Hypericin-photodynamic therapy leads to interleukin-6 secretion by HepG2 cells and their apoptosis via recruitment of BH3 interacting-domain death agonist and caspases.

Photodynamic therapy (PDT) has emerged as a capable therapeutic modality for the treatment of cancer. PDT is a targeted cancer therapy that reportedly leads to tumor cell apoptosis and/or necrosis by facilitating the secretion of certain pro-inflammatory cytokines and expression of multiple apoptotic mediators in the tumor microenvironment. In addition, PDT also triggers oxidative stress that directs tumor cell killing and activation of inflammatory responses. However, the cellular and molecular mechanisms underlying the role of PDT in facilitating tumor cell apoptosis remain ambiguous. Here, we investigated the ability of PDT in association with hypericin (HY) to induce tumor cell apoptosis by facilitating the induction of reactive oxygen species (ROS) and secretion of Th1/Th2/Th17 cytokines in human hepatocellular liver carcinoma cell line (HepG2) cells. To discover if any apoptotic mediators were implicated in the enhancement of cell death of HY-PDT-treated tumor cells, selected gene profiling in response to HY-PDT treatment was implemented. Experimental results showed that interleukin (IL)-6 was significantly increased in all HY-PDT-treated cells, especially in 1 µg/ml HY-PDT, resulting in cell death. In addition, quantitative real-time PCR analysis revealed that the expression of apoptotic genes, such as BH3-interacting-domain death agonist (BID), cytochrome complex (CYT-C) and caspases (CASP3, 6, 7, 8 and 9) was remarkably higher in HY-PDT-treated HepG2 cells than the untreated HepG2 cells, entailing that tumor destruction of immune-mediated cell death occurs only in PDT-treated tumor cells. Hence, we showed that HY-PDT treatment induces apoptosis in HepG2 cells by facilitating cytotoxic ROS, and potentially recruits IL-6 and apoptosis mediators, providing additional hints for the existence of alternative mechanisms of anti-tumor immunity in hepatocellular carcinoma, which contribute to long-term suppression of tumor growth following PDT.

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.

Cofactors for low serum albumin levels among HIV-infected individuals in Southern India.

BACKGROUND AND AIMS: Malnutrition and low serum albumin among human immunodeficiency virus (HIV)-infected individuals are cofactors for HIV disease progression. The present study aimed to identify the proportion of HIV-infected individuals with low serum albumin and the possible cofactors among highly active antiretroviral therapy (HAART) experienced and HAART naïve individuals. METHODS: A total of 835 HIV-infected individuals (HAART-experienced, HAART-naïve) were included in the study. RESULTS: Of the 835 individuals, 44.6% had normal (4.2-5.2 g/dL) and 55.4% had abnormal (<4.2 g/dL) albumin levels. The abnormal group had significantly lower body mass index (BMI) compared with the normal group (P = .02). Among those with abnormal albumin, 388 (84%) were HAART experienced compared with 239 (64%) with normal albumin (P < .001). Among the abnormal group, 259 (55.9%) had CD4 count <200 cells/mL as compared with 124 (33.3%) in the normal group (P < .001). CONCLUSIONS: CD4 count and lower were the major cofactors for low serum albumin among HIV-infected individuals. Therefore, serum albumin would be a useful biochemical test for HIV disease in resource-limited settings.

Ethnic variation in certain hematological and biochemical reference intervals in a south Indian healthy adult population.

BACKGROUND: We established the biochemical and hematological reference intervals among a south Indian healthy adult population attending an HIV referral centre in Chennai, southern India. METHODS: In a cross sectional study, 213 study subjects (129 male and 84 female) were studied between March and August 2005. All of the parameters were analyzed using standard hematological and biochemical techniques. RESULTS: Certain biochemical (viz. total bilirubin, alanine transaminase, albumin, creatinine, total protein, lipid profile, creatine phosphokinase, uric acid and lactate) and hematological (mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration and lymphocyte levels) parameters presented higher upper limits. In addition, the upper limits of white blood cell count, platelet count, hematocrit, red blood cell count and hemoglobin level were low in comparison to the currently reported ranges. CONCLUSION: Ethnic variation in reference intervals was observed in certain biochemical and hematological analytes in a south Indian adult population.

