A pilot study on the effects of curd (dahi) & leaf protein concentrate in children with protein energy malnutrition (PEM).

BACKGROUND & OBJECTIVE: In protein-energy malnutrition (PEM) there is a significant impairment of immunity, both cell-mediated and humoral, which may be reversed with nutritional rehabilitation. With the use of probiotics like curd (dahi) and micronutrient-rich leaf protein concentrate (LPC), this immune recovery may be hastened. This study was conducted to assess the impact of supplementation of curd and LPC on nutritional status, and immunity as assessed by anthropometry, haemoglobin, ferritin levels, T- cell subpopulation and C-reactive protein (CRP), in children suffering from PEM. METHODS: Eighty moderate to severely malnourished children (1-5 yr) were randomized to receive either curd or LPC in addition to WHO recommended two-step diet over 15 days. Nutritional, immunological and haematological parameters were measured before and after supplementation and compared within the groups. RESULTS: The change in weight, haemoglobin level and CD4:CD8 T-cell subpopulation was significant in both the groups after supplementation. Response of CRP was blunted in PEM. Serum ferritin decreased significantly after supplementation in both groups. INTERPRETATION & CONCLUSION: Curd and LPC when added to diet of malnourished children, may have therapeutic value by accelerating immune recovery. More studies need to be done on a larger sample to confirm these findings.

Phenotypic and genotypic detection of ESBL mediated cephalosporin resistance in Klebsiella pneumoniae: emergence of high resistance against cefepime, the fourth generation cephalosporin.

OBJECTIVES: Cephalosporins belonging to second and third generation are commonly used in India for the treatment of Klebsiella pneumoniae. Report on resistance among K. pneumoniae strains to second and third generation cephalosporins are on rise in this country, which has been attributed to emergence of strains expressing extended-spectrum beta-lactamases (ESBLs). The aim of this study was to evaluate the in vitro susceptibility of K. pneumoniae to broad-spectrum cephalosporins particularly to cefepime, a recently introduced fourth generation cephalosporin in relation to ESBL production. METHODS: This study has been carried out in two phases among K. pneumoniae strains isolated between October 2001 and September 2002 (phase I, before marketing of cefepime in India) and between August 2003 and July 2004 (phase II, after marketing of cefepime in India). Minimum Inhibitory Concentration (MIC) was determined by a commercial strip containing gradient of antimicrobials (Strip E-test). Detection for ESBL production was carried out by DDST, E-test ESBL and PCR. RESULTS: Antimicrobial resistance profile of K. pneumoniae strains to five cephalosporins as analyzed by WHONET 5 identified 15 different resistance profiles among the 108 phase I isolates, ranging from resistance to none (19.44%) to all the five cephalosporin (8.33%) and eight different resistance profiles among the 99 phase II isolates, ranging from resistance to none (9.1%) to all the five cephalosporins (36.4%). Among the 108 phase I isolates a total of 71 (65.72%) and out of 99 phase II isolates, a total of 87 (88.0%) could be identified as ESBL producers. Among the isolates, regardless of the phase of the isolation, those characterized by production of ESBL showed overall higher frequency of resistance to cephalosporins (range 19.7-85.9% and 51.7-100% in phase I and phase II, respectively) compared to those for ESBL non-producers (range 0-13.5% and 0-25% in phase I and phase II, respectively). Ten randomly selected isolates from the most common resistance phenotypes probably belonged to a single strain as evident by MIC patterns, genotypic characterization and resistance profile to non-cephalosporin group of antimicrobials thereby pointing out the possibility of an outbreak. CONCLUSIONS: PCR may be regarded as a reliable method for detection of ESBL since in addition to the strains that could be identified as ESBL producers by DDST and E-test ESBL; PCR could demonstrate ESBL production among additional 32 strains (15 in phase I and 17 in phase II). Continued uses of cephalosporin group appear to be a potential risk factor for emergence of ESBL producing K. pneumoniae strains. In addition, as noted in the present study, the rise of resistance to cefepime that has been introduced recently in this country for therapeutic use could be of concern.

