In vitro Antioxidant and Anti-inflammatory Activities of the Flower Extracts of Hibiscus vitifolius L

Objective: To determine the antioxidant and anti-inflammatory activities of the solid powder extracted from the ethyl acetate fraction of the flower Hibiscus vitifolius L. Methods: The flower extract assessed for antioxidant activity using the 1,1–diphenyl-2-picryl-hydrazile (DPPH) radical scavenging assay and the reduced power assay was performed using the Ferric Reducing Capacity (FRC) assay. In vitro anti-inflammatory activity was assessed using human peripheral blood mononuclear cells (PBMC) induced by lipopolysaccharide (LPS) to test the production method of nitric oxide (NO). Results: The solid powder extracted from the ethyl acetate fraction of the flower Hibiscus vitifolius L showed good antioxidant activity in the scavenging DPPH radicals and the FRC assay compared to the standard sample. This powder sample also showed good anti-inflammatory activity in cell viability (LPS induced PBMC) assay and nitric oxide (NO) assay. Conclusion: These results suggest that the powder sample extracted from the ethyl acetate fraction of the flower Hibiscus vitifolius L has substantial antioxidant and anti-inflammatory activity.

Assessment of Leuconostoc spp. invasion in standing sugarcane with cracks internode

Aim: The post-harvest deterioration is most vexing problems of sugar industry and these losses due to Leuconostoc spp. cause enormous depreciation in cane tonnage and recovery. Thus, the objective of this study was to assess the invasion of Leuconostoc spp. in sugarcane. Methodology: Single internode is divided in three layers, viz., rind, first half and second half. Leuconsotoc spp. was grown and identified on specific growth media using juice from three portions of both the internodes (with cracks and without cracks) separately. Total Leuconostoc load, bacterial characterization, total soluble solids (TSS), reducing sugars (RS) and dextran were also determined. Results: The results revealed that cracked portions were incurred with Leuconostoc, however, rind layer had highest load followed by first half than the second half. More RS, high TSS and dextrans were quantified in cracked internode than without crack internode. Rind layer reported to have high RS, TSS and dextrans than the other portions. Interpretation: Canes possessing cracks had higher invasion of Leuconostoc spp., RS and dextran content than the ones lacking cracks leading to higher loss in sucrose content. Thus, there is a need to control these bacteria in sugarcane, especially in standing canes.

Mitochondrial DNA content of peripheral blood mononuclear cells in ART untreated & stavudine/zidovudine treated HIV-1-infected patients

Background & objectives: Nucleoside reverse transcriptase inhibitors (NRTIs) are known to cause mitochondrial toxicity. This study was done to estimate mitochondrial DNA (mtDNA) content of peripheral blood mononuclear cells (PBMCs) among human immunodeficiency virus (HIV) infected, NRTI treated and antiretroviral therapy (ART)-naïve patients and evaluate the utility of mtDNA content as a biomarker of mitochondrial toxicity. Methods: mtDNA content in PBMCs of 57 HIV-infected ART untreated and 30 ART treated with stavudine (d4T) or zidovudine (AZT) containing regimen were compared against 24 low-risk healthy controls (LoRHC). Results: There was a significant (P=0.01) reduction in mtDNA content among HIV-infected (104; 80-135) compared to LoRHC (127; 110-167), and it was the same in both the treated (104.8; 88-130) and untreated patients (104.7; 78-142). mtDNA significantly (P=0.014) declined in ART treated patients symptomatic for toxicity (97; 74-111) than the asymptomatic patients (128; 103- 153). Interpretation & conclusions: mtDNA depletion in PBMCs was evident among HIV-infected individuals on ART. Moreover, as mtDNA content was reduced among the patients symptomatic for toxicity than the asymptomatic in both the HIV-infected groups, the current study supports mtDNA content of PBMCs to serve as a biomarker of mitochondrial dysfunction induced by NRTI and HIV. Longitudinal studies with a large sample need to be done to confirm these findings.

