Immune response against M protein-conserved region peptides from prevalent group A Streptococcus in a North Indian population

BackgroundGroup A streptococci (GAS) cause infections with a high prevalence in most developing countries. A GAS vaccine under trial that is based on the amino-terminus of the M protein provides type-specific immunity, and hence seems ineffective in India because of heterogeneous emm types. However, the conserved C-terminal region of the M protein protects against multiple serotypes. In this paper, the immune response generated against the conserved C-repeat region of the M protein was checked in an Indian population to establish their vaccine candidature.MethodsWhen screened for GAS, patients with pharyngitis, rheumatic fever/rheumatic heart disease (RF/RHD), and invasive disease showed heterogeneous emm types, out of which five prevalent types (1-2, 11, 49, 75 and 112) were selected for the study. The C-terminal region of their M proteins showed conserved C1-, C2-, and C3-repeats. The C1-repeat was more diverse and had two different J14-like sequences. Peptides to these C-terminal regions (J14.1 and J14-R6) were designed. Antibodies against these peptides were analyzed using the sera of 130 GAS-infected volunteers.ResultsSerum antibodies were significantly higher in patients with acute rheumatic fever, RHD, and invasive disease than in patients with pharyngitis or the healthy controls. The serum antibodies to these peptides was higher in teenagers and adults than in children.ConclusionResults showed an association between streptococcal disease progression and the age-related development of immunity to the conserved regions. Hence, these peptides could be considered protective in impeding streptococcal infections worldwide

HIV Infection and Host Genetic Mutation among Injecting Drug-users of Northeastern States of India

A community-based cross-sectional study was conducted among injecting drug-users (IDUs) of the northeastern states of India to understand the host genetic factors that confer resistance to HIV infection. The study aimed at assessing the existence and magnitude of genetic mutations of chemokine receptors, such as CCR2-64I, CCR-5 D-32, and SDF-1-3‘A, that are known to confer resistance to HIV infection and progression of disease in some set-ups. In total, 711 IDUs from Manipur, Mizoram, Nagaland, and Meghalaya were sampled for the study. The selected participants were interviewed to study their sociodemography, risk behaviours, and risk perceptions after obtaining their verbal informed consent. The interview was followed by collection of about 5 mL of blood samples by an unlinked anonymous method for studying genetic mutation and HIV infection. All the blood samples were transported to and processed at the clinical medicine laboratory of the National Institute of Cholera & Enteric Diseases, Kolkata, India. The genetic mutations were detected by polymerase chain reaction (PCR) and the restriction fragment length polymorphism (RFLP) assay techniques. The study revealed that 328 (46.1%) IDUs were aged 20–29 years, 305 (42.9%) were aged 30–39 years, and only two (0.3%) were aged above 49 years. The rate of HIV seropositivity varied widely among the IDUs living in different northeastern states that ranged from 4.5% to 61%. There was not a single IDU with CCR5 homozygous mutation. Mutated genes of CCR2-64I and SDF-1-3'A were detected in the frequencies of 49% and 23% respectively in them. The rate of HIV seropositivity in IDUs having CCR2 mutant gene was 27% (n=94) and without mutation was 27% (n=98). Similarly, HIV seropositivity in IDUs with and without SDF1 mutation was 28% (n=46) and 27% (n=146) respectively. Both the differences were not statistically significant. A CCR5 homozygous mutation is known to be the most prominent marker that confers resistance against HIV infection. The absence of CCR5 mutant gene in this population suggests that they do not have any additional protection against HIV infection. Analysis also revealed that, although mutation of CCR2 and SDF1 was present in this population, it did not confer any additional resistance against HIV. This indicates that the IDUs of northeastern India are not additionally protected against HIV infection through genetic mutation and are, therefore, vulnerable to acquire HIV infection due to high-risk behaviour and other related factors

HIV Infection and Host Genetic Mutation among Injecting Drug-users of Northeastern States of India

