drank heavily, moderately, or not at all, based on the Cahalan classification system.Their findings suggested that initiation of moderate to heavy alcohol use and incurring hangovers during adolescence may adversely influence neurocognitive functioning. For females, more drinking days in the past year predicted a greater reduction in performance on visuospatial tasks, in particular visuospatial memory, from baseline to follow-up. For males,vertical farming greenhouse a tendency was seen for more hangover symptoms in the year before follow-up testing to predict a relative worsening of sustained attention.Another set of studies demonstrated that youths who drank heavily exhibited greater brain activation while viewing alcohol advertisements than while viewing ads for nonalcoholic beverages.

Adolescents are exposed to alcohol advertising materials on a daily basis in many countries. As studies in adults with AUD have shown atypical responses to alcohol related materials,Tapert and colleagues used fMRI analyses to determine whether similar response patterns existed in adolescents who drink. The study included 15 adolescents ages 14 to 17 with AUD and 15 demographically similar adolescents who drank infrequently. The participants were shown pictures of alcoholic and nonalcoholic beverage advertisements during neuroimaging. Adolescents with histories of heavy drinking showed greatly enhanced neural activation while viewing the pictures of alcoholic beverages compared with pictures of nonalcoholic beverages. The extent of alcohol-related activation was greatest for those with the highest levels of monthly alcohol intake . In contrast, youth with limited drinking histories showed similar levels of activation while viewing the two beverage picture types. These results demonstrated pronounced alcohol cue reactivity in heavy drinking teens, particularly in reaction to alcohol advertising materials. Studies examining longer-term impacts of adolescent alcohol misuse have yielded mixed results. Some studies reported a maturing-out without significant consequences in adulthood, while others found ongoing effects on mental health, physical health, and social functioning, as well as higher levels of alcohol use and AUD.Analyses using data from the National Longitudinal Alcohol Epidemiologic Survey determined that 40% of those initiating alcohol use before age 15 were diagnosed with AUD at some point in their lives compared to only 10% of those who delayed the onset of drinking until age 21 or later.The first study of adolescents to assess the association between age of adolescent drinking onset and neurocognitive performance found that earlier age of drinking onset predicted poorer performance on tasks requiring psychomotor speed and visual attention. Similarly, an earlier age of onset of regular drinking predicted poorer performances on tests of cognitive inhibition and working memory.This study suggested that early onset Several studies have reported that the associations between alcohol and brain structure and function differ by sex, especially in adolescents engaging in binge drinking. While not conclusive across the literature, female adolescents who engaged in binge drinking appeared to show effects such as blunted activation in frontal, temporal, and cerebellar cortices compared to females who did not drink, whereas male adolescents who engaged in binge drinking showed the opposite activation pattern.Female adolescents ages 15 to 17 meeting criteria for AUD showed larger prefrontal cortex volumes than female controls, while male adolescents with AUD had smaller prefrontal cortex volumes than male controls.

A similar finding was observed for white matter.Adolescence is the peak time for both onset of substance misuse and emergence of mental illness, including anxiety disorders, bipolar disorder, major depression, eating disorders, and psychosis.The National Survey on Drug Use and Health estimated that 20% of adolescents had a mental illness that persisted into adulthood.Moreover, adolescents with a past-year major depressive episode were more likely to be current binge alcohol users .However, it remains unclear how comorbid mental health problems contribute to and exacerbate the neurobiological effects of alcohol misuse.4 Frontal and temporal cortical thinning may predict increased vulnerability to development of adolescent depression. In the NCANDA sample of 692 adolescents without a history of depression, the 101 youth who transitioned into depression were found at study baseline to have thinner cortices in the superior frontal cortex, precentral and postcentral regions, and superior temporal cortex, beyond effects attributable to age and sex.Childhood trauma and post-traumatic stress symptoms have been shown to confer increased risk for adolescent and adulthood AUD, mental illness, and physical health problems.Youth with trauma exposure showed thinner frontal cortices, and those with chronic post-traumatic stress disorder had smaller orbital frontal cortices and less superior posterior cortical and cerebellar gray matter volume.These observations indicate that trauma may be associated with structural brain aberrations. NCANDA has also examined the relationship between childhood trauma and subsequent adolescent alcohol use.In a sample of 392 NCANDA participants, adverse childhood event history was linked to greater self-reported executive dysfunction spanning four annual follow-ups. Greater childhood trauma also was linked to less connectivity in sensorimotor and cognitive control networks at baseline. This reduced connectivity explained the relationship between executive dyscontrol and subsequent increased frequency of adolescent binge drinking .Sleep patterns change substantially during adolescence and emerging adulthood.Lack of sleep, going to sleep relatively late, and large weekend-weekday sleep differences all are risk factors for alcohol use in adolescents and young adults.Similarly, in the NCANDA sample, sleep difficulties in adolescence predicted later substance use problems.The reverse has also been seen, with acute and chronic alcohol intake altering sleep structure and electroencephalography patterns in older adolescents and adults.NCANDA will continue to longitudinally examine whether these changes remain evident into adulthood and how alcohol use influences sleep neurobiology.

Co-use of multiple substances may influence the relationship between alcohol use and neural integrity. For example, during a spatial working memory task, adolescents with co-occurring AUD and cannabis use disorder showed less inferior frontal and temporal neural activation but a greater medial frontal response compared to adolescents with AUD alone.Co use of alcohol with cannabis also may adversely influence executive functioning.Given the high rates of co-occurring alcohol and other substance use during adolescence,future well-powered studies will benefit from detailed analyses of various combinations of substances of abuse on neural and neurocognitive outcomes. In adults with AUD, improvements in attention and concentration, reaction time, and memory are generally seen after 2 to 8 weeks of abstinence;however, executive functioning, processing speed, visuospatial,vertical farming supplies and verbal skills appear more resistant to recovery,and spatial processing deficits may persist for years.Younger adults tend to recover more quickly and completely than older adults .As mentioned previously, preliminary evidence suggested that adolescent heavy drinkers showed greater response to alcohol cues,more emotional reactivity and poorer distress tolerance,and poorer visuospatial performance compared with adults.These effects remitted after a month of abstinence, indicating that some deficits are linked to alcohol intake and may be transitory. However, executive dysfunction and negative mood states did not remit within 4 weeks of abstinence, suggesting that these differences may have predated the onset of heavy drinking or may take more time to recover. As reported by Infante et al., cortical gray matter volume decreases were greater in proximity to reported drinking episodes in a dose response manner, suggesting a causal effect and raising the possibility that normal growth trajectories may recover with alcohol abstinence.However, other studies have suggested that impaired visuospatial functioning following adolescent AUD persisted even after reduced levels of use.Problematic substance use trajectories among people living with HIV during the COVID-19 pandemic are poorly understood . Addressing this omission is critical for several reasons. The onset of the COVID-19 pandemic in the United States drastically disrupted people’s daily routines, livelihoods, and outlets for social participation . Recommended transmission mitigation strategies, like limiting social contact and physical distancing, required people to adapt their personal and professional lives to comply with support infection control mandates . Although critical for mitigating the spread of COVID-19, these strategies have also adversely affected the general public’s mental health . Researchers have suggested that the social circumstances arising from the COVID-19 pandemic may have contributed to increases in psychosocial stress , problematic alcohol and recreational drug use, and limited access to mental health support, harm reduction, and substance use treatment .