The advantage of topographic prominence-adopted filter for the detection of short-latency spikes of retinal ganglion cells

Electrical stimulation through retinal prosthesis elicits both short and long-latency retinal ganglion cell (RGC) spikes. Because the short-latency RGC spike is usually obscured by electrical stimulus artifact, it is very important to isolate spike from stimulus artifact. Previously, we showed that topographic prominence (TP) discriminator based algorithm is valid and useful for artifact subtraction. In this study, we compared the performance of forward backward (FB) filter only vs. TP-adopted FB filter for artifact subtraction. From the extracted retinae of rd1 mice, we recorded RGC spikes with 8×8 multielectrode array (MEA). The recorded signals were classified into four groups by distances between the stimulation and recording electrodes on MEA (200-400, 400-600, 600-800, 800-1000 µm). Fifty cathodic phase-1(st) biphasic current pulses (duration 500 µs, intensity 5, 10, 20, 30, 40, 50, 60 µA) were applied at every 1 sec. We compared false positive error and false negative error in FB filter and TP-adopted FB filter. By implementing TP-adopted FB filter, short-latency spike can be detected better regarding sensitivity and specificity for detecting spikes regardless of the strength of stimulus and the distance between stimulus and recording electrodes.

Effect of Stimulus Waveform of Biphasic Current Pulse on Retinal Ganglion Cell Responses in Retinal Degeneration (rd1) mice

A retinal prosthesis is being developed for the restoration of vision in patients with retinitis pigmentosa (RP) and age-related macular degeneration (AMD). Determining optimal electrical stimulation parameters for the prosthesis is one of the most important elements for the development of a viable retinal prosthesis. Here, we investigated the effects of different charge-balanced biphasic pulses with regard to their effectiveness in evoking retinal ganglion cell (RGC) responses. Retinal degeneration (rd1) mice were used (n=17). From the ex-vivo retinal preparation, retinal patches were placed ganglion cell layer down onto an 8x8 multielectrode array (MEA) and RGC responses were recorded while applying electrical stimuli. For asymmetric pulses, 1st phase of the pulse is the same with symmetric pulse but the amplitude of 2nd phase of the pulse is less than 10 microA and charge balanced condition is satisfied by lengthening the duration of the pulse. For intensities (or duration) modulation, duration (or amplitude) of the pulse was fixed to 500 micros (30 microA), changing the intensities (or duration) from 2 to 60 microA (60 to 1000 micros). RGCs were classified as response-positive when PSTH showed multiple (3~4) peaks within 400 ms post stimulus and the number of spikes was at least 30% more than that for the immediate pre-stimulus 400 ms period. RGC responses were well modulated both with anodic and cathodic phase-1st biphasic pulses. Cathodic phase-1st pulses produced significantly better modulation of RGC activity than anodic phase-1st pulses regardless of symmetry of the pulse.

A rapid and low-cost microscopic observation drug susceptibility assay for detecting TB and MDR-TB among individuals infected by HIV in South India.

Background: The converging epidemics of HIV and tuberculosis (TB) pose one of the greatest public health challenges of our time. Rapid diagnosis of TB is essential in view of its infectious nature, high burden of cases, and emergence of drug resistance. Objective: The purpose of this present study was to evaluate the feasibility of implementing the microscopic observation drug susceptibility (MODS) assay, a novel assay for the diagnosis of TB and multi-drug-resistant tuberculosis (MDR-TB) directly from sputum specimens, in the Indian setting. Materials and Methods: This study involved a cross-sectional, blinded assessment of the MODS assay on 1036 suspected cases of pulmonary TB in HIV-positive and HIV-negative patients against the radiometric method, BD-BACTEC TB 460 system. Results: Overall, the sensitivity, specificity, positive predictive value, and negative predictive value of the MODS assay in detecting MTB among TB suspected patients were 89.1%, 99.1%, 94.2%, 95.8%, respectively. In addition, in the diagnosis of drug-resistant TB, the MODS assay was 84.2% sensitive for those specimens reporting MDR, 87% sensitivity for those specimens reporting INH mono-resistance, and 100% sensitive for specimens reporting RIF mono-resistance. The median time to detection of TB in the MODS assay versus BACTEC was 9 versus 21 days (P < 0.001). Conclusion: Costing 5 to 10 times lesser than the automated culture methods, the MODS assay has the potential clinical utility as a simple and rapid method. It could be effectively used as an alternative method for diagnosing TB and detection of MDR-TB in a timely and affordable way in resource-limited settings.

