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.

Evaluation of alcohol use pattern among OST patients using alcohol biomarkers: Report from community clinics in India.

Background: Opioid-dependent patients undergoing opioid substitution therapy (OST) consume alcohol in a hazardous pattern which adversely affects their treatment outcome. This study aims to measure alcohol biomarkers to screen for secondary alcohol use in OST patients. Methods: A pilot study was planned to measure alcohol biomarkers (AST, ALT, GGT, and CDT) to assess alcohol use in OST patients from three community clinics. The biomarkers were categorized based on the reported frequency of alcohol use. The association of the biomarkers with the frequency of alcohol consumption was determined using the post hoc (Mann-Whitney) test. Results: Forty-five patients with a mean (SD) age of 37.04 (10.7) years were included in the study. Alcohol intake was reported in daily, weekly, and monthly patterns by 22, 63, and 16% of the patients, respectively. High levels of ALT, GGT, and CDT were measured in patients with daily use of alcohol. Serum CDT levels significantly differentiate daily and weekly use from monthly consumption of alcohol. Conclusions: Alcohol biomarkers significantly predict the pattern of alcohol use among OST patients. These results can be prudent in low-resource community clinics to improve the overall outcomes of OST in India.

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.

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.

Use of Filter Paper to Measure Alcohol Biomarkers among Opioid-Dependent Patients on Agonist Maintenance Treatment: A Community-Based Study.

BACKGROUND: Harmful Alcohol use is frequent among opioid dependents patients undergoing agonist maintenance treatment. The objective assessment of harmful alcohol use can be done using laboratory measures of serum biomarkers. For community-based patients, there is often a requirement of an alternative method due to lack of onsite laboratory services. The aim of the study was to examine filter paper as a matrix to measure serum biomarkers of harmful alcohol use. METHODS: The initial phase involved standardization of the filter-paper-based assay. Conditions were optimised for extraction and estimation of alcohol biomarkers (Aspartate Aminotransferase; AST, Alanine Aminotransferase; ALT, Gamma Glutamyl transferase; GGT and Carbohydrate Deficient Transferrin; CDT) from the filter paper. For clinical validation, serum samples were collected from community clinics. Biomarker levels obtained from both the methods were correlated using linear regression analysis. Limits of agreement between the two methods was estimated using the Intraclass Correlation Coefficient (ICC). RESULTS: The extraction of enzymes (AST, ALT and GGT) from filter paper was carried out using the substrate buffer available with the reagent kit (Randox, UK). CDT was readily extracted from filter paper using deionised water. Serum biomarker levels measured from samples collected from community clinics correlated well with filter paper extracted levels (ICC 0.97-0.99). More than 90% of alcohol biomarker levels were recovered from the filter paper matrix using this method. CONCLUSION: Filter paper has the potential to be used as a matrix to objectively measure alcohol biomarkers among opioid-dependent patients from community settings lacking onsite laboratory facilities.

Development and clinical evaluation of a dried urine spot method for detection of morphine among opioid users.

BACKGROUND: In recent years, drug testing in body fluids has gained popularity for validating self-reported drug use. The storage and transportation of urine specimens is a major concern for remote areas where the facilities for performing drug abuse testing are lacking. AIMS AND OBJECTIVES: The aim of the present study was to develop an efficient method for testing opiate in dried urine spots (DUS) and to evaluate its clinical applicability. MATERIALS AND METHODS: The methodology involved optimization of conditions for extraction, recovery, short-, and long-term stability (room temperature, 4°C,-20°C) for detection of opiate from dried urine spots. Further, the extraction efficiency from dried urine spots was compared with the conventional drug testing methodology. The screening was done by using enzyme-linked immunosorbent assay technique, and confirmation was achieved by gas chromatography equipped with nitrogen phosphorus detector. RESULTS: Deionized water was found to be a suitable extracting solvent compare to bi-carbonate buffer (pH 9.2) and saline. Primary screening was achieved by 2 punches taken from a 20-µl (diameter 1.3 cm) spotted urine samples, whereas confirmation was achieved by 2 complete circles each of 20 µl sample volume. The recovery was found to be 99.41% in water. No sign of significant degradation was seen among all storage conditions. CONCLUSIONS: In the current study, DUS has achieved the same level of precision and reproducibility as that of standard methods used for drug testing in urine. Hence, the DUS sampling appears to have potential to detect opiate among drug users in a clinical setting.

Evaluation of Dried Urine Spot Method to Screen Cotinine among Tobacco Dependents: An Exploratory Study.