These predictions warrant prioritized attention for PLHIV given this population’s disproportionate vulnerability to adverse outcomes, even in pre-pandemic times . Many studies have investigated problematic alcohol and recreational drug use trajectories in PLHIV, particularly in the contexts of stress, coping, and sexual risk-taking . Prior studies have found that problematic alcohol consumption and recreational drug use commonly co-occur with other adverse psychosocial conditions, including depressive symptoms, loneliness, and inadequate social support . Problematic alcohol, heavy marijuana, and recreational drug use are linked to sub-optimal HIV outcomes and quality of life, including sexual risk-taking, impaired cognitive function, non-adherence to HIV care recommendations, weakened immunity, and premature mortality . Yet, few studies have investigated whether problematic alcohol, marijuana, and recreational drug use have changed since the COVID-19 pandemic in PLHIV. Our study’s objective was to compare the short-term trajectories in binge drinking, marijuana, and recreational drug use spanning preCOVID-19 pandemic to early pandemic time points among a prospective cohort of PLHIV. We hypothesized that substance use would increase during the pandemic. Additionally, we hypothesized that depressive symptoms and loneliness would be linked to increased substance use behaviors and social support would be associated with decreased substance use behaviors. We argue that psychosocial conditions and a lack of social support at the start of the pandemic may serve as proxies for prevalent adversity in participants’ lives and their potential to avoid negative trajectories. We developed group-based trajectory models in SAS 9.4 to identify clusters of individuals who shared similar trajectories in binge drinking, daily marijuana use, and recreational drug use over time. We used the multi-variable mixture modeling macro, PROC TRAJ, which employs maximum likelihood to estimate model parameters and handles missing values .Following these iterative processes, we evaluated improved model fit based on Bayesian Information Criterion , the size of each group, and the probability of membership in a specific group; specifically lower BIC values, > 5% of the sample in each group, and an average probability of ≥ 0.70 for participants in each trajectory group . After finding the best-fit number of group trajectories, we adjusted each trajectory for the best shape . We iteratively compared models with increasing numbers of groups, omitting and adding parameters based on BIC values to assess improvements in goodness-of-fit. Based on the group-based trajectory analyses, we derived a categorical variable representing trajectory group membership for each substance use behavior to test their associations with other co-factors. Fisher’s exact tests were used for categorical variables and student’s tests for continuous variables. We tested bivariate and multivariate logistic regression models to estimate the association between trajectory group membership and co-factors. Finally, we tested generalized linear mixed models using repeated measures for each substance use outcome to further confirm our results. We did not assume any structured error correlations due to limited time points; therefore, we modeled our data using unstructured correlation matrices. We included time along with co-factors used in the group-based trajectory models as independent variables. We also tested an interaction of sex assigned at birth by time to account for potential cohort effects. We allowed only the intercept to vary between subjects and the regression slopes as fixed. The current study is among the first to examine short-term problematic alcohol, marijuana, and recreational drug use trajectories during the beginning of the COVID-19 pandemic in a large, prospective cohort of PLHIV in the United States. We observed two-level, short-term, group based trajectories that distinguished participants as non-binge drinkers versus binge drinkers, non- or infrequent marijuana users versus daily marijuana users, and users versus non-users of recreational drugs.

Measuring drug use in epidemiological research studies poses challenges

A mammalian embryo can successfully implant in the uterus only after it has become a blastocyst. This consists of a small cluster of cells called the inner cell mass, which eventually develops into the fetus, and a thin layer of outer cells called trophectoderm, which gives rise to the placenta . When cells in the trophectoderm become ready to implant, their metabolic rate increases and they develop a repertoire of surface molecules that allows them to adhere to the uterine epithelium. But for this interaction to occur, the uterus must first mature to a receptive state. The process of synchronization between blastocyst and uterus involves a complex network of membrane-associated molecules and soluble signals, including steroid hormones, growth factors and lipid media tors. In a series of landmark studies, Dey and colleagues have established that cannabinoid receptors and their endoge nous activator, anandamide, are integral components of this network. Before it becomes receptive, the uterus contains exceedingly high levels of anandamide—higher, in fact, than anywhere else in the body. As the tissue progresses into receptivity, anandamide concentra tions drop, suggesting that a lower cannabinoid tone creates a friendlier environment for implantation. In keeping with this idea, activation of cannabinoid recep tors on the trophectoderm inhibits blasto cyst development and implantation. This simple scenario does not explain all the available data, however. If the only role of the endogenous cannabinoid system were to prevent implantation, mutant mice that do not express cannabinoid receptors should be more fertile than wild-type animals. Yet the opposite is true: mutant ani mals actually have 40% fewer pregnancies than do wild-type ones. Another inconsistency is that anandamide improves implan tation when administered at very low doses,vertical farming system lower than those needed to impair it. How can these seemingly opposite effects be explained?

Dey and coworkers addressed this question using either freshly isolated mouse blastocysts or, when they needed larger quantities of tissue, trophectoderm cells in culture. The researchers found that anandamide activates cannabinoid receptors in the trophectoderm to initiate two distinct signaling cascades in a dose-dependent manner. At a low concentration , anandamide stimulated the extracellular signal–regulated protein kinase ; at a higher concentration , anan damide reduced calcium entry by closing voltage-operated N-type calcium channels. These events had opposite functional con sequences, such that blastocysts treated with 7 nM anandamide became competent for implantation, whereas those treated with 28 nM anandamide did not. Thus, the uterus may be able to titrate its anandamide production to either promote or arrest embryo implantation . These results confirm and extend the regulatory function of the endogenous cannabinoid system in female fertility. At the same time, they raise two potentially serious public health issues. The first derives from clinical reports that link defects in anandamide breakdown to increased risk of spontaneous abortion. These studies found that expression of fatty acid amide hydrolase, the enzyme that catalyzes the intracellular degradation of anandamide, was reduced in lymphocytes obtained from women who had had mis carriages, compared with those who had not. It is unclear, however, whether the mechanism underlying these miscarriages is related to the ability of anandamide to stop implantation . The second issue stems from the fact that a large number of women of childbearing age—4.9%, according to a recent survey10—regularly smoke marijuana. How might this drug affect their fertility? If the same signaling system described in mice were present in humans, one might expect fertility problems among heavy marijuana users. Although there is no clear indication at present that marijuana impairs fertility, this possibility should certainly be revisited in the light of the current animal data.The first infectious human retrovirus to be identified, before we even imagined the devastation of HIV, was human T-lymphotropic virus type 1 1. HTLV-1 causes disease—mainly adult T-cell lymphoma or leukemia or HTLV-1-associated myelopa thy—in only small fraction of infected individuals.

