Soluble Enolase 1 of Candida albicans and Aspergillus fumigatus Stimulates Human and Mouse B Cells and Monocytes.

Because of the growing numbers of immunocompromised patients, the incidence of life-threatening fungal infections caused by Candida albicans and Aspergillus fumigatus is increasing. We have recently identified enolase 1 (Eno1) from A. fumigatus as an immune evasion protein. Eno1 is a fungal moonlighting protein that mediates adhesion and invasion of human cells and also immune evasion through complement inactivation. We now show that soluble Eno1 has immunostimulatory activity. We observed that Eno1 from both C. albicans and A. fumigatus directly binds to the surface of lymphocytes, preferentially human and mouse B cells. Functionally, Eno1 upregulated CD86 expression on B cells and induced proliferation. Although the receptor for fungal Eno1 on B lymphocytes is still unknown, the comparison of B cells from wild-type and MyD88-deficient mice showed that B cell activation by Eno1 required MyD88 signaling. With respect to infection biology, we noted that mouse B cells stimulated by Eno1 secreted IgM and IgG2b. These Igs bound C. albicans hyphae in vitro, suggesting that Eno1-induced Ab secretion might contribute to protection from invasive fungal disease in vivo. Eno1 also triggered the release of proinflammatory cytokines from monocytes, particularly IL-6, which is a potent activator of B cells. Together, our data shed new light on the role of secreted Eno1 in infections with C. albicans and A. fumigatus. Eno1 secretion by these pathogenic microbes appears to be a double-edged sword by supporting fungal pathogenicity while triggering (antifungal) immunity.

Enolase 1 of Candida albicans binds human CD4+ T cells and modulates naïve and memory responses.

To obtain a better understanding of the biology behind life-threatening fungal infections caused by Candida albicans, we recently conducted an in silico screening for fungal and host protein interaction partners. We report here that the extracellular domain of human CD4 binds to the moonlighting protein enolase 1 (Eno1) of C. albicans as predicted bioinformatically. By using different anti-CD4 monoclonal antibodies, we determined that C. albicans Eno1 (CaEno1) primarily binds to the extracellular domain 3 of CD4. Functionally, we observed that CaEno1 binding to CD4 activated lymphocyte-specific protein tyrosine kinase (LCK), which was also the case for anti-CD4 monoclonal antibodies tested in parallel. CaEno1 binding to naïve human CD4+ T cells skewed cytokine secretion toward a Th2 profile indicative of poor fungal control. Moreover, CaEno1 inhibited human memory CD4+ T-cell recall responses. Therapeutically, CD4+ T cells transduced with a p41/Crf1-specific T-cell receptor developed for adoptive T-cell therapy were not inhibited by CaEno1 in vitro. Together, the interaction of human CD4+ T cells with CaEno1 modulated host CD4+ T-cell responses in favor of the fungus. Thus, CaEno1 mediates not only immune evasion through its interference with complement regulators but also through the direct modulation of CD4+ T-cell responses.

Potential of Capparis decidua plant and eggshell composite adsorbent for effective removal of anionic dyes from aqueous medium.

The presence of hazardous dyes in wastewater poses significant threats to both ecosystems and the natural environment. Conventional methods for treating dye-contaminated water have several limitations, including high costs and complex operational processes. This study investigated a sustainable bio-sorbent composite derived from the Capparis decidua plant and eggshells, and evaluated its effectiveness in removing anionic dyes namely tartrazine (E-102), methyl orange (MO), and their mixed system. The research examines the influence of initial concentration, contact time, pH, adsorbent dosage, and temperature on the adsorption properties of anionic dyes. Optimal removal of tartrazine (E-102), methyl orange (MO), and their mixed system was achieved at a pH of 3. The equilibrium was achieved at 80 min for MO and mixed systems, and 100 min for E-102. The adsorption process showed an exothermic nature, indicating reduced capacity with increasing temperature, consistent with heat release during adsorption. Positive entropy values indicated increased disorder at the solid-liquid interface, attributed to molecular rearrangements and interactions between dye molecules and the adsorbent. Isotherm analysis using Langmuir, Freundlich, Temkin, and Redlich-Peterson models revealed that the Langmuir model best fit the experimental data. The maximum adsorption capacities of 50.97 mg/g, 52.24 mg/g, and 56.23 mg/g were achieved for E-102, MO, and the mixed system under optimized conditions, respectively. The pseudo-second-order kinetic model demonstrated the best fit, indicating that adsorption occurs through physical and chemical interactions such as electrostatic attraction, pore filling, and hydrogen bonding. Hence, the developed bio-sorbent could be a sustainable and cost-effective solution for the treatment of anionic dyes from industrial effluents.

