Primary HIV Drug Resistance among Recently Infected Cases of HIV in North-West India.

BACKGROUND: Antiretroviral treatment may lead to the emergence of HIV drug resistance, which can be transmitted. HIV primary drug resistance (PDR) is of great public health concern because it has the potential to compromise the efficacy of antiretroviral therapy (ART) at the population level. OBJECTIVE: To estimate the level of primary drug resistance among recently infected cases of HIV in 6 ART centres of North-Western India from September 2014 to June 2016. METHODS: The level of primary drug resistance was studied among 37 recently infected HIV cases identified by Limiting antigen (Lag) avidity assay based on modified Recent Infection Testing Algorithm (RITA). The reverse transcriptase region of HIV-1 pol gene (1-268 codons) was genotyped. The sequences were analyzed using the Calibrated Population Resistance (CPR) tool of Stanford University HIV drug resistance (DR) database to identify drug resistance. RESULTS: Among 37 isolates studied, 6 (16.2%) samples showed primary drug resistance (PDR) against reverse transcriptase (RT) inhibitor. The proportion of primary drug resistance was 22.2% (2/9) among female sex workers, 14.3% (1/7) among men having sex with men, and 14.3% (3/21) among injecting drug users. Observed mutations were K219R, L74V, K219N, and Y181C. Injecting drug user (IDU) has showed resistance to either nucleoside/nucleotide reverse transcriptase inhibitors (NRTI) or nonnucleotide reverse transcriptase inhibitors (NNRTI). CONCLUSION: Resistance to either NRTI or NNRTI among the recently is a new challenge that needs to be addressed. The fact that both Y181C isolates are IDUs is important and represents 2/21 (~10%) NNRTI drug resistance. Surveillance for primary drug resistance (PDR) needs to be integrated into next generation of HIV surveillance as access to ART is increasing due to introduction of test and treat policy.

Modified Release Biodegradable Polymeric Microspheres of Stavudine: Cell Viability, Cellular Uptake, Hemolysis Studies and In Vivo Pharmacokinetics.

Family of retroviruses which replicates through the use of the reverse transcriptase enzyme or the enzyme needed to convert RNA to DNA for replication bears Human Immunodeficiency Virus (HIV). It causes irreversible destruction of the immune system leading to the occurrence of opportunistic infections and malignancies. The eradication of HIV is highly unlikely as the cells of the mononuclear phagocyte system (MPS) besides CD4 T lymphocytes are the specific hosts for HIV which need to be targeted even after extended blood plasma profile of antiviral drug to maintain viral suppression and reduced disease progression. Aiming the current goal, biodegradable polymeric microspheres of PLGA 50: 50 (RESOMER(®) 505H) were developed and evaluated. These polymeric particles encapsulating Stavudine (d4T) exhibited nearly 100% cell viability during cytotoxicity studies in comparison to pure d4T. The histological studies have revealed the in vivo biocompatibility while hemolysis studies certified the liability of formulation to be used parenteraly exhibiting no significant hemolytic toxixicty. The in vivo pharmacokinetics has shown the extended drug release from microsphere matrix upto a month. The calculated AUCtotal for d4T loaded polymeric microspheres was found to be 3341.656 µM h/ml; which was nearly 54 times than the total AUC of free d4T injected subcutaneously to the control group of animals; exhibiting the stable d4T concentration in blood avoiding fluctuation of the same indicating decreased probabilities of development of resistance against the treatment. Combination of targeted and subcutaneous administration of d4T will not only provide the stable and extended release of drug but also eradicate the hidden HIV hosted by macrophages. The concomitant regimen will potentially enhance the therapeutic efficacy with patient compliance; renewing new hopes for complete cure and improved quality of life in the AIDS patient.

Slipped capital femoral epiphysis associated with vitamin D deficiency: a series of 15 cases.

