Quantitative evaluation of AgNORs in bone tumours.

AIMS: Primary tumours of bone present a significant diagnostic and therapeutic challenge at times. Silver stained nucleolar organiser regions (AgNORs) have been widely used in a variety of tissues but with a limited study on bone tumours. Our study was aimed at the evaluation of AgNOR count in various neoplastic lesions of bone. METHODS: : One hundred biopsies of bone lesions were included in this study. Five samples of foetal lumbar vertebrae obtained from foetal autopsies were taken as control. The study included 58 males and 42 females with age ranging from 5 to 70 years. Fifty-two cases were malignant while 48 were benign in nature. Silver staining for nuclear organiser regions was performed according to one-step silver staining technique in these cases. NORs seen as black dots were counted in the nuclei of 100 cells. RESULTS: Our study revealed that the mean count was highest in malignant lesions (4.00+/-0.69) compared with benign lesions (2.16+/-0.43) and normal bone (1.32+/-0.14). Statistically, the AgNOR count showed a significant difference (P<0.001) in all these lesions. CONCLUSIONS: The results of the current study revealed that malignant lesions had a greater mean AgNOR count than benign tumours and the normal bone. Thus, quantification of AgNORs strongly correlates with the type as well as aggressiveness of the bone tumour and is diagnostically useful in tumour differentiation.

A 16-year study of HIV seroprevalence and HIV-related diseases in a teaching tertiary care hospital in India.

A total of 183,912 persons were screened from September 1986 to May 2002 in and around Haryana, out of whom 1178 were reactive (0.64%) for anti-HIV antibodies. The overall incidence of HIV-1 in the seropositive patients was 98.5%, HIV-2 alone was 0.8% and 0.7% had a mixed infection with both HIV-1 and 2. High seropositivity (22.8%) was observed among the relatives of these HIV-positive individuals. The heterosexual route (78%) remained the predominant mode of transmission. Among the jail inmates only one individual out of 1306 (1986 to 1993) was found to be positive. Tuberculosis (46.7%) was the most common opportunistic infection in these seropositive patients. Significant titres of Venereal Disease Research Laboratory tests were observed in 8.8% seropositive patients, 7.9% were positive for hepatitis B surface antigen and only two patients were suffering from chancroid. Forty-six of these HIV-seropositive patients had already died. In order to cope with this epidemic, new models of care and cost-effective preventive measures are needed.

Fine needle aspiration cytology in diagnosis of tuberculous lymphadenitis.

Fine needle aspiration cytology and culture were done on 200 patients with clinical diagnosis of tuberculous lymphadenitis. Cytological changes consistent with tuberculosis were seen in 124 and 100 of these showed growth of Mycobacterium tuberculosis. Atypical mycobacteria were not isolated from any