The virus, however, cuts a wide swath, infecting about 10–20 million people worldwide by spreading from person to per son through infected cells in semen, blood and breast milk. Despite years of effort, the identity of the receptor that facilitates the spread of HTLV-1 has remained elusive. In a recent issue of Cell, Manel et al. provide evidence that GLUT-1, a ubiquitous glucose transport protein, is a receptor for HTLV-1 . Most retroviral receptors have been cloned by transferring a library of sequences from a cell line that is permissive for entry, and therefore expresses the receptor, into one that is nonpermissive. A marked version of the virus can then be used to specifically select cells with the receptor gene from cells that acquired irrelevant cellular sequences. HTLV-1 infects most cell lines but is found primarily in lymphocytes, including lymphoma cells, in infected patients. The paucity of nonpermissive cell lines, as well as difficulties in making high titers of marked virus, has foiled attempts at conventional receptor cloning strategies. For these reasons, Manel et al. used a clever deductive approach that relied on several clues to identify a receptor candidate. The first clue was that cells expressing the HTLV-1 envelope protein, which binds the receptor and initiates entry, showed perturbations in lactate and glucose metabolism— namely, delayed acidification of the cell culture medium. This observation implicated a receptor involved in lactate or glucose transport. Because many known retroviral receptor proteins are transporters, a trans port protein has always been a strong con tender for the HTLV-1 receptor. Another crucial clue came from HTLV-1 envelope binding studies, which suggested that the receptor was expressed very early after lymphocyte activation, and that its temporal pattern of expression was distinct from that of many other activation markers. Together, these clues led the authors to GLUT-1, a member of a family of glucose transport proteins. The tactic of finding a receptor to meet the authors’ criteria, rather than letting the virus select the receptor in a functional screen, carries with it a considerable burden of proof. The authors used several lines of evidence to support their case. Binding studies demonstrated a direct interaction between the HTLV-1 envelope and GLUT-1, but not other related transport proteins. Down modulating GLUT-1 led to reduced HTLV-1 binding and infection, whereas increasing GLUT-1 expression restored these outcomes.

Finally, the authors provided data that GLUT-1 is a receptor for a related, relatively avirulent retrovirus, HTLV-2, fulfilling predictions from previous studies. Collectively, the results pro vide a compelling case for GLUT-1 as an HTLV-1 receptor .Cambodia has the highest HIV prevalence of any Asian country, and over the last decade has experienced the most serious HIV/AIDS epidemic in Southeast Asia. Heterosexual contact is the major route of HIV trans mission, and female sex workers remain the group at highest risk. Although crucial progress has been made in reducing risky sexual behavior, including widespread condom use and promotion of reduced number of sexual partners, HIV prevalence among FSW remains high, ranging from 11% to 26%. Poverty, low literacy, a high prevalence of sexually transmitted infections,cannabis vertical farming and a highly mobile work force are contributing factors to the epidemic. Recent research has also identified drug use and, in particular, amphetamine-type stimulant use as a serious emerging problem associated with HIV risk among FSW, which threatens to reverse downward trends in HIV infection rates in the region. Amphetamine-type stimulants include a range of syn the ticpsycho stimulants, including methamphetamine, amphetamine, and ecstasy, which can be injected, smoked, or taken orally. Effects of these drugs include euphoria, alertness, arousal, increased libido, increased sympathetic nervous responses,and perceived increases in confidence, energy and physical strength. In Cambodia, a pill form of methamphetamine known as “yama” is widely produced, trafficked, and used. The tablets generally contain about 25% methamphetamine. “Crystal” is generally about 85% metham phetamine and more addictive. Although yama pills are swallowed, both forms are usually melted and the vapors inhaled, resulting in rapid neurologic effects. Use of ATS has been associated with elevated HIV risk behavior in many countries and in several population sub groups. The United Nations Office on Drugs and Crime reports that use of these drugs is widespread in Asia and increasing rapidly in Cambodia. In Cambodia, ATS are highly available both in pill and crystalline form and are generally ingested or smoked; injection use is uncommon. The Cambodia National Authority for Combating Drug Abuse estimated that 70% of all drug users in Cambodia use ATS. The drug accounts for the majority of all drug seizures by authorities, and, in pill form, has been ranked as the leading drug of abuse for the past nine years with consistent increases since 2006, at which time it was estimated that 30,000 tablets of yama were consumed orally or smoked there daily. Use is particularly high among vulnerable populations, including FSW, men who have sex with men , and street children.Self-reported measures of drug use have the advantage of being noninvasive and permit evaluation over longer time periods compared with bio chemical assessments. However, study participants may misrepresent drug use due to social desirability bias, stigma, poor recall, poorly worded questions, or poorly worded response categories in surveys and interviews, all of which could result in mis-classification of measured exposures. Although studies have shown that the use of Audio Computer-Assisted Self Interview increases reporting of sensitive and stigmatized behaviors, research suggests that the validity of self-reported drug use varies by population, race/ ethnicity, mental health, and drug treatment status, although not by gender.

Accuracy has varied in studies of arrestee populations but have been reported as higher in groups sampled in emergency department and STI clinics. Those that report more frequent drug use, compared to infrequent use, are more likely to self-report recent drug use. Urine toxicology assessments provide sensitive and valid measures of many drug types; but some, like ATS, are restricted to a short time frame due to rapid metabolization. The detection window may also depend on the physical condition of the individual , route of drug ingestion , frequency of use, and drug-related factors such as purity. To explore the validity of self-reported ATS use among young FSW in Phnom Penh, Cambodia, we com pared self-reported ATS use with results from concurrently collected urine toxicology tests. We also examine whether sociodemographic, sex-work venue, and HIV status were associated with validity of self-reported ATS use.The Young Women’s Health Study was a prospective study of HIV and ATS use among 15–29 year-old women engaged in sex work in a diversity of settings in Phnom Penh, Cambodia. Data for this cross-sectional validity assessment was collected at the second quarterly study visit women attended. Both self-reported measures and urine toxicology testing for ATS were assessed. The YWHS-2 was led by HIV-research and HIV-prevention specialists from the Cambodian National Center for HIV, AIDS, Dermatology, and STDs , the Cambodian Women’s Development Association , the University of California San Francisco in the United States, and the Kirby Center at the University of New South Wales in Australia.Young women at high risk of HIV infection were the target study population. Inclusion criteria were age 15–29 years, understanding of spoken Khmer, Cambodian ethnicity, reporting of at least two different sexual partners in the last month or engaging in transactional sex within the last three months, plans to stay in the Phnom Penh area for 12 months, being biologically female, and being able to provide voluntary informed consent. Study methods have been described previously. In brief, trained field assistants from the CWDA recruited women from community locations, provided study information, and obtained group informed consent. Women who consented were then seen by appointment at the YWHS-2 clinic site; free transportation was provided. Participants received US $5 and condoms at each study visit. Contact information was collected to facilitate participant tracking and maximize follow-up. Women were asked to enroll for a one-year study with quarterly study visits. Data collection for this validity assessment occurred in November 2009 at the second study visit . All study visits included administration of a structured questionnaire in Khmer by trained inter viewers who queried participants about sociodemo graphic characteristics, health care, occupational and sexual risk exposures, alcohol and self-reported ATS use as well as testing for HIV and ATS using blood and urine samples, respectively. The second visit was used for this cross-sectional analysis because questions about past 48-hour drug use were added to the questionnaire starting with that visit; thus, it was the first available time point for comparison and validation of self-reported use and urine testing.

