Period Of Abstinence Research Articles

Cannabinoids detected in exhaled breath condensate after cannabis use

Cannabinoids can be detected in breath after cannabis use, but different breath matrices need to be explored as studies to date with filter-based devices that collect breath aerosols have not demonstrated that breath-based measurements can reliably identify recent cannabis use. Exhaled breath condensate (EBC) is an unexplored aqueous breath matrix that contains condensed volatile compounds and water vapor in addition to aerosols. EBC was collected from participants both before and at two time points (0.7 ± 0.2 h and 1.7 ± 0.3 h) after observed cannabis use. Eleven different cannabinoids were monitored with liquid chromatography tandem mass spectrometry. Five different cannabinoids, including Δ9-tetrahydrocannabinol (THC), were detected in EBC collected from cannabis users. THC was detected in some EBC samples before cannabis use, despite the requested abstinence period. THC was detected in all EBC samples collected at 0.7 h post use and decreased for all participants at 1.7 h. Non-THC cannabinoids were only detected after cannabis use. THC concentrations in EBC samples collected at 0.7 h showed no trend with sample metrics like mass or number of breaths. EBC sampling devices deserve further investigation with respect to modes of cannabis use (e.g, edibles), post use time points, and optimization of cannabinoid recovery.

The Survival Benefit of Reabstinence After Harmful Alcohol Use Following Early Liver Transplant for Severe Alcohol-Associated Hepatitis: A Multicenter ACCELERATE Study.

Early (i.e., without mandated period of abstinence) liver transplant (LT) for alcohol-associated hepatitis is the fastest-growing indication for LT in the United States and Europe. Harmful alcohol use after LT is associated with poor outcomes, but the distinction of establishing abstinence after return to drinking (i.e., reabstinence) is understudied. This study aims to characterize the survival outcomes of achieving reabstinence after post-LT harmful alcohol use. We analyzed early LT recipients from 12 US LT centers between 2006 and 2021. Post-LT alcohol use was characterized as harmful using criteria of "binge" (≥5 [men] or ≥4 [women] drinks in < 24 hours) or "frequent" (≥4 days in one week) by interview or phosphatidylethanol >20 ng/mL. Reabstinence was defined as ≥12 consecutive months without harmful alcohol use after harmful alcohol use. Among 347 LT recipients (64% male, median age 43, median Model for End-Stage Liver Disease-Sodium score 38) with median post-LT follow-up of 2.2 years (interquartile interval 1.1-3.6), 276 (80%) recipients had no evidence of harmful alcohol use, 35 (10%) recipients had reabstinence, and 36 (10%) recipients had continued harmful alcohol use without reabstinence. Five-year predicted survival, adjusted for age, sex, and Model for End-Stage Liver Disease-Sodium score, was lowest among LT recipients with continued harmful alcohol use (77%), but similar among those with no harmful use (93%) and reabstinence (94%). Achieving reabstinence after post-LT harmful alcohol use is associated with similar 5-year post-LT survival compared with those without evidence of post-LT harmful alcohol use. Our findings highlight the importance of early detection and treatment of post-LT alcohol use.

Neurocognitive and clinical profile of male patients with substance use disorder in early remission phase with and without comorbid depression

Substance Use Disorder (SUD) represents one of the most frequent conditions worldwide which commonly coexists with major depressive disorder (MDD). This comorbidity (SUD + MDD) is one of the most prevalent with patients showing certain social and clinical characteristics that could lead to a worsening of their cognitive performance. However, despite these particularities, only a few studies have addressed the possible differences in cognitive performance between patients with SUD + MDD compared with those with SUD-only patients. Therefore, the aim of this study is to examine the clinical and cognitive profile of patients with SUD + MDD vs. SUD-only who are in early remission phase. For this purpose, 271 male patients underwent a clinical and neuropsychological assessment (SUD + MDD group: N = 101; SUD-only group: N = 170). Results indicated that SUD + MDD patients showed worse cognitive performance than SUD in visuospatial reasoning, verbal memory and learning, recognition, and processing speed even after a 3-month period of abstinence. Furthermore, these patients exhibited more self-reported prefrontal symptoms, as well as worse social and clinical conditions. This study indicates that the neurocognitive and clinical profile of patients with SUD + MDD could represent a risk since their characteristics have been associated with poorer recovery and prognosis. Our results could be helpful in clinical practice highlighting the need for cognitive remediation strategies in these populations, providing information that would allow the implementation of more appropriate treatments and preventive strategies.

P-004 Impact of short abstinence versus testicular sperm on sperm DNA fragmentation: a systematic review and meta-analysis of prospective studies