The use of immunomodulators as an adjunct to antituberculous chemotherapy in non-responsive patients with osteo-articular tuberculosis.

We studied 51 patients with osteo-articular tuberculosis who were divided into two groups. Group I comprised 31 newly-diagnosed patients who were given first-line antituberculous treatment consisting of isoniazid, rifampicin, ethambutol and pyrazinamide. Group II (non-responders) consisted of 20 patients with a history of clinical non-responsiveness to supervised uninterrupted antituberculous treatment for a minimum of three months or a recurrence of a previous lesion which on clinical observation had healed. No patient in either group was HIV-positive. Group II were treated with an immunomodulation regime of intradermal BCG, oral levamisole and intramuscular diphtheria and tetanus vaccines as an adjunct for eight weeks in addition to antituberculous treatment. We gave antituberculous treatment for a total of 12 to 18 months in both groups and they were followed up for a mean of 30.2 months (24 to 49). A series of 20 healthy blood donors served as a control group.Twenty-nine (93.6%) of the 31 patients in group I and 14 of the 20 (70%) in group II had a clinicoradiological healing response to treatment by five months. The CD4 cell count in both groups was depressed at the time of enrolment, with a greater degree of depression in the group-II patients (686 cells/mm(3) (sd 261) and 545 cells/mm(3) (sd 137), respectively; p < 0.05). After treatment for three months both groups showed significant elevation of the CD4 cell count, reaching a level comparable with the control group. However, the mean CD4 cell count of group II (945 cells/mm(3) (sd 343)) still remained lower than that of group I (1071 cells/mm(3) (sd 290)), but the difference was not significant. Our study has shown encouraging results after immunomodulation and antituberculous treatment in non-responsive patients. The pattern of change in the CD4 cell count in response to treatment may be a reliable clinical indicator.

Pediatric HIV infection.

OBJECTIVE: The present cross sectional study was undertaken to study clinical profile of HIV infection in children in Northern India. METHODS: 64 children from newborn to eighteen years, presenting for confirmation of diagnosis of HIV infection or monitoring of CD4-CD8 counts in confirmed cases, were evaluated. Children were categorized as per CDC classification of Pediatric HIV. The diagnosis was confirmed by serological tests or PCR assay. CD4-CD8 counts were done by FACS Count. RESULTS: Majority of the children were between 18 months to 5 years. Adolescents comprised 24% of the case. 51.5% children were infected through the mode of mother to child transmission. 39% of the case was transfusion-mediated. Unsafe medical injections probably contributed to 6.2% and heterosexual promiscuity led to 3.1% cases. Clubbing, not described in Indian studies so far, was seen in 9.3% cases. CONCLUSIONS: HIV infection is a chronic childhood disease extending into adolescence, and contaminated blood and unsafe medical injections are still important routes of HIV transmission in India.

Oropharyngeal carriage of Candida species in HIV-infected patients in India.

The present investigation represents the first study of oropharyngeal carriage of Candida and other yeasts in HIV-infected patients in India. One hundred and fifty HIV-positive patients were investigated by culturing their swish samples on plates of CHROMagar Candida. Ninety-eight patients (65.3%) were positive for Candida and four (2.7%) were positive for other yeasts. Among them, the first Indian C. dubliniensis isolate has been recovered. Molecular typing of selected C. albicans isolates by AP-PCR revealed two major genotypes based on the banding patterns. The susceptibilities of 30 Candida isolates to five antifungal agents including the new triazole voriconazole were determined in a micro-dilution test, according to the NCCLS protocol M 27. All the 22 C. albicans isolates were susceptible to five antimycotic agents (flucytosine, amphotericin B, fluconazole, voriconazole and itraconazole) except one isolate (VPCI-122), which was resistant to flucytosine (MIC > or = 64 mg l-1). The azole-resistant isolates reported here endorse the role of antifungal susceptibility testing whenever antifungal treatment with azoles is planned.