Slipped capital femoral epiphysis (SCFE) is uncommon in India and we routinely look for associated metabolic or endocrine abnormalities. In this study we investigated a possible association between vitamin D deficiency and SCFE. All children presenting with SCFE during the study period had their 25-hydroxyvitamin D levels measured as part of an overall metabolic, renal and endocrine status evaluation, which included measurement of body mass index (BMI). Vitamin D status was compared with age-, gender- and habitat-matched controls with acute trauma or sepsis presenting to our emergency department. A total of 15 children (12 boys and three girls) with a mean age of 13 years (sd 1.81; 10 to 16) presented for treatment for SCFE during a two-year period beginning in January 2010. Renal and thyroid function was within the normal range in all cases. The mean BMI was 24.9 kg/m(2) (17.0 to 33.8), which was significantly higher than that of the controls (p = 0.006). There was a statistically significant difference between the mean values of 25-hydroxyvitamin D in the children with SCFE and the controls (11.78 ng/ml (SD 5.4) versus 27.06 ng/ml (SD 5.53), respectively; p < 0.001). We concluded that, along with high BMI, there is a significant association of vitamin D deficiency and SCFE in India.

Magnetic properties of planar nanowire arrays of Co fabricated on oxidized step-bunched silicon templates.

Planar nanowire (NW) arrays of Co grown on oxidized step-bunched Si(111) templates exhibit room temperature ferromagnetic behaviour for wire widths down to 25 nm. Temperature and thickness dependent magnetization studies on these polycrystalline NW arrays show that the magnetic anisotropy of the NW array is dominated by shape anisotropy, which keeps the magnetization in-plane with easy axis parallel to the wires. This shape related uniaxial anisotropy is preserved even at low temperatures (10 K). Thickness dependent studies reveal that the magnetization reversal is governed by the curling mode reversal for thick wires whereas thinner wires exhibit a more complex behaviour which is related to thermal effects and size distribution of the crystal grains that constitute the NWs.

Comparison between acetylator phenotype and genotype polymorphism of n-acetyltransferase-2 in tuberculosis patients.

BACKGROUND: Isoniazid (INH) is one of the most important drugs of antitubercular treatment regime, and in some cases it causes hepatotoxicity. It is metabolized by hepatic N-acetyltransferase-2 (NAT2). AIM: To compare whether both methods, i.e., genotype NAT2 and phenotype test of measuring serum INH levels, are useful to identify acetylator status of patients on antitubercular treatment (ATT). METHODS: A total of 251 tuberculosis (TB) patients on standard treatment were followed up to 6 months for this study. NAT2 genotype was assessed by PCR with restriction fragment length polymorphism (RFLP) whereas serum INH levels were measured by fluorometry. RESULTS: Of the 251 patients, 50 (19.9%) developed ATT-induced hepatotoxicity. By phenotypic estimation, in the hepatotoxicity group, 17/50 (34%) were slow acetylators whereas 33/50 (66%) were fast acetylators. Genotypically, 19/50 (38%) were slow acetylators and 31/50 (62%) fast acetylators. By phenotypic analysis, in non-hepatotoxicity group, 46/201 (22.9%) were slow acetylators and 155/201 (77.1%) fast acetylators. By genotypic analysis, 30/201 (14.9%) were slow acetylators and 171/201 (85%) fast acetylators. Overall, slow acetylators (25.1%) measured phenotypically were not significantly different from slow acetylators (19.5%) measured genotypically. CONCLUSION: This study suggests that the acetylator status of TB patients can be detected by phenotypic method as efficaciously as by genotypic method. Therefore, phenotypic method can replace genotypic method to determine acetylating status as phenotypic method is simple and inexpensive.

A forgotten disease reminds itself with a rare complication.

Diagnosed cases of sexually transmitted diseases (STD) represent tip of the iceberg and Donovanosis in one of them. Donovanosis, in most cases is obvious clinically, but rely for its confirmation on the demonstration of donovan bodies in histological sections and cytological preparation. In an extremely rare setting, this disease may get complicated by the development of squamous cell carcinoma. We report this occurrence in an 18-year-old girl to review the currently forgotten status of donovanosis amongst the STDs and the poor outcome of the disease if left untreated.