Abstract Study question Does testicular sperm result in lower sperm DNA fragmentation (SDF) compared to ejaculated sperm after short abstinence? Summary answer Testicular sperm exhibited significantly lower SDF than ejaculated sperm post-short abstinence. What is known already Optimal sperm DNA integrity is vital for fertilization and embryo health. Research shows testicular sperm typically exhibit lower SDF than ejaculated sperm with a standard abstinence period in the same individuals. Additionally, evidence indicates reducing abstinence to &amp;lt; 2 days may decrease SDF, presenting a simpler, more cost-effective strategy than surgical retrieval. Yet, no studies have directly compared the efficacy of shorter abstinence against testicular sperm extraction for lowering SDF. Study design, size, duration Meta-analysis of 16 randomized controlled and prospective observational studies including four on testicular sperm (TESA or TESE, called TS) and twelve on short abstinence ejaculation. The meta-analysis followed MOOSE guidelines, scrutinizing databases including Cochrane Library, Web of Science, Embase, MEDLINE(R) and PUMBED up to November 16, 2023. The analysis was conducted using RevMan. The observational studies’ methodological quality was assessed using the Newcastle-Ottawa Scale, and the overall evidence quality was evaluated following the GRADE criteria. Participants/materials, setting, methods To compare short ejaculation duration and TESA (are not directly compared in the literature) for SDF levels, we analyzed relevant data from studies of each method. We adjusted the participant numbers in the TS group by 1/3 and included each TS study 3 times, to perform a comparison against the short duration studies which were in a ratio of 1:3. This approach maintained an unaltered cumulative subject count for the meta-analysis of TS studies. Main results and the role of chance A total of 641 patients were included, comprising 120 and 521 patients with SDF measurements following TS and ejaculation after a short abstinence period, respectively. The studies had varied inclusion criteria, with not all patients having an initial elevated SDF. Some studies had incomplete details on age and other demographics. However, the mean± SD age of 93 TS patients was 38.15± 5.48 years vs. 37.7± 6.0 years of 444 short abstinence patients, demonstrating no significant difference (P = 0.544). Short abstinence durations ranged from 1 to 48 hours. Diverse DNA fragmentation tests were used: TUNEL assay in three testicular sperm studies, SCD assay in one, and in the short abstinence group, 4 used TUNEL and 6 used SCD assays, along with one each using SCSA and Halosperm. The mean± SD SDF was lower in the TS group than in the short abstinence group (Mean difference -9.48, 95%CI -12.45 to -6.52, P &amp;lt; 0.001, I2= 85%). Sensitivity analysis revealed that no single study significantly influenced the results. Employing the GRADE criteria, the initial assessment categorized the overall quality of evidence as low due to the observational nature of the acquired data. All studies were of medium to high quality. Limitations, reasons for caution Our meta-analysis, the first to compare two distinct methods for reducing SDF, while navigating through challenges of diverse study designs, variable patient characteristics, and differing DNA fragmentation tests. Despite these limitations, it marks a significant step in a field where direct, head-to-head comparisons have been absent, offering crucial insights. Wider implications of the findings This study suggests testicular sperm may be better than ejaculated sperm for improving SDF in infertility cases. Direct comparisons are needed, before deeming short abstinence less effective. Future research should directly compare reproductive outcomes using both methods. Trial registration number not applicable

P-372 Reproductive ability in survivors of childhood, adolescent, and young adult testicular tumors

Abstract Study question There is little comprehensive fertility evaluation in testicular tumors. Summary answer Our study provides evidence that sexual dysfunction and infertility represent the major persisting side effects with testicular tumor patients. What is known already Testicular cancer, although accounting for 1% of all cancers, is the most common cancer in men of reproductive potential with a peak incidence in men between the ages of 15-35, with more than 95% of the patients being cured.There are many researches on semen quality in testicular cancer. Study design, size, duration We performed a retrospective study by collecting 126 records of testicular cancer patients who attempted sperm banking in the Human Sperm Bank, West China Second University Hospital, Sichuan University between 2011 and 2022. Participants/materials, setting, methods Semen samples were collected by masturbation after 2–7 days of sexual abstinence. While a few of the patients had a longer period of sexual abstinence than one week, due to emergency treatment (data not shown). All samples were allowed to liquefy at 37 °C for 60 min and were then assessed according to World Health Organization protocols (WHO, 1999, 2010). Main results and the role of chance Semen quality and hormone levels before chemotherapy/radiotherapy In our study, we compare semen samples before and after orchiectomy, no differences in semen analysis were observed between pre-and post-orchiectomy groups. Semen quality after cancer treatment: type of treatment Total sperm number worsened significantly at T3(T6, 6 months after the end of treatment), T6 compared to baseline (T0, before beginning cancer treatment) and returned to near-baseline values at T24 in Carboplatin 2 cycles and EP/BEP 2-4 cycles groups. Moreover, Serum LH and FSH was increased at T3, T6 compared at T0. Sexual Functioning The BFSI (Brief Male Sexual Function Inventory) is a validated self-report measure of sexual functioning. We compared BSFI scores before and after orchiectomy, no differences were observed in sexual drive, sexual erection, sexual ejaculation, DEE (drive, erection, ejaculation) problems and sexual satisfaction between pre-and post-orchiectomy groups. Chemotherapy/radiation group significantly fewer sexual satisfaction problems compared with untreatment/surgery group. Fertility outcome Fertility data were available for 88 patients. 65 (73.9%) had no children, for various reasons (young age, financial reasons, marital status), 6 (6.8%) already had at least one child prior to their cancer diagnosis. Seventeen patients (19.3%) wanted children, but only 11 of these succeeded in achieving fatherhood, through natural fertility (8 patients) or ART (3 patients). Limitations, reasons for caution Small sample size, and the follow-up was not long enough. Wider implications of the findings We analyzed semen parameters, hormone levels, sexual functioning, fertility outcome for testicular cancer patient, both before diagnosis and after treatment, with the aim to provide practical recommendations. Trial registration number not applicable’