SSR identification and phylogenetic analysis in four plant species based on complete chloroplast genome sequences.

The effective utilization of traditional Chinese medicine (TCM) has been challenged by the difficulty to accurately distinguish between similar plant varieties. The stability and conservation of the chloroplast genome can aid in resolving genotypes. Previous studies using nuclear sequences and molecular markers have not effectively differentiated the species from related taxa, such as Machilus leptophylla, Hanceola exserta, Rubus bambusarum, and Rubus henryi. This study aimed to characterize the chloroplast genomes of these four plant species, and analyze their simple sequence repeats (SSRs) and phylogenetic positions. The results demonstrated the four chloroplast genomes consisted of 152.624 kb, 153.296 kb, 156.309 kb, and 158.953 kb in length, involving 124, 130, 129, and 131 genes, respectively. They also contained four specific regions with mononucleotide being the class with the most members. Moreover, these repeating types of SSR were various in individual class. Phylogenetic analysis showed that M. leptophylla was clustered with M. yunnanensis, and H. exserta was confirmed as belonging to the family Ocimeae. Additionally, R. bambusarum and R. henryi were grouped together but differed in their SSR features, indicating that they were not the same species. This research provides evidence for resolving species and contributes new genetic information for further studies.

Simultaneous adsorption of methylene blue and amoxicillin by starch-impregnated MgAl layered double hydroxide: Parametric optimization, isothermal studies and thermo-kinetic analysis.

Textile and pharmaceutical effluents contain significant amounts of dyes and antibiotics, which pose a serious threat to the ecosystem when discharged directly. Therefore, they should be treated by facile treatment techniques using low-cost materials. Layered double hydroxide (LDH) and its hybrids have emerged as robust and economic adsorbents for water treatment. Herein, magnesium/aluminum LDH and its starch-based composite were synthesized by a co-precipitation technique. The physicochemical features of the developed adsorbents were thoroughly characterized using various analytical tools. The developed materials were tested for the eradication of methylene blue (MB) and amoxicillin (AMX) in batch mode adsorption by varying operating conditions. Adsorption performance depends on the solution's pH. Under optimum adsorption conditions of pH 11, adsorbent dosage of 50 mg/L, and treatment time of 120 min, starch-impregnated MgAl-LDH exhibited maximum MB and AMX adsorption capacities of 114.94 and 48.08 mg/g, respectively. The adsorption mechanism states that hydrogen bonds and weak van der Waals forces are responsible for the removal of pollutants by the developed materials. Moreover, equilibrium and kinetic studies revealed that the removal of dye and antibiotic followed the Freundlich and Langmuir models with the pseudo-second-order reaction kinetics, respectively. The spent adsorbents were regenerated using 0.1 M HCl (for MB) and methanol (for AMX) eluent, and reusability studies ensured that the developed adsorbents retained their performance for up to four consecutive adsorption/desorption cycles. MgAl-LDH and its starch-based hybrid could thus be used to effectively remove organic contaminants from wastewater streams on a commercial scale.

Association between HIV clinic caseloads and viral load suppression in New York City.

The relationship between HIV patient caseload and a clinic's ability to achieve viral load suppression (VLS) in their HIV patient population is not understood. The New York City Department of Health and Mental Hygiene (NYCDOHMH) administered a survey to clinics providing HIV care to people living with HIV (PLWH) in NYC in 2016. Clinics were stratified by quartiles of HIV patient caseload and dichotomized by whether ≥85% (n = 36) or <85% (n = 74) of their patients achieved VLS. Multivariable logistic regression adjusted for confounders of age, sex, ethnicity, and race. Provider to patient ratios (PPR) were calculated for each clinic as staffing full time equivalents per 100 HIV patients.