Meconium peritonitis presenting as abdominal calcification: three cases with different pathology.

Intra-abdominal calcification is uncommon in newborns and has several causes of which meconium peritonitis is the most frequent. Three neonates with intra-abdominal calcification as a complication of meconium peritonitis are presented. The types of meconium peritonitis were cystic, meconium pseudocyst and meconium ascites. Two required surgical intervention. Meconium peritonitis should be considered in newborns with intra-abdominal calcification.

Prevalence of multidrug-resistant tuberculosis among newly diagnosed cases of sputum-positive pulmonary tuberculosis.

BACKGROUND & OBJECTIVES: The prevalence of multidrug-resistant tuberculosis (MDR-TB) is increasing throughout the world. Although previous treatment for TB is the most important risk factor for development of MDR-TB, treatment-naοve patients are also at risk due to either spontaneous mutations or transmission of drug-resistant strains. We sought to ascertain the prevalence of MDR-TB among new cases of sputum-positive pulmonary TB. METHODS: This was a prospective, observational study involving newly diagnosed cases of sputum-positive pulmonary tuberculosis diagnosed between 2008 and 2009 carried out in New Delhi, India. All sputum-positive TB cases were subjected to mycobacterial culture and first-line drug-susceptibility testing (DST). MDR-TB was defined as TB caused by bacilli showing resistance to at least isoniazid and rifampicin. RESULTS: A total of 218 cases of sputum-positive pulmonary tuberculosis were enrolled between 2008 and 2009. Of these, 41 cases had negative mycobacterial cultures and DST was carried out in 177 cases. The mean age of the patients was 27.8 ± 10.2 yr; 59 patients (27%) were female. All patients tested negative for HIV infection. Out of 177 cases, two cases of MDR-TB were detected. Thus, the prevalence of MDR-TB among newly diagnosed pulmonary tuberculosis patients was 1.1 per cent. INTERPRETATION & CONCLUSIONS: MDR-TB prevalence is low among new cases of sputum-positive pulmonary TB treated at primary care level in Delhi. Nation-wide and State-wide representative data on prevalence of MDR-TB are lacking. Efforts should be directed towards continued surveillance for MDR-TB among newly diagnosed TB cases.

Prevalence of multidrug-resistant tuberculosis among category II pulmonary tuberculosis patients.

BACKGROUND & OBJECTIVES: Multidrug-resistant tuberculosis (MDR-TB) has emerged as a significant global health concern. The most important risk factor for the development of MDR-TB is previous anti-tuberculosis therapy. Category II pulmonary TB includes those patients who had failed previous TB treatment, relapsed after treatment, or defaulted during previous treatment. We carried out this study to ascertain the prevalence of MDR-TB among category II pulmonary TB patients. METHODS: This was a cross-sectional, descriptive study involving category II pulmonary TB patients diagnosed between 2005 and 2008. All sputum-positive category II TB cases were subjected to mycobacterial culture and drug-susceptibility testing (DST). MDR-TB was defined as TB caused by bacilli showing resistance to at least isoniazid and rifampicin. RESULTS: A total of 196 cases of sputum-positive category II pulmonary tuberculosis patients were included. Of these, 40 patients (20.4%) had MDR-TB. The mean age of MDR-TB patients was 33.25 ± 12.04 yr; 9 patients (22.5%) were female. Thirty six patients showed resistance to rifampicin and isoniazid; while 4 patients showed resistance to rifampicin, isoniazid and streptomycin. The prevalence of MDR-TB among category-II pulmonary tuberculosis patients was 20.4 per cent. INTERPRETATION & CONCLUSIONS: The prevalence of MDR-TB in category II TB patients was significant. However, nation-wide and State-wide representative data on prevalence of MDR-TB are lacking. We stress the importance of continuous monitoring of drug resistance trends, in order to assess the efficacy of current interventions and their impact on the TB epidemic.

Giant intramuscular calcifying aponeurotic fibroma of gluteus maximus: case report.