P-086 Consequences of elevated levels of DNA fragmentation and its influence on pregnancy – A multicentric cohort analysis

Abstract Study question Does an abnormal DNA Fragmentation Index impact the clinical outcomes of patients undergoing IVF treatment? Summary answer Statistical differences were seen in biochemical, implantation, clinical, and ongoing pregnancy rate in sperms with high and low DFI scores with autologous and donated oocytes. What is known already Sperm chromatin integrity is associated with embryo development and clinical outcomes of an IVF cycle. DNA Fragmentation Index (DFI) is a diagnostic tool for the detection of spermatozoa with damaged chromosomes in the ejaculate. DNA fragmentation is a significant cause of day 3 embryo arrest and the development of poor-quality embryos. The interventions associated with elevated levels of DFI are ICSI, lifestyle changes and antioxidant therapies. However, the published data regarding the potential of embryo development with high DFI is limited to the sample size thus, encouraging a need for more research in this area. Study design, size, duration The study is population-based and included 768 male patients who underwent DFI score detection across 44 centers at Indira IVF Hospital Private Limited from January 2023 to September 2023. The abstinence period for all the patients was recorded during the DFI detection. After the DFI assessment patients were classified into normal DFI score ( &amp;lt; =15%) (n = 203) and abnormal DFI score (&amp;gt;15%) (n = 565). Participants/materials, setting, methods The retrospective cohort analysis was performed on our databank on the male patients regardless of their age (21-50), ethnicity, and medical history. DFI was calculated using a Sperm Chromatin Dispersion assay. Patients were categorized into two groups: Normal DFI ( &amp;lt; =15%) and Abnormal DFI (&amp;gt;15%). The characteristics of population along with clinical outcomes were evaluated after transferring embryos fertilized with normal DFI sperm and abnormal DFI sperm. The data was analyzed and estimated by SPSS software. Main results and the role of chance Our analyses discovered that embryos fertilized with sperms of normal DFI score have significantly enhanced the likelihood of implantation compared with embryos fertilized with sperms of abnormal DFI (50.27% v/s 44.40%; p = 0.047). Simultaneously, early pregnancy was evaluated by determining the values of beta-HCG in blood, also known as biochemical pregnancy rate (BPR). The embryos fertilized with sperms of normal DFI score significantly enhanced the BPR compared with embryos fertilised with sperms of abnormal DFI (67% v/s 59.1%; p = 0.048). Clinical Pregnancy Rate (CPR) defined as presence of a heartbeat of gestational sac observed during the 6-week scan. Furthermore, it was evaluated that the CPR showed a significant enhancement in embryos fertilized with sperms of normal DFI score in comparison with embryos fertilized with sperms of abnormal DFI (65.5% v/s 57.9%; p = 0.034). However, it was noticed that male DFI score had no impact on the miscarriage rate of embryos inseminated with normal DFI score and abnormal DFI score (7.52% v/s 7.34%; p = 0.687). Moreover, the pregnancy was tracked, and it was revealed that embryos inseminated with sperms of normal DFI score significantly enhanced the ongoing pregnancy till 12 weeks compared with embryos inseminated with sperms of abnormal DFI score (60.6% v/s 53.6%; p = 0.039). Limitations, reasons for caution Embryos are known to have a self-correct mechanism to correct the presence of mosaicism in the embryos. Hence, the oocyte quality also affects the outcomes of the IVF cycle. Therefore, using a high grade of the oocyte for the patients with abnormal DFI score can be useful. Wider implications of the findings Our data suggest that abnormal DFI have an impact on the clinical outcomes of an IVF cycle. Therefore, examining the levels of DNA Fragmentation is utmost imperative for a successful pregnancy. Trial registration number not applicable

P-083 Short ejaculatory abstinence time of 2 hours results in higher pregnancy outcomes compared to the recommended abstinence period of 2-7 days