Illness perception about hepatitis C virus infection: a cross-sectional study from Khyber Pakhtunkhwa Pakistan.

BACKGROUND: Hepatitis C virus (HCV) infection is a debilitating chronic health problem and can be fatal if left untreated. Illness perceptions are self-manifested beliefs that influence the ability of individuals to cope with their disease and perceive it as manageable or threatening condition. Limited evidence is available from low resource settings regarding patient perception about HCV. In this study, we aimed to assess the perception of individuals with HCV, the impact of their sociodemographic and clinical characteristics on their HCV perception, and its link to patient-oriented treatment outcomes. METHODS: A cross-sectional survey was undertaken enrolling individuals with HCV who attended Hepatitis C clinics at two hospitals of Khyber Pakhtunkhwa, Pakistan. Illness perception was measured using Brief Illness Perception Questionnaire (BIPQ). Descriptive statistics, Kruskal Wallis tests and Mann Whitney U tests were performed to study patient sociodemographic and clinical characteristics and to analyze the questionnaire results. Multivariable linear regression was used to assess determinants associated with perception scores. RESULTS: Participants represented poor HCV perception and their overall mean BIPQ score was 43.35, SD = 13.15. Participants had a low degree of understanding about their illness (mean coherence score = 2.92, SD = 1.85). Individuals with more than four years, compared to less than one year, of estimated HCV infection were more likely to view that their illness would continue (mean timeline score = 6.27, SD = 2.50 versus 5.36, SD = 2.53; respectively, p < 0.01). Similarly, individuals with hepatic cirrhosis, compared to without, were more likely to attribute symptoms to their disease (mean identity score = 5.48, SD = 2.14 versus 4.89, SD = 2.38; respectively, p = 0.04). Female participants reported higher degrees at which the illness affected them emotionally (i.e., emotional representation) and lower coherence about HCV than males (p = 0.04 and 0.006, respectively). Individuals who did not achieve sustained virological response 24 weeks after treatment with interferon-based therapy, compared to treatment naïve individuals, reported lower trust in being successfully treated with newer anti-HCV agents (i.e., direct acting antivirals) (p = 0.029). However, multivariable linear regression revealed that no sociodemographic or clinical determinants were associated with a higher BIPQ score (i.e., more threatening, or negative perceptions). CONCLUSION: Individuals with HCV in Pakistan generally report threatening or negative views about HCV infection. Lack of trust in treatment efficacy was also apparent, especially in those who experienced failed anti-HCV treatments in the past. Healthcare professionals should consider these perceptions when treating individuals with HCV to optimize their compliance by aligning their perception with the high effectiveness of current anti-HCV therapies.

Protocol paper: kainic acid excitotoxicity-induced spinal cord injury paraplegia in Sprague-Dawley rats.