A community-based cross-sectional study was conducted among injecting drug-users (IDUs) of the northeastern states of India to understand the host genetic factors that confer resistance to HIV infection. The study aimed at assessing the existence and magnitude of genetic mutations of chemokine receptors, such as CCR2-64I, CCR-5 D-32, and SDF-1-3`A, that are known to confer resistance to HIV infection and progression of disease in some set-ups. In total, 711 IDUs from Manipur, Mizoram, Nagaland, and Meghalaya were sampled for the study. The selected participants were interviewed to study their sociodemography, risk behaviours, and risk perceptions after obtaining their verbal informed consent. The interview was followed by collection of about 5 mL of blood samples by an unlinked anonymous method for studying genetic mutation and HIV infection. All the blood samples were transported to and processed at the clinical medicine laboratory of the National Institute of Cholera & Enteric Diseases, Kolkata, India. The genetic mutations were detected by polymerase chain reaction (PCR) and the restriction fragment length polymorphism (RFLP) assay techniques. The study revealed that 328 (46.1%) IDUs were aged 20-29 years, 305 (42.9%) were aged 30-39 years, and only two (0.3%) were aged above 49 years. The rate of HIV seropositivity varied widely among the IDUs living in different northeastern states that ranged from 4.5% to 61%. There was not a single IDU with CCR5 homozygous mutation. Mutated genes of CCR2-64I and SDF-1-3\u2019A were detected in the frequencies of 49% and 23% respectively in them. The rate of HIV seropositivity in IDUs having CCR2 mutant gene was 27% (n=94) and without mutation was 27% (n=98). Similarly, HIV seropositivity in IDUs with and without SDF1 mutation was 28% (n=46) and 27% (n=146) respectively. Both the differences were not statistically significant. A CCR5 homozygous mutation is known to be the most prominent marker that confers resistance against HIV infection. The absence of CCR5 mutant gene in this population suggests that they do not have any additional protection against HIV infection. Analysis also revealed that, although mutation of CCR2 and SDF1 was present in this population, it did not confer any additional resistance against HIV. This indicates that the IDUs of northeastern India are not additionally protected against HIV infection through genetic mutation and are, therefore, vulnerable to acquire HIV infection due to high-risk behaviour and other related factors

Inhibition of NO2, PGE2, TNF-α, and iNOS EXpression by Shorea robusta L.: An Ethnomedicine Used for Anti-Inflammatory and Analgesic Activity

This paper is an attempt to evaluate the anti-inflammatory and analgesic activities and the possible mechanism of action of tender leaf extracts of Shorea robusta, traditionally used in ailments related to inflammation. The acetic-acid-induced writhing and tail flick tests were carried out for analgesic activity, while the anti-inflammatory activity was evaluated in carrageenan-and dextran- induced paw edema and cotton-pellet-induced granuloma model. The acetic-acid-induced vascular permeability, erythrocyte membrane stabilization, release of proinflammatory mediators (nitric oxide and prostaglandin E2), and cytokines (tumor necrosis factor-α, and interleukins-1β and -6) from lipopolysaccharide-stimulated human monocytic cell lines were assessed to understand the mechanism of action. The results revealed that both aqueous and methanol extract (400 mg/kg) caused significant reduction of writhing and tail flick, paw edema, granuloma tissue formation (P < 0.01), vascular permeability, and membrane stabilization. Interestingly, the aqueous extract at 40 μg/mL significantly inhibited the production of NO and release of PGE2, TNF-α, IL-1β, and IL-6. Chemically the extract contains flavonoids and triterpenes and toxicity study showed that the extract is safe. Thus, our study validated the scientific rationale of ethnomedicinal use of S. robusta and unveils its mechanism of action. However, chronic toxicological studies with active constituents are needed before its use

Evolution from invasive arterial puncture to a venous access for cerebral angiography: "Cath Lab to CT suite"

Background: Digital subtraction angiography (DSA) is considered as the gold standard in the evaluation of intracranial aneurysms. This study was undertaken to evaluate the effectiveness of computed tomogram angiography (CTA) in the detection and accurate characterization of intracranial aneurysms in suspected cases of nontraumatic subarachnoid hemorrhage. The importance of three-dimensional volume rendering of the intracranial vasculature and it′s used as an aid in improving diagnostic capabilities with regards to intracranial aneurysms in multi-detector computed tomography angiography (MDCTA), was stressed upon. This study also tried to probe whether MDCTA alone can be used in detection and treatment of intracranial aneurysms in emergency situations. Materials and Methods: Suspected cases of nontraumatic acute subarachnoid hemorrhage, over an 18 months period, underwent CTA in 16-slice-computed tomography suite. Fifty cases where CTA demonstrated intracranial aneurysms were studied. A set protocol of three-dimensional reconstruction was followed. Comparison of findings of MDCTA with surgical notes was performed. DSA was done in ambiguous cases. Results: Aneurysm was confidently diagnosed by CTA in 48 cases, and further confirmed on surgery. In doubtful cases, DSA was performed and then diagnosed as aneurysm. Thus, the sensitivity of CTA is diagnosing aneurysm is 96.6%, with a specificity of 100%. Conclusions: Digital subtraction angiography is an invasive, relatively costly, procedure to be done by highly skilled personnel with serious complication rate of 1%. This can be replaced by MDCTA, which is noninvasive, cost effective and easy to perform, and DSA can be reserved for doubtful or difficult cases. Following a set protocol of three-dimensional reconstruction helps in reducing errors

Oncogenic human papilloma virus and cervical pre-cancerous lesions in brothel-based sex workers in India