A cross sectional serological study of the co-infection of hepatitis B virus, hepatitis C virus and human immunodeficiency virus amongst a cohort of IDUs at Delhi.

A cross-sectional study was undertaken to find out co-prevalence of various infectious markers like Human Immunodeficiency Virus (HIV), Hepatitis B virus (HBV), Hepatitis C virus (HCV) and Syphilis infection amongst a cohort of injecting drug users (IDUs) in the city of Delhi. A total of 246 IDUs were enrolled during the 3 months period of the study. The results revealed a high prevalence of the viral markers studied i.e., HBV-39.59%, HCV-36.45%, HIV-36.99% and Syphilis-6.09%. A single marker infection was detected amongst 9.14% for HBV, 8.37% for HCV, 4.87% for HIV and 0.83% for Syphilis in samples tested for multiple markers. All the four markers could be detected in 1.76%. Amongst 11.16% and 27.9% of these samples, three and two markers respectively could be detected. The study revealed the problem of IV drug use and high prevalence of infectious markers including HIV in certain populations of Delhi and emphasizes the need for relevant interventions in these localised pockets.

Immune response in HIV-1-infected children with thalassaemia given a primary course of DPT vaccine before acquiring HIV-1 infection.

The effect of HIV infection on immune response to diphtheria and tetanus primary immunisation was investigated in 24 HIV-1-positive multi-transfused (MT) children with thalassaemia and compared with 48 HIV-1-negative MT thalassaemic children and 36 HIV-1-negative non-transfused (NT) children in the community. Diphtheria and tetanus antibody levels in the HIV-1-positive MT group were comparable with the two HIV-negative groups. The proportions of children with antibody titres below the protective level (i.e. <0.01 IU/ml) for antidiphtheria antibodies were 20.8, 16.6 and 16.6%, and 12.5, 12.5 and 13.9% for anti-tetanus antibodies in the three groups, respectively. On the other hand, delayed-type hypersensitivity (DTH) response to diphtheria and tetanus antigens was significantly depressed in the HIV-1-positive group compared with the HIV-negative controls. The mean percentages of both mature (CD20+) and immature (CD10+) B-cell counts were significantly higher in the HIV-1-positive group than in the HIV-negative MT and NT groups (p<0.05). Levels of serum immunoglobulins and spontaneously secreted immunoglobulins were significantly higher in the HIV-1-positive group compared with both HIV-negative groups. The HIV-1-positive group showed a mean (SD) IL-6 of 52.9 (28.8) pg/ml compared with 23.7 (12.1) pg/ml and a detection rate of 54.2% in the HIV-negative MT group, and 23.6 (8.2) pg/ml and a 50% detection rate in the HIV-negative NT group. The IL-2 level was significantly lower (p<0.05) in the HIV-1-positive group [41.7% detection rate and mean (SD) 28.8 (17.1) pg/ml] than in the HIV-negative MT and NT groups [75% and 83.3% detection rates and mean (SD) 57.2 (42.3) pg/ml and 99.3 (51.1) pg/ml, respectively]. During follow-up for 3 years, the frequency of major infections was significantly higher in the HIV-1-positive group than in the other two groups. Acute pneumonia and acute sinusitis were the predominant infections regardless of HIV status while primary bacteraemia, osteomyelitis, pyogenic meningitis and septic arthritis were common in the HIV-1-positive group. We conclude that, in HIV-1-infected children pre-immunised with DPT, DTH response to diphtheria and tetanus antigens might be more reliable than anti-diphtheria and anti-tetanus antibody levels in predicting susceptibility to major bacterial infections.

Disease progression markers during asymptomatic phase of HIV-1 infected children with unimpaired CD4+ cell values: evaluation of repeat CD4+ cell evaluation vs. other immunological parameters.