BACKGROUND: Excitotoxicity-induced in vivo injury models are vital to reflect the pathophysiological features of acute spinal cord injury (SCI) in humans. The duration and concentration of chemical treatment controls the extent of neuronal cell damage. The extent of injury is explained in relation to locomotor and behavioural activity. Several SCI in vivo methods have been reported and studied extensively, particularly contusion, compression, and transection models. These models depict similar pathophysiology to that in humans but are extremely expensive (contusion) and require expertise (compression). Chemical excitotoxicity-induced SCI models are simple and easy while producing similar clinical manifestations. The kainic acid (KA) excitotoxicity model is a convenient, low-cost, and highly reproducible animal model of SCI in the laboratory. The basic impactor approximately cost between 10,000 and 20,000 USD, while the kainic acid only cost between 300 and 500 USD, which is quite cheap as compared to traditional SCI method. METHODS: In this study, 0.05 mM KA was administered at dose of 10 µL/100 g body weight, at a rate of 10 µL/min, to induce spinal injury by intra-spinal injection between the T12 and T13 thoracic vertebrae. In this protocol, detailed description of a dorsal laminectomy was explained to expose the spinal cord, following intra-spinal kainic acid administration at desired location. The dose, rate and technique to administer kainic acid were explained extensively to reflect a successful paraplegia and spinal cord injury in rats. The postoperative care and complication post injury of paraplegic laboratory animals were also explained, and necessary requirements to overcome these complications were also described to help researcher. RESULTS: This injury model produced impaired hind limb locomotor function with mild seizure. Hence this protocol will help researchers to induce spinal cord injury in laboratories at extremely low cost and also will help to determine the necessary supplies, methods for producing SCI in rats and treatments designed to mitigate post-injury impairment. CONCLUSIONS: Kainic acid intra-spinal injection at the concentration of 0.05 mM, and rate 10 µL/min, is an effective method create spinal injury in rats, however more potent concentrations of kainic acid need to be studied in order to create severe spinal injuries.

MicroRNA regulation of vascular smooth muscle cells and its significance in cardiovascular diseases.

Cardiovascular disease (CVD) is among the leading causes of death worldwide. MicroRNAs (miRNAs), regulatory molecules that repress protein expression, have attracted considerable attention in CVD research. The vasculature plays a big role in CVD development and progression and dysregulation of vascular cells underlies the root of many vascular diseases. This review provides a brief introduction of the biogenesis of miRNAs and exosomes, followed by overview of the regulatory mechanisms of miRNAs in vascular smooth muscle cells (VSMCs) intracellular signaling during phenotypic switching, senescence, calcification, and neointimal hyperplasia. Evidence of extracellular signaling of VSMCs and other cells via exosomal and circulating miRNAs is also presented. Lastly, current drawbacks and limitations of miRNA studies in CVD research and potential ways to overcome these disadvantages are discussed in detail. In-depth understanding of VSMC regulation via miRNAs will add substantial knowledge and advance research in diagnosis, disease progression, and (or) miRNA-derived therapeutic approaches in CVD research.

Drug Resistance in Hepatitis C Virus: Future Prospects and Strategies to Combat It.

Induction of highly pathogenic hepatitis C virus (HCV) causes chronic hepatitis round the world. This virus is easily prone to developing resistance against antiviral drugs because of two viral polymerases that do not possess the proofreading and overlapping reading frame abilities. There is more than one explanation for how this virus builds up resistance against antiviral drug treatments. Assays are now available to detect HCV-resistant variants, based on phenotypic and genotypic assays, and next generation sequencing. But these assays are of a little value at baseline, because they are not influential enough for making therapeutic decisions in HCV patients. Moreover, HCV monitoring is now an essential part of clinical practice. Special patients, such as those with thalassemia, renal transplant due to renal failure, and the patients undergoing hemodialysis, are at higher risk for acquiring this infection. Management of HCV infection in these patient groups is complicated by multiple side effects, including flu-like symptoms, neutropenia, fever, and neuropsychiatric disorders, thus limiting the use of ribavirin and coexisting iron overload. In HCV patients suffering from depression, the treatment may be discontinued because of some defects in neurochemical pathways caused by interferon, which can enhance the level of depression in these patients. In addition, obesity has been found to be a marker of failure of HCV treatment. There will be many resistance tolerant HCV treatment options available in the near future.

Spinal Cord Injury: Pathophysiology, Multimolecular Interactions, and Underlying Recovery Mechanisms.

Spinal cord injury (SCI) is a destructive neurological and pathological state that causes major motor, sensory and autonomic dysfunctions. Its pathophysiology comprises acute and chronic phases and incorporates a cascade of destructive events such as ischemia, oxidative stress, inflammatory events, apoptotic pathways and locomotor dysfunctions. Many therapeutic strategies have been proposed to overcome neurodegenerative events and reduce secondary neuronal damage. Efforts have also been devoted in developing neuroprotective and neuro-regenerative therapies that promote neuronal recovery and outcome. Although varying degrees of success have been achieved, curative accomplishment is still elusive probably due to the complex healing and protective mechanisms involved. Thus, current understanding in this area must be assessed to formulate appropriate treatment modalities to improve SCI recovery. This review aims to promote the understanding of SCI pathophysiology, interrelated or interlinked multimolecular interactions and various methods of neuronal recovery i.e., neuroprotective, immunomodulatory and neuro-regenerative pathways and relevant approaches.