Summary: A community-based cross-sectional study was conducted in brothel-based sex workers of West Bengal, Eastern India, to determine their oncogenic human papillomavirus (HPV) status and the presence of pre-cancerous lesions. A total of 229 sex workers from three districts of West Bengal participated in the study. All the study participants were interviewed with the aid of a pre-tested questionnaire to determine their sociodemographics, risk behaviour and risk perceptions after obtaining informed verbal consent. The interview was followed by collection of cervical cells from all participants using a disposable vaginal speculum and cervical cytobrush. Oncogenic HPV DNA was detected by real-time polymerase chain reaction (PCR). A simultaneous Papanicolaou test (‘Pap smear’) was performed to detect cervical cytological abnormalities. Overall, the prevalence of oncogenic HPV was found to be 25% (58/229) among the studied population. A subset (n = 112) of the sample was tested separately to determine the existence and magnitude of HPV genotypes 16 and 18. The results showed that genotype 16 was prevalent in 10% (11/112), genotype 18 in 7% (8/112) and both genotype 16 and 18 in 7% (8/112). The HPV prevalence rate showed a decreasing trend with age, being 71.4% in the 10–19 years age group, 32.3% in the 20–29 years age group, 18.3% in the 30–39 years age group and 2.5% in the ≥40 years age group (statistically significant differences, P ≤ 0.00001). Considering the duration of sex work, oncogenic HPV prevalence was found to be 55% (n = 21) and 19% (n = 35) in sex workers with a sex working duration of ≤1 year and >1 year, respectively. This difference was found to be statistically significant both by univariate and multivariate analysis. In this study, it was observed that sex workers with an average number of daily clients of six or more had an HPV prevalence of 67% (n = 6), those with four to five clients had a prevalence of 45% (n = 9), those with two to three clients had a prevalence of 30% (n = 34) and those with one or less clients had a prevalence of 10% (n = 9) (statistically significant differences, P = 0.00003). Multivariate analysis showed a statistical association only with a duration of sex work of ≤1 year [odds ratio (OR) = 3.3; 95% confidence interval (CI) 1.4–7.6] and daily income of Rupees (Rp) ≥101 (OR = 2.5; 95% CI 1.3–5). Regarding pre-cancerous lesions, 2 of 229 sex workers showed the presence of a low-grade squamous intraepithelial lesion along with high-risk HPV. Thus, 1% of the studied population suffer from a pre-cancerous lesion caused by high-risk HPV. This study concludes that young sex workers are particularly vulnerable to high-risk HPV, similar to human immunodeficiency virus (HIV). The observation of older sex workers relatively free from HPV supports the view of acquired immunity against HPV, which needs to be studied in-depth further. There is a need for a suitable community-based intervention programme targeted towards sex workers, with special reference to younger sex workers, for control and prevention of HPV and cervical cancer. Vaccination against HPV for newly entrant sex workers may be an important component for a successful intervention programme. Keywords: Human papillomavirus, Oncogenic HPV, Sex workers, India, Risk factors, Cervicitis, Cervical pre-cancerous lesion, ST

Anti-herpes virus activities of bioactive fraction and isolated pure constituent of <it>Mallotus peltatus</it>: an ethnomedicine from Andaman Islands

Abstract Background Viral infections, particularly the infections caused by herpes simplex virus (HSV), represent one of the most serious public health concerns globally because of their devastating impact. The aim of this study was to evaluate the antiviral potential of methanolic crude extract of an ethnomedicine Mallotus peltatus, its active fraction and pure compound, against HSV-1 F and HSV-2 G. Result The cytotoxicity (CC50, the concentration of 50% cellular toxicity), antiviral effective concentration (EC50, the concentration required to achieve 50% protection against virus-induced cytopathic effect), plaque reduction and the selectivity index (SI, the ratio of CC50 and EC50) was determined. Results showed that the crude methanolic extract of M. peltatus possessed weak anti-HSV activity. In contrast, the active fraction A and isolated ursolic acid from fraction A exhibited potent antiherpesvirus activity against both HSV-1 (EC50 = 7.8 and 5.5 μg/ml; SI = 22.3 and 20) and HSV-2 (EC50 = 8.2 and 5.8 μg/ml, and SI = 21.2 and 18.97). The fraction A and isolated ursolic acid (10 μg/ml) inhibited plaque formation of HSV-1 and HSV-2 at more than 80% levels, with a dose dependent antiviral activity, compared to acyclovir. The time response study revealed that the anti-HSV activity of fraction A and isolated ursolic acid is highest at 2–5 h post-infection. Moreover, the time kinetics study by indirect immunofluorescence assay showed a characteristic pattern of small foci of single fluorescent cells in fraction A- treated virus infected cells at 2 h and 4 h post-infection, suggesting drug inhibited viral dissemination. Further, the PCR study with infected cell cultures treated with fraction A and isolated ursolic acid at various time intervals, failed to show amplification at 48–72 h, like acyclovir treated HSV-infected cells. Moreover, fraction A or isolated ursolic acid showed no interaction in combination with acyclovir. Conclusion This study revealed that bioactive fraction A and isolated ursolic acid of M. peltatus has good anti-HSV activity, probably by inhibiting the early stage of multiplication (post-infection of 0–5 h), with SI value of 20, suggesting its potential use as anti-HSV agents.</p