Raha syrup attenuates the morphine withdrawal-induced locomotor sensitisation by increasing the cerebrospinal fluid serotonin in rats receiving the opium tincture maintenance treatment.

Morphine withdrawal increases locomotor sensitisation, relapse and impair regulation of serotonin system. We evaluated the effectiveness of Raha syrup on the cerebrospinal fluid (CSF) serotonin levels following locomotor sensitisation in morphine-withdrawn rats receiving the opium tincture (OT). Morphine withdrawal rats gavaged daily with OT and Raha syrup (for 30 days) and then challenged with morphine and evaluated for locomotor activity and CSF serotonin levels before morphine challenge and 2 weeks after cessation of treatment. Raha syrup attenuated locomotor activity, increased the CSF serotonin after morphine challenge, and continued 2 weeks after cessation of treatment in rats receiving OT. Whereas, rats receiving OT alone after morphine challenge exhibited a relative decrease in locomotor activity, without changing CSF serotonin. Raha syrup attenuated locomotor sensitisation in morphine-withdrawn rats receiving OT probably by increasing serotonin. Therefore, administration of Raha syrup along with OT may benefit to treatment relapse in addicts receiving OT maintenance treatment.

A machine learning approach for computation of cardiovascular intrinsic frequencies.

Analysis of cardiovascular waveforms provides valuable clinical information about the state of health and disease. The intrinsic frequency (IF) method is a recently introduced framework that uses a single arterial pressure waveform to extract physiologically relevant information about the cardiovascular system. The clinical usefulness and physiological accuracy of the IF method have been well-established via several preclinical and clinical studies. However, the computational complexity of the current L2 optimization solver for IF calculations remains a bottleneck for practical deployment of the IF method in real-time settings. In this paper, we propose a machine learning (ML)-based methodology for determination of IF parameters from a single carotid waveform. We use a sequentially-reduced Feedforward Neural Network (FNN) model for mapping carotid waveforms to the output parameters of the IF method, thereby avoiding the non-convex L2 minimization problem arising from the conventional IF approach. Our methodology also includes procedures for data pre-processing, model training, and model evaluation. In our model development, we used both clinical and synthetic waveforms. Our clinical database is composed of carotid waveforms from two different sources: the Huntington Medical Research Institutes (HMRI) iPhone Heart Study and the Framingham Heart Study (FHS). In the HMRI and FHS clinical studies, various device platforms such as piezoelectric tonometry, optical tonometry (Vivio), and an iPhone camera were used to measure arterial waveforms. Our blind clinical test shows very strong correlations between IF parameters computed from the FNN-based method and those computed from the standard L2 optimization-based method (i.e., R≥0.93 and P-value ≤0.005 for each IF parameter). Our results also demonstrate that the performance of the FNN-based IF model introduced in this work is independent of measurement apparatus and of device sampling rate.

Projection of healthcare demand in Germany and Switzerland urged by Omicron wave (January-March 2022).

In January 2022, after the implementation of broad vaccination programs, the Omicron wave was propagating across Europe. There was an urgent need to understand how population immunity affects the dynamics of the COVID-19 pandemic when the loss of vaccine protection was concurrent with the emergence of a new variant of concern. In particular, assessing the risk of saturation of the healthcare systems was crucial to manage the pandemic and allow a transition towards the endemic course of SARS-CoV-2 by implementing more refined mitigation strategies that shield the most vulnerable groups and protect the healthcare systems. We investigated the epidemic dynamics by means of compartmental models that describe the age-stratified social-mixing and consider vaccination status, type, and waning of the efficacy. In response to the acute situation, our model aimed at (i) providing insight into the plausible scenarios that were likely to occur in Switzerland and Germany in the midst of the Omicron wave, (ii) informing public health authorities, and (iii) helping take informed decisions to minimize negative consequences of the pandemic. Despite the unprecedented numbers of new positive cases, our results suggested that, in all plausible scenarios, the wave was unlikely to create an overwhelming healthcare demand; due to the lower hospitalization rate and the effectiveness of the vaccines in preventing a severe course of the disease. This prediction came true and the healthcare systems in Switzerland and Germany were not pushed to the limit, despite the unprecedentedly large number of infections. By retrospective comparison of the model predictions with the official reported data of the epidemic dynamic, we demonstrate the ability of the model to capture the main features of the epidemic dynamic and the corresponding healthcare demand. In a broader context, our framework can be applied also to endemic scenarios, offering quantitative support for refined public health interventions in response to recurring waves of COVID-19 or other infectious diseases.