Responses of morphological, physiological, and biochemical characteristics of maize (Zea mays L.) seedlings to atrazine stress.

Atrazine is a synthetic herbicide applied to control broadleaf weeds in different crops. In many parts of the world, atrazine is mainly applied for controlling weeds in maize fields. However, studies on the possible adverse effects of atrazine on maize crop can hardly be found in literature. The present study was therefore conducted to evaluate the effect of atrazine on different characteristics of maize seedlings including germination, growth, chlorophyll contents, soluble sugars, proteins and proline levels, ions accumulation, cell viability, and cell injury. In addition, the effects of atrazine on reactive oxygen species (ROS) accumulation and antioxidant enzymes activities in maize seedlings were estimated. It was found that at high concentration, atrazine slightly but significantly inhibited seed germination and growth of maize seedlings. Light-harvesting pigments (chlorophylls a and b, and total carotenoids) exhibited a higher sensitivity to atrazine and were negatively impacted by atrazine at doses above 50 ppm. Atrazine caused an increase in soluble sugars at all tested doses and decrease in soluble proteins at the highest tested dose. Exposure of maize seedlings to atrazine resulted in an increased cell injury and decreased cell viability. Atrazine did not affect the concentration of Na+, K+, and Ca2+ ions in maize seedlings to any greater extent; however, some minor changes were observed in some cases. An increase in the stress marker, proline, was found upon exposure to atrazine. The observed effects of atrazine in maize seedlings can be attributed to oxidative stress as revealed by an increase in H2O2 content and higher activities of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) enzymes in atrazine-treated seedlings. The present investigation concludes that atrazine has the potential to adversely affect germination and growth of maize seedlings by inducing oxidative stress that causes increased cell injury and decreased cell viability as well as impairs the concentration of light-harvesting pigments.

Pesticide-induced oxidative stress and antioxidant responses in tomato (Solanum lycopersicum) seedlings.

Excessive use of pesticides can adversely affect the growth of non-target host plants in different ways. Pesticide-induced stress can affect non-target plants through elevated levels of reactive oxygen species (ROS) responsible for detrimental effects on cell metabolism, biochemical and other physiological activities. In response to oxidative stress, plant activates antioxidant defense system consisting of both enzymatic and non-enzymatic components. In the present investigation, three commonly used pesticides, emamectin benzoate, alpha-cypermethrin and imidacloprid, were assessed for causing oxidative stress in tomato. The oxidative damage induced by these pesticides at five different concentrations i.e. 1/4X, 1/2X, recommended application dose (X), 2X and 4X in the root and shoot tissues of tomato plant/seedlings were evaluated. Following pesticide exposure for 35 days, cell viability, cell injury, total soluble sugar (TSS) and total soluble proteins (TSP) were measured. Antioxidant activities were estimated by measuring activity levels of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) peroxidase (POD), ascorbate peroxidase (APX) and proline. Hydrogen peroxide (H2O2) levels were analysed as ROS, lipid peroxidation was measured in term of thiobarbituric acid reactive substances (TBARS) as membrane damage caused by ROS was also assessed. Analysis of the data revealed that pesticides application at higher concentrations significantly elevated ROS levels and caused membrane damage by the formation of TBARS, increased cell injury and reduced cell viability both in root and shoot tissues compared with non-treated plants. Moreover, a gradual decrease in the levels of TSS and TSP was observed in plants subjected to increasing doses of pesticides. To cope with pesticide-induced oxidative stress, a significant increase in levels of antioxidants was observed in the plants exposed to higher doses of pesticides. Shoot tissues responded more drastically by producing higher levels of antioxidants as compared to root tissues indicating the direct exposure of shoots to foliar application of pesticides. Taken together, these results strongly suggested that the application of pesticides above the recommended dose can provoke the state of oxidative stress and can cause oxidative damages in non-target host plants.

