Human papillomaviruses other than 16, 18 and 45 are the major high risk hpv genotypes amongst women with abnormal cervical smear cytology residing in kuwait: implications for future vaccination strategies

Objective:The study was undertaken to determine the prevalence of different high risk HPV [HR-HPV] genotypes amongst women residing in Kuwait with epithelial abnormalities in cervical smears and to detect any difference in the distribution of these genotypes between Kuwaiti and Non-Kuwaiti women or between the cytological diagnosis groups

Materials and methods:Thinprep specimens from women with epithelial abnormalities on cervical smears were subjected to Aptima HR-HPV assay and those found to be HR-HPV positive were genotyped using the Aptima HPV genotyping assay into three groups [i] HPV 16, [ii] HPV 18/45 and [iii] others

Results: HR-HPV was found in 229 [30.57%] of the 749 cases with epithelial abnormalities. Of the 116 cases subjected to further genotyping, the non-16, 18, 45 genotype accounted for the most prevalent group accounting for 62.93% of the cases, followed by HR-HPV 16 [31.04%] cases and HPV 18/45 [6.03%] of cases. There was no significant difference between Kuwaiti and non-Kuwaiti women nor was any difference noted between the various cytological diagnosis group

Conclusion:Most HR-HPV infections amongst women residing in Kuwait with epithelial abnormalities are due to HPV types other than the 16, 18 and 45. As HPVs 16 and 18 are considered to be the most prevalent HR-HPV genotypes worldwide, causing invasive cancer, the findings of this study is significant from an epidemiological standpoint. It should also be taken into account before undertaking any HPV vaccination program since the available HPV vaccines protect against HR-HPVs 16 and 18 only

Comparison of cefoxitin disc diffusion test, oxacillin screen agar, and PCR for mecA gene for detection of MRSA.

Cefoxitin is a potent inducer of the mecA regulatory system. It is being recommended for detection of methicillin resistance in Staphylococcus aureus (MRSA) when using disk diffusion testing. The aim of our study was to evaluate the efficacy of cefoxitin disc diffusion test to characterize MRSA and compare it with oxacillin agar screening and detection of mecA gene by PCR. MATERIALS AND METHODS: Fifty strains of S. aureus isolated from clinical samples were used in the study. Routine antibiotic susceptibility testing was performed including oxacillin disk. Oxacillin screen agar plates with 4% NaCl and 6 microg/ml of oxacillin were inoculated and interpreted as per standard guidelines. Cefoxitin disc diffusion test was performed using 30 microg disc and zone sizes were measured. PCR for amplification of the mecA gene was performed. RESULTS: Out of the 50 isolates, 28 were found to be methicillin resistant by oxacillin disc diffusion test, 30 were resistant by oxacillin screen agar method, and 32 were resistant with cefoxitin disc diffusion. For these 32 isolates mecA gene was positive. CONCLUSION: Results of cefoxitin disc diffusion test is in concordance with the PCR for mecA gene. Thus, the test can be an alternative to PCR for detection of MRSA in resource constraint settings.

Divergent strains of human immunodeficiency virus type 1 circulating in India, subtyped by heteroduplex mobility assay.

Genomic variations in HIV-1 represent a major problem in understanding disease progression, studying drug resistance and developing effective vaccines. Heteroduplex Mobility Assay (HMA) was used for analyzing HIV-1 subtypes resulting from genetic similarity or divergence of C2 -V3 -V5 region of envelope gene between HIV-1 strains obtained from clinical samples in a tertiary care center at Pune. DNA from the PBMCs of infected individuals was amplified by nested PCR. Heteroduplexes were then formed by denaturing DNA from the unknowns with DNA from the reference strains. The results were analyzed by polyacrylamide gel electrophoresis. Out of 177 samples analyzed, 170 were of subtype C (96%). Four samples were found to be of subtype B (2.2%); in three samples, no definitive assignment of subtype was possible by HMA and these perhaps could be circulating recombinant forms (CRFs) of HIV-1. These findings may have significant implications toward development of a candidate vaccine for India.

Prevalence of extended-spectrum beta-lactamases among Escherichia coli and Klebsiella pneumoniae isolates in a tertiary care hospital.