An alternative to real experiments is to have students collect data in a virtual environment

Greater thalamic response to risky reward versus loss feedback in PSUs is consistent with research demon strating that thalamic BOLD signals are linked to relapse in cocaine-dependent individuals . Thalamus acts as a relay center for the brain by sending sensory information to insula for further interoceptive processing ; hypo activation to loss may reflect differences in relay and integration of information during decision making. With respect to baseline characteristics, DSUs endorsed higher baseline levels of state depression than PSUs, which may have affected RGT performance given that individuals with depression tend to be risk averse . However, given that mean scores for DSUs are substantially below the Beck Depression Inventory threshold for clinical depression [in nonclinical populations, scores above 20 indicate depression ; it is unlikely that DSUs performed in a manner consistent with samples with depression.Across OSUs, lower frontal, temporal, parietal, insular, and thalamic BOLD signals during risky decisions compared with safe decisions predicted greater future marijuana use . These regions are considered important for executive functions such as inhibitory control, working memory, and attention as well as for being relay centers for integrating information critical for decision making . Therefore, blunted responses in these regions while making choices between risky and safe options may predispose young adults to repeatedly choose marijuana consumption despite potential negative consequences . While cumulative marijuana uses between study visits was related to baseline BOLD patterns, lack of relationship between cumulative interim stimulant use and baseline BOLD signal suggests that while adose–response effect may exist between brain activation and marijuana use,outdoor vertical farming the relationship between brain activation and stimulant use may be better defined through a categorical perspective that includes accompanying clinical sympto mology.

Although PSUs and DSUs used marijuana at significantly high rates , groups did not differ categorically in marijuana abuse/dependence frequency. In contrast, stimulant use in and of itself might not be related to brain differences unless it is accompanied by clinical problems, suggesting that a categorical perspective is a more useful way to conceptualize differences.This study has several unique strengths, including its longitudinal design, use of a model previously applied to chronic stimulant users, and assessment of substance use from both categorical and dimensional perspectives . However, this study is limited by our sample’s significant co-use of marijuana and the categorical criteria that prioritized differences as a function of SUD over marijuana use disorder given that PSUs and DSUs did not differ on baseline/interim marijuana use. In addition, although SUD has been associated with greater incidence of psychiatric illness , lack of clinical symptom measures collected at follow-up hinders our ability to determine whether mental health symptoms affected interim substance use. We are also limited by an inability to evaluate the RGT from a trial by-trial perspective to determine whether BOLD response patterns translate into future behavior or are affected by the preceding trial; due to the limited number of separate 40 and 80 trials, it would not be possible to obtain sufficient statistical power to conduct such an analysis.Project work in a statistics class is a valuable tool for giving students a context to the data they are using and a motivation for learning statistical reasoning . Projects also give the students an appreciation of the practical issues involved in carrying out experiments and collecting data, an outcome encouraged by Higgins . However there are also practical issues in implementing student projects in a curriculum, particular in large classes. For example, our biomedical students are very keen to use friends and family as subjects in their statistics experiments, raising ethical concerns, while other students want to use equipment and resources that are beyond the scope of an introductory statistics course. The result has been that the students often end up doing simplistic experiments which may actually reinforce a trivial view of the role of data analysis in science.

Such environments are particularly useful in helping students understand issues in experimental design by giving them more complex settings than they would have access to in a real experiment, as in the industrial process and greenhouse simulations of Darius et al. or the virtual vaccination trial of Duchateau et al. . In this paper we present an online environment, the Island, where students can conduct studies involving virtual human subjects. Again the first aim of the environment is to engage the students in thinking about the design of the experiment given a statistical question of interest. The second aim is to provide them with data from their design that they can then use in learning statistical methodology. The Island involves two main simulations running at different timescales. In Section 2 we give an overview of the historical simulation that forms the basis of the Island population while in Section 3 we describe the simulation that runs in the present to allow students to conduct experimental studies. In both sections we will outline the design of the corresponding simulations and their role in supporting learning. We emphasize this in Section 4 with four examples of student engagement with the Island. An initial motivation for this system was the need for a virtual environment where students could collect data for addressing questions in epidemiology. While it is easy to generate some simulated data for a randomized clinical trial, for example, our belief is that thinking about issues in epidemiology requires access to a population that can be studied more deeply. Important requirements include the ability to consider the ancestors of particular individuals, to determine whether diseases have a genetic component, and to explore geographic relationships between individuals in order to look for infective characteristics. For these reasons our aim was to simulate a population over time in a spatial context. The Islanders live in 39 villages that range in population from just 26 to 2,292 and provide environmental effects in the simulation of the Islanders. Figure 1 shows samples of the Islander images that are included in the interface, created using the approach discussed in Bulmer and Engstrom . The Island map and other features of the interface can be seen online using the link in Section 1.1. Within each village the Islanders then live within houses. Many live in families while others live as couples or alone.

Again there are environmental effects tied to houses,vertical farming benefits such as an increased chance of taking up smoking if other people in the house already smoke. The location of the house is also important since various diseases are linked to particular parts of the Island and transmissible diseases are spread based on distance between individuals. Sampling an Islander at random is deliberately a difficult task. Each village does have a hall with records of births, deaths and marriages but Islanders do move around during their lives. To choose an Islander from the interface as a subject for a study the student needs to select their village and then select their house and then select the individual within the house. With variable numbers of houses and individuals in each of these layers the naive solution for choosing a ‘random’ sample will likely be biased. The design of the Island simulation is that it should run in real time. We continue the historical simulation, updating the population at the start of each month. In this way the underlying environment changes slowly over time with some Islanders dying and new ones being born. In December 2010 the Islanders even settled a new village that previously had not existed in the simulation.There is a fundamental tension in our design. We want the simulation to be realistic since we believe that will help students engage with the virtual environment. However there is a point at which realism becomes counterproductive towards our aims of engaging students in the role of statistical reasoning in scientific inquiry. For example, suppose we included a cause of death called Lung Cancer and made it so that Islanders with higher smoking levels were more likely to die from this disease. Students could collect data on smoking history and cause of death and look for this relationship but if they found an association it would probably not be surprising or interesting to them since it is the outcome they expect. They will not have discovered anything new by conducting their study. Instead of using real names for diseases we have thus tried to use poetic names wherever possible. These include Summer’s Pain , Diego’s Cough, Ruin and Jungle Sickness. One of these four is indeed modelled on lung cancer, including the association with smoking history, but now it is a more open question for students to explore. For example, what data do you need to collect to distinguish between these conditions and how can you convince somebody that you have identified ‘lung cancer’ on the Island? The virtual population described so far can be viewed as a framework for adding further simulations. The histories and images of the population give a broad context for experimental investigation. Returning to the original aim of this paper, we can thus enable students to obtain the various benefits of project work by adding the kinds of tasks which would provide the appropriate data to help learn statistical thinking. We have used this environment with an introductory statistics course for science students with around 1,200 students per year. They are asked to complete an experimental project to “demonstrate your understanding of the statistical methodology you have learned in the course”. One particular advantage of the online environment is that students can conduct their experimental work quite quickly towards the end of the semester. This means that they can have a clear statistical method in mind rather than the all too common practice of just collecting data without any plan of how to analyze it. Section 4 will show some examples of student work but we begin in this section with an overview of the design and mechanics of the experimental environment.A key feature of this innovation has been the involvement of students in its creation. From the outset we planned a two-phase curriculum design process for developing and using the Island in experimental projects. The first phase involved an assessment task where students had to prepare a research proposal with the Islanders as their subjects. For each student proposal that required an addition to the Island we began by searching for existing research on the topic. This gave plausible ranges for response variables as well as suggesting relationships that might be included in the simulation. As before there is tension between reality and fantasy here. Making the simulated processes perfectly match reality would be technically difficult and, as with the smoking and lung cancer example, may not actually be desirable. We felt it was important to keep students on their statistical toes by omitting some associations that they might expect to find while adding some other associations that would surprise them, though we did keep this at a low level. A better alternative for the long term is to add tasks that are somehow native to the Island. For example, in the first phase we added the fictional Dalpa Leaves and allowed Islanders to “chew lime-soaked dalpa leaves for ten minutes”. This was added as a control for chewing lime-soaked coca leaves but dalpa leaves were given their own effects that students could study independently in the future. The simulations used to generate the data that students observe involve a wide range of approaches. Our own earlier systems to simulate data for statistical exercises were based on standard models, such as using a linear model to generate outcomes based on parameters to which random Normal variability was added. In contrast the Island relies heavily on differential equations to capture changes in physiology at a more basic level and then links these with various statistical models as needed. While the population simulation moves in monthly steps, the experimental simulation updates at 30-second intervals. At this level each student has their own copy of the Island, tied to their login, so that changes they make to Islanders through experimental treatments are independent of changes made by other students. Similarly, such changes made by the students do not affect the underlying historical simulation. For example, a student can never kill an Islander through their actions since this would mean that they would then need a separate timeline in the historical simulation. Students conduct experiments by selecting an Islander and then allocating a task for them.