Abstract Study question What is the impact of a short abstinence (2h) compared to the recommended (2-7 days) ejaculatory abstinence time on embryo utilization rate and pregnancy outcome? Summary answer A short abstinence of 2 hours results in similar embryo utilization rates, but in reduced sperm DNA fragmentation and higher pregnancy rates. What is known already Semen parameters exhibit variability among and within individuals, and ejaculatory abstinence time (EA) is a known factor contributing to intra-individual variation. Despite the challenge of providing a specific recommendation for the optimal timeframe, recent studies suggested that a shorter abstinence is likely to decrease DNA fragmentation levels in semen and improve pregnancy and live birth rates following assisted reproductive technology. At present, ESHRE recommends a more limited range than the WHO manual, advising an ejaculatory abstinence of 3 to 4 days. Study design, size, duration This prospective proof-of-concept sibling oocyte study (n = 100 cycles), conducted between January 2018 and June 2023, required two sperm samples per patient. The initial sperm sample was produced on the day of oocyte retrieval following the requested abstinence period of 2-7 days. The second sperm sample was collected 2 hours later. Sibling oocytes were randomly allocated to injection with the first or second sample. Sibling oocytes of each patient were cultured under the same conditions. Participants/materials, setting, methods Inclusion criteria were ≥ 6 mature oocytes (MII), fresh ejaculates with concentration &amp;gt;1x106/ml, ICSI with subsequent blastocyst culture. Endpoints were pregnancy outcome, fertilization, blastocyst development, utilization rate, sperm concentration/motility and DNA fragmentation index (DFI) measured by TUNEL. Statistical analysis was done taking into account female age and clustering. A total of 104 couples consented to provide two samples on the day of oocyte retrieval and were enrolled. Main results and the role of chance Average abstinence periods of 98:48h±20:01 vs 02:05h±00:18 were observed. Sperm concentration (x106/ml), progressive forward motility and DFI, were 46.53±39.6 vs 37.0±37.1 (p &amp;lt; 0.001), 52.14%±20.1 vs 49.79%±20.4 (p = 0.152) and 6.76%±7.0 vs 5.37%±7.2 (p = 0.024), respectively. Of the 1282 collected oocytes, 634 were injected with the first sperm sample and 648 with the ‘2h EA sample’. Fertilization rates per MII (79.3% vs 80.7%, p = 0.154) and good-quality day 3 embryos (76.7% vs 77.6%, p = 0.276) were similar between both groups. From the 93 patients with blastocyst culture (with average female age of 33.9±5.0 years; average male age of 36.4±6.6 years), 1173 oocytes were stratified for injection: 580 (first sample) and 593 (second sample), respectively. Fertilization rates (81.4% vs 82.3%, p = 0.074) and good-quality day 5 blastocysts (46.2% vs 49.5%, p = 0.534) did not differ between the groups. Utilization rate per inseminated (30.5% vs 35.8%, p = 0.126) or fertilized (37.5% vs 43.4%, p = 0.153) oocyte was similar between the groups. Single fresh and frozen embryo transfers were compared between embryos generated after the requested (2-7 days; n = 91) and short (2h; n = 94) abstinence. Pregnancy rate was significantly higher after short abstinence: 49.5% (45/91) vs 62.8% (59/94); p = 0.01. Ongoing pregnancy rate was 31.9% (29/91) and 40.4% (38/94); p = 0.10. Limitations, reasons for caution This is a prospective proof-of-concept study, not powered to detect differences in utilization and pregnancy outcomes. Not all patient are capable of producing two sperm samples within 2 hours from each other. Wider implications of the findings The results indicate a significant, yet not relevant difference in DNA fragmentation index (DFI) but a relevant difference in pregnancy rate between the two groups. If increased live birth rates are obtained after short abstinence, recommendations concerning EA might need adaptation. Trial registration number NCT03323892

P-099 The development of a novel sperm washing medium, enriched with serotonin as the active constituent, with the objective of enhancing sperm motility and viability

Abstract Study question Is it possible to develop a novel product that increases the success rate of treating male infertility through assisted reproductive techniques? Summary answer Sperm washing media, aiming to improve sperm quality, enhance sperm motility, and generally support male reproductive health in the field of assisted reproductive techniques. What is known already Infertility impacts 15% of all couples globally, with 50% of infertility cases attributed solely to male factors. Reduced semen motility is identified as a primary factor directly associated with infertility. The utilization of supplementation to enhance overall sperm quality has gained increasing popularity worldwide. The sperm preparation process in Assisted Reproductive Technology (ART) is crucial for successful fertilization, with sperm washing mediums playing a critical role in enhancing efficiency. Sperm-washing media is a specialized solution utilized in assisted reproductive technologies such as intrauterine insemination and in vitro fertilization to remove the seminal fluid and debris for sperm to undergo capacitation. Study design, size, duration In this study, 200 normozoospermic men were included, and their semen samples were analyzed over an 18-month period following the guidelines outlined by the World Health Organization (2010) to assess the efficacy of serotonin, selenium, zinc, L-Carnitine, and Coenzyme Q10 supplementation on sperm motility, viability, ROS and DNA damage. Participants/materials, setting, methods Sperm samples obtained from 200 volunteer men undergoing infertility assessments were subdivided into three samples.The sperm samples underwent preparation using the swim-up technique. The study involved the examination of samples in three distinct washing media: (1) Origio, (2) Irvine Scientific, (3) a prototype (serotonin, selenium, zinc, L-carnitine, and Coenzyme Q10). Each medium was analyzed for motility, apoptosis, reactive oxygen species (ROS), and DNA damage. Main results and the role of chance It is found that mean basal total sperm counts are 38 million. After sperm washing, mean total sperm counts of Origio®, Irvine® and Prototype are 15 million respectively (p &amp;gt; 0.05). As total progressive motility is % 40 before washing, it is increased to 62%,56% and 64% after washing with Origio®, Irvine® and Prototype, respectively (p &amp;gt; 0.05). Annexin V staining is used for detection of apoptosis in sperm samples. On average, early apoptosis over the total sperm count(TSC) per subgroups are 34.67% for Prototype, 33.17% for Origio® and 30.96% for Irvine®, whereas late apoptosis ratios are found significantly lower in Prototype compared to Origio and Irvine; 8.7%, 28.26% and 27.74%, respectively. Ratio of necrosis over TSC per subgroups is 3.15% for Prototype, 13.49% for Origio® and 18.78% for Irvine® whereas viability ratios are as 53.48%, 25.08%, 22.52%, respectively.(p &amp;lt; 0.05). ROS levels in semen after washing are significantly lower for Prototype (5.9%, 10.29%, and 18.22%, Prototype, Origio® and Irvine®, respectively) (p &amp;lt; 0.05). Limitations, reasons for caution Semen samples from males aged 28 to 45, who are unable to conceive, do not smoke, and have not undergone any surgical interventions, are collected from patients after a sexual abstinence period of 3-4 days. Wider implications of the findings We propose that our serotonin-activated washing media for sperm preparation can serve as a suitable choice for selecting high-quality sperm in assisted reproductive technology programs. These molecules may enhance the success of assisted reproduction techniques when present in sperm preparation media. Trial registration number SerotoneX