The availability of a marker that could predict the course of disease progression in HIV-infected individuals would be of considerable relevance during the asymptomatic stage in order to undertake timely prophylactic measures. A prospective study was undertaken in a group of 42 children suffering from thalassemia major with HIV-1 infection to assess the status of immune parameters, such as peripheral CD4+ T lymphocyte (CD4+ cell) percentage, delayed type of hypersensitivity (DTH) response to recall antigens, detection rate and levels of p24 antigen, and levels of beta-2 microglobulin and cytokines in serum. All were assessed at an interval of 2 years during the asymptomatic period, (baseline and follow-up assessments) in relation to the development of AIDS defining illness within a follow-up period of 3 years. No difference could be observed in the mean CD4+ cell percentage at baseline between those who progressed subsequently to develop AIDS within the follow-up period (progressors) and those who did not (non-progressors). However, at the point of follow-up assessment the progressor group showed significantly lower CD4+ cell percentage compared to the non-progressor group (33 +/- 4.9 vs. 22 +/- 5.6; p < 0.05), although in the progressor group there was no correlation of the baseline and follow-up CD4+ cell percentage with the duration of the AIDS-free interval. However, in the progressor group there was a strong negative correlation between the rate of decline in CD4+ cell percentage and subsequent duration of the AIDS-free interval (r = -0.859). Analysis of additional immune parameters at baseline revealed that the progressor group, despite having CD4+ cell values comparable to non-progressors, showed impaired DTH response (number and total induration of positive responses being 2.0 +/- 1.23 and 6.2 +/- 1.4 in the former group vs. 3.2 +/- 0.76 and 12.6 +/- 3.80 in the later group; p < 0.05 for both the parameters), and elevated levels (mg/l) of serum beta-2 microglobulin (2.92 +/- 0.89 vs. 1.38 +/- 0.43; p < 0.05). The serum cytokine profile at baseline in the progressor group showed a T helper type-2 (Th2) dominant pattern, i.e. elevation of interleukin-4 (IL-4) and interleukin-10 (IL-10) levels with decreased levels of interleukin-2 (IL-2) and gamma interferon (gamma-IFN) compared to the non-progressor group that showed a T helper type-1 (Th1) dominant profile, i.e., elevation of IL-1 and gamma-IFN level with decreased levels of IL-4 and IL-10 (p < 0.05 for all four cytokines). The present study points out that rate of decline rather than single point of assessment of CD4+ cell values is a more reliable predictor for disease progression in HIV-1 infected children. In addition, parameters such as DTH response, serum levels of beta-2 microglobulin and serum cytokine profile, may provide valuable predictors of subsequent fall in CD4+ cell value.

Synergistic immunosuppression by candida in HIV infection: a cytokine based analysis.

Candida is a common opportunistic pathogen in HIV infection and is regarded a signal infection for progression to AIDS. Cytokine imbalances between Th1/Th2 groups have been described in both candida and HIV infections. A study was undertaken to assess the role of candida in furthering immunosuppression in HIV infection based on cytokine levels and CD4 cell counts. 30 Indian subjects were enrolled; 10 HIV positive patients with and 10 without mucosal candidiasis and 10 age matched controls. Th1 cytokines; interleukin (IL) 2, IL 12 and interferon (IFN) gamma, Th2 cytokines; IL 4, IL 6, IL 10 and tumor necrosis factor (TNF) alpha with CD 4 cell counts were estimated using ELISA in all subjects. CD4 cell counts were reduced in both patient groups as compared to controls; significantly more in patients with both HIV and candida infections. There was a decrease in Th1 cytokine levels in all patients; lower levels of Th1 cytokines were seen in patients with both infections. Among the Th2 cytokines, there was a significant increase in the levels of IL 6, IL 10 and TNF alpha in both patient groups; IL 10 and TNF alpha values were significantly raised in patients with dual HIV and candida infections as compared to the other patients. There was no difference in IL 4 values across the subject groups. A positive correlation between CD4 cell counts and Th1 cytokine levels and a negative correlation with Th2 cytokines were noted; these were stronger in patients with both HIV and candidiasis. Thus, there was a Th1/Th2 cytokine imbalance with CD4 cell count reduction in all HIV infected patients, which was more pronounced in patients with both infections. It can be concluded that, owing to the depressed CD4 cell count and Th1 response and increased Th2 cytokines in patients with both candidiasis and HIV as compared to patients with only HIV candidiasis may have a synergistic immunosuppressive effect with HIV in patients with dual infections.