Ecotoxicological evaluation of two anti-dandruff hair shampoos using Lemna minor.

Hair shampoos, a mixture of various organic and organic compounds, are commonly used personnel care products. Since shampoos are used in almost every household and beauty shop, their ingredients are common components of domestic and municipal wastewater. However, studies on the effect of shampoos to aquatic plants can hardly be found in literature. Therefore, the present study was conducted to investigate the phytotoxic effects of two commonly used anti-dandruff shampoos (named here AD 1 and AD 2) using Lemna minor as a biotest organism. For toxicity assessment, frond number, fresh and dry biomass, and light-harvesting pigments (chlorophyll a, b and total carotenoids) of Lemna were used as end points. Five different concentrations (0.001, 0.01, 0.1, 1, and 5%) of each shampoo were tested in comparison to the control. At lower concentrations of shampoos, some minor and non-significant stimulatory effects were observed in some parameters, but at concentrations above 0.01% both the shampoos significantly inhibited almost all parameters in Lemna. The EC50 values obtained for frond number were 0.034 and 0.11% for AD 1 and AD 2, respectively. The fresh biomass gave EC50 values of 0.07 and 0.066% for AD 1 and AD 2, respectively. Based on the preset study, it can be speculated that shampoo contamination at higher concentrations in water bodies can be a threat to aquatic organisms. This study can be used as a baseline to further investigate shampoo toxicity using other species and to explore the mechanism of shampoo toxicity in aquatic plants.

Effectiveness of muscle energy technique on cervical range of motion and pain.

Every individual in his/her life experiences cervical pain at some stage which may restrict daily activities. Various approaches are available for the management of cervical pain, which include surgical, pharmacological and physical therapy. Different Manual therapy techniques are used for the treatment of neck pain. Muscle energy techniques and active isolated stretching are under consideration. This case series study was conducted at physical therapy department of North West General Hospital, Peshawar from August 2015 to January 2016 to find out the effectiveness of muscles energy techniques on cervical range of motion and pain. A total of 20 patients suffering neck pain, both genders having age of 25 - 50 years, cervical ROM limitation and muscles spasm were included in the study. Patients were treated by muscles energy technique (MET). The patient's outcome measures were Inclinometer and Visual Analogue Scale (VAS). Data was analyzed using SPSS version 20. The mean age was 32.3±6.53 years. The patient treated with muscles energy technique showed clinically improvement in the range of motion and pain. Results showed that pre and post treatment differences were statistically significant for cervical flexion (0.001), cervical extension (0.001), cervical right side rotation (0.001), cervical left side rotation (0.001), cervical right side bending (0.001), and cervical left side bending (0.01). Paired t-test finding for the pain showed statistically significant difference (0.005). It was concluded that Muscles Energy Technique is effective in the treatment of restricted range of motion and cervical pain.

Modulation of Wnt signaling pathway by hepatitis B virus.

Hepatitis B virus (HBV) has a global distribution and is one of the leading causes of hepatocellular carcinoma. The precise mechanism of pathogenicity of HBV-associated hepatocellular carcinoma (HCC) is not yet fully understood. Viral-related proteins are known to take control of several cellular pathways like Wnt/ß-catenin, TGF-ß, Raf/MAPK and ROS for the virus's own replication. This affects cellular persistence, multiplication, migration, alteration and genomic instability. The Wnt/FZD/ß-catenin signaling pathway plays a significant role in the pathology and physiology of the liver and has been identified as a main factor in HCC development. The role of ß-catenin is linked mainly to the canonical pathway of the signaling system. Progression of liver diseases is known to be accompanied by disturbances in ß-catenin expression (mainly overexpression), with its cytoplasmic or nuclear translocation. In recent years, studies have documented that the HBV X protein and hepatitis B surface antigen (HBsAg) can act as pathogenic factors that are involved in the modulation and induction of canonical Wnt signaling pathway. In the present review we explore the interaction of HBV genome products with components of the Wnt/ß-catenin signaling pathway that results in the enhancement of the pathway and leads to hepatocarcinogenesis.