P-046 Ejaculatory abstinence: is there any association with embryo developmental competence? A Retrospective Study in time-lapse incubators

Abstract Study question Is ejaculatory abstinence (EA) associated with fertilization and blastocyst development in ICSI cycles with embryo culture in time-lapse incubators? Summary answer There is no association between length of EA and fertilization, blastocyst development or morphology. What is known already Semen analysis is pivotal in medically assisted reproduction (MAR), serving as a significant indicator of male partners’ health. The period of EA is a relevant factor that can affect sperm quality. Numerous studies described EA association with clinical outcomes, while its impact on embryological results remains inconclusive. Recent meta-analyses, despite a conflicting literature, highlighted positive effects of shorter abstinence periods. A recent study suggested that prolonged abstinence might reduce fertilization rate in ICSI cycles without time-lapse imaging, thereby reducing usable zygotes’ rate Study design, size, duration Retrospective study including all oocytes (N = 7164) from 1148 ICSI cycles (N = 902 couples) cultured in time-lapse incubators at a single IVF center between January 2020 and December 2022. Only non-donor fresh gametes were included (Mean Maternal age: 38.5±4 years; Paternal Age: 41±5.6 years). The primary embryological outcomes were fertilization rate and type, and blastocyst development per inseminated oocyte and per 2PN-zygote. Secondary outcomes were euploidy, day of blastocyst development and Gardner’s morphology. Participants/materials, setting, methods Semen analysis was conducted according to WHO. All oocytes were cultured up to day7 in EmbryoScope or GERI incubators with continuous media and refresh in day5 (if needed). If requested, PGT-A was performed to assess full-chromosome uniform aneuploidies. Univariate and multivariate logistic regressions were conducted. Possible confounders were assessed among ovarian stimulation, maternal, paternal, sperm, and IVF cycles characteristics. Associations were confirmed with Generalized estimating equations (GEE) to account for repeated measurements among oocytes’ cohort. Main results and the role of chance Logistic regressions and GEE were conducted to assess associations between possible confounders and blastocyst development per inseminated metaphase-II oocyte. Maternal age, sperm concentration, and motility A+B were associated. The same confounders were reported on blastocyst development among 2PN-zygotes. The days of EA (1-8 days) did not show any association with fertilization (p = 0.198) or blastocyst development per metaphase-II oocyte (p = 0.495). No association was reported with abnormal fertilization, neither combining all abnormalities in a single group, nor clustering them as 1PN, 3PN, 3.1PN, and multi-PN. An increasing trend was reported in blastocyst development among 2PN-zygotes according to the length of EA (1-day: 17/37,45.9%; 2-days: 190/424,44.8%; 3-days: 1405/2574,54.6%; 4-day: 746/1410,52.9%; 5-days: 450/827,54.4%; 6-days: 70/133,52.6%; 7-days: 46/101,45.5%; 8-days: 85/162,52.5%), but this was not significant in a multivariate logistic regression (p = 0.887). Length of EA showed no association with euploidy (p = 0.424), nor with day of development (p = 0.966) and Gardner’s inner cell mass (p = 0.204) or trophectoderm (p = 0.428) morphology. Limitations, reasons for caution This study is inherently limited by its retrospective design. The limited number of cycles included was balanced using time-lapse incubators to improve the accuracy of fertilization check and benefit from an undisturbed embryo culture environment. Artificial Intelligence will be implemented to automate developmental timings’ annotation and objectify embryo quality assessment. Wider implications of the findings The length of EA seems not associated with embryological outcomes, adding to the controversy on this topic. Here, we included only ICSI cycles with blastocyst culture conducted in time-lapse to enhance the quality of the evidence. Larger datasets and diverse settings are now required, especially enriched with poor prognosis patients. Trial registration number not applicable

Sex differences in behavior, cognitive, and physiological recovery following methamphetamine administration

Intact executive functions are required for proper performance of cognitive tasks and relies on balance of excitatory and inhibitory (E/I) transmission in the medial prefrontal cortex (mPFC). Hypofrontality is a state of decreased activity in the mPFC and is seen in several neuropsychiatric conditions, including substance use disorders. People who chronically use methamphetamine (meth) develop hypofrontality and concurrent changes in cognitive processing across several domains. Despite the fact that there are sex difference in substance use disorders, few studies have considered sex as a biological variable regarding meth-mediated hypoactivity in mPFC and concurrent cognitive deficits. Hypofrontality along with changes in cognition are emulated in rodent models following repeated meth administration. Here, we used a meth sensitization regimen to study sex differences in a Temporal Order Memory (TOM) task following short (7 days) or prolonged (28 days) periods of abstinence. GABAergic transmission, GABAA receptor (GABAAR) and GABA Transporter (GAT) mRNA expression in the mPFC were evaluated with patch-clamp recordings and RT-qPCR, respectively. Both sexes sensitized to the locomotor activating effects of meth, with the effect persisting in females. After short abstinence, males and females had impaired TOM and increased GABAergic transmission. Female rats recovered from these changes after prolonged abstinence, whereas male rats showed enduring changes. In general, meth appears to elicit an overall decrease in GABAAR expression after short abstinence; whereas GABA transporters are decreased in meth female rats after prolonged abstinence. These results show sex differences in the long-term effects of repeated meth exposure and suggest that females have neuroprotective mechanisms that alleviate some of the meth-mediated cognitive deficits.