Evaluation of two indigenous rapid and two ELISA assays for the diagnosis of HIV infection India.

Purpose: Human immunodeficiency virus (HIV) diagnostic tests are being used extensively in India. However, the evaluation data on these assays are very limited. The present study evaluates indigenous HIV test kits manufactured in India. Materials and Methods: A total of 200 characterised specimens were assayed with Comb AIDS - RS Advantage HIV 1+2 Immunodot Test, Enzaids HIV 1+2 ELISA test, Enzaids Duet HIV Antigen+antibody ELISA test and Signal HIV Flow Through HIV 1+2 test kits. Performance characteristics of these assays were calculated. Results: Sensitivity, specificity, positive predictive value, negative predictive value and efficiency of all the assays were 100% except for Signal HIV Flow Through HIV 1+2 test kit. The specificity, positive predictive value and efficiency of the Signal HIV Flow Through HIV 1+2 test kit were 98.9%, 98.9% and 99.4%, respectively. The Enzaids Duet HIV kit was found to be extremely sensitive in detecting p24 Ag with the sensitivity of 1.5 pg/mL. Conclusions: To conclude, selection of better diagnostic assay is very much important to resolve discrepancies in HIV diagnosis. All these assays under evaluation in this report have got excellent performance characteristics and much suitable to use in serial testing algorithms in use for resources limited settings.

Establishment of T-lymphocyte subset reference intervals in a healthy adult population in Chennai, India.

Estimation of CD4+ T-lymphocytes continues to be an important aspect for monitoring HIV disease progression and response to antiretroviral therapy. Most of the diagnostic laboratories often rely on western text books for CD4+ T-lymphocyte reference values, which could, often be unreliable for usage in local settings. Therefore, we attempted to establish the reference values for T-lymphocyte subsets among healthy adults in a cross-sectional study carried out at the YRG Centre for AIDS Research and Education (YRG CARE) in Chennai, south India, in 213 (84 female and 129 male) healthy, HIV-1/2 seronegative adults as volunteers. Whole blood specimens were processed for CD4+, CD8+ T-lymphocyte estimation and haematological parameters. The established range of CD4+ T-lymphocyte counts for men and women were 383-1347 cells/microl (mean 865 and median 845 cells/microl) and 448-1593 cells/microl (mean 1021 and median 954 cells/microl), respectively. Women had significantly higher absolute CD4+ Tlymphocyte counts (P<0.001) and CD4+:CD8+ T-lymphocyte ratio as compared to men. The established normal range of CD4+ T-lymphocyte % was 21-59 (mean 40.2 and median 40.1). The influence of age was not observed in any of the parameters except CD4+/CD8+ T-lymphocyte ratio with the >45 yr age group. Further studies with greater sample size may be required to define the staging of HIV disease in relation to the normal CD4 T-lymphocyte count in the general population.

Comorbidities among HIV-infected injection drug users in Chennai, India.

BACKGROUND & OBJECTIVE: HIV-infected injection drugs users (IDUs) are known to have high rates of co-infections. A few reports exist on comorbidities among HIV-infected IDUs in India. We carried out a retrospective study to analyse data on comorbidities in India and treatment challenges faced when treating HIV-infected IDUs in India. METHODS: A retrospective chart review of 118 HIV-infected IDUs who accessed care at the YRG Centre for Substance Abuse-Related Research, Chennai, between August 2005 and February 2006 was done. Demographic, laboratory and clinical information was extracted from medical records. Descriptive demographic and clinical characteristics and distributions of comorbidities across CD4 cell count strata were analysed. RESULTS: All IDUs were male with a median age of 35.5 yr. The majority were married with average monthly income less than INR 3000 per month. The prevalence of hepatitis B and C infections were 11.9 and 94.1 per cent, respectively. Other common co-morbidities included oral candidiasis (43.2%), tuberculosis (33.9%), anaemia (22.9%), lower respiratory tract infections (16.1%), cellulitis (6.8%), herpes zoster (9.3%) and herpes simplex (9.3%). Among participants with CD4+ < 200 cells/microl, the prevalence of TB was 60 per cent. INTERPRETATION & CONCLUSION: IDUs in Chennai were commonly co-infected with HBV, HCV and tuberculosis, complicating use of antiretroviral and anti-tuberculous therapy. The current regimens available for the management of HIV and TB in India may need to be re-assessed for IDUs given the potential for increased rates of hepatotoxicity.