Descriptive statistics for covariates were calculated by AUD/remission status

The bivariate association of each covariate with AUD/remission status was tested using multi-nomial regression analysis with persistent AUD as the reference category. Concordance rates for AUD/remission status in related and unrelated pairs were calculated. Relatives’ AUD/remission status was the dependent variable in a multi-variable multinomial logistic regression, with relatives’ persistent AUD as the reference category and non-abstinent and abstinent remission as the two outcome categories. The primary independent variables were proband non-abstinent and abstinent remission; their association with relatives’ remission status was adjusted for the covariates listed above. The interactions of proband non-abstinent and abstinent remission with the dummy variable representing related pairs were tested one at a time in the fully adjusted regression to determine whether the association of probands’ AUD/remission status with relatives’ AUD/remission status varied in related and unrelated pairs. The final regression was calculated separately in related and unrelated pairs. The Huber–White robust variance estimator was used to adjust for the clustering of family data. Data sets and variables were created using SAS statistical software, version 9.2. Analyses were performed using Stata Statistical Software, version 14.2.This study explicitly modeled abstinent and non-abstinent remission in probands who were recruited from AUD treatment programs and in their first-degree family members with life-time AUDs to test for familial associations of remission in high-risk families and to define a phenotype which can be used to explore associations of remission with potentially heritable characteristics. Results showed that individuals who were related to an abstinent proband were more than three times as likely to be abstinent themselves, compared to individuals related to a proband with persistent AUD; this association was not significant in unrelated pairs. The significant association of probands’ with relatives’ abstinent remission in related but not in unrelated proband-relative pairs suggests there are familial influences on abstinent remission which may be due to genetic or familial environmental factors.

The familial association of abstinent remission in this sample selected for high-risk for AUDs has not been observed previously. The association of abstinence in one family member with abstinence in another stands in contrast to a host of null findings regarding familial influences on remission from other studies in population-based,vertical farming tower high-risk and clinical samples using a variety of definitions of remission. The current analyses used an explicit abstinent and non-abstinent remission phenotype, distinct from AUDs and consistent with the idea that the distribution of risks for development of, and for remission from, AUDs may not lie on the same continuum. Our results suggest that there may be genetic or familial environmental influences on abstinent remission and demonstrate that departing from the more common risk factor-to-remission comparisons within families may indeed prove useful. When remission is the target phenotype, remission in all family members should be measured explicitly, rather than measuring it as an outcome only in target subjects but not in their relatives. This will facilitate the examination of potentially heritable characteristics underpinning abstinent outcomes, such as social responsiveness, that may increase the likelihood of remission, as well as the investigation of family environments associated with remission from AUDs. Much more work will need to be conducted to identify heritable traits that may be related to abstinent remission and to probe for mediators and moderators of their effect. In addition to potentially heritable effects on abstinent remission, another explanation for the current findings might rest with a social contagion model, or the spread of behavior within a family due to social proximity. Analysis of large social networks from a population-based study indicated that both heavy drinking and abstinence clustered in networks, and also that the heavy drinking or abstinence of relatives and friends at one time-point were associated with changes in the subject’s alcohol consumption, to heavier drinking or abstinence, at a subsequent time-point. The same may be true within families affected by severe AUDs, where abstinence in one person may influence another family member with an AUD to try to quit drinking. This possibility is consistent with evidence that abstinence is the most stable form of remission among individuals with severe AUDs. If older family members with life-time AUD are abstinent as younger family members are developing alcohol problems, it is possible that younger members, if they recognize severe problems in themselves, may look to older members for direction or example, or that older members may recognize problems in younger members and intervene.

In fact, analysis of twin data showed that the variance associated with treatment-seeking for alcohol problems was accounted for primarily by familial influences, with 41% of the variance due to genetics, 40% due to shared environment, and just 19% to unique environment. In the current study, all probands had by definition been treated, which precluded examination of familial associations for treatment-seeking; however, abstinent relatives had the highest rates of treatment seeking in the sample, suggesting an association of relatives’ with probands’ treatment-seeking. More than 40% of probands and relatives were remitted in this high-risk sample, with abstinence the most common type of remission in probands and abstinent and non abstinent remission equally common in relatives. An earlier study in the COGA sample found that more than 50% of all subjects with life-time alcohol dependence reported periods of abstinence lasting 3 months or more, with 16.1% reporting abstinence of 5 or more years. Similar to the relatives in the current study, abstainers were older than individuals who never abstained, had a greater number of life-time symptoms and were more likely to have sought formal treatment and to have attended self-help groups. Other sampling frames also show similarities to the current data. Abstinent individuals with life-time AUD from population-based data had more AUD symptoms than remitted non-abstinent individuals. In a national sample of individuals self-identified as ‘in recovery’, abstainers compared to non-abstainers were older, more likely to have received professional treatment and to have attended self-help meetings, and had significantly more life-time alcohol dependence symptoms. These similarities across a range of samples suggest that individuals who become abstinent, regardless of sampling frame, represent a severe end of the AUD continuum. In the current study, abstinence may represent a common end-point for individuals with severe AUD. It is possible that non-abstinent remitters will become abstinent for a period, or periods, of time. Given that nearly half of abstinent relatives in the current study had been remitted for 10 or more years, abstinence may indeed represent an end-point for subjects who remit from severe AUDs.Despite efforts to improve mental health over the last 60 years, suicide remains a critical public health concern worldwide.Suicide was the second leading cause of death globally in 2012 among individuals aged 15–29years,with an estimated 80%–90% of suicide deaths attributable to mental health or substance use disorders.

