A K Rai

Hybrid Capture 2 Assay Based Evaluation of High-Risk HPV Status in Healthy Women of North-East India

BACKGROUND High risk HPV (HR-HPV) testing has been recommended as an effective tool along with cytology screening in identification of cervical intraepithelial lesions (CINs) and prevention of their progress towards invasive cervical cancer. The aim of this study was to assess the HR-HPV DNA status by Hybrid Capture 2 (HC2) assay in healthy asymptomatic women of North-East India. MATERIALS AND METHODS This study examined cervical cell samples of forty three (n=43) healthy women by HC2 assay. A High Risk HPV DNA kit (Qiagen) was used which can detect 13 high risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68. RESULTS The mean relative light units (RLU) for samples was in the range of 141-5, 94, 619. HR-HPV DNA was confirmed in 16% (7/43) of participant women samples. Among demographic and clinical parameters, menstrual irregularity (p=0.039) and infection history (p=0.028) has shown statistically significant differences between the HR-HPV-positive and negative groups. In the HR-HPV positive group, two women were confirmed for CINs after colposcopy and histopathologic examination. CONCLUSIONS We suggest that there may be an association between irregular menstruation and infection history of the urogenital tract with HR-HPV DNA prevalence in North-East Indian asymptomatic women. HC2 assay can be a valuable tool for HR-HPV screening.

Alcohol and Tobacco Increases Risk of High Risk HPV Infection in Head and Neck Cancer Patients: Study from North-East Region of India

Background Human papilloma virus (HPV) associated Head and Neck Cancers (HNCs) have generated significant amount of research interest in recent times. Due to high incidence of HNCs and lack of sufficient data on high-risk HPV (hr-HPV) infection from North -East region of India, this study was conceived to investigate hr-HPV infection, its types and its association with life style habits such as tobacco, alcohol consumption etc. Methods A total of one hundred and six primary HNC tumor biopsy specimens were collected. These samples were analyzed for hr-HPV DNA (13 HPV types) using hybrid capture 2 (HC2) assay and genotyping was done by E6 nested multiplex PCR (NMPCR). Results The presence of hr-HPV was confirmed in 31.13% (n = 33) and 24.52% (n = 26) of the HNC patients by nested multiplex PCR (NMPCR) and HC2 assay respectively. Among hr-HPV positive cases, out of thirteen hr- HPV types analyzed, only two prevalent genotypes, HPV-16 (81.81%) followed by HPV-18 (18.18%) were found. Significant association was observed between hr-HPV infection with alcohol consumption (p <0.001) and tobacco chewing (p = 0.02) in HNC cases. Compared to HPV-18 infection the HPV-16 was found to be significantly associated with tobacco chewing (p = 0.02) habit. Conclusions Our study demonstrated that tobacco chewing and alcohol consumption may act as risk factors for hr-HPV infection in HNCs from the North-East region of India. This was the first study from North-East India which also assessed the clinical applicability of HC2 assay in HNC patient specimens. We suggest that alcohol, tobacco and hr- HPV infection act synergistically or complement each other in the process of HNC development and progression in the present study population.

Distinct Involvement of 9p21-24 and 13q14.1-14.3 Chromosomal Regions in Raw Betel-Nut Induced Esophageal Cancers in the State of Meghalaya, India