Nitric oxide dependent killing of mycobacterium tuberculosis by human mononuclear phagocytes from patients with active tuberculosis.

In this study we have demonstrated that nitric oxide, the product of the arginine dependent pathway of human mononuclear phagocytes effectively kills the M.tuberculosis in-vitro. The release of reactive nitrogen intermediates was triggered by incubation with various proinflammatory cytokines namely IFN gamma,TNF-alpha and IL-1R. We have earlier shown that human mononuclear phagocytes can be induced to release nitric,oxide (NO) radicals which can kill tumour cells. In the present communication, by using colony forming assays we demonstrated that human mononuclear phagocytes can effectively kill M.tuberculosis by using a NO dependent pathway. Treatment of mononuclear phagocytes with L-arginine resulted in markedly increased killing activity whereas, by using NGMMA, an analogue of L-arginine, the cidal activity could be brought down to the basal level. These results clearly suggest that cytokines, particularly IFN-gamma, induced NO release and its reactive product with oxygen radical, peroxynitrite, could play an important role in the killing of M. tuberculosis by human mononuclear phagocytes. A significant production of interleukin-4 and interleukin-10, by the ex-vivo matured, untreated macrophages from the active tuberculosis patients indicate that regulation of cytokine network to encourage in situ/local production of nitric oxide may be useful in the management of pulmonary tuberculosis.

Evaluation of epidemiological and serological predictors of human immunodeficiency virus type-1 (HIV-1) infection among high risk professional blood donors with western blot indeterminate results.

BACKGROUND: Indeterminate pattern of results in Western blot (WBI) for human immunodeficiency virus type-1 (HIV-1) infection may represent early HIV-1 infection or may be non-specific in origin. This issue can be resolved by follow up testing upto at least 6 months resulting in psychological distress as well as in high drop out rates among those undergoing investigation pointing out the need for additional parameters that could help in determining the status of HIV-1 infection at the time of initial testing itself in individuals with WBI pattern. OBJECTIVE: The objectives of the present study were: (i) to determine the frequency of HIV-1 infected individuals in a group of professional donors showing WBI patterns in initial testing on the basis of follow up serological studies; (ii) to find out if any HIV related epidemiological or serological characteristics recorded at the time of initial testing could be considered as predictor for HIV-1 infection in WBI specimens; and (iii) to evaluate two alternative serodiagnostic strategies for HIV-1 infection viz. multiple EIAs based on different antigen preparations/principles and a line immunoassay (LIA) employing recombinant antigens in resolving status of HIV-1 infection in specimens showing WBI results at initial testing. STUDY DESIGN: Professional donors with WBI patterns belonging to EIA reactive and EIA nonreactive groups were subjected to study of epidemiological profile, prevalence of sexually transmitted diseases (STD) markers and follow up serological testing for HIV-1 at 6, 12, 24 and 48 weeks intervals to record any seroconversion. The initial and follow up specimens from the donors with initial WBI results were subjected to two EIAs (one based on dot immunoassay using synthetic HIV-1 antigens and other based on microwell EIA using recombinant HIV-1 proteins) as well as to LIA. RESULTS: Professional donors with initial WBI results, from the EIA reactive group had higher proportion of unmarried individuals (90.3%), with history of heterosexual promiscuity (75%) and visit to STD clinics (36.1%) compared with the WBI donors from the EIA nonreactive group (72.7, 42.4 and 12.1%, respectively, P values < 0.001). Prevalence of antitreponemal antibodies was higher in the former group (16.7%) compared with the later group (1.5%, P value < 0.002). Seroconversion was recorded in 4 (7.3%) out of 55 EIA reactive WBI donors from the EIA reactive group that were characterised by high optical density (OD) values in EIA, 'p24 only' pattern of band in WB and positivity by LIA at the time of initial testing. LIA was found to be more reliable test compared with combination of EIAs to determine status of HIV-1 infection in WBI specimens at the time of initial testing. CONCLUSION: The present study points out that parameters like history of heterosexual promiscuity, prevalence of STD markers, high OD values in screening EIA, 'p24' only pattern of bands in WB and positivity by LIA could have individual predictive values for HIV-1 infection in specimens showing WBI pattern of results at initial testing.