Results from Canton Grisons of Switzerland suggest repetitive testing reduces SARS-CoV-2 incidence (February-March 2021).

In February 2021, in response to emergence of more transmissible SARS-CoV-2 virus variants, the Canton Grisons launched a unique RNA mass testing program targeting the labour force in local businesses. Employees were offered weekly tests free of charge and on a voluntary basis. If tested positive, they were required to self-isolate for ten days and their contacts were subjected to daily testing at work. Thereby, the quarantine of contact persons could be waved.Here, we evaluate the effects of the testing program on the tested cohorts. We examined 121,364 test results from 27,514 participants during February-March 2021. By distinguishing different cohorts of employees, we observe a noticeable decrease in the test positivity rate and a statistically significant reduction in the associated incidence rate over the considered period. The reduction in the latter ranges between 18 and 50%. The variability is partly explained by different exposures to exogenous infection sources (e.g., contacts with visiting tourists or cross-border commuters). Our analysis provides the first empirical evidence that applying repetitive mass testing to a real population over an extended period of time can prevent spread of COVID-19 pandemic. However, to overcome logistic, uptake, and adherence challenges it is important that the program is carefully designed and that disease incursion from the population outside of the program is considered and controlled.

Smart investment of virus RNA testing resources to enhance Covid-19 mitigation.

A variety of mitigation strategies have been employed against the Covid-19 pandemic. Social distancing is still one of the main methods to reduce spread, but it entails a high toll on personal freedom and economic life. Alternative mitigation strategies that do not come with the same problems but are effective at preventing disease spread are therefore needed. Repetitive mass-testing using PCR assays for viral RNA has been suggested, but as a stand-alone strategy this would be prohibitively resource intensive. Here, we suggest a strategy that aims at targeting the limited resources available for viral RNA testing to subgroups that are more likely than the average population to yield a positive test result. Importantly, these pre-selected subgroups include symptom-free people. By testing everyone in these subgroups, in addition to symptomatic cases, large fractions of pre- and asymptomatic people can be identified, which is only possible by testing-based mitigation. We call this strategy smart testing (ST). In principle, pre-selected subgroups can be found in different ways, but for the purpose of this study we analyze a pre-selection procedure based on cheap and fast virus antigen tests. We quantify the potential reduction of the epidemic reproduction number by such a two-stage ST strategy. In addition to a scenario where such a strategy is available to the whole population, we analyze local applications, e.g. in a country, company, or school, where the tested subgroups are also in exchange with the untested population. Our results suggest that a two-stage ST strategy can be effective to curb pandemic spread, at costs that are clearly outweighed by the economic benefit. It is technically and logistically feasible to employ such a strategy, and our model predicts that it is even effective when applied only within local groups. We therefore recommend adding two-stage ST to the portfolio of available mitigation strategies, which allow easing social distancing measures without compromising public health.

Morphine dependence and withdrawal-induced changes in mouse Sertoli cell (TM4) line: Evaluation of apoptotic, inflammatory and oxidative stress biomarkers.

Chronic morphine exerts deleterious effects on testicular function through either suppression of germ cells or somatic including Sertoli cells, probably through the activation of inflammatory, oxidative, and apoptosis biomarkers. Thus, the present study aimed to investigate whether the damaging effects of morphine dependence were reversed by the spontaneous morphine withdrawal or incubation with methadone and/or naloxone in Sertoli (TM4) cells using an in- vitro cell model of morphine dependence. Morphine dependence in TM4 cells was induced by increasing daily doses of morphine for 10 days and then maintained for two weeks in 5 µM. The cAMP levels were measured for an evaluation of morphine dependence. The cell viability and inflammatory, oxidative, apoptosis biomarkers, and glial cell-derived neurotrophic factor (GDNF) were measured after the end of treatment following the incubation of cells with methadone and naloxone and spontaneous withdrawal from morphine. We found that morphine dependence decreased cell viability, GDNF level and increased the levels of pro-oxidant, pro-inflammatory, and apoptotic biomarkers in TM4 cells, while spontaneous withdrawal from morphine and by naloxone decreased the levels of the biomarkers of pro-inflammatory and apoptotic in TM4 cells. Also, despite the low levels of pro-inflammatory factors following morphine withdrawal by methadone, it increased the cleaved/pro-caspase3 ratio in TM4 cells. This study showed that morphine dependence increased apoptosis probably via oxidative stress and inflammation pathways in TM4 cells. Also, it seems likely that spontaneous and naloxone withdrawal have beneficial consequences in the treatment of morphine dependence than methadone therapy, although they may require longer incubation periods.