BACKGROUND Raw betel nut (RBN) chewing is an important contributing factor for esophageal squamous cell carcinoma (ESCC), although associated genomic changes remain unclear. One difficulty in assessing the effects of exclusively RBN induced genetic alterations has been that earlier studies were performed with samples of patients commonly using tobacco and alcohol, in addition to betel-quid. Both CDKN2A (at 9p21) and Rb1 gene (at 13q14.2) are regarded as tumor suppressors involved in the development of ESCC. Therefore, the present study aimed to verify the RBNs ability to induce ESCC and assess the involvement of CDKN2A and Rb1 genes. METHODS A panel of dinucelotide polymorphic markers were chosen for loss of heterozygosity studies in 93 samples of which 34 were collected from patients with only RBN-chewing habit. Promoter hypermethylation was also investigated. RESULTS Loss in microsatellite markers D9S1748 and D9S1749, located close to exon 1β of CDKN2A/ARF gene at 9p21, was noted in 40% ESCC samples with the habit of RBN-chewing alone. Involvement of a novel site in the 9p23 region was also observed. Promoter hypermethylation of CDKN2A gene in the samples with the habit of only RBN-chewing alone was significantly higher (p=0.01) than Rb1 gene, also from the samples having the habit of use both RBN and tobacco (p=0.047). CONCLUSIONS The data indicate that the disruption of 9p21 where CDKN2A gene resides, is the most frequent critical genetic event in RBN-associated carcinogenesis. The involvement of 9p23 as well as 13q14.2 could be required in later stages in RBN-mediated carcinogenesis.

Expression of aberrant CD markers in acute leukemia: A study of 100 cases with immunophenotyping by multiparameter flowcytometry

BACKGROUND Acute leukemia is a heterogenous disease having diverse phenotypes. Immunophenotyping by flowcytometry is essential for diagnosis of myeloid and lymphoid subtypes. Aberrant phenotype incidence is controversial and dissimilar results have been reported by different groups. OBJECTIVES Purpose of the study was to determine the incidence of aberrant phenotypes in North East Indian patients with acute leukemia. METHODS We analysed a total of 100 cases (AML = 36, ALL = 61, MPAL = 3) by multiparametric flow cytometry using an acute panel of monoclonal antibodies (MoAbs). The MoAbs were selected to identify differentiation-associated antigens of both myeloid and lymphoid lineages. RESULTS Aberrant phenotypes were found in 21 (58.3%) cases of AML, 36 (59.2%) cases of B-ALL and 6 (66.7%) cases of T-ALL. CD7 was the most frequent lymphoid associated antigen found in 33% of AML cases while CD117 was the myeloid antigen most frequently detected in ALL (54%) cases. Aberrant expression of CD 117 is highly significant by Fischers exact test (P< 0.0001). CONCLUSION We conclude that aberrant phenotypes are present in a great majority of acute leukemia patients of North East India. Future studies will be directed to correlate of these markers with prognosis and therapeutic response.

Prevalence of Major Insect Pests and Predators of Pigeon Pea (Cajanus cajan L.) in Middle Gujarat

A survey was conducted on pigeon pea crop in Panchmahals district of Gujarat. A total of 3 talukas were surveyed to record major pigeon pea pests namely Spiny brown bug, Clavigralla gibbosa Spinola, Tur pod bug, Riptortus pedestris Fb., Green stink bug, Nezara viridula Linn., Aphids, Aphis craccivora Koch, Aphis fabae Scopoli, Cow bugs, Otinotus oneratus W., Oxyrachis tarandus F., Scale insect, Ceroplastodes cajani Maskell, Icerya purchasi Maskell, Mealybugs, Phenacoccus solenopsis Tinsley, Paracoccus marginatus W.& G., Coccidohystrix insolita Green, Leaf hoppers, Empoasca kerri Pruthi, Plume moth, Exelastis atomosa Walshingham, Pod borer, Helicoverpa armigera Hubner, Spotted pod borer, Maruca testulalis Geyer, Hairy caterpillars, Spilosoma obliqua Walker, Leaf webbers/folders, Grapholita critica Meyr., Blue butterfly, Lampides boeticus Linn., Blister beetles, Mylabris pustulata Thumberg, Ash/Grey weevil, Myllocerus undecimpustulatus Faust, Thrips, Megalurothrips usitatus Bagnall, Eriyophid mite, Aceria cajani Channa., Pod fly, Melanogromyza obtusa Malloch, and Termite, Odontotermes spp., Out of these pests, 9 insect species each were observed as regular and sporadic pests whereas other 2 as major and 5 as minor pests respectively. Total number of 25 insect pests species and 8 species of predators were recorded on pigeon pea crop during the period of study. The predators recorded mainly belonged to orders Coleoptera, Hymenoptera, Dictyoptera, Araneida and Neuroptera.