An efficient LC-QTOF-mass spectrometry method for monitoring nal-trexone compliance in urine of opioid-dependent subjects.

Naltrexone (NTX) is an orally effective opiate antagonist used in maintenance treatment for opiate dependence. Its utility is limited by the patient's noncompliance. The study aimed to develop an efficient method for the detection of NTX in urine by LC-QTOF-mass spectrometry (MS) and its application to NTX compliance in opioid-dependent subjects. Sample preparation included a dilution step and direct injection to LC-QTOF-MS. Chromatographic separation was achieved with a C-18 column using a mixture of mobile phase 0.1 percent formic acid in water and 0.1 percent formic acid in 95 percent methanol. The calibration curve was linear in the range 1-100 ng/mL with a correlation coefficient higher than 0.996. Precision and accuracy were acceptable, and the recovery efficiency range was 80-85 percent. The current LC-QTOF-MS method is simple, precise, sensitive, and can be used for monitoring NTX compliance among opioid-dependent subjects in a clinical setting.

Disability and quality of life in patients with treatment-resistant schizophrenia on long- term clozapine therapy.

Objectives: To assess disability and quality of life (QOL) in treatment resistant schizophrenia (TRS) on long term clozapine therapy and assess their correlation with positive, negative and cognitive symptoms. Methodology: Disability and QOL in forty patients with TRS (as per modified Kane's criteria) were assessed using World Health Organization Disability Assessment Schedule 2.0 and World Health Organization Quality of Life-BREF. Scale for assessment of positive symptoms, scale for assessment of negative symptoms and Addenbrooke's cognitive examination-III were used to assess positive, negative and cognitive symptoms. Medication adherence rating scale assessed medication adherence. Results: Disability and QOL correlated significantly with medication adherence, negative and cognitive symptoms but not with positive symptoms. Subgroup analysis revealed significant difference between medication adherence (good vs poor) and cognitive (impairment vs non-impairment) groups. Conclusion: Negative and cognitive symptoms, and medication adherence correlated with disability and QOL.

Effectiveness of smoking cessation intervention in opioid-dependent male subjects on buprenorphine maintenance treatment: An open-label trial.

Background: High prevalence (more than 80%) rates of tobacco smoking have been found both in, opioid-dependent subjects and among opioid-dependent subjects on opioid substitution treatment (OST) with buprenorphine or methadone. Aim: We aimed to explore the efficacy of combined nicotine replacement therapy (NRT) and individual counseling (IC) when compared to NRT alone in subjects on OST with buprenorphine. Methods: This study was carried out in a tertiary medical care center. It was an open-label randomized clinical trial. A total of 57 buprenorphine maintained smokers were recruited and randomized into two groups. They were assigned nicotine gum for 4 weeks plus either (1) a baseline IC session, and a second IC session after 1 week, or (2) simple advice to quit. In the first group, 31 subjects received NRT with IC and in the second group, 26 subjects received NRT plus simple advice to quit. The primary outcomes of this study were seven days point prevalence abstinence, biochemically confirmed by carbon monoxide (CO) breath analyzer, and reduction in smoking (mean no. of cigarettes or bidis/day). The smoking behavior during the 4 weeks follow-up period was assessed by the timeline follow-back (TLFB) method and confirmed by the CO breath analyzer. Results: The group of subjects who received NRT with IC showed higher rates of smoking cessation at the end of treatment (51%) as compared to the NRT and simple advice group where smoking cessation rates were around 8% (P < 0.001). Conclusion: A multi-component approach (pharmacotherapy and counseling) enhances treatment outcomes and enhances rates of abstinence from smoking.

Serum BDNF Levels Among Patients with Alcohol Dependence, Depression and Alcohol Dependence with Comorbid Depression - A Comparative Study.