Connecting self-report and instrumental behavior during incubation of food craving in humans.

Incubation of craving is a phenomenon describing the intensification of craving for a reward over extended periods of abstinence from reinforcement. Animal models use instrumental markers of craving to reward cues to examine incubation, while human paradigms rely on subjective self-reports. Here, we characterize an animal-inspired, novel human paradigm that showed strong positive relationships between self-reports and instrumental markers of craving for favored palatable foods. Further, we found consistent nonlinear relationships with time since last consumption and self-reports, and preliminary patterns between time and instrumental responses. These findings provide a novel approach to establishing an animal-inspired human model of incubation.

Changes of attentional bias in patients with alcohol use disorder during abstinence: A longitudinal study

BackgroundAlcohol Use Disorder (AUD) is linked to an attentional bias towards alcohol-related cues (e.g. images, smells), which acquire incentive properties and promote continued consumption. MethodWe investigated how the general and alcohol attentional bias evolved longitudinally in AUD patients along two periods of abstinence: t = 0 (baseline, 1–3 months of abstinence) and t = 1 (follow-up; 6 months of abstinence), as well as their relationship with alcohol-related variables. General and alcohol-specific attentional bias were evaluated by the Classic and the Alcohol Stroop tests (neutral and alcohol conditions) in abstinent AUD patients and controls. ResultsAt t = 0, the AUD group exhibited both general and alcohol-specific attentional biases, with greater effect in the general bias. At t = 1, alcohol-specific attentional bias decreased specifically in the AUD group and reached control levels (with interference index levels increasing from 1-3 months to 6 months). However, general attentional bias showed a trend toward improvement but it did not significantly change through abstinence process (linear mixed models, controlling for age, BMI, sex and education). ConclusionsIn AUD patients, general and alcohol attentional biases exhibit different trajectories during abstinence, with the attentional bias toward alcohol improving significantly throughout this process whereas general attentional bias is maintained.

Dynamic regulation of CeA gene expression during acute and protracted abstinence from chronic binge drinking of male and female C57BL/6J mice

While there are numerous brain regions that have been shown to play a role in this AUD in humans and animal models, the central nucleus of the amygdala (CeA) has emerged as a critically important locus mediating binge alcohol consumption. In this study, we sought to understand how relative gene expression of key signaling molecules in the CeA changes during different periods of abstinence following bouts of binge drinking. To test this, we performed drinking in the dark (DID) on two separate cohorts of C57BL/6J mice and collected CeA brain tissue at 1 day (acute) and 7 days (protracted) abstinence after DID. We used qRTPCR to evaluate relative gene expression changes of 25 distinct genes of interest related to G protein-coupled receptors (GPCRs), neuropeptides, ion channel subunits, and enzymes that have been previously implicated in AUD. Our findings show that during acute abstinence CeA punches collected from female mice had upregulated relative mRNA expression of the gamma-aminobutyric acid receptor subunit alpha 2 (Gabra2), and the peptidase, angiotensinase c (Prcp). CeA punches from male mice at the same time point in abstinence had upregulated relative mRNA encoding for neuropeptide-related molecules, neuropeptide Y (Npy) and somatostatin (Sst), as well as the neuropeptide Y receptor Y2 (Npyr2), but downregulated Glutamate ionotropic receptor NMDA type subunit 1 (Grin1). After protracted abstinence, CeA punches collected from female mice had increased mRNA expression of corticotropin releasing hormone (Crh) and Npy. CeA punches collected from male mice at the same timepoint had upregulated relative mRNA expression of Npy2r, Npy, and Sst. Our findings support that there are differences in how the CeA of male and female mice respond to binge-alcohol exposure, highlighting the need to understand the implications of such differences in the context of AUD and binge drinking behavior.

Non-Invasive Biomarkers for Differentiating Alcohol Associated Hepatitis from Acute Decompensation in Patients with ALD.

Alcohol-associated hepatitis (AH) is the most severe form of alcohol-related liver disease. The natural course of alcohol-related liver disease is influenced by heavy alcohol consumption and abstinence periods. Differentiating between AH and decompensated cirrhosis (DC) could be extremely challenging in clinical practice due to clinical and bioclinical similarities. The severity of AH is made on bioclinical grounds, the severe form necessitating corticotherapy treatment. Liver biopsy is still the standard of care for establishing the diagnosis in atypical presentations. The pathogenesis of AH is an interplay between gene expression, cytokine dysregulation, the immune system and the gut microbiota. Non-invasive tests are increasingly and widely used for the purpose of early diagnosis and reliable prognostication. The non-invasive tests are emerging in concordance with disease pathogenesis. In this review, we describe the non-invasive tools that can distinguish AH from DC. We outline the available cut-offs and their performance in diagnosis and prognosis, as well as in assessing the treatment response to corticotherapy. Promising circulating biomarkers like keratin 18, microRNAs and sphingolipids will be in the review.