Sexually-transmitted disease (STD) markers in multitransfused children in relation to human immunodeficiency virus type-1 (HIV-1) infection: impact of STD markers in blood donors.

Association of serological markers for various sexually transmitted diseases (STDs) with Human Immunodeficiency Virus type 1 (HIV-1) infection among heterosexually promiscuous blood donors is well recognized. However, possibility of similar association among children receiving transfusion from these donors has not been adequately explored. Study on the association of some STD markers with HIV-1 infection in multitransfused (MT) children showed that both HIV-1 positive and HIV-1 negative groups of children had significantly higher prevalence of hepatitis B virus (HBV) markers, i.e. HBsAg (23 and 30 per cent), anti-HBs (46 and 57 per cent), anti-HBc (18 and 23 per cent), antibodies to cytomegalovirus of IgM class (36 and 37 per cent) and IgG class (72.7 and 70 per cent), IgG antibodies to Herpes Simplex Virus type-2 (23 and 23 per cent) compared to control group. However, seropositivity for HIV-1 infection could not be attributed as a risk factor for any of these markers in the group of MT children. On the contrary, significant association of anti HBc, anti HCV, CMV IgG and antitreponemal antibody with HIV-1 infection could be recorded in the group of blood donors. It appeared that due to low prevalence of HIV-1 infection among blood donors in India, both HIV-1 positive and HIV-1 negative groups of MT children received transfusion from HIV negative donors predominantly resulting in a comparable prevalence of STD markers in both the groups of MT children due to cumulative effect of transfusion from HIV-1 negative donors.

Analysis of HIV seropositive thalassemic children for antibodies specific to Aspergillus fumigatus by luminescent immunoassay.

The applicability of luminescent immunoassay (LIA) in serodiagnosis of fungal infections in multitransfused (MT) thalassemic children seropositive for human immunodeficiency virus (HIV) was investigated. Thirty-one sera samples from HIV infected pediatric patients with thalassemia receiving repeated blood transfusions were analysed for the presence of antibodies specific to Aspergillus fumigatus by LIA. The LIA was standardized using well defined antigens of A. fumigatus. Ten out of 31 (32.2%) of the MT-HIV positive patients were found to have anti-Aspergillus antibodies in their sera by LIA. The ELISA could detect A. fumigatus specific antibodies in 25.8% (8 out of 31) of the patients. Thus, 20% more number of patients turned to be positive for aspergillosis by LIA as compared to ELISA. The difference was found to be statistically significant (p < 0.005). Of the MT-HIV negative patients only 1 out of 33 (3%) showed A. fumigatus specific antibodies by LIA and ELISA both. In age and sex matched control group (n = 25) none of the patients was found to be positive for antibodies to A. fumigatus. LIA was found to have better discriminatory value indicating, thereby, its utility in diagnosis of aspergillosis in compromised patients.

Prevalence of transfusion associated infections in multitransfused children in relation to mandatory screening of HIV in donated blood.