The study aimed to assess and compare the serum brain derived neurotrophic factor (BDNF) levels in patients with alcohol dependence, depression and alcohol dependence with comorbid depression. Three groups each of 30 alcohol-dependent, depressive and alcohol-dependent with comorbid depressive patients seeking treatment were included. BDNF levels were estimated, and scales were administered to assess severity of alcohol dependence (using severity of alcohol dependence questionnaire, SADQ) and depressive symptoms (using Hamilton depression rating scale, HDRS). The mean BDNF value in ADS, depression and ADS with comorbid depression group was 16.4 ng/mL, 14.4 ng/mL and 12.29 ng/mL respectively, and the differences were statistically significant. In ADS group and ADS with comorbid depression groups significant negative association existed between BDNF and SADQ scores (r = -0.371, p = .043 and r = -0.0474, p = .008 respectively). There were significant negative association between BDNF and HDRS scores in depression and comorbid ADS and depression group (r = -0.400, p = .029 and r = -0.408, p = .025 respectively). The BDNF level was significantly lower in the ADS with comorbid depression group and was associated with severity of dependence and depression across the groups.

Does ethyl glucuronide in hair correlate with alcohol consumption? A comparative study with other traditional biomarkers among individuals with alcohol dependence syndrome.

AIM: Ethyl glucuronide (EtG) in hair is a direct biomarker proven to be useful for the detection of chronic excessive alcohol use. This study investigated the association of self-reported alcohol consumption with traditional biomarkers: GGT, AST, ALT, CDT, and MCV in blood and a direct biomarker, hair EtG, in a total of 122 patients with alcohol dependence syndrome. The diagnostic accuracy of the biomarkers to differentiate heavy from non-heavy drinkers was also evaluated. METHOD: GGT, AST, and ALT in serum were measured by Automated Chemistry Analyzer, MCV in blood was measured by Haematology Analyzer, serum CDT was analyzed by ELISA, and EtG in hair was evaluated by gas chromatography-mass spectrometry. The association between the biomarkers and the amount of alcohol consumed (self-reported) was determined using Spearman's rank correlation. RESULTS: All participants showed EtG level above the cut-off (0.03 ng/mg). Hair EtG showed a statistically significant linear and positive correlation with the amounts (in grams) of alcohol consumed (r = 0.60; p < 0.001). No correlation was observed among the traditional biomarkers and the quantity of alcohol consumed. Also, EtG showed an excellent receiver operating characteristic (ROC) curve (98%) with good sensitivity (85%) and specificity (60%) to classify heavy drinkers among individuals with alcohol dependence syndrome. CONCLUSION: Hair EtG can be helpful to estimate retrospective alcohol consumption in long-term chronic alcohol consumption cases. Hair EtG also provides a reliable diagnostic test to detect heavy drinkers among individuals with alcohol dependence syndrome.

An efficient method for simultaneous detection of Pheniramine, Pentazocine and cotinine in urine by Gas Chromatography in De-addiction program.

Background: Nonmedical use of prescription drugs for recreational purposes is a major health problem that raises high concerns for public health. Recently, several laboratory studies have reported the misuse of pentazocine, an agonist-antagonist opioid in combination with antihistamines in opioid addicts. Illicit self-administration of prescription drugs has been increasingly reported in India. Urinalysis as an adjunct to self-report plays a key role in providing additional information in the treatment of drug users. This paper aims to discuss a simple, convenient, and rapid capillary column gas-liquid chromatography method for simultaneous detection of pentazocine, pheniramine, and cotinine in urine. Methods: The sample was extracted with chloroform and isopropanol (3:1,v/v) and evaporated to dryness. After reconstitution with methanol, it was directly subjected to gas chromatography analysis. Method performance was evaluated and validated in terms of sensitivity, precision, the limit of detection (LOD), and the limit of quantification (LOQ). Findings: The linearity obtained was in the range of 50-1000 ng/ml with a correlation coefficient (r) above 0.999 for each drug. Good LOQ (50ng/ml) was obtained with each drug. Also, the developed method was effective in analyzing samples from patients with suspected abuse of these drugs. Conclusion: The technique was found to be simple, robust, sensitive, and precise in the simultaneous analysis of drugs (pentazocine, pheniramine, and cotinine). This method was proved to be useful and cost-effective in treating and monitoring patients seeking help for addiction in clinical settings.

Exploring the clinical variables among drug-dependent HIV-Positive patients: Implications for treatment strategies.