Rotation from methadone to buprenorphine using a micro-dosing regime in patients with opioid use disorder and serious mental illness: A case series.

Inducting buprenorphine from methadone has traditionally involved initial opioid withdrawal, with risk of mental state deterioration in patients with serious mental illness (SMI). Micro-dosing of buprenorphine, with small incremental doses, is a novel off-label approach to transitioning from methadone and does not require a period of methadone abstinence. Given the limited literature about buprenorphine microdosing, we aimed to evaluate the feasibility and safety of inducting buprenorphine in a series of patients on methadone with SMI. For this retrospective case series, we reviewed the records of 16 patients with SMI at a Melbourne addiction treatment centre, from January 2021 to July 2022, who transitioned via micro-dosing, from high-dose methadone (>30 mg) to buprenorphine and depot-buprenorphine. Psychiatric diagnoses, mental state, other substance withdrawal, transfer success, transition time, opioid withdrawal symptoms and overall patient experience were collected via objective and subjective reporting. Methadone to buprenorphine transfer was completed by 88% of patients. Mental health measures remained stable with the exception of mildly increased anxiety. Median transfer time was 6.5 days for inpatients, 9 days for mixed setting and 10 days for outpatients. Most patients (93%) rated their experience 'manageable' reporting mild withdrawal symptoms. One patient met study criteria for precipitated withdrawal. This retrospective case series provides evidence that the use of a micro-dosing buprenorphine induction for methadone to buprenorphine transitions, including to depot-buprenorphine, has negligible risk, is tolerated by patients with SMI and is unlikely to precipitate an exacerbation of their mental illness.

Cannabidiol Protects against the Reinstatement of Oxycodone-Induced Conditioned Place Preference in Adolescent Male but Not Female Rats: The Role of MOR and CB1R.

Cannabidiol (CBD), a phytocannabinoid, appeared to satisfy several criteria for a safe approach to preventing drug-taking behavior, including opioids. However, most successful preclinical and clinical results come from studies in adult males. We examined whether systemic injections of CBD (10 mg/kg, i.p.) during extinction of oxycodone (OXY, 3 mg/kg, i.p.) induced conditioned place preference (CPP) could attenuate the reinstatement of CPP brought about by OXY (1.5 mg/kg, i.p.) priming in adolescent rats of both sexes, and whether this effect is sex dependent. Accordingly, a priming dose of OXY produced reinstatement of the previously extinguished CPP in males and females. In both sexes, this effect was linked to locomotor sensitization that was blunted by CBD pretreatments. However, CBD was able to prevent the reinstatement of OXY-induced CPP only in adolescent males and this outcome was associated with an increased cannabinoid 1 receptor (CB1R) and a decreased mu opioid receptor (MOR) expression in the prefrontal cortex (PFC). The reinstatement of CCP in females was associated with a decreased MOR expression, but no changes were detected in CB1R in the hippocampus (HIP). Moreover, CBD administration during extinction significantly potentialized the reduced MOR expression in the PFC of males and showed a tendency to potentiate the reduced MOR in the HIP of females. Additionally, CBD reversed OXY-induced deficits of recognition memory only in males. These results suggest that CBD could reduce reinstatement to OXY seeking after a period of abstinence in adolescent male but not female rats. However, more investigation is required.

After a period of forced abstinence, rats treated with the norepinephrine neurotoxin DSP-4 still exhibit preserved food-seeking behavior and prefrontal cortex fos-expressing neurons

AimsRelapse is a common characteristic of compulsive behaviors like addiction, where individuals tend to return to drug use or overeating after a period of abstinence. PFC (prefrontal cortex) neuronal ensembles are required for drug and food-seeking behaviors and are partially regulated by Norepinephrine (NE). However, the contributions of neuromodulators, such as the adrenergic system, in food-seeking behavior are not fully understood. Main methodsTo investigate this, we trained male and female rats to press a lever in an operant chamber to obtain banana-flavored food pellets for ten days. We then administered DSP-4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride), a neurotoxin that diminishes norepinephrine levels in the brain. The rats were kept in their home cages for ten more days before being returned to the operant chambers to measure food-seeking behavior. Key findingsDespite receiving DSP-4, the PFC neuronal ensembles measured by Fos and food-seeking behavior did not differ between groups, but rather sex. SignificanceAlthough both NE and Fos expressing neurons are implicated in food-seeking, they do not seem to be involved in a cue-contextual induced re-exposure response.

Brain Function and Marijuana: Are there Changes after Abstinence of Consumption? Preliminary Results.