Significant gaps remain in empirical research examining suicidality among marginalised populations. Marginalised women, such as sex workers who are street involved or use drugs, experience disproportionately high levels of social and health-related risks and harms, including stigma, discrimination and violence as a result of dynamic structural drivers including poverty, criminalisation and racism. While sex workers are a diverse population working from indoor in-call and out-call venues to street-based settings, previous studies high light substantial unmet mental health needs of more marginalised and street-involved sex workers. Studies among street-based sex workers and those who use drugs underscore the associations of social exclusion,vertical indoor farming depression and post-traumatic stress disorder with suicidality.Research demonstrates greater risk for suicidality among those with a history of trauma and among street-involved sex workers who report histor ical experiences of violence and childhood abuse.Furthermore, indigenous women are vastly over-repre sented among street-based sex workers in North America and face devastating and multi-generational effects of trauma and socioeconomic dislocation as a result of colonialism, racialised policies and displacement from land and home communities.Various biological, interpersonal and socio-structural factors contribute to our understanding of suicidal behaviours.While evidence has demonstrated that some forms of cognitive behavioural therapy and pharmacological interventions may reduce suicidality, the literature is hampered by publication bias and significant heterogeneity of strategies and outcome measures.Due to ethical challenges and limitations to studying suicide and its proxies , there remains a paucity of evidence from randomised controlled trials to support the efficacy of prevention interventions.Researchers have largely focused on examining suicidality outcomes , which may not be fully generalisable to understanding suicide or accurately evaluating treatment approaches.Furthermore, stigma continues to hinder research and reporting of suicidality.There remains an urgency to better understand pathways to suicidality, with literature highlighting the need for innovative psycho logical and psychosocial treatments and tailored inter vention approaches for key marginalised populations.Given the complex aetiological pathways to suicide and limited effectiveness of well-established evidence-based interventions to reduce the burden of suicidality, the US National Institute of Mental Health has called for innovative research on suicide prevention and treatment for suicidality.A number of psychedelic drug therapies are being revisited following a 40-year hiatus in research into their potential for the treatment of depression, anxiety, PTSD, eating disorders and addiction.Psychedelic drugs include the classic serotonergic psychedelics or ‘hallucinogens’ lysergic acid diethylamide , psilocybin, dimeth yltryptamine and mescaline, as well as the ‘enactogen’ or ‘empathogen’ methylenedioxymethamphetamine ,all of which are being investigated in clinical/preclinical studies for their neuropharmacological functions and potential as adjuncts to psychotherapy.While renewed interest in psychedelic medicine is challenged by various funding and methodological and legal impediments, the emerging evidence indicating improved outcomes for some individuals suffering from mental health and addiction issues has generated new scientific inquiry and an imposing obligation to advance this research.Recent observational studies in the USA demonstrate significant associations between life time psychedelic use and reduced recidivism and intimate partner violence among populations of prison inmates and reduced psychological distress and suicidality among the general adult population.Despite the multifaceted structural and social inequities that shape poor mental health burden among marginalised and street-involved sex workers, there remains a paucity of data on suicide rates and research that system atically examines factors that potentiate or mitigate suicidality among sex workers, particularly in the global north. Some evidence suggests that psychedelic drug use may be protective with regard to suicidality and is associated with significant improvements in psychological well-being and reductions in depression and anxiety in clinical settings,yet existent research is characterised by large gaps. Given the urgency of addressing and preventing suicide and calls for prioritising innovative interventions, this study aimed to longitudinally investigate whether life time psychedelic drug use is associated with a reduced incidence of suicidality among a community-based prospective cohort of marginalised women. We postulated that psychedelic drug use would have an independent protective effect on suicidality over the study period.Data for this study were drawn from a large, community-based, prospective cohort of women sex workers initiated in 2010, known as An Evaluation of Sex Workers Health Access . Eligibility criteria for study participants included cisgender or transgender women, 14 years of age or older, who exchanged sex for money within the last 30 days. AESHA participants completed interviewer-administered questionnaires and HIV/sexually transmitted infection /hepatitis C virus serology testing at enrolment and biannually. Experiential staff are represented across interview, outreach and nursing teams, including coordinators with substantial community experience. Participants were recruited across Metro Vancouver using time–location sampling and community mapping strategies, with day and late-night outreach to outdoor sex work locations , indoor sex work venues and online. Weekly outreach by experiential staff is conducted to over 100 sex work venues by outreach/nursing teams operating a mobile van, with regular contact as well as encouraging drop-in to women only spaces at the research office, contributing to an annual retention rate of >90% for AESHA participants. The main interview questionnaire elicits responses related to sociodemographics , the work environment , client characteristics , intimate partners , trauma and violence and comprehensive injection and non-injection drug use patterns. The clinical questionnaire relates to overall physical, mental and emotional health, and HIV testing and treatment experiences to support education, referral and linkages with care. The research team works in close partnership with the affected community and a diversity of stakeholders and regularly engages in knowledge exchange efforts. AESHA is monitored by a Community Advisory Board of over 15 sex work, women’s health and HIV service agencies, as well as representatives from the health authority and policy experts, and holds ethical approval through Providence Health Care/University of British Columbia Research Ethics Board.

Youth with worse HIV disease severity are more likely to engage in substance use

A GWAS of opioid response in a Japanese sample found that the C allele of rs2952768, which is in an LD block with rs7591784 , was significantly associated with greater postoperative opioid analgesic requirements, as well as lower reward dependence in healthy volunteers, lower risk of polydrug use in volunteers with methamphetamine dependence, alcohol dependence, and eating disorders, and increased expression of CREB1 in human postmortem brains . The G allele in rs7591784 was associated in our study with lower pretreatment methamphetamine use and better treatment outcomes, both suggestive of less severe methamphetamine use disorder, and as rs7591784 and rs2952768 are strongly linked, our results provide support for the previous association between the C allele of rs2952768 and lower severity of methamphetamine use disorder observed in the Japanese GWAS. Whether rs7591784 directly effects CREB expression or function is not known, but our results and previous studies suggest that variability in CREB signaling and subsequent changes in methamphetamine-induced gene expression may influence clinical severity of methamphetamine use problems and success in quitting methamphetamine and that the CREB signaling pathway may be a target for the development of medications to treat methamphetamine use disorder. Phosphodiesterase inhibitors modulate signaling via the CREB pathway via increases in cAMP and ibudilast, a nonselective phosphodiesterase inhibitor,vertical farming company is in clinical development for methamphetamine use disorder . Previous GWAS found SNPs in CDH13 to be among the most significant SNPs associated with a diagnosis of methamphetamine dependence and with the subjective response to amphetamine among healthy volunteers . None of the SNPs related to CDH13 in our study were significantly associated with methamphetamine use frequency following Bonferroni correction. The lack of significant association in our study may be due to the different phenotypes examined in the previous GWAS compared to the current study that examined methamphetamine use frequency in a treatment-seeking sample or may be due to limited power to detect SNPs with small effect size in our small sample. Methamphetamine use frequency as well as results of our SNP analyses differed greatly between males and females.