Extended-spectrum beta-lactamases (ESBLs) continue to be a major problem in clinical setups the world over, conferring resistance to the expanded-spectrum cephalosporins. Knowledge about their prevalence is essential to guide towards appropriate antibiotic treatment. The aim of the present study is to determine the prevalence of ESBL producers among Escherichia coli and Klebsiella pneumoniae isolates at a tertiary care institution. A total of 357 clinical isolates comprising E. coli (n = 181) and K. pneumoniae (n = 176) were recovered from various clinical samples over a period of six months from April to September 2006. Antibiogram profile of these isolates was determined to commonly used antibiotics, along with screening for ESBL production by the screening test as recommended by the Clinical Laboratory Standards Institute (CLSI). Isolates which showed positive results with screening test were shortlisted for confirmatory tests of ESBL production. Two tests were performed: phenotypic confirmatory test with combination disk and the minimum inhibitory concentration (MIC) reduction test. Out of 357 isolates of E. coli and K. pneumoniae screened for ESBL production, 120 were found to be potential ESBL producers. Of these, 80 isolates were confirmed to be ESBL producers. Thus the prevalence of ESBL-producing isolates of E. coli and K. pneumoniae was found to be 22% (80 out of 357). This was significantly lower than the data available from other hospitals.

Tuberculous brain abscess in a patient with HIV infection.

Tuberculous Brain Abscess (TBA) is a rare manifestation of CNS tuberculosis. Only a few cases have been reported in literature. A twenty six year old male presented with high grade fever, throbbing headache and altered sensorium. Examination revealed neck stiffness and papilloedema. His chest X-ray showed evidence of healed pulmonary tuberculosis. MRI Brain showed a well circumscribed hyper intense lesion in the left parietal region with perilesional edema and mass effects. Stereotactic aspiration of the abscess yielded frank creamy pus. PCR for Mycobacterium tuberculosis MPB 64 was positive which confirmed the lesion to be of tuberculous etiology. Patient responded well to four-drug regimen of antitubercular treatment.

Cytodiagnosis of anaplastic astrocytoma with metastasis to the cerebrospinal fluid in a neonate--a case report.

A case of left cerebral hemispheric anaplastic astrocytoma with metastasis to the cerebrospinal fluid in a 50-day-old male child diagnosed on cytology and later confirmed by histological examination is reported.

Cytologic diagnosis of Enterobius vermicularis eggs in an enterocutaneous fistula.

A middle-aged female underwent a laparotomy for suspected ovarian cancer and developed a discharging sinus in the right iliac fossa. Smears of the discharge showed helminthic eggs which were characterised as those of Enterobius Vermicularis. The possibility of an enterocutaneous fistula was suggested which was subsequently confirmed during a relook laparotomy.

Nucleolar organiser regions and morphologic subtypes of squamous cell carcinoma of cervix.

Nucleolar organiser regions (NOR) are chromosomal segments coding for ribosomal RNA. The number of nucleolar organiser regions correlates with cellular proliferation and has a diagnostic and prognostic value in neoplastic diseases. The aim of this study was to study the morphologic subtypes of squamous cell carcinoma of the uterine cervix in biopsy and hysterectomy specimens and to determine the AgNOR counts in these morphologic subtypes. One hundred and seventy-nine cases of squamous cell carcinoma of the uterine cervix were classified into three morphological subtypes namely large cell non keratinising squamous cell carcinoma (113), keratinising squamous cell carcinoma (64) and small cell carcinoma (2). A correlation of histologic typing on biopsy and hysterectomy specimens was done in 142 cases with a complete correlation in 90.5%. Fifty-eight of these tumours were stained with silver nitrate to demonstrate Nucleolar organiser regions (AgNORS). The mean AgNOR count was higher (3.66) in keratinising squamous cell carcinoma, which is known to have a worse prognosis as compared to 3.04 in large cell non keratinising carcinoma and (3.45) in the two cases of small cell carcinoma of the uterine cervix.

Cytology of cystitis cystica glandularis of the bladder--a case report.

The cytologic findings of cystitis cystica glandularis--a form of proliferative cystitis which can be mistaken for a tumor clinically and radiologically have hardly been recorded. We describe a case of cystitis cystica where the bladder washings showed a spectrum of findings which were later corroborated on biopsy.

Paragangliomas--a diagnostic dilemma on fine needle aspirates.

Paragangliomas are uncommon tumours arising from the neural crest and are widely distributed throughout the body. Because of the morphologic diversity of these tumours fine needle aspirate (FNA) diagnosis is not very accurate in identifying these neoplasms. Review of FNA from seven paragangliomas (three extra-adrenal, three carotid body and one multicentric in location) was conducted to find any diagnostic criteria. On initial morphologic diagnosis five of the seven paragangliomas were considered malignant (four undifferentiated and one adenocarcinoma). Review of these cases in our series made us wiser. Pleomorphic nuclei, prominent nucleoli, clumped chromatin with finely granular cytoplasm and occasional intranuclear cytoplasmic inclusions were characteristically seen and these were probably the cause for misdiagnosis. This lesion must be entertained in an aspirate from the neck or retroperitoneum which contains cells showing marked nuclear pleomorphism with naked nuclei and have a tendency to spindle or from acini. To confirm these cases argyrophilic granules were demonstrated in two of the four cases studied. However, neuron specific enolase and calcitonin were found to be non-contributory. Ultrastructural examination was helpful and done in one case showed abundant neurosecretory granules.