The legalization of cannabis use and false claims about the plant Cannabis sativa to be considered a pharmaceutical product have been found to increase consumption, lower risk perception, and lead to more health problems, without reducing criminal activity. Brain function, typically assessed by neuropsychological tests, shows abnormalities with acute marijuana use, but inconsistent results have been published after abstinence, with a maximum follow-up of 28 days. Our previous research, using neuropsychological tests and brain perfusion single photon emission computed tomography (neuroSPECT), demonstrated consistent abnormalities in brain function among schoolchildren who consume marijuana compared to their non-consuming peers. The aim of this study is to investigate whether brain function changes in 20 adult marijuana users after 6 months of abstinence. Comparison of neuropsychological tests (Rey Complex Figure; Porteus Maze; Four subtests of WAIS-IV Intellectual Tests; STROOP; D2) and perfusion neuroSPECT (functional images), obtained in relation to recent consumption and after 6 months of serial drug-screening test confirmed abstinence. In a one-year period (2020-2021) only five compliant participants were recruited. The COVID-19 pandemic was a limiting factor. Preliminary results of neuropsychological tests, functional brain perfusion images and limited statistical analysis are presented. The results of the neuropsychological tests of the three subjects who completed the abstinence period so far show some improvement in working memory and attention after abstinence. NeuroSPECT shows disorganized hypoperfusion of variable severity in relation to recent consumption, involving areas associated with cognitive function such as the posterior cingulate and temporal lobes, in our five initially enrolled patients, when compared to a normal database. Of these, only two participants have already been re-evaluated with neuroSPECT after 6 months of abstinence, one of whom showed some improvement on the post-abstinence images. We analyze the methodological challenges of this research, including the pandemic, to incorporate the appropriate corrections in the next phase of our investigation. Our final findings may provide clinicians and users with information about the long-term effects of marijuana use on brain function.

Effects of long and short ejaculatory abstinence on sperm parameters: a meta-analysis of randomized-controlled trials.

This study aimed to investigate the effects of ejaculatory abstinence on sperm parameters. This analysis was registered in PROSPERO (CRD42023472124). We performed a search on PubMed using the following text terms: (("sperm parameters" OR "sperm analysis" [Mesh]) AND ("sperm DNA fragmentation" OR "DNA fragmentation" [Mesh]) AND ("sexual abstinence" [Mesh] OR "abstinence")) and an advanced search in Scopus using the terms ("sperm parameters" OR "sperm parameters" OR "DNA fragmentation") AND ("abstinence"). The sperm parameters that were investigated were sperm volume, total sperm motility, progressive sperm motility, sperm concentration, sperm morphology, and sperm DNA fragmentation (SDF). A two-day cut-off as a "short" or "long" abstinence period has been defined. Thirteen studies published between 2013 and 2022 were included in this meta-analysis. A total of 2,315 patients, ranging from 6 to 836 from each cohort, were enrolled in the study. We showed that longer abstinence time was associated with greater sperm concentration (mean difference [MD]: 8.19; p <0.01), sperm volume (MD: 0.96; p <0.01), and higher SDF (MD: 3.46; p <0.01), but lower progressive sperm motility (MD: -1.83; p <0.01). Otherwise, no statistically significant difference was observed in patients with longer vs. shorter abstinence times regarding total sperm motility (MD: -1.83; p = 0.06). Meta-regression analysis showed that days of abstinence were positively and linearly related to sperm concentration (slope: 3.74; p <0.01) and SDF (slope: 0.65; p = 0.044). According to our data, short ejaculatory abstinence is associated with better sperm quality. Indeed, a higher percentage of progressive sperm motility and lower levels of SDF have been reported in a short abstinence cohort. In contrast, the long abstinence group reported a higher sperm concentration. https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42023472124.

Restoring Cardiac Health: Abstinence Leads to Recovery from Alcohol-Induced Cardiomyopathy

Chronic drinking can lead to alcohol-induced cardiomyopathy (ACM), a disease currently without a cure. Using a mouse model of ACM, we observed significant cardiac dysfunction after 30 days. Our study’s goal was to investigate the potential restoration of cardiac function following a period of abstinence. Our hypothesis is that abstinence can lead to an observable recovery in overall cardiac function. C57BL/6J mice were given the Lieber-DeCarli liquid diet with 5% ethanol for 30 days, with two ethanol binges on days 10 and 30 (5 g/kg). After the 30-day ethanol exposure, the mice were transitioned to an ethanol-free diet for an additional 30 days. Left ventricular catheterization and echocardiography were conducted at the 30- and 60-day timepoints. 30 days of ethanol consumption led to significant decreases in both systolic and diastolic function. However, following a period of alcohol abstinence, there was a marked improvement. Specifically, the 30-day ethanol mice exhibited increases in stroke work and dP/dt max after 30 days of abstinence; stroke work increased from 1119 ± 89 mmHg*μL to 2032 ± 139 mmHg*μL (p &lt; 0.0001), and dP/dt max increased from 8054 ± 664 mmHg/s to 11967 ± 449 mmHg/s (p &lt; 0.001). Diastolic function also improved; dP/dt min from -7711 ± 561 mmHg/s to -10202 ± 594 (p &lt; 0.01). We also found a significant increase in load-independent cardiac contractility, i.e. the slope of end systolic pressure volume relation: 6.89 ± 0.6 mmHg/μL to 10.59 ± 1.57 mmHg/μL (p &lt; 0.05). In conclusion, abstinence after chronic ethanol exposure can lead to a restoration of cardiac function in C57BL/6J mice. This model parallels limited clinical data on abstinence and will allow us to study the underlying mechanisms responsible for the restoration of function. R21AA029747 (JG, CB), F30AA030472 (JE). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.