Dynamic modelling to identify mitigation strategies for the COVID-19 pandemic.

Relevant pandemic-spread scenario simulations can provide guiding principles for containment and mitigation policies. We devised a compartmental model to predict the effectiveness of different mitigation strategies with a main focus on mass testing. The model consists of a set of simple differential equations considering the population size, reported and unreported infections, reported and unreported recoveries, and the number of COVID-19-inflicted deaths. We assumed that COVID-19 survivors are immune (e.g., mutations are not considered) and that the virus is primarily passed on by asymptomatic and pre-symptomatic individuals. Moreover, the current version of the model does not account for age-dependent differences in the death rates, but considers higher mortality rates due to temporary shortage of intensive care units. The model parameters have been chosen in a plausible range based on information found in the literature, but it is easily adaptable, i.e., these values can be replaced by updated information any time. We compared infection rates, the total number of people getting infected and the number of deaths in different scenarios. Social distancing or mass testing can contain or drastically reduce the infections and the predicted number of deaths when compared with a situation without mitigation. We found that mass testing alone and subsequent isolation of detected cases can be an effective mitigation strategy, alone and in combination with social distancing. It is of high practical relevance that a relationship between testing frequency and the effective reproduction number of the virus can be provided. However, unless one assumes that the virus can be globally defeated by reducing the number of infected persons to zero, testing must be upheld, albeit at reduced intensity, to prevent subsequent waves of infection. The model suggests that testing strategies can be equally effective as social distancing, though at much lower economic costs. We discuss how our mathematical model may help to devise an optimal mix of mitigation strategies against the COVID-19 pandemic. Moreover, we quantify the theoretical limit of contact tracing and by how much the effect of testing is enhanced, if applied to sub-populations with increased exposure risk or prevalence.

Microinjection of the BDNF receptor antagonist ANA-12 into the nucleus accumbens and medial-prefrontal cortex attenuates morphine-induced reward memory, and alterations of BDNF levels and apoptotic cells in rats.

This study was designed to examine the effects of intra- nucleus accumbens (NAc) of BDNF receptor antagonist ANA-12 on the acquisition and expression and intra- medial-prefrontal cortex (mPFC) of ANA-12 on the extinction and reinstatement of morphine-induced conditioned place preference (CPP) and also BDNF levels and apoptotic neurons in the NAc and mPFC of rats. In this study, adult male Wistar rats (200-250 g) were used. Two separate cannulas were inserted bilaterally into the NAc and/or mPFC. ANA-12 (3 µg/0.5 µl/side) was injected into the NAc and/or mPFC to evaluate the rewarding effects of morphine using a CPP paradigm. Then, the levels of BDNF and apoptotic in the NAc and mPFC were assessed at the end of each treatment phase using ELISA and TUNEL methods, respectively. All of vehicle-treated rats following morphine CPP showed the increase of BDNF levels and apoptotic neurons in the NAc and mPFC. ANA-12 significantly attenuated the acquisition and expression of morphine-induced CPP, BDNF levels and apoptotic neurons in the NAc during the acquisition, but not the expression phase. Also, ANA-12 significantly facilitated the extinction, but no effect on reinstatement of morphine CPP, and decreased BDNF levels and apoptotic neurons in the mPFC during the extinction, but not the reinstatement. We conclude that blocking TrkB with ANA-12 showed therapeutic effects on morphine-associated reward memory and neuronal death in the NAc and mPFC induced by morphine CPP. Thus, the BDNF-TrkB signaling may be important in the acquisition, expression, extinction, but not the reinstatement of morphine CPP.