Fine needle aspiration cytology, sonography and radionuclide scanning in solitary thyroid nodule.

One hundred and ninety three consecutive patients with solitary thyroid nodule (STN), with a mean age of 36.0 +/- 12.8 years and male to female ratio of 5.6:1, were studied. Ninety five percent of patients came from iodine deficient regions. Seventy two percent presented for local neck swelling, 12.4% for hyperthyroid state and in 7.7% STN was discovered incidentally. Scintigraphically, 77.7% of nodules were cold, 12.4% hot and 8.5% warm. Sonographic evaluation did not reveal any characteristic echotexture diagnostic of malignancy, but detected clinically nonpalpable accessory nodules in 20.6% of patients. Fine needle aspiration cytology was positive for malignancy in 6.2% of patients. Features of follicular and Hurthle cell neoplasm were seen in 12.9% of aspirates. Eighty eight (45.6%) STN were resected surgically. Histologically, there was one false positive and one false negative aspirate and a case of parathyroid adenoma on aspiration proved to be parathyroid carcinoma on histology.

Extramedullary involvement in acute lymphoblastic leukemia and its relation to therapy.

Extramedullary involvement (EMI) of various body sites was studied by doing CSF cytology anu FNAC of testes and lymph nodes in 82 children with acute lymphoblastic leukemia (ALL). Forty-five patients had received no therapy (group I) and 37 had been diagnosed and treated earlier and were now in remission (36) or relapse (group II). EMI was documented in 48.9% and 32.4% of patients of group I and II respectively. EMI was seen more frequently in patients with FAB-L 2 subtype (66.6%) as compared to FAB-L1 subtype (34.6%) in group I. Lymph nodes and testes were commonly involved in group I and II patients respectively. Successful remission after complete chemotherapy was achieved in 82.5% of group I patients. However, successful remission rates were 95.2% in patients without any evidence of EMI versus 68.4% in those with EMI. These differences were also observed when response to therapy was analysed separately for patients with FAB-L1 and FAB-L2 subtypes.

Diagnostic accuracy of immediate interpretation of fine needle aspirates.

A total of 197 fine needle aspirates (FNA) from various body sites was subjected to immediate interpretation to determine its diagnostic accuracy. For the entire series, the immediate interpretation revealed a sensitivity of 97 per cent, specificity of 96 per cent, a positive predictive value (PV) of 92 per cent, a negative predictive value of 98 per cent and an efficiency of the test of 96 per cent. The false positive rate was 1.5 per cent and false negative rate 0.5 per cent. The diagnostic accuracy of immediate interpretation was comparable for aspirates from different sites. Our results indicate that immediate interpretation of FNA is ideal for determining the adequacy of material and providing a rapid basic diagnosis. This technique could offer useful substitute to frozen sections particularly when facilities for the latter are not available.

Pulmonary blastoma.

Pulmonary blastoma is a rare tumour of the lung. We report three cases of this tumour. In all three cases, a possibility of blastoma was considered on aspiration cytology of the tumours. The relevant literature including its pathogenesis is reviewed.

Monoclonal antibody MAb C-50 in buccal mucosa cancer cells.

Staining patterns of cancerous and normal tissues from 20 cases of proven buccal cancer and ten control individuals respectively to monoclonal antibody MAb C-50 were studied. Nine of 20 patients and 4 of 10 control individuals showed positive immunofluorescence respectively. There was no correlation between clinical stage of disease and MAb C-50 staining nor any significant difference between the ulcerative or exophytic growth groups. One section with T4 lesion of buccal cancer with normal salivary tissue showed positive immunofluorescence of normal salivary cells. In our study the presence of receptor for MAb C-50 on buccal mucosa cancer cells was found to be statistically insignificant.

The role of fine needle aspiration cytology of breast lumps in the management of patients.

The accuracy of diagnosis with fine needle aspiration cytology (FNAC) was assessed in 3230 patients with breast lumps between 1978 and 1986. The results for sensitivity, specificity, positive predictive value and negative predictive value were 97.6, 99.4, 99.5 and 97.2 per cent respectively; the overall accuracy was 98.4 per cent. Results reveal that a positive cytologic report by an experienced cytopathologist need not wait for histologic confirmation and the patient can be given definitive treatment based on the cytology report.