BACKGROUND AND OBJECTIVES: Assessment of cotinine, a metabolite of nicotine in body fluids, is an important approach for validating the self-report among tobacco users. Adaptation of assays on dried urine spots (DUSs) has advantages of ease of collection, transportation, minimal invasiveness, and requirement of small volume. The aim of the present study was to develop an efficient method for testing cotinine in DUSs and evaluating its clinical applicability. METHODS: This involved optimization of conditions for detection, recovery, and stability of cotinine from dried urine, spotted on filter paper. Enzyme-linked immunosorbent assay was used for screening, whereas confirmation was done by gas chromatography. For clinical applicability, urine samples of tobacco users were tested. RESULTS AND INTERPRETATION: Water was found to be a suitable extracting solvent as compared to carbonate-bicarbonate buffer (pH 9.2) and saline. Screening was achieved by two punches taken from a 20 µl (diameter 1.3 cm) spotted urine samples, and confirmation was achieved by five complete circles each of 20 µl sample volume. The recovery was found to be 97% in water. Limit of detection for the method was found to be 100 ng/ml. No signs of significant degradation were found under all storage conditions. All the urine samples of tobacco users were found to be positive by a conventional method as well as DUSs, and the method proved to be efficient. CONCLUSIONS: DUS samples are a useful alternative for biological monitoring of recent nicotine use, especially in developing countries where sample logistics could be an important concern.

Association of ankyrin repeats & kinase domain containing 1 (ANKK1) gene polymorphism with co-morbid alcohol & nicotine dependence: A pilot study from a tertiary care treatment centre in north India.

BACKGROUND & OBJECTIVES: The frequently encountered co-morbidity of alcohol dependence (AD) with nicotine dependence (ND) increases the risk for various diseases. Ankyrin repeats and kinase domain containing 1 (ANKK1) gene polymorphism is reported to be associated with both ND and AD. This study was undertaken to investigate the possible association of alcohol and tobacco use variables with ANKK1 polymorphism in co-morbid alcohol- and nicotine-dependent treatment seekers visiting a tertiary care centre in north India. METHODS: Seventy nine male participants (18-65 yr old) fulfilling diagnostic criteria for ND and AD were included in the study. The socio-demographic data, along with alcohol and tobacco use profile, was recorded and ANKK1 profiling was carried out. Both the allele groups, A1 and A2, were compared with respect to demographic and substance dependence profile. Univariate binary logistic regression analysis was performed to determine the risk of high nicotine and alcohol consumption with genotype. RESULTS: The A1 carrier group (n=33) reported a significantly higher amount of alcohol and tobacco consumed per day. The scores on parameters of ND were found to be significantly higher in this group. The logistic regression analysis revealed that participants with A1 genotype were 2.5 times more likely to report higher amount of alcohol and nicotine consumption than A2 carriers. INTERPRETATION & CONCLUSIONS: The study provides an indication for the association of ANKK1 polymorphism in the form of higher substance consumption among alcohol dependent smokers, who are A1 carriers and thus may require higher attention of the treatment provider.

Dried urine spots for detection of benzodiazepines.

BACKGROUND AND AIM: Benzodiazepines (BZD) are widely prescribed to substance users. However, the nonmedical use of prescription BZD often leads to abuse and dependence. Therefore, it is important to detect BZD among substance users seeking treatment. The aim of the present study was to develop an efficient method for testing BZD on dried urine spot (DUS) and evaluating its clinical applicability. METHODS: This involved optimization of conditions for the detection, recovery, and stability of BZD from dried urine, spotted on filter paper. Enzyme linked immuno-sorbent assay was used for screening whereas confirmation was done by gas chromatography. For clinical applicability, urine samples of BZD users were tested. RESULTS: The recovery was found to be 99.7% in de-ionized water from 20 µl spotted urine samples. Limit of detection, inter-day and intra-day CV were found to be 100 ng/ml, 4.22% and 3.83%, respectively. BZD were found stable in DUS for 3 weeks at room temperature, and for 3 months at 4°C and -20°C. All the urine samples of benzodiazepine users were found positive by conventional method as well as the DUS method. CONCLUSION: DUS method proved to be efficient for BZD testing with advantages of ease of collection, transportation, minimal invasiveness and small sample volume. It offers a useful alternative for BZD testing especially in developing countries where logistics of sample collection and transportation could be an important concern.

Profile of nicotine use among alcohol dependent patients visiting a tertiary care center in north India.