Saffron (Crocus sativus L.) stigma reduces symptoms of morphine-induced dependence and spontaneous withdrawal in rats.

Background: Chronic morphine induces physical and psychological dependence signs. Saffron (Crocus sativus L.) stigma has been shown to have anxiolytic, antidepressant, and antinociceptive properties and to alleviate naloxone-precipitated withdrawal signs.Objectives: Therefore, this study was designed to examine the effects of saffron aqueous extract on the severity of physical-psychological dependence, voluntary morphine consumption, and the cerebrospinal fluid (CSF) serotonin levels following locomotor sensitization in morphine-dependent rats and in rats undergoing morphine withdrawal.Materials: Adult male rats were treated with morphine (10 mg/kg, sc twice daily) for 10 days. Rats received saffron extract (60 mg/kg, ip) daily, during the induction of morphine dependence and/or withdrawal. Then, rats were tested for spontaneous withdrawal signs, anxiety using the elevated plus-maze, depression using sucrose preference test, and voluntary morphine consumption using a two-bottle choice paradigm, and then challenged with morphine (1 mg/kg, ip) to evaluate of locomotor sensitization and CSF serotonin levels.Results: The results showed saffron extract during induction of morphine dependence decreased the severity of withdrawal signs (P = .05), while it had no effect on anxiety and depression-like behaviors. Saffron extract during morphine withdrawal exhibited an increase in the percentage (or ratio) of open/total arm entries (P = .017), higher levels of sucrose preference (P = .0001), a lower morphine preference ratio (P = .02) and also, a decrease in locomotor activity (P = .004) and an increase in the CSF serotonin levels (P = .041) in rats challenged to morphine.Conclusions: Saffron extract may exert a protective effect against morphine-induced behavioral sensitization in rats, probably through increasing serotonin levels.

Environmental Enrichment Ameliorates Psychological Dependence Symptoms and Voluntary Morphine Consumption in Morphine Withdrawn Rats Under Methadone Maintenance Treatment.

INTRODUCTION: Previous studies have shown that physical and psychological dependence and the vulnerability to relapse are still present during MMT. Thus, this study examined whether Enriched Environment (EE) would attenuate anxiety, depressive, and obsessive-compulsive-like behaviors, as well as voluntary morphine consumption following Methadone Maintenance Treatment (MMT) in morphine withdrawn rats. METHODS: The rats were injected bi-daily doses (10 mg/kg, 12-h interval) of morphine for 14 days. Then, the rats were reared in a Standard Environment (SE) or EE for 30 more days during morphine withdrawal, simultaneous with receiving MMT. The rats were tested for anxiety (the Elevated Plus Maze [EPM]) and depression (Sucrose Preference Test [SPT]), Obsessive-Compulsive Disorder (OCD) as grooming behavior, and voluntary morphine consumption using a Two-Bottle Choice (TBC) paradigm. RESULTS: The findings revealed that EE experience in morphine withdrawn rats under MMT significantly increased the EPM open-arm time and higher sucrose preference than SE rats. Also, we found that the EE decreased the self-grooming behavior and morphine preference ratio in morphine withdrawn rats receiving MMT compared to the SE group. CONCLUSION: We conclude that exposure to EE decreased methadone-induced anxiety, depressive and OCD-like behaviors, and voluntary morphine consumption in morphine withdrawn rats under MMT. Thus, the EE seems to be one of the strategies for reducing MMT-induced behavioral dysfunction and the risk of relapse induced by morphine withdrawal.

Swimming exercise during morphine abstinence in parents-to-be attenuated morphine-induced conditioned place preference and locomotor sensitization only in male rat offspring.