This study explored the profile of HIV positive patients seeking treatment at a tertiary care addiction treatment facility. A retrospective study was done to collet detailed information on clinical characteristics: drug use (type, age of initiation, duration), general medical condition and past treatment history. The study included 138 patients with mean (SD) age 30.2 (8.3) years. Opioid dependence with injecting drug use (IDU) was diagnosed in 97% of the patients. The median age of injecting onset was 24.5 years (IQR 20-31 years). The most frequently injected substances were pheniramine (60.1%) and buprenorphine (59.4%). Past treatment seeking was reported by 57% patients and interestingly they were less likely to present any medical condition (2 =69.611, p < 0.001). Variability in the age of onset of drug use indicates the need for broad based approach to prevent IDU and motivation to seek treatment may lead to better health conditions.

Effect of chronic opioid use on the hematological and inflammatory markers: A retrospective study from North India.

Background: Chronic opioid use affects biological functioning implicating the hematopoietic and immune system. It may alter various hematological parameters and inflammatory markers. This study aimed to assess the association of opioid dependence with the hematological parameters and inflammatory markers in the Indian population. Methods: A retrospective chart review was done among opioid dependent (ODS) males and healthy controls (HC) who visited the center's laboratory between Jan 2017 and Dec 2018 for hematological investigations. Clinical records reviewed for opioid use details like type, duration, and route of administration. The hematological profile presented as Mean or median. Mann-Whitney U test was used to compare the hematological parameters between the cases and controls. Results: The study included 191 ODS patients and 123 controls. Among ODS patients, a significant decrease in the levels of hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin and an increase in RBC count and lymphocytes was observed when compared to controls. The inflammatory markers, Neutrophil-Lymphocyte Ratio (NLR) and Platelet-Lymphocyte Ratio, were significantly lower among ODS. Longer duration of opioid use leads to increased NLR among ODS patients. Opioid use by injection did not alter any of the hematological parameters compared to non-injection drug use. Conclusion: Chronic opioid use has a significant effect on the hematopoietic cells. Opioid use for longer durations increases the inflammatory markers suggesting underlying infections.

Urinalysis based assessment of compliance and drug use patterns in patients prescribed tramadol: A cross-sectional study from a tertiary care centre.

BACKGROUND: Strict adherence to pharmacological dosage regimens is a prerequisite to the success of most treatments, particularly for patients in drug abuse programs. The compliance of tramadol, an atypical non-scheduled narcotic analgesic, using objective method has not been adequately studied in an Indian setting. AIM: To evaluate the compliance and pattern of drug use among opioid-dependent subjects prescribed tramadol based on urinalysis. METHOD: Fifty male opioid-dependent patients (ICD 10), seeking treatment at a tertiary de-addiction treatment centre of North India on tramadol prescription for atleast past four weeks were included. Self-reported substance use was recorded using semi-structured proforma. Ten ml of urine was collected for the assessment of compliance of tramadol of other substance use (morphine, buprenorphine, dextropropoxyphene, pentazocine, cannabis, benzodiazepines, pheniramine). All these drugs were analyzed using the immunoassay-based Cassette test and Gas Chromatography in human urine. RESULT: Mean age of the participants was 42.8 years and the mean duration of opioid use was 15.9years. The urine specimen of all subjects tested positive for tramadol. Urinalysis revealed benzodiazepines, cannabis, and pheniramine to be the most common substances of use in this population. It was seen that agreement of self-reporting and urine test results was good for morphine (κ = 0.558) and cannabis (κ = 0.312) and was poor for buprenorphine, pentazocine, and pheniramine. CONCLUSION: The study demonstrates the continued use of several illicit or non-prescribed medications in a medication-assisted opioid treatment population. The results affirm the reliability of urinalysis as an adjunct for testing compliance in such a population.

Serum Corticotrophin Releasing Factor (CRF) and its correlation with stress and craving in detoxified opioid-dependent subjects.

Corticotrophin Releasing Factor (CRF) might be suitable as biological measure of stress as it is implicated directly in both central neurological and endocrine stress-response. The study aims to compare serum CRF levels and perceived stress in opioid-dependent subjects (n = 53) with non-using controls (n = 47) and to correlate them with general and instantaneous craving (in cases only). Perceived stress score and serum CRF levels were significantly higher among the users. No significant correlation with craving was found. The significant difference in serum CRF levels indicate feasibility of measuring CRF levels in peripheral fluids and asserts its role as biochemical measure of stress.

Association of Serotonergic Pathway Gene Polymorphisms With Behavioral Parameters in Patients With Opioid Dependence.