Toxic Metal Pollution in Pakistan and Its Possible Risks to Public Health.

Environmental pollution has increased many folds in recent years and in some places has reached levels that are toxic to living things. Among pollutant types, toxic heavy metals and metalloids are among the chemicals that pose the highest threat to biological systems (Jjemba 2004). Unlike organic pollutants, which are biodegradable, heavy metals are not degraded into less hazardous end products (Gupta et al. 2001). Low concentrations of some heavy metals are essential for life, but some of them like Hg, As, Pb and Cd are biologically non-essential and very toxic to living organisms. Even the essential metals may become toxic if they are present at a concentration above the permissible level (Puttaiah and Kiran 2008). For example, exposure to Zn and Fe oxides produce gastric disorder and vomiting, irritation of the skin and mucous membranes. Intake of Ni, Cr, Pb, Cd and Cu causes heart problems, leukemia and cancer, while Co and Mg can cause anemia and hypertension (Drasch et al. 2006). Similarly, various studies indicated that overexposure to heavy metals in air can cause cardiovascular disorders (Miller et al. 2007; Schwartz 2001), asthma (Wiwatanadate and Liwsrisakun 2011), bronchitis/emphysema (Pope 2000), and other respiratory diseases (Dominici et al. 2006).

TOTAL LYMPHOCYTE COUNT AS A SUBSTITUTE TO CD4 COUNT IN MANAGEMENT OF HIV INFECTED INDIVIDUALS IN RESOURCE LIMITED SOCIETY.

BACKGROUND: Pakistan is a resource limited society and gold standard parameters to monitor HIV disease activity are very costly. The objective of the study was to evaluate total lymphocyte count (TLC) as a surrogate to CD4 count to monitor disease activity in HIV/AIDS in resource limited society. METHODS: This cross sectional study was carried out at HIV/AIDS treatment centre, Pakistan Institute of Medical Sciences (PIMS), Islamabad. A total of seven hundred and seventy four (774) HIV positive patients were enrolled in this study, and their CD4 count and total lymphocyte count were checked to find any correlation between the two by using Spearman ranked correlation coefficient. Results: The mean CD4 count was (434.30 +/- 269.23), with minimum CD4 count of (9.00), and maximum of (1974.00). The mean total lymphocyte count (TLC) was (6764.0052 +/- 2364.02) with minimum TLC (1200.00) and maximum TLC was (20200.00). Using the Pearson's correlation (r) there was a significant and positive correlation between TLC and CD4 count. (r2=0.127 and p=0.000) at 0.01 level. CONCLUSION: Our study showed a significant positive correlation between CD4 count and total lymphocyte count (TLC), so TLC can be used as a marker of disease activity in HIV infected patients.

Anti-retroviral drugs compliance in intravenous and non intravenous drug abusers.