This study was designed to examine the effects of swimming exercise during morphine abstinence in parents-to-be before mating on morphine-induced conditioned place preference (CPP) and locomotor sensitization in the pubertal male and female rat offspring. Male and female Wistar rats were injected with bi-daily doses (10 mg/kg, 12 h intervals) of morphine for 14 days. The exercising rats exposed to a regular swimming exercise (45 min/d, five days per a week) during 30 days of morphine abstinence before mating. Then, the pubertal male and female rat offspring were tested for morphine-induced CPP and locomotor sensitization (using the open field). The results showed that the pubertal male offspring of the morphine-abstinent parents-to-be exhibited an increase in CPP to morphine and locomotor activity after morphine challenge than the offspring from the control group. While, swimming exercise in morphine-abstinent parents-to-be decreased CPP score and locomotor activity in the pubertal male offspring than control offspring. Thus, exposure to swimming exercise in morphine-abstinent parents-to-be before mating may exert a protective effect against morphine-induced reward and locomotor sensitization in their pubertal offspring which may prevent the vulnerability of the first generation to drug abuse following opiate-addicted parents before mating.

BDNF receptor antagonism during the induction of morphine dependence exacerbates the severity of physical dependence and ameliorates psychological dependence in rats.

This study examined the effects of systemic administration of the TrkB receptor antagonist (ANA-12) during induction of morphine dependence on the severity of physical and psychological dependence and the cerebrospinal fluid (CSF) BDNF levels in morphine-dependent and withdrawn rats. Rats became morphine-dependent by increasing daily doses of morphine for 7 days, along with ANA-12 injection. Then, rats were tested for the severity of physical dependence on morphine (spontaneous withdrawal signs), anxiety-like (the elevated plus maze), depressive-like (sucrose preference test) behaviors after spontaneous morphine withdrawal. Also, the CSF BDNF levels were assessed 2 h after the last dose of morphine and day 13 after morphine withdrawal in morphine-dependent and withdrawn rats. We found that the morphine withdrawal signs were significantly higher in morphine dependent rats receiving ANA-12 on days of 5-7 after morphine withdrawal, also ANA-12 exacerbated overall dependence severity. While, the percentage of time spent in the open arms and sucrose preference were higher in morphine-dependent rats receiving ANA-12 than morphine-dependent rats receiving saline. Also, the ANA-12 injection decreased the CSF BDNF levels following morphine dependence, while increased it after morphine withdrawal. We conclude that the ANA-12 exacerbated the severity of physical morphine dependence but attenuated the anxiety/depressive-like behaviors in morphine-dependent and withdrawn rats. Also, ANA-12 injection was able to reverse the changes in the CSF BDNF levels. Therefore, ANA-12 is not more likely to complete treatment for opiate addiction.

Enriched Environment Ameliorates Cognitive Deficits and Locomotor Sensitization in Morphine-Withdrawn Rats Receiving Methadone Maintenance Treatment.

OBJECTIVE: This study was designed to examine whether enriched environments (EE) would attenuate object recognition and spatial learning and memory deficits and locomotor sensitization induced by methadone maintenance treatment (MMT) in morphine-withdrawn rats. METHODS: Male Wistar rats (170 ± 10 g) were injected with bi-daily doses (10 mg/kg, 12-h intervals) of morphine for 14 days. Rats receiving MMT were reared in the standard environment (SE) or EE during 30 days of morphine withdrawal. Then, the rats were tested for object recognition (the object recognition memory test, ORMT) and spatial learning and memory (the water maze) and then challenged with morphine (1 mg/kg, i.p.) and evaluated for locomotor activity (open-field box). RESULTS: The results revealed that the dependent/saline/EE (D/Sal/EE) and D/methadone/EE (D/Meth/EE) rats exhibited significant preference for the new object (p = 0.006 and p = 0.049), spent more time in the target zone (p = 0.045 and p = 0.005) on the water maze, and displayed a lower level of distance traveled (p = 0.002 and p = 0.0001) compared to their control groups reared in SE. CONCLUSIONS: We conclude that exposure to EE could ameliorate the object recognition and spatial memory deficits and also decrease locomotor sensitivity in morphine-withdrawn rats receiving MMT. Thus, EE may be beneficial in the treatment of addiction during MMT.

Effect of crocin on morphine tolerance and serum BDNF levels in a rat model of neuropathic pain.