HIV-1 drug resistance among untreated patients in India: Current status.

HAART has dramatically improved survival and quality of life among people living with HIV and AIDS globally. However, drug resistant mutations of HIV are a great challenge to the benefits of HAART. Antiviral resistance can be mediated either by changes in the molecular target of therapy (the primary mechanism observed in HIV-1) or in other viral proteins that indirectly interfere with a drug's activity. Drug resistant mutations easily evolve in the presence of sub-optimal adherence. With the introduction of generic HAART, there has been a steep increase in the number of patients put on HAART in India. It should also be noted that since most patients pay for medications out of their own pockets, interruptions in therapy due to monetary constraints are not uncommon. There is little information on HIV drug resistance in resource constrained settings like India where the predominant circulating HIV-1 sub-type is C. The transmissibility of drug-resistant forms of the virus is also a major concern especially when formulating treatment guidelines. This article reviews published data available on the patterns of HIV-1 drug resistance among treatment naïve in India.

Methylation of E-cadherin and hMLH1 genes in Indian sporadic breast carcinomas.

Hypermethylation of promoter regions leading to inactivation of tumor suppressor genes is a common event in the progression of several tumor types. We have employed a novel restriction digestion based multiplex PCR assay to analyse the methylation status of promoter regions of tumor suppressor genes (p16, hMLH1, MGMT and E-cadherin) in sporadic breast carcinomas of Indian women. The present results indicated the absence of hypermethylation in promoter region of p16 and MGMT genes. However, 6 of the 19 (31.6%) sporadic breast carcinomas showed hypermethylation in the promoters of two of the genes analysed; three in hMLH1 and another three in E-cad. Since our earlier studies have shown lack of genetic alterations such as missense mutations and deletions in the tumor associated genes-p16, ras and p14ARF in sporadic breast tumors, the epigenetic alterations of the two genes reported in the present study could be of interest and might be among the events in the genesis/progression of sporadic breast carcinomas.

Usefulness of an observational database to assess antiretroviral treatment trends in India.

BACKGROUND: Human immunodeficiency virus (HIV) observational cohorts, which are established worldwide, support comparative studies across different regions. They have played an important role in developing international and country-specific HIV treatment and care guidelines. We describe the YRG CARE Chennai HIV observation database (YCHOD) and highlight its utility in monitoring trends in antiretroviral treatment use and HIV disease outcomes in India. METHODS: The baseline characteristics, time trends in antiretroviral treatment, trends in incidence of acquired immune deficiency syndrome (AIDS)-defining illness and mortality following the introduction of highly active antiretroviral therapy (HAART) in India were assessed using YCHOD. RESULTS: Till January 2005, 7647 HIV-positive patients had registered in YCHOD. A majority of the patients were men (69%) and had a mean age of 32 years. At baseline, 14% had an existing AIDS-defining illness. Among patients who required therapy by the WHO criteria, 14% initiated antiretroviral therapy (ART) in 1996 and 35% in 2000. Since the dramatic cost reduction of generic HAART in 2001, there has been an increase in the proportion of patients receiving ART to 57% in 2004. In patients who were started on HAART, the incidence of at least one AIDS-defining illness was 2% in 2001, and this decreased to 0.48% in 2004. CONCLUSION: We feel observational cohorts are useful as a surveillance tool for monitoring trends in treatment and disease progression. Standardized observational data collected systematically on HIV-infected individuals will help to assess the cost-effectiveness of ART and in planning ART strategies for India.

HIV-1 western blot assay: What determines an indeterminate status?