Any change in risk behavior related to acquisition of human immunodeficiency virus (HIV) infection is likely to reduce simultaneously the risk for other agents transmitted through identical routes. A study carried out in the city of Delhi, India on the load of transfusion associated infections among multitransfused (MT) children in relation to mandatory screening of HIV infection in donated blood indicated unchanged prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV) and hepatitis D virus (HDV) infections among the group of MT children transfused after the implementation of mandatory screening of HIV infections in blood banks, i.e. post-implementation period (prevalence of HBV, HCV and HDV being 32.8%, 31.3% and 1.6% respectively) compared to a group of MT children transfused over a similar duration before the implementation of mandatory screening i.e. pre-implementation period (prevalence of HBV, HCV and HDV being 28.1%, 26.6% and 1.6% respectively). However, reduction could be recorded in the prevalence of IgM and IgG classes of antibodies to both CMV and HSV-2 infections among MT children receiving transfusion during the post-implementation period (prevalence of 3.1% and 37.1% for CMV IgM and CMV IgG respectively; prevalence of 3.1% and 25% for HSV-2 IgM and HSV-2 IgG, respectively) compared to the group of MT children transfused in the pre-implementation period (prevalence of 15.6% and 56.3% for CMV IgM and CMV IgG respectively; prevalence of 18.8% and 45.2% for HSV-2 IgM and HSV-2 IgG, respectively). These reductions were statistically significant (p values < 0.02 and < 0.05 for CMV IgM and CMV IgG; p values < 0.01 and < 0.02 for HSV-2 IgM and HSV-2 IgG respectively). These observations were in accordance with the recorded reduction in the prevalence of CMV and HSV-2 infections and unaltered prevalence of HBV, HCV and HDV infections in the group of donors donating blood during the post-implementation period compared to those donating in the pre-implementation period. Study of epidemiological risk factors among blood donors showed a change in behavior towards safer sex practice with only 13.0% of donors in the post-implementation period having history of sex with one or more female commercial sex workers during their donation periods compared to 41.5% of donors in the pre-implementation period having similar history (p < 0.001). However no change could be recorded in the proportion of donors donating at frequency higher than the permissible guidelines among the two groups. The present study points out nosocomial transmission as well as limitations in the existing guidelines for screening of infectious agents in blood banks as possible incriminating factors towards acquisition of hepatitis virus infections in blood donors as well as in MT children.

Colposcopy as a tool for detection of human papillomavirus infection of uterine cervix in the setting of high prevalence of gynaecologic infections.

A cross sectional study involving 257 women from the Maternal and Child Health Centre (MCH) in Delhi was initiated for screening clinically, cytologically, colposcopically, and microbiologically for several gynaecologic infections. Eighty percent of the women had one or more gynaecologic infections and 31.1% had three or more infections. Cytology revealed changes suggestive of condyloma in 3 (1.2%) women only, while colposcopic examination suggested HPV changes in 117 (45.5%) women. A very high proportion of colposcopically detected lesions (78.6%) had evidence of HPV related changes in histology. The specificity of these lesions were further confirmed by Pan HPV DNA in-situ hybridisation, when 84% of the colposcopy is a valuable tool for detecting subclinical HPV lesions in a setting with high prevalence of gynaecologic infections.

Profile of antigen-specific antibody response detectable by western blot in relation to diagnostic criteria for human immunodeficiency virus type-1 (HIV-1) infection.