BACKGROUND: Use of tobacco among alcohol dependent population is quite frequent. This co-morbidity increases the risk for various diseases. Understanding the pattern of tobacco use with co-morbid alcohol use may help in planning appropriate prevention/treatment strategies. The study aimed at examining the profile and pattern of nicotine use among alcohol dependent patients visiting a tertiary care treatment center in North India. MATERIALS AND METHODS: Male patients fulfilling diagnostics and statistical manual of mental disorder fourth edition, criteria for nicotine and alcohol diagnostics and statistical dependence, attending the out-patient department of the tertiary care treatment center were recruited after obtaining informed consent. The socio-demographic profile, drug use history, nicotine associated health problems and general health problem were recorded. Motivation to stop tobacco use was assessed qualitatively using the direct questions about their interest and intentions to quit. RESULTS: A total of 150 subjects were included in the study. The mean age of the study sample was 37.6 ± 10.44 years. Tobacco was reported as the gateway drug in 90% of the cases. Exclusive bidi use reported in 42% of the subjects. Mean duration of bidi and co-morbid alcohol use was higher than cigarette or smokeless tobacco use. Self-reported health problems associated with nicotine use and general health was reported by 41% and 39% of the subjects. Unsuccessful past quit attempts was present in 85% cases. More than 90% of subjects remained interested in quitting the tobacco use. An increased liver enzyme (aspartate transaminase, alanine transaminase and gamma-glutamyl transferase) were observed in 43, 32 and 47% of the cases. CONCLUSION: The results suggest the nicotine and alcohol dependent patients represent a separate population requiring higher attention from the treating physician.

A descriptive study of clinical, hematological, and biochemical parameters of inhalant users seeking treatment at a tertiary care center in India.

BACKGROUND: Inhalant (or solvent) abuse is the purposeful inhalation of vapors or gases, intended to produce pleasurable psychoactive effects. There is a dearth of Indian studies on inhalant users. AIM: The present study aimed to describe the socio-demographic, clinical, and psychosocial characteristics of inhalant users visiting a Tertiary Care Center in North India. MATERIALS AND METHODS: The study was a retrospective chart review for 50 inhalant users who sought treatment for the first time from the center over a period of 2 years. All patients seeking treatment for inhalant use at the center were evaluated by a psychiatrist. RESULTS: Mean age of the sample was 17.16±4.09 years and majority comprised of children and adolescents (72.2%). There were only three girls (6%). Majority comprised of school drop-outs (82%), from lower socio-economic status (80%). Mean age of initiation of first substance was 14.13±4.27 years and inhalants were first drugs for 38%. Duration of inhalant use ranged between 1 month and 7.5 years. Use was mostly uninterrupted, and 88% were dependent users. Correction fluid was the commonest product, used by huffing or sniffing. A large majority (86%) had used at least one other substance besides inhalants, and 8% reported involvement in high-risk sexual behaviors. Comorbid psychiatric disorder was seen in 8% of sample. Positive family history was observed in 30% of the sample. The mean hemoglobin of the sample was 11.88±0.60, with low hemoglobin in 25% of users. Neutrophils, lymphocytes, eosinophils, and monocytes were elevated beyond normal in 10.8%, 6.5%, 15.2%, and 7.5%, respectively. There was no evidence of leucopenia. Bilirubin and serum glutamic pyruvic transaminase was elevated in 6.6% and 13% of inhalant users, respectively. CONCLUSION: The study adds to the limited date available on the treatment-seeking inhalant users from Indian settings. There is a need to examine the pattern of inhalant use in larger samples, across multiple sites in a prospective manner.

Analysis of the stability of urea in dried blood spots collected and stored on filter paper.

The ability to use dry blood spots (DBSs) on filter paper for the analysis of urea levels could be an important diagnostic tool for areas that have limited access to laboratory facilities. We developed a method for the extraction and quantification of urea from DBSs that were stored on 3M Whatman filter paper and investigated the effect of long-term storage on the level of urea in DBSs. DBSs of 4.5 mm in diameter were used for our assay, and we determined the urea levels in blood using a commercially available enzymatic kit (UV GLDH-method; Randox laboratories Ltd., UK). The DBSs on filter discs were stored at 4℃ or at 37℃ for 120 days. The mean intra- and inter-assay coefficient of variance for our method of urea extraction from dried blood was 4.2% and 6.3%, respectively. We collected 75 fresh blood samples and compared the urea content of each fresh sample with the urea content of DBSs taken from corresponding fresh blood samples. Regression analysis reported a regression coefficient (r) value of 0.97 and a recovery of urea from dried spots was 102.2%. Urea concentrations in DBSs were stable for up to 120 and 90 days when stored at 4℃ and 37℃, respectively. Our results show that urea can be stored and quantitatively recovered from small volumes of blood that was collected on filter paper.