BACKGROUND: The Western blot assay is the gold standard for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). However, indeterminate Western blot reactivity to HIV-1 proteins may occur in individuals, who may not be infected with HIV. AIM: This retrospective study was aimed to determine the diagnostic value of the interpretation criteria in relation to commercial kits for HIV-1 diagnosis. METHODS AND MATERIALS: A total of 556 serum/plasma specimens collected from high-risk population attending our HIV clinic from 2000-2004 were tested by three different western blot kits: NEW LAV BLOT I (n=244), HIV BLOT 2.2; (n=112), Genetic Systems HIV-1 (n=237). And the results of western blot strips were analyzed using the various interpretation criteria: WHO/NACO, CDC/ ASTPHLD, ARC, FDA, CRSS and JHU. Some specimens were run on more than one kit. RT-PCR assay was performed on 5 specimens, which were indeterminate with LAV BLOT I. RESULTS: The discrepancy in LAV BLOT I positive results were between 157(64)-176(72), and indeterminate results were between 44(18) to 63(25). No such variations were observed in genetic systems. There are some HIV negative (by PCR) specimens were indeterminate in LAV BLOT I revealing the kit more sensitive and less effective for diagnostic purpose. CONCLUSION: The genetic systems kit is superior to other kits we analyzed and its results are concordant with HIV-1 PCR results. To report, the choice of western blot commercial kit is paramount important than the use of particular interpretation criteria for the diagnosis of HIV-1.

A comparative study on idiopathic pulmonary fibrosis and secondary diffuse parenchymal lung disease.

BACKGROUND AND AIMS: Clinical characteristics of patients diagnosed to have Diffuse parenchymal lung disease (DPLD) were evaluated in this study. DESIGN AND SETTING: Retrospective evaluation, a tertiary care center in South India. MATERIAL AND METHOD: Subjects diagnosed to have DPLD over a five-year period were included in this study. Data pertained to clinical characteristics and lab parameters were obtained. STATISTICAL CONSIDERATIONS: t- test for Mean values and chi-square test for comparing proportions were used. RESULTS: There were 73 eligible patients included for evaluation. Secondary cause for DPLD was diagnosed in 40 (55%) and idiopathic pulmonary fibrosis (IPF) was diagnosed in 33 (45%). The mean age was 45+/-11 and 53+/-10 years, of these 5 (12%) and 17 (52%) were male subjects in the secondary DPLD and IPF group respectively. The mean age, dyspnoea, cough, clubbing and crepitations were noted to be higher in patients with IPF as compared to patients with secondary DPLD. Fifty patients were followed up for a mean of 13 months (28 secondary DPLD and 18 IPF). Follow up data was available in 46 patients. Of these subjects prednisone alone was initiated in 24 subjects and combination with azathioprine in 22. Subjective improvement in symptoms was noted in 29/46 (63%), 19 with secondary DPLD and 10 with IPF. CONCLUSION: symptoms and signs were noted more frequently with IPF, subjective improvement to treatment was noted in 63% and the best response was noted among patients diagnosed to have sarcoidosis. A prospective trial is needed to study the long term prognosis and therapeutic response among Indian patients.

Regression of attained milestones in an HIV infected infant.

We report a case of an 18-month-old child with regression of attained developmental milestones as a manifestation of HIV encephalopathy. This is the first such report in Indian literature. Commencing antiretroviral therapy in this child resulted in arrest of further regression of milestones. This alerts pediatricians to be aware that early developmental delay and regression may be a presenting manifestation of HIV infection in a child.

Multiple cranial nerve palsy in an HIV-positive patient.

We report the case of a 32 -year- old HIV-positive Indian male who presented with sixth nerve (bilateral), ninth, tenth and twelfth nerve palsies; cerebellar and posterior column involvement. CT scan showed gyriform enhancement in the right occipital lobe and nodular leptomeningeal enhancement in the left frontal lobe. Cytomegalovirus serology was positive and the patient was treated as presumed CMV. HIV can present with multiple cranial neuropathy and varied neurological involvement.

Panophthalmitis due to rhizopus in an AIDS patient: a clinicopathological study.

Various opportunistic infections in the eye have been reported earlier in AIDS. We report a case of panophthalmitis in an AIDS patient where the eviscerated tissue on histopathologic and microbiologic examination showed the fungus Rhizopus.

Immune recovery vitritis presenting as panuveitis following therapy with protease inhibitors.

Immune reconstitution in acquired immunodeficiency syndrome (AIDS) patients on highly active anti-retroviral therapy (HAART) with cytomegalovirus (CMV) retinitis manifested as posterior segment intraocular inflammation has been reported. We report an adult HIV-positive Indian male with clinically inactive CMV retinitis who developed panuveitis with hypopyon. This was related to immune recovery mediated by combination anti-retroviral treatment, including protease inhibitors.