None of the three SNPs that were nominally significant in males, including rs7591784, approached significance in females suggesting that although the female sample size was relatively small, the lack of significant associations for these SNPs in females is unlikely to be due to limited power in females alone. Previous studies in rodents have found sex differences in methamphetamine pharmacokinetics , methamphetamine-induced plasma corticosterone levels , methamphetamine-related neurotoxicity , and methamphetamine self-administration with female rats acquiring methamphetamine self-administration faster, self-administering more methamphetamine, and exhibiting higher rates of methamphetamine reinstatement than male rats . In humans, female methamphetamine users have a higher risk of Parkinson’s disease , greater reductions in hippocampal volume and higher prevalence of physiologic dependence symptoms compared to male methamphetamine users and these biological or other psycho social differences may have a greater influence on methamphetamine use frequency in females than the SNPs examined here. Interestingly, amphetamine-induced CREB-mediated transcription differs dramatically between male and female mice in the nucleus accumbens, ventral tegmental area, amygdala, and locus coeruleus with greater CREB-meditated gene transcription following amphetamine in females suggesting that the significant association between rs7591784 and methamphetamine-related phenotypes observed in our study in males but not females may be due to underlying sexual dimorphism in the CREB signaling pathway. The one SNP that was nominally associated with methamphetamine use frequency assuming an additive model in females, rs163030, was associated with caudate volume in a GWAS and rs163030 may influence methamphetamine use frequency in females by altering structure or functioning of the caudate, a brain region implicated in impulsivity and methamphetamine addiction . Additional studies investigating sex differences in the biological and social influences on methamphetamine addiction are warranted. This study has several limitations. The sample size is small and the power to detect an association between a candidate SNP and methamphetamine use frequency with a small effect size is limited.

As a result the study is subject to false negative results. Also, numerous findings from candidate gene studies have failed to replicate and results from this study are preliminary and require replication in an independent sample prior to making any conclusions. To mitigate this risk, we emphasized selection of candidate SNPs that had previously been associated with methamphetamine-relevant phenotypes in GWAS. Our study did not genotype rs2709386, which was most strongly associated with opioid sensitivity in the previous Japanese GWAS, and although rs2709386 and rs7591784 are highly linked, future studies are necessary to determine which SNP is more strongly associated with methamphetamine use and treatment outcomes. Lastly, the sample was drawn participants of several methamphetamine pharmacotherapy clinical trials and results from a treatment-seeking sample may not be generalizable to methamphetamine uses as a whole. In summary, we found an association between rs7591784 near CREB1 and pretreatment methamphetamine use, an important indicator of disease severity and predictor of subsequent treatment outcomes, as well as methamphetamine use during treatment independent of pretreatment methamphetamine use in males but not females. Replication of this result in independent samples is necessary but our results combined with previous research suggest that variability in CREB signaling may influence severity of methamphetamine use disorder as well as success in quitting methamphetamine with outpatient treatment and that medications targeting the CREB pathway such as the non selective phosphodiesterase inhibitor ibudilast may be effective treatments for methamphetamine use disorder. Future studies should examine the role of CREB-related polymorphisms and the associated epigenetic changes on response to treatment for methamphetamine use disorder and whether these biological influences on methamphetamine use differ between males and females.Worldwide, it is estimated that there are over three million youth living with HIV globally, with the majority of youth acquiring HIV perinatally . Youth with perinatally-acquired HIV may show cognitive deficits as well as developmental delay even among those with reconstituted immunologic and virologic status, making PHIV a common infectious cause of perinatally-acquired developmental disability globally . Combination antiretroviral therapy for children with PHIV has resulted in substantial improvements in health with survival beyond childhood and reductions in morbidity and mortality .

Early HIV infection, immune activation, and viral persistence during a critical period of development may be especially detrimental to developing brains in youth with PHIV . Brain development is an extended process that begins prenatally and continues throughout the first two decades of life,indoor vertical farming with increased sensitivity to experience during the first year of life in pathways responsible for sensory, language and higher order cognitive development . Adolescence is also a crucial developmental window marked by a period of rapid brain maturation via synaptic pruning and myelination. White matter volume increases while grey matter volume decreases , with parietal grey matter reduction prominent before adolescence, followed by dorsal, mesial, and orbital frontal grey matter reduction during and after adolescence . Studies including neuroimaging combined with cognitive evaluation allow for an in vivo characterization of how HIV and cART may mediate brain development . In adults, studies of post-mortem tissue and in vivo neuroimaging combined with cognitive testing have revealed atrophy in cortical and subcortical structures that is related to HIV severity and cognitive performance . Still, the effects of early HIV infection on the underlying brain in adolescents with PHIV have not been well-characterized . Adolescent brains are also subject to environmental influences, including substance use. Neuroimaging and neuropsychological studies in youth who use substances have found structural brain abnormalities, including grey matter volume reductions, as well as cognitive dysfunction .Thus, to carefully study effects of PHIV on youth treated with cART, it is important to account for substance use. We present one of the first studies to investigate the impact of HIV severity and coincident substance use on regional and total brain volumes and their association with cognition in PHIV youth. Other studies on grey matter volumes in PHIV do not focus on regional grey matter or substance use or are in PHIV populations with varying clinical characteristics from our cohort . Since PHIV youth often exhibit global cognitive functioning, working memory, and processing speed deficits , we hypothesized that frontal and parietal regions, regions important for higher-order cognitive functioning, would show volume reduction as compared to typically-developing, HIV unexposed and uninfected youth and smaller volumes would be associated with worse cognitive performance. We also hypothesized that HIV disease severity and substance use would be associated with reduced cortical grey matter volumes among adolescents with PHIV. 40 PHIV youth from a single site participating in the Adolescent Master Protocol study of the NIH Pediatric HIV/ AIDS Cohort Study network were recruited. Institutional review board approvals were obtained. Parents, legal guardians, or youth aged 18 years or older provided written informed consent; minors provided written assent. A control group of 334 typically developing, HIV-unexposed and uninfected youth was generated using frequency-matching for sex and age from the Pediatric Imaging, Neurocognition, and Genetics study Magnetic Resonance Imaging database from five sites . Of note, information regarding alcohol and drug use was not collected in the PING cohort. Total grey matter and 10 a priori cortical ROIs volumes were identified as primary outcomes. The remaining 74 ROIs were analyzed as secondary outcomes. Descriptive statistics and graphical methods were used to confirm the normality assumption for volume measures. Volumes were compared between PHIV youth and HIV-unexposed and uninfected youth using linear regression models. Results were reported for models with and without adjusting for age at scan, sex, race, caregiver education attainment, annual household income , and intracranial volume. Caregiver education was classified as high school education and below vs. greater than high school. Annual household income was classified as $30,000 and below vs. greater than $30,000. Percent change in volume as compared to HIV-unexposed and uninfected youth was calculated based on adjusted means.For evaluating associations of volume with cognitive functioning, we considered the 10 primary ROIs as well as total grey matter. Linear regression was used to evaluate associations between ROI volume as well as total grey matter and working memory, processing speed, and cognitive proficiency indices, adjusting for sex and age at scan. Three sets of sensitivity analyses were conducted: 1) including substance use in models of associations of brain volumes with HIV measures for grey matter volumes; 2) adjusting for total grey matter volume in models evaluating associations between brain volumes and substance use for secondary ROIs; 3) including substance use, sex, and age at scan in analyses evaluating associations of brain volumes with cognition. As described previously for other neuroimaging studies in this cohort , for PHIV youth, the mean interval between scanning and assessment of recent disease markers was 1.8 months with 83% of recent VL measures within three months prior to scanning. Mean interval between scanning and cognitive testing was 3.8 months . All but 2 participants completed cognitive assessments within 1 year of neuroimaging, with 31 within 3 months. One fourth of PHIV youth reported tobacco use, 35% alcohol, and 35% marijuana use. 13 youth reported both alcohol and marijuana use, and 8 youth reported use of tobacco, alcohol, and marijuana. Due to small numbers reporting illicit drug use beyond marijuana , this measure was not considered further in statistical analyses . There was no difference in substance use by race. Adjustment for substance use in models evaluating associations between primary ROI volumes and HIV disease severity as well as cognitive function did not alter findings. Similarly, adjustment for total grey matter volume in models evaluating associations of HIV severity and substance use measures with secondary ROI volumes did not alter findings, with no associations identified based on a FDR level of 0.10. Positive associations observed between primary brain volumes and cognitive function persisted after adjustment for substance use. Bilateral superior frontal volumes positively correlated with CPI, WMI, and PSI after adjustment for substance use.The current study is one of the first to examine relationships among PHIV infection, disease severity, and substance use on the brain grey matter and cognitive outcomes in PHIV youth. We found that PHIV youth had reduced total grey matter volume as well as reduced volumes in the rostral middle frontal, postcentral, precentral, and superior parietal gyri, compared to similarly-aged HIV-unexposed and uninfected youth. These patterns persisted after adjusting for sex, age at scan, race, caregiver education attainment, annual household income, and intracranial volume.