BACKGROUND: Intravenous drug abuse is often associated with poor adherence to anti-retroviral drugs in HIV/AIDS. Very few studies in Pakistan have determined implications of intravenous drug abuse on anti-retroviral drug compliance in HIV/AIDS patients. The objectives of the study were to assess and compare the adherence to anti-retroviral drugs in intravenous drug users (IDUs) and non-intravenous drug users (NIDUs) and to determine various factors influencing the adherence to anti-retroviral (ARV) drugs in HIV positive IDUs in HIV treatment centre Pakistan Institute of Medical Sciences (PIMS), Islamabad. METHODS: This descriptive observational study was carried out at HIV/AIDS treatment and care centre PIMS, Islamabad. A total of 162 HIV positive male (81 IDU and 81 NIDU) were enrolled in this study. They were followed over a period of five years from 2008-2012. ARV drug compliance and anti-tuberculosis treatment (ATT) outcome in IDUs and NIDUs were assessed using standard outcome parameters. RESULTS: Among IDUs Hepatitis C was positive in 63 (77.77%) cases and negative in 18 (22.22%) cases. In NIDUs hepatitis C was positive in 5 (6.17%) and negative in 76 (93.82%) (p=0.000). In IDUs Pulmonary tuberculosis was present in 61 (75.30%) patients and in NIDUs it was present in 52 (64.19%) (p=0.171). Regarding ATT outcome, amongst IDUs 41 (50.61%) lost to follow up, 16 (19.75%) were compliant to treatment and 4 (4.93%) were transferred out. In NIDUs, 2 (2.46%) patients were lost to follow-up, 38 (46.91%) remained compliant to treatment and 6 (7.40%) were transferred out (p=0.000). Regarding end status of ARVs, in IDUs, 48 (59.25%) were lost to follow-up, 1 (1.23%) was defaulter, 16 (19.75%) were compliant to treatment, 8 (9.87%) were transferred out and 8 (9.87%) expired. In NIDUs, 73 (90.12%) were compliant to treatment, 5 (6.17%) expired, 2 (2.46%) were lost to follow-up. CONCLUSIONS: Due to various socioeconomic and clinical factors, compliance to ARVs in IDUs is poorer as compared to NIDUs. The factors recognized are illiteracy, poor socioeconomic status, unemployment and various comorbidities (Hepatitis B, C and Tuberculosis) which are more prevalent in IDUs.

The Impact of Calcium and Vitamin D Supplementation Prior to Thyroidectomy on Mitigating Postoperative Hypocalcemia.

Background Hypocalcemia remains the most frequent complication after thyroidectomy. It can either be transient or permanent, and patients often find it unpleasant due to its association with prolonged hospitalization. The objective of this study was to determine the role of preoperative calcium and vitamin D supplementation in preventing hypocalcemia after subtotal/total thyroidectomy. Material and methods This quasi-experimental study was conducted at the Department of General Surgery, Ayub Teaching Hospital, Abbottabad, Pakistan, from January 2023 to July 2023. We utilized non-probability purposive sampling. Patients undergoing total thyroidectomy were recruited and followed until discharge to ascertain outcomes. They were divided into two groups: Group A received vitamin D (200,000 IU) one week preoperatively as single intramuscular dose and calcium (1 gm) orally two times daily for one week preoperatively, while Group B served as the control. Venous blood samples were taken post-surgery, on the second and seventh day and at day 30 to assess hypocalcemia. Chi-square test was employed, comparing hypocalcemia in both groups with a p-value ≤0.05 considered significant. Results One hundred thirty-two patients underwent subtotal/total thyroidectomy, with 46.2% (n = 61) being male and 53.8% (n = 71) female. The mean age of these patients was 37.12 ± 6.22 years, ranging from 20 to 50 years, and 57.6% (n = 76) were aged over 35 years. More than half (55.3%, n = 73) hailed from rural areas, while 4.7% (n = 59) resided in urban locales. Among the patients, 15.9% (n = 21) had a history of diabetes, and 24.2% (n = 32) were hypertensive. The mean body mass index (BMI) was 23.32 ± 2.41 kg/m2, with 7.6% (n = 10) classified as obese. The mean preoperative serum calcium level was 9.87 ± 1.07 mg/dL. Postoperatively, the mean serum calcium level was 8.74 ± 0.83 mg/dL. Both Group A (preoperative vitamin D and calcium supplementation) and Group B (control) demonstrated comparable baseline characteristics before undergoing thyroidectomy. The incidence of postoperative hypocalcemia was notably lower in Group A, with only 4.5% (n = 3) experiencing this complication. By contrast, 24.2% (n = 16) of patients in Group B, the control group, developed hypocalcemia (P = 0.001). Conclusion Our study supports the use of preoperative calcium and vitamin D supplementation in patients undergoing thyroidectomy to combat hypocalcemia. The treated group showed significantly lower hypocalcemia compared to the untreated group B. We recommend preoperative calcium and vitamin D supplementation for all thyroidectomy patients to reduce related morbidities and hospitalization duration.