BACKGROUND: Variability in the profile of antigen-reactive bands in Western blot for serodiagnosis of human immunodeficiency virus (HIV) infection may result in disagreement regarding interpretation of positive result, due to lack of consensus in the interpretive criteria laid down by various organisations. OBJECTIVES: The objectives of this study were (i) to find out the extent of disagreement over various criteria regarding interpretation of positivity in Western blot and (ii) to review the discordance by retesting the discordant specimens using recombinant antigens as well as by performing repeat Western blot in follow-up specimens. STUDY DESIGN: A total of 467 specimens from high-risk groups, diagnosed positive for HIV type-1 (HIV-1) infection by the criteria of at least one of the five organisations, viz. Association of State and Public Health Laboratories Directors (ASTPHLD), Consortium for Retrovirus Serology (CRSS), American Red Cross (ARC) and World Health Organisation (WHO), were analysed to find out the extent of discordance between various criteria for interpretation of Western blot positivity. The discordant specimens were subjected to line immunoassay (LIA) using recombinant antigens. Also, follow-up Western blots were performed in case of discordant specimens at 6, 12 and 24 weeks intervals. RESULTS: We observed that criteria laid down by ASTPHLD, CDC and CRSS scored all the specimens as positive while ARC and WHO criteria scored 13 (2.8%) and 18 (3.8%) of specimens, respectively, as negatives which were detected as positives by other criteria (discordant specimens). The gp41 reactive band was the most frequently missing band, being undetectable in 11.6% of specimens while bands reactive to p24, p31, gp120 and gp160 could not be recorded in 1.9%, 9.4% and 3.2% and 1.5.% of specimens, respectively. Testing of the discordant specimens with recombinant antigen preparation and with repeat Western blot in follow-up specimens collected at 6, 12 and 24 weeks demonstrated all bands undetectable in initial Western blot, except 25% of gp41 reactive bands. CONCLUSIONS: It is felt that before selecting any criterion for Western blot positivity, it should be evaluated in the local population at risk for HIV-1 infection with additional or follow-up tests.

Further evaluation of alternative strategy for HIV testing in India.

A total 320 sera from groups at risk for HIV were evaluated by two (198 specimens) or three (122 specimens) screening tests for confirmatory anti-HIV testing in comparison to Western Blot as gold standard. Sera positive by both screening tests showed 100% correlation with Western Blot although with a false positivity rate of 3.2%. In specimens positive by 1st screening test but negative by the second, (considered negative for anti-HIV antibody as per WHO algorithm), 8.7% were found to be Western Blot positive showing the serious problem of false negativity of the proposed WHO algorithm. Employing the system of three screening test systems did not provide additional benefit for the specimens positive by initial two screening tests since all of them were positive by third test also. However, the study involving three screening tests substantiated the need for Western Blot in discordant specimens (i.e. positive by first test but negative by second), since in this group one out of 22 (4.6%) such specimens were Western Blot positive. Considering the serious consequences of both false positive and false negative results, it is felt that alternative strategy of confirmatory anti-HIV serology, although economical may not be suitable substitute for Western Blot in India at this juncture when the prevalence of HIV infection is relatively low.

Blood, bone marrow and splenic lymphocyte subset profiles in Indian visceral leishmaniasis.

We examined the lymphocyte subsets in peripheral blood, bone marrow and spleen of 11 patients with acute visceral leishmaniasis (VL) and 9 with chronic VL before and after 8 weeks of antileishmanial therapy. On admission, the CD4 cell count was depressed in the peripheral blood of acute and chronic VL cases as compared to the value in 10 normal control subjects. In contrast, CD4 cell counts were higher in the bone marrow in acute and chronic cases, and in splenic aspirates of chronic cases only, compared to normal values. The peripheral blood CD8 cell count, while normal in acute cases, was uniformly low in chronic cases. Counts of CD8 cells were also low in bone marrow of acute and chronic cases, as well as in splenic aspirates of chronic cases only. All these differences were significant (P < 0.05). After treatment, the CD4 cell count in the peripheral blood increased, but decreased in bone marrow and splenic aspirates. The CD8 cell count remained unaltered in the peripheral blood but increased significantly (P < 0.05) in bone marrow and spleen. The results suggest that in VL the peripheral blood picture may not reveal the actual T cell subset profile in the reticuloendothelial system. The changes in CD8 cell counts in the bone marrow and spleen seem to be independent, and are probably influenced by antileishmanial therapy.