Laboratory profiles of treatment-seeking subjects with concurrent dependence on cannabis and other substances: a comparative study.

BACKGROUND: Cannabis is one of the most widely used illicit drugs in India and worldwide. It is considered to have a minimal effect on physical health. OBJECTIVES: The aim of this study was to compare the laboratory profiles of treatment-seeking patients who were cannabis dependent, and drug users who concurrently use other substances, with non-users. MATERIALS AND METHODS: Medical records of patients, whose urine was tested for the detection of cannabis within the last year, were considered for the study. The inclusion criteria for the study group were; co-morbid diagnosis of cannabis dependence according to DSM-IV TR criteria, positive urine drug screen for cannabis, and at least one biochemical or hematological examination report during the treatment period. The subjects who underwent all of the above mentioned tests, but who were negative for any psychoactive substance with no past or current history of substance use, were placed in the control group. RESULTS: A total of 51 subjects fulfilled the inclusion criteria for the study group and 30 subjects were considered as controls. There was no significant difference found between the demographic profiles of the subject and control groups. The mean duration of cannabis use in the patients was 9.53 ± 8.06 years. Serum levels of; bilirubin, SGOT (serum glutamic oxaloacetic transaminase), SGPT (serum glutamic pyruvic transaminase), total protein, alkaline phosphatase, ESR, and eosinophil counts, were raised in; 13.7%, 15.6%, 33.3%, 17.6%, 37.2%, 75% and 5.8% of subjects, respectively. The relative monocyte count was lower than normal in 92% of cases. Physical complaints were reported in 98% of subjects. The two groups showed significant differences in serum alkaline phosphatase [t (79) = 6.5, P ≤ 0.01], TLC [t (79) = 2.36, P = 0.03] and hemoglobin levels [t (79) = 5.50, P ≤ 0.01]. CONCLUSIONS: Abnormal laboratory parameters were observed in patients with cannabis dependence. The study emphasizes the need for regular physical examinations and laboratory investigations for cannabis users.

Apo E genotyping from blood stored on filter paper.

BACKGROUND & OBJECTIVES: Dried blood spotted on to filter paper has been found suitable for a large number of studies. In tropical countries with varying temperature conditions the use of dried blood needs to be validated. We carried out this study to assess the use of blood spotted filter paper as a transport system to study genotyping of Apo E gene. METHODS: Fifty five patients visiting Cardiothoracic Neuroscience Centre (CNC) OPD at the All India Institute of Medical Sciences (AIIMS), New Delhi, and referred for lipid investigations to Cardiac Biochemistry Laboratory were selected at random. Blood was spotted on to Whatman 3 MM filter paper, dried and stored at room temperature. Genomic DNA was extracted and genotyping was carried out at the end of 0, 3 and 12 months. The study was further validated using samples collected on to filter paper from four centres and stored for eight years at room temperature. The temperature and humidity conditions of the centre varied widely. RESULTS: Fifty five samples collected on to filter paper showed exact match of the genotyping when compared to fresh blood. In dried blood samples collected and stored for 1 yr at room temperature DNA extraction and apo E genotyping was done successfully. INTERPRETATION & CONCLUSIONS: The present results showed the feasibility of using dried blood samples on filter paper for apo E genotyping in tropical temperature. The findings need to be validated on a large sample before being recommended for use.

Use of filter paper stored dried blood for measurement of triglycerides.

Adaptation of assays on dried blood has advantages of ease of collection, transportation, minimal invasiveness and requirement of small volume. A method for extraction and estimation of triglyceride from blood spots dried on filter paper (Whatman no. 3) has been developed. A single dried blood spot containing 10 muL blood was used. Triglyceride was efficiently extracted in methanol from blood dried on filter paper by incubation at 37 degrees C for two hours with gentle shaking. For the estimation, a commercially available enzymatic method was used. Blood spot assays showed mean intra and inter assay coefficient of variance of 6.0% and 7.4% respectively. A comparison of paired whole blood spots and plasma samples (n = 75, day 0) gave an intraclass correlation of 0.96. The recovery was 99.6%. The dried blood triglyceride concentrations were stable for one month when the filter discs were stored at room temperature (16-28 degrees C). Storage of filters at 4 degrees C extended the stability and triglycerides could be quantitatively recovered after 3 months of storage.