How Much Does Vertical Farming Cost

The cost of vertical farming can vary significantly depending on various factors such as the scale of the operation, the technology and equipment used, the location, and the crops being grown. Here are some key cost considerations in vertical farming:

  1. Infrastructure and Facility Costs: The initial investment in setting up a vertical farm includes costs for acquiring or constructing a suitable facility, such as a building or warehouse. This may involve retrofitting the space with necessary systems like lighting, climate control, irrigation, and vertical growing structures. The costs can vary greatly based on the size and complexity of the operation.
  2. Lighting Systems: Vertical farms typically use artificial lighting, such as LED grow lights, to provide the necessary light spectrum for plant growth. The cost of lighting systems depends on the size of the farm, the type and quality of lights used, and the specific lighting requirements of the crops being grown. LED lighting can be energy-efficient but may have a higher upfront cost compared to traditional lighting options.
  3. Climate Control and HVAC: Vertical farms require precise control over temperature, humidity, and ventilation to create optimal growing conditions. The cost of climate control systems, including heating, ventilation, and air conditioning (HVAC), depends on the size of the facility and the level of automation and sophistication required.
  4. Growing Systems and Equipment: Vertical farming often utilizes hydroponic or aeroponic systems to grow plants in a soilless environment. The cost of these systems varies based on the scale, complexity, and type of technology employed. This includes costs for nutrient delivery systems, pumps, sensors, automation equipment, and monitoring systems.
  5. Operational Expenses: Ongoing operational expenses in vertical farming include costs for electricity, water, nutrients, labor, and crop inputs such as seeds or seedlings. These costs will depend on the size of the operation, the efficiency of the systems, and local utility rates.
  6. Research and Development: Depending on the level of innovation and technology used, vertical farming may involve research and development costs for optimizing growing techniques, developing proprietary systems, or customizing technologies.

It is challenging to provide a specific cost estimate for vertical farming as it can vary widely depending on the factors mentioned above. Large-scale commercial vertical farms with advanced technology and automation can require significant upfront investment, while smaller-scale operations or vertical farming at home can be more affordable. Conducting a detailed feasibility study, considering the specific requirements of your operation, and consulting with experts in the field can help provide a more accurate cost estimation for your vertical farming project.

When Did Vertical Farming Start

Vertical farming as a concept has been around for several decades, but its modern form began to take shape in the early 2000s. The idea of growing plants vertically, stacked in layers or on vertical surfaces, emerged as a potential solution to address various challenges in agriculture, such as land scarcity, climate limitations, and food security concerns.

One of the pioneers in vertical farming is Dickson Despommier, a professor of environmental health sciences at Columbia University. Despommier popularized the concept of vertical farming in 1999 with his book “The Vertical Farm: Feeding the World in the 21st Century.” His book laid out the vision and potential benefits of growing crops in multilevel indoor environments using hydroponics or aeroponics.

Following Despommier’s work, vertical farming gained more attention and traction in the early 2000s. Companies and entrepreneurs began exploring and implementing vertical farming systems, experimenting with different technologies and designs to maximize productivity and efficiency. Advancements in LED lighting, hydroponic systems, and automation technologies also contributed to the growth and development of vertical farming.

Around the mid-2000s, the first commercial vertical farming ventures started to emerge. These early pioneers focused on leafy greens and herbs as the primary crops due to their fast growth and suitability for controlled indoor environments. As the industry evolved, vertical farming expanded to include a wider range of crops, including strawberries, tomatoes, peppers, and even vine crops like cucumbers.

Since then, vertical farming has continued to gain momentum globally. Numerous vertical farms have been established in various countries, utilizing innovative techniques and technologies to improve efficiency, sustainability, and crop quality. The industry is constantly evolving, with ongoing research and development driving advancements in vertical farming practices.

Overall, while the concept of growing plants vertically can be traced back further, the modern era of vertical farming began to take shape in the early 2000s with the work of Dickson Despommier and subsequent efforts by researchers, entrepreneurs, and companies to explore and develop this innovative farming approach.

How Long to Dry Cannabis on Drying Rack

The drying process for cannabis after harvest typically takes around 7 to 14 days, depending on various factors such as humidity, temperature, and the desired moisture content. When using a drying rack, here are some general guidelines to follow:

  1. Hang the buds: After harvesting the cannabis plants, trim the excess leaves and branches, leaving only the buds. Hang the buds upside down on the drying rack. This allows for proper airflow around the buds.
  2. Ideal environment: The drying environment should have a humidity level of around 45-55% and a temperature of approximately 20-24°C (68-75°F). These conditions help prevent mold or mildew growth and preserve the quality of the buds.
  3. Check for dryness: During the drying process, regularly monitor the buds for moisture content. Gently squeeze a small bud between your fingers to assess its dryness. It should feel slightly dry and crispy on the outside but still have some moisture inside.
  4. Slow drying: It’s important to dry the cannabis buds slowly to ensure proper curing and preservation of cannabinoids and terpenes. Rapid drying can result in a harsh taste and reduced potency. Avoid using excessive heat or forced air, as this can accelerate the drying process and negatively impact the final product.
  5. Patience: Drying cannabis properly requires patience. It’s better to dry the buds slowly over a slightly longer period than to rush the process. This allows for a more controlled and even drying, resulting in better quality buds.
  6. Final moisture content: The drying process is considered complete when the buds reach a moisture content of around 10-15%. To determine this, use a hygrometer or moisture meter to measure the moisture levels in the buds. Once the desired moisture content is reached, the buds are ready for the curing process.

Remember, these guidelines are general and can vary depending on the specific strain, environmental conditions, and personal preferences. It’s essential to closely monitor the drying process and make adjustments as needed to achieve the desired results. Properly dried and cured cannabis will have improved flavor, potency, and overall quality.