Background and aims Opioid dependence is a chronic, relapsing substance use disorder with a multifactorial etiology, including a genetic component. Serotonin pathway gene polymorphisms have been an important focus of research for psychiatric disorders, including substance use disorders. This study aimed to identify the association of serotonin pathway gene polymorphisms with self-harm, depressive symptoms, impulsiveness, and aggression in patients with opioid dependence. Method The study group comprised 366 subjects with opioid dependence and 200 healthy volunteers. Patients were assessed for a history of self-harm, depressive symptoms, impulsiveness, and aggression using standard tools. Genomic DNA was used for genotyping of four polymorphisms - 5-HTTLPR, STin2 VNTR, TPH1 A218C, and TPH2 G703T. Results The short allele of 5-HTTLPR polymorphism showed a significant difference between cases (59.8%) and controls (40.1%) (p=0.001) and revealed an association with age at opioid dependence (p=0.033). There was a borderline significance of the short allele of 5-HTTLPR with the duration of opioid use (p=0.061) and non-plan impulsivity (p=0.076), suggesting a role of 5-HTTLPR in the susceptibility of opioid dependence. The other markers did not differ between cases and controls. However, the STin2A polymorphism revealed a significant association with anger scores, which may indicate its role in aggressive behavior. Conclusions The present study, the first of its kind, suggests an association of 5-HTTLPR polymorphism with opioid dependence and STin2A polymorphism with aggressive behavior among opioid-dependence patients, signifying the role of these markers in our patient population.

Influence of catechol-O-methyltransferase enzyme gene polymorphism on alcohol and tobacco consumption in North Indian treatment seeking population.

BACKGROUND: The co-occurrence of alcohol and tobacco dependence is frequently witnessed in treatment settings. It is a challenge for clinicians to treat such patients due to their powerful biological association. AIM: The study is aimed to assess the relationship of Catechol-O-methyltransferase (COMT) Val158Met polymorphism with substance intake among individuals who are dependent on both alcohol and tobacco. MATERIALS AND METHODS: A cross-sectional study involving patients coming to the outpatient department was planned. Brief information on their sociodemographic and substance use profile was recorded. Genotyping of COMT Val158Met was carried out using established polymerase chain reaction-restriction fragment length polymorphism method. The COMT genotyping was classified based on the presence or absence of Met allele using the dominant model. Descriptive statistics, Chi-square test, Mann-Whitney test, and Binary logistic regression analysis were performed to analyze the data. RESULTS: The study included 104 alcohol and nicotine co-dependent subjects. More than eighty percent of the participants were educated above secondary level, married, and employed. The allele frequencies of met and Val were found to be 0.23 and 0.77, respectively. Forty percent of the participants reported tobacco-related health problems. The odds of consuming alcohol and nicotine were four times high among Met allele carriers. While the Fagerström test for nicotine dependence and heaviness of smoking index scores were up to four and eight times higher among met allele (odds ratio 4.3 and 8.9, respectively). CONCLUSION: Patients carrying Met allele are reported to consume higher amounts of alcohol and tobacco and were likely to score high among measures of nicotine dependence. Thus met allele carriers needs additional attention for a successful treatment outcome.

Co-administration of nalbuphine attenuates the morphine-induced anxiety and dopaminergic alterations in morphine-withdrawn rats.

INTRODUCTION: The classical effects of exogenous opioids, such as morphine, are predominantly mediated through µ-opioid receptors. The chronic use of morphine induces anxiety-like behavior causing functional changes in the mesolimbic dopaminergic system. The mixed µ/κ-agonist, nalbuphine, used either as an analgesic or as an adjuvant with morphine, produces different and opposite effects. However, whether nalbuphine can be used to antagonize morphine-induced anxiety and dopaminergic alterations is not fully known. OBJECTIVE: This study aimed to compare acute and chronic effects of nalbuphine on morphine-induced anxiety and dopaminergic alterations in rats. METHODS: Male adult Wistar albino rats were made opioid-dependent by administering increasing doses of morphine (5-25 mg/kg; i.p.; b.i.d.). Withdrawal was induced by naloxone (1 mg/kg, i.p.), 4 h after the last morphine injection. Anxiety-like behavior was measured using Activity Monitor (Coulbourn Instruments, Inc. USA). Thereafter, the animals were sacrificed and the brain dissected out and the level of cAMP and the transcriptional and translational expression of TH was measured. Nalbuphine was co-administered with morphine, acutely and chronically, at various doses (0.1, 0.3, 1.0, 3.0 mg/kg, i.p.). RESULTS: Morphine-dependent rats showed a significant higher anxiety and cAMP levels and a significant decrease in the expression of TH. Co-administration of chronic doses of nalbuphine attenuates the higher anxiety, cAMP levels, and upregulates the TH expressions; however, the acute nalbuphine treatment does not attenuate the morphine-induced side effects. CONCLUSION: Therefore, nalbuphine might have an important role in attenuating the anxiety and the effects of the dopaminergic pathway and may have potential in the treatment of opioid addiction.