High proportion of isosporiasis among HIV-infected patients with diarrhea in southern India.

We investigated 245 diarrheal stool specimens from HIV-positive subjects between January 2003 and December 2006 to determine the etiological role of coproparasites. Parasitic etiology was observed in 91 (37.1%) cases. Isospora belli (26.1%) was the most common parasite followed by Entameba histolytica/dispar (3.3%), Cryptosporidium spp. (2.9%), Giardia intestinalis (1.6%), and Strongyloides stercoralis (1.2%). Interesting trends of significant increase in the number of cases of I. belli and decline in Cryptosporidium spp. were observed during the study period.

Hydrothorax in association with Scopulariopsis brumptii in an AIDS patient in Chennai, India.

Over the past decade, an increasing number of opportunistic fungal infections have been reported in immunocompromised subjects. Microascus spp. and their anamorphs Scopulariopsis spp. have been recovered from a wide geographical range. We report a case of Scopulariopsis brumptii in a 27-year-old man with AIDS presenting with breathlessness, pericardial effusion and hydrothorax in Chennai, southern India, in February 2007. Because of respiratory arrest, the patient was intubated. However, the patient developed obstructive shock and died due to cardiac dysfunctions. This report underlines the need for a direct, intensive approach to investigations in immunocompromised patients, especially those with AIDS.

Prevalence of aflatoxin B1 in liver biopsies of proven hepatocellular carcinoma in India determined by an in-house immunoperoxidase test.

Hepatocellular carcinoma (HCC) is the fourth leading cause of cancer-related death in the world. The incidence of HCC in India is reportedly low and varies from 0.2 to 1.9 %. Aflatoxins, secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus, are potent human carcinogens implicated in HCC. The prevalence of aflatoxin B1 (AFB1) as co-carcinogen was analysed using an in-house immunoperoxidase test in 31 liver biopsies and 7 liver-resection specimens from histopathologically proven HCC, and in 15 liver biopsies from cirrhosis patients (control group). Serum was tested for hepatitis B and C serological markers using commercial assays, and for AFB1 using an in-house ELISA with a sensitivity of approximately 1 ng ml(-1) for AFB1. In spite of positive AFB1 immunostaining in HCC cases, all serum specimens, from both HCC and the control groups, were AFB1-negative. There were 18 (58.1 %) HCC cases that revealed AFB1 in liver biopsies; 68.8 % (n=11) of non-B non-C hepatitis cases with HCC and 46.1 % (n=6) of the hepatitis B surface-antigen-positive subjects were positive for AFB1. Out of the two hepatitis B/hepatitis C virus co-infected cases, one was positive for AFB1. Of seven tumour-resection samples, six were positive for AFB1. Only one case revealed AFB1 in the non-tumour area of the resected material. Thus AFB1 staining was significantly associated with tumour tissue (P=0.03). Aflatoxins proved to have a significant association with HCC in this peninsular part of the subcontinent. The impact seems to be a cumulative process, as revealed by the AFB1 deposits in HCC liver tissue, even though the serum levels were undetectable.

Evaluation of the growth inhibitory activities of Triphala against common bacterial isolates from HIV infected patients.