Calcifying aponeurotic fibroma (CAF) is a rare soft tissue tumour which occurs mainly in children and adolescents. It usually involves the distal portion of the extremities, i.e. the hands and feet. A 2-year-old child with a large CAF is presented. The case was unusual in view of its large size (10 × 7 × 5 cm), dense calcification in such a young child, and because it was situated in the substance of gluteus maximus muscle. There has been no recurrence at 3-year follow-up.

Severe thrombocytopenia secondary to tubercular lymphadenitis.

A 5-year-old girl presented with a left axillary lymph node mass associated with generalised petechiae and mucosal bleeding and was diagnosed as tubercular lymphadenitis associated with immune thrombocytopenia. She responded well to anti-tubercular therapy. Tuberculosis should be considered in the differential diagnosis of lymphadenopathy and thrombocytopenia.

To compare CSF adenosine deaminase levels and CSF-PCR for tuberculous meningitis.

This study was planned to compare the adenosine deaminase (ADA) levels and polymerase chain reaction (PCR) in cerebrospinal fluid (CSF) as a rapid method to diagnose tuberculosis meningitis (TBM). Fifty-four adult patients with suspected TBM and 37 controls were included in this study. The median ADA level was 21U/L of most likely TBM, 14U/L of unconfirmed TBM and 5U/L of controls. PCR for Mycobacterium tuberculosis was positive in 12 out of 27 most likely TBM cases, 5 out of 27 unconfirmed TBM cases and 3 out of 37 controls. Using a cut off level of >10U/L, CSF-ADA had a sensitivity of 92.5% and specificity of 97% for the diagnosis of TBM. PCR for M. tuberculosis had a sensitivity of 44.5% and specificity 92% in the most likely TBM cases. This study shows that CSF-ADA is a more sensitive indicator than PCR for the diagnosis of M. tuberculosis.

Diagnostic efficacy of adenosine deaminase levels in cerebrospinal fluid in patients of tubercular meningitis: A comparison with PCR for Mycobacterium Tuberculosis.

BACKGROUND: The rapid diagnosis of Tubercular meningitis (TBM) is fundamental to clinical outcome. The key to diagnosis lies in Cerebrospinal fluid (CSF) analysis and radiological investigations. There are numerous lacunae in the confirmation of diagnosis of TBM from CSF. PURPOSE: The aim of present study was to compare the efficacy of CSF adenosine deaminase (ADA) level assays and Polymerase chain reaction (PCR) for Mycobacterium tuberculosis (M. tuberculosis) in the diagnosis of TBM. METHODS: Fifty four adult patients with suspected TBM and 37 controls were included in the study and CSF analyzed for ADA and PCR for M. tuberculosis. The cases were subdivided into definite (5), highly probable (22), probable (22) and possible TBM (5) as per previously validated criteria. The first two were grouped as "most likely" TBM (27) and last two as "unconfirmed" TBM (27). RESULTS: The mean ADA of the "most likely" TBM was 29±24, "unconfirmed" TBM was 21 ± 15 and controls were 4.8±2.2 U/L. The ADA levels correlated with CSF proteins, absolute lymphocyte count and the staging of the disease. Using a cut off level of >L10 U/L, CSF ADA had a sensitivity of 92.5% and specificity of 97%. PCR for M. tuberculosis was positive in 12 out of 27 "most likely" TBM cases, 5 out of 27 "unconfirmed" TBM cases and 3 out of 37 controls. PCR for M. tuberculosis had a sensitivity of 44.5% and specificity of 92% in the "most likely" TBM cases. CONCLUSIONS: ADA is a rapid, inexpensive and sensitive test in the diagnosis of TBM. It is more sensitive than AFB smear and culture. PCR is another rapid test in the diagnosis of TBM with a good specificity, even in those patients already on presumptive anti-tuberculous treatment. However, despite the sensitivity and specificity of CSF ADA, it should be corroborated with AFB smear and CSF PCR.

Histoplasmosis: cytodiagnosis and review of literature with special emphasis on differential diagnosis on cytomorphology.