Impact of Chronic Alcohol and Opioid Dependence on Biochemical Parameters: A Retrospective Case Control Study from a Tertiary Care Treatment Center in North India.

BACKGROUND: Assessment of biochemical parameters can help in the comprehensive management of patients with substance use disorders (SUDs). The aim of this study was to analyse the biochemical parameters of patients with alcohol and opioid dependence at an addiction treatment facility. METHODS: This retrospective study analysed the investigation reports of male patients (aged 18 to 70 years) who visited outpatient department (OPD) with primary diagnosis as opioid dependence syndrome (ODS) or alcohol dependence syndrome (ADS). The data included liver function tests (LFTs), kidney function tests (KFTs), and electrolyte tests conducted in the laboratory in a span of one year. FINDINGS: The study included 713 ADS, 654 ODS, and 227 controls. The ADS group showed significant elevations in mean values of bilirubin, aspartate transaminase (AST), alanine transaminase (ALT), and gamma-glutamyl transferase (GGT) as compared to other groups. A significant decrease in albumin levels in ADS group and raised potassium levels in ODS group was observed. De Ritis ratio above threshold (AST/ALT > 2.0) alone and along with raised GGT levels was observed among 11.3% and 9.7% of patients with ADS, respectively (P < 0.001). Electrolyte abnormalities were present in about 20.0% of patients with ADS and ODS as compared to 8.4% among controls (P < 0.001). CONCLUSION: LFT and electrolyte abnormalities are frequently observed in patients with alcohol and opioid dependence. De Ritis ratio along with raised GGT levels significantly denotes ADS group. These results merit attention in the course of clinical care of alcohol and opioid-dependent patients.

Brain-derived neurotrophic factor (BDNF) levels in first-episode schizophrenia and healthy controls: A comparative study.

BACKGROUND: Abnormalities in brain development and plasticity have been associated with the pathophysiology of schizophrenia. The role of brain-derived neurotrophic factor (BDNF) in schizophrenia is the recent area of interest because it regulates neurogenesis. The current study aimed to assess and compare serum BDNF levels between first-episode schizophrenia patients and healthy controls, and evaluate its correlation with the socio-demographic and clinical variables. METHODOLOGY: It was a cross-sectional comparative study for the assessment of serum BDNF levels between patients with first-episode schizophrenia (N=50) and healthy controls (N-50) conducted in the Department of Psychiatry at a tertiary care public hospital attached to a medical school in North India. Participants were assessed for the socio-demographic parameters, nicotine dependence, and clinical details using structured scales. Serum BDNF level estimated using the sandwich ELISA technique. The comparison between the groups was done by using a Student t-test or chi-square test. Spearman correlation was performed between mean BDNF scores and demographic or illness variables in both first-episode schizophrenia and healthy control groups. RESULTS: There was a significantly lower mean score of total serum BDNF levels in first-episode schizophrenia patients as compared to controls (8.44 ± 1.54 vs 10.44 ± 2.04; t = 5.52, p < 0.001; 95% CI = 1.28-2.71). The total FTND scores for smokeless tobacco use were negatively correlated to BDNF levels among healthy controls (r=-0.30, p=0.03) as well as in the first-episode schizophrenia group (r=-0.32, p= 0.04). None of the other illness-related variables were correlated to serum BDNF values in the first episode schizophrenia group. CONCLUSION: Individuals with first-episode schizophrenia have lower serum BDNF levels than healthy controls. The illness-related factors such as duration of untreated psychosis or psychopathology were not correlated with BDNF levels. Thus abnormal signaling of BDNF can lead to abnormal brain functioning which can make an individual more susceptible to schizophrenia.