The isolation of microbial agents less susceptible to regular antibiotics and the rising trend in the recovery rates of resistant bacteria highlights the need for newer alternative principles. Triphala has been used in traditional medicine practice against certain diseases such as jaundice, fever, cough, eye diseases etc. In the present study phytochemical (phenolic, flavonoid and carotenoid) and antibacterial activities of aqueous and ethanol extracts of Triphala and its individual components (Terminalia chebula, Terminalia belerica and Emblica officinalis) were tested against certain bacterial isolates (Pseudomonas aeruginosa, Klebsiella pneumoniae, Shigella sonnei, S. flexneri, Staphylococcus aureus, Vibrio cholerae, Salmonella paratyphi-B, Escherichia coli, Enterococcus faecalis, Salmonella typhi) obtained from HIV infected patients using Kirby-Bauer's disk diffusion and minimum inhibitory concentration (MIC) methods. T. chebula was found to possess high phytochemical content followed by T. belerica and E. officinalis in both aqueous and ethanol extracts. Further, most of the bacterial isolates were inhibited by the ethanol and aqueous extracts of T. chebula followed by T. belerica and E. officinalis by both disk diffusion and MIC methods. The present study revealed that both individual and combined aqueous and ethanol extracts of Triphala have antibacterial activity against the bacterial isolates tested.

Bacterial etiology of diabetic foot infections in South India.

BACKGROUND: Foot infections are a frequent complication of patients with diabetes mellitus, accounting for up to 20% of diabetes-related hospital admissions. Infectious agents are associated with the worst outcomes, which may ultimately lead to amputation of the infected foot unless prompt treatment strategies are ensued. The present study sought to reveal the bacterial etiology of diabetic foot ulcers in South India, the diabetic capital of India. METHODS: A 10-month-long descriptive study was carried out to analyse the aerobic and anaerobic bacterial isolates of all patients admitted with diabetic foot infections presenting with Wagner grade 2-5 ulcers. Bacteriological diagnosis and antibiotic sensitivity profiles were carried out and analysed using standard procedures. RESULTS: Diabetic polyneuropathy was found to be common (56.8%) and gram-negative bacteria (57.6%) were isolated more often than gram-positive ones (42.3%) in the patients screened. The most frequent bacterial isolates were Pseudomonas aeruginosa, Staphylococcus aureus, coagulase-negative staphylococci (CONS), and Enterobacteriaceaes. Forty-nine cultures (68%) showed polymicrobial involvement. About 44% of P. aeruginosa were multi-drug-resistant, and MRSA was recovered on eight occasions (10.3%). Bacteroides spp. and Peptostreptococcus spp. were the major anaerobic isolates. CONCLUSIONS: Our study supports the viewpoint put forth by previous South Indian authors that the distribution of gram-negative bacteria (57.6%) is more common than that of gram-positive ones (42.3%) and it is contrary to the viewpoint that diabetic foot infections are frequently monomicrobial. Furthermore, recovery of multi-drug-resistant P. aeruginosa isolates is of serious concern, as almost no one has reported the same from the South Indian milieu.

Colonization of mycoplasma in the upper respiratory tract of AIDS patients with pulmonary symptoms in Chennai, India.

BACKGROUND & OBJECTIVE: Mycoplasmas have been implicated in causing minor to severe respiratory infections in man. Mycoplasmas are considered to act as cofactors in patients with AIDS. A preliminary study was conducted to isolate mycoplasmas from sputum specimens of AIDS patients and non-HIV patients with underlying pulmonary symptoms and signs. METHODS: A total of 130 sputum samples (100 from AIDS patients and 30 from non-HIV) were cultured on standard pleuropneumonia-like organisms (PPLO) glucose agar up to 3 wk. The plates were examined for the presence of fried-egg colonies characteristic of Mycoplasma. Subsequently the plates were stained using Diene's stain. Sputum specimens from the AIDS patients were also screened for other bacterial pathogens. RESULTS: Mycoplasmas were detected from 36 (36%) of the AIDS patients and only 5 (16.6%) of the non HIV control individuals with underlying pulmonary symptoms. Data on the detection rates of other microorganisms from the AIDS cases were also analysed. INTERPRETATION & CONCLUSION: This preliminary study provided supportive evidence that mycoplasma colonized in upper respiratory tract of individuals with AIDS to a larger extent than that of the non HIV subjects with pulmonary symptoms. Further studies need to be done to characterize mycoplasma isolates to species level.