BACKGROUND: Human infection with Histoplasma capsulatum runs the gamut from asymptomatic to disseminated disease. In immunocompromised patients, a tiny inoculum can lead to widespread disseminated infection. Early diagnosis and initiation of treatment is therefore important. OBJECTIVE: To review the cases of histoplasmosis diagnosed on fine needle aspiration cytology (FNAC) and to discuss the clinical presentation, associated inflammatory response, load of organisms and differential diagnosis on cytomorphology in these cases. METHODS: Retrospective review of seven cases of histoplasmosis at a tertiary-care centre during the period from 1998 to 2009 was performed. Clinical presentation along with cytomorphological features were studied and discussed in detail. RESULTS: The mean age of patients was 48.6 years and six out of seven were male. History of immunodeficiency (HIV) was available in five cases. Six patients presented with peripheral and/or abdominal lymphadenopathy. One patient had nodular shadows in both lungs and two also had skin lesions. On cytological smears, a variable load of uniform round to oval, about 2-4 microm in diameter, budding yeasts were seen intracellularly (within histiocytes) as well as extracellularly. In one case (HIV positive), these organisms were also seen within neutrophil polymorphonuclear leucocytes. In two cases, an inflammatory response in the form of epithelioid cell granulomas along with multinucleated giant cells was seen. CONCLUSIONS: FNAC is a reliable tool to recognize infection with H. capsulatum in tissues. This infection can cause a variable inflammatory response, which should be considered while reporting on such cases.

High frequency of loss of allelic integrity at Wilms' tumor suppressor gene-1 locus in advanced breast tumors associated with aggressiveness of the tumor.

BACKGROUND: The product of Wilms' tumor suppressor gene (WT1), a nuclear transcription factor, regulates the expression of the insulin-like growth factor (IGF) and transforming growth factor (TGF) systems, both of which are implicated in breast tumorigenesis and are known to facilitate angiogenesis. In the present study, WT1 allelic integrity was examined by Loss of Heterozygosity (LOH) studies in infiltrating breast carcinoma (n=60), ductal carcinoma in situ (DCIS) (n=10) and benign breast disease (n=5) patients, to determine its possible association with tumor progression. METHODS: LOH at the WT1 locus (11p13) as determined by PCR-RFLP for Hinf1 restriction site and was subsequently examined for its association with intratumoral expression of various growth factors i.e. TGF-beta1, IGF-II, IGF-1R and angiogenesis (VEGF and Intratumoral micro-vessel density) in breast carcinoma. RESULTS: Six of 22 (27.2%) genetically heterozygous of infiltrating breast carcinoma and 1 of 4 DCIS cases showed loss of one allele at WT1 locus. Histologically, the tumors with LOH at WT1 were Intraductal carcinoma (IDC) and were of grade II and III. There was no correlation in the appearance of LOH at WT1 locus with age, tumor stage, menopausal status, chemotherapy status and lymph node metastasis. The expression of factor IGF-II and its receptor, IGF-1R was significantly higher in carcinoma having LOH at WT1 locus. A positive correlation was observed between the TGF-beta1, VEGF expression and IMD scores in infiltrating carcinoma. CONCLUSIONS: The current study indicates that the high frequency of loss of allelic integrity at Wilms' tumor suppressor gene-1 locus in high-graded breast tumors is associated with aggressiveness of the tumor.

Bi-substitution-induced magnetic moment distribution in spinel Bi(x)Co(2-x)MnO(4) multiferroic.

We report the near-edge x-ray absorption spectroscopy (NEXAFS) at the Co/Mn L(3,2) edge and oxygen K edge of the well-characterized Bi-substituted Co(2)MnO(4) multiferroic samples. The evolution of peak features in NEXAFS spectra of the Co/Mn L(3,2) edge and O K edge show the Bi-induced redistribution of magnetic cations (Co/Mn). The variation in valence states of Co and Mn in all the substituted compositions is consistent with the observed ferrimagnetic behaviour of the samples. Magnetization data show the decrease in molecular field complementing the ferrimagnetism. The role of Bi in the enhancement of magnetic interactions as well as the appearance of ferroelectricity in Bi(x)Co(2-x)MnO(4) (0≤x≤0.3) is discussed.

Electron field emission from graphitic nanoflakes grown over vertically aligned carbon nanotubes.

Single step growth of self-assembled graphitic nanoflakes (GNF) over carbon nanotubes (CNT) on iron coated silicon(100) substrates is reported. These nanostructures were grown by varying the deposition time in a microwave plasma enhanced chemical vapor deposition reactor using acetylene, hydrogen and argon as reactant gases. Scanning electron microscope (SEM) studies of the deposited carbon films revealed that with increase in deposition time from 3 minutes to 6 minutes, the surface topography of the films transformed from one dimensional cylindrical nanostructure to flat-shaped two-dimensional nanoflakes. Carbon film deposited for 5 minutes showed improved surface coverage as compared to films deposited for 6 minutes i.e., surface area of the CNT film covered with nanoflakes increased as compared to carbon film deposited for higher durations. Field emission studies of films deposited at 5 minutes and 6 minutes showed increase in turn-on field, required for electron emission, from 2.7 V/microm to 2.9 V/microm respectively. However, such a combination of one dimension carbon and two dimension carbon may prove useful in applications where high surface area films are required.

Experimentally induced various inflammatory models and seizure: understanding the role of cytokine in rat.

BACKGROUND: The mechanism of epileptogenesis is not well established. There is higher incidence of seizures among patients with chronic inflammatory disease. Cytokines are rapidly induced in the brain after a variety of stimuli including inflammation. Aim of this study was to produce various inflammatory models and seizure to understand the role of TNFalpha in above mentioned models. MATERIALS AND METHODS: A total of 54 male rats were included in the study. Animals were divided into 3 groups of colitis, arthritis, and cotton wool granuloma. Each group had 3 subgroups of control, model and treatment. At the end of 3 days in colitis, 17 days in arthritis and 7 days in cotton wool granuloma groups a subconvulsive dose of PTZ (40 mg/kg i.p) was injected to note seizure onset and seizure score. Brain samples were subjected to DNA fragmentation testing. Presence of inflammation was confirmed by morphology and histology. Plasma and brain TNFalpha levels were measured. RESULTS: The models of colitis, arthritis and CWG were effectively produced as evidenced by morphology and histology scores (p<0.001). Seizure onset was reduced and grade was increased (p<0.001). Thalidomide reduced the morphological, histological (p<0.002), DNA fragmentation and seizure grade (p<0.001) while increased seizure onset (p<0.001) in the arthritis group. TNFalpha levels in both plasma and brain were reduced following thalidomide treatment (p<0.002) in arthritis group. There were no significant findings in colitis or cotton wool granuloma groups. CONCLUSION: Inflammation was associated with decreased threshold to PTZ induced seizure. Thalidomide is effective in reducing the extent of arthritis as well as reducing the seizure scoring and increasing seizure onset in the adjuvant arthritis group. Thalidomide was also effective in reducing TNFalpha levels thus contributing to its antiepileptic activity.

Structural investigations of mode of action of drugs: structure and conformation of a novel peptidyl nucleoside antibiotic chryscandin hydrochloride pentahydrate.

The structure and conformation of antibiotic chryscandin, a structural analog of the terminus 3'-aminoacyladenylate moiety of tRNA, has been determined. The crystals belong to orthorhombic space group P2(1)2(1)2(1). The structure was refined to an R value of 0.065 for 1872 reflections. The structure and conformation has been compared with that of puromycin. The sugar pucker is different from that of puromycin, thus creating different orientation of peptide group with respect to nucleoside. All the water molecules are involved in hydrogen bonding. The crystal is also stabilized by stacking of adenine bases and p-methoxyphenyl rings. The results will be helpful in understanding structure-biological activity relationships, identification of inhibitors of metallopeptidases, and how chryscandin interacts with ribosomal units.