BACKGROUND: Chronic use of morphine treatment for neuropathic pain leads to morphine-induced analgesic tolerance. Crocin contained in Crocus sativus L., exerts anti-inflammatory and analgesic effects. This study examined the effects of crocin on morphine tolerance and serum BDNF levels on neuropathic pain induced by chronic constriction injury (CCI) in rats. METHODS: CCI model of neuropathic pain was done in male Wistar rats (200-250 g). Rats were treated with crocin (15 or 30 mg/kg, intraperitoneally) alone or simultaneously with morphine (10 mg/kg, subcutaneously) during or after induction of CCI. Pain behavioral responses including mechanical allodynia and thermal hyperalgesia were measured from days of 15-27 after CCI. Then, rats were evaluated for serum BDNF levels on days 14 and/or 27. RESULTS: We found that morphine tolerance developed after the induction of neuropathic pain. The injection of crocin (15 and 30 mg/kg) was able to enhance analgesic effect of morphine by reduction of mechanical allodynia on days 15-27 post-surgery in CCI rats. While preemptive administration of crocin at a lower dose (15 mg/kg) maintained the analgesic effect of morphine. Morphine injection and/or co-administration with crocin (15, 30 mg/kg) decreased serum BDNF levels in CCI rats. CONCLUSION: These findings indicate that crocin may have a therapeutic effect to maintain morphine analgesic efficacy and also to prevent the development of morphine tolerance in neuropathic pain, but probably not through BDNF.

Impact of different intensities of forced exercise on deficits of spatial and aversive memory, anxiety-like behavior, and hippocampal BDNF during morphine abstinence period in male rats.

Forced exercise can alleviate cognitive-behavioral deficits in an experimental model of addiction. However, the effects of different intensities of forced exercise in improving behavioral, cognitive and biochemical deficits during morphine abstinence period are not well investigated. Thus, the current work examined the effects of different loads of forced exercise on cognition functions, anxiety behavior and BDNF changes in the hippocampus, and prefrontal cortex (PFC), and also serum levels of BDNF and corticosterone during the abstinent period in male rats. Animals received morphine injections (10 mg/kg, twice a day) for 10 consecutive days. Then, the animals were exposed to a 4-week forced exercise training program under low, moderate or high intensities (30 min per session on 5 days a week), which accompanied by behavioral and biochemical tests. In Experiment 1, anxiety-like behaviors using elevated plus maze (EPM), and light/dark box (L/D box) were examined. In Experiment 2, cognitive functions using T-maze alteration and passive avoidance tasks were tested, which accompanied by BDNF measurements in the hippocampus and PFC. In Experiment 3, serum levels of BDNF and corticosterone following the termination of forced exercise regimen were measured. Morphine-abstinent animals exhibited anxiogenic -like behaviors in the EPM, but not L/D box. They also exhibited impaired T-maze alternation performance and passive avoidance memory, and a decline in hippocampal BDNF, but not PFC. Forced exercise at a moderate intensity alleviated anxiety, cognitive and BDNF defects in morphine-abstinent animals. The high load exercise enhanced serum levels of corticosterone in both saline and morphine groups. Thus, regular moderate forced exercise may be beneficial in preserving cognitive and mood functions in male addicts during the abstinent period and drug rehabilitation.

Voluntary exercise and estradiol reverse ovariectomy-induced spatial learning and memory deficits and reduction in hippocampal brain-derived neurotrophic factor in rats.

Ample evidences have demonstrated the beneficial effects of physical exercise on cognitive functions such as learning and memory. It is well established that female sex hormones have an important role in regulating learning and memory. This study was designed to investigate the effects of voluntary exercise and estrogen replacement on learning and memory deficits and reduction in hippocampal brain derived neurotrophic factor (BDNF) levels induced by ovariectomy. Ovariectomized rats were given daily vehicle or 17 ß-estradiol (20 µg/kg) and allowed to freely exercise in a running wheel over the course of 2 weeks. After this period, they were trained and tested on a water-maze spatial task for 5 consecutive days, followed by a probe test one day later. At the end of the behavioral tests, all animals were decapitated and their hippocampal levels of BDNF were measured. Ovariectomy impaired spatial learning and memory and reduced hippocampal BDNF levels. Exercise significantly improved performance during both training and the retention of the water-maze task and increased hippocampal BDNF. Exercise, 17 ß-estradiol and their combination recovered the impairing effects of ovariectomy on learning and memory performance. The combined treatment did not produce stronger effect than either exercise or 17 ß-estradiol alone. Our findings provide an important evidence about positive influences of regular exercise and estrogen treatment against cognitive and BDNF deficits induced in ovariectomized rats, an experimental model of menopause.

Age- and sex-related changes in the severity of physical and psychological dependence in morphine-dependent rats.

Gender- and age-dependent effects on the severity of morphine dependence are still controversial. The aim of this study was to investigate the effects of age and sex on the severity of physical and psychological dependence in morphine-dependent rats. The adult/aged male and female Wistar rats were chronically treated with bi-daily doses (10 mg/kg, at 12 h intervals) of morphine for 14 days. Then, rats were tested for the severity of physical dependence on morphine (spontaneous withdrawal signs), anxiety-like (the elevated plus maze), depressive-like (sucrose preference test) and grooming behaviors after spontaneous morphine withdrawal. We found that the morphine withdrawal signs decreased after 3 and 7 days of withdrawal in female and male rats respectively, while there was no significant difference in overall dependence severity between the two sexes or ages. Also, we found that the withdrawal of morphine led to increased anxiety, depression and obsessive-compulsive behavior in the D (dependent)/Adult male and female rats. Also, the D/aged female and male rats exhibited a reduction in depressive-like behavior than the D/Adult rats. Moreover, the D/female rats exhibited a decreased obsessive-compulsive behavior in both age groups than male rats. We conclude that age has no effect on the duration of withdrawal from morphine and overall dependence severity. While, the duration of withdrawal from morphine was lower in female than male rats. Our results showed a sex difference on the duration of morphine withdrawal and an age difference in the expression of psychological dependence on morphine. Thus, therapeutic strategies may be different for opiate-dependent individuals in physical and psychological dimensions.

Effect of enriched environment during adolescence on spatial learning and memory, and voluntary consumption of morphine in maternally separated rats in adulthood.

This study was designed to examine the effect of environmental enrichment (EE) during adolescence on spatial learning and memory and voluntary morphine consumption in maternally separated (MS) male and female rats in adulthood. Male Wistar rats were allowed to mate with female virgin Wistar rats. Pups were separated from the dams daily for 180 min during postnatal days 2-14. All pups were weaned on day 21. The pups of both sexes were reared in a standard (SE) or enriched (EE) environment during postnatal days 21-50. Then, adulthood rats were tested for spatial learning and memory (Morris Water Maze), and voluntary consumption of morphine using a two-bottle choice paradigm (TBC). We found that the MS/SE rats showed longer escape latencies to find the platform on the third (the male) and fourth (the female) days of training than No MS/SE rats. Also, exposure to EE shortened the latency to escape in the male and female MS rats as training progressed than MS/SE rats. Moreover, the No MS/EE and MS/EE male rats spent significantly more time in the target zone compared with the SE control groups in the probe test. We also found that voluntary morphine consumption was higher in the male and female MS/SE than No MS/SE rats, while it was lower in the male and female MS/EE rats. The present results have shown that EE treatment may have potential therapeutic application for the prevention of the development of drug addiction and recovery from cognitive deficits following neonatal MS during adulthood.

Environmental Enrichment Attenuates Morphine-Induced Conditioned Place Preference and Locomotor Sensitization in Maternally Separated Rat Pups.

INTRODUCTION: This study investigated the effect of the environmental enrichment during adolescence on morphine-induced Conditioned Place Preference (CPP) and locomotor sensitization in maternally separated male and female rat pups. METHODS: Male Wistar rats were allowed to mate with female virgin Wistar rats. Pups were separated from them 3 hours per day during 2-14 days postnatal. All pups were weaned at 21 Postnatal Day (PND) and reared in standard environment or enriched environment from 21 to 50 PND with litter-mates of the same sex. The CPP and behavioral sensitization to morphine were assessed by an unbiased place conditioning paradigm and open filed method. RESULTS: The results showed that the maternal separation enhanced morphine-induced CPP in both sexes, locomotor sensitization in male pups and tolerance to morphine-induced motor activity in female pups during adolescence. While, male and female pups reared in an EE exhibited a decrease in morphine-induced CPP, locomotor sensitization and tolerance induced by maternal separation compared to their control pups. CONCLUSION: Access to enriched environment during adolescence may have a protective effect against morphine-induced reward, locomotor sensitization and tolerance in adolescent male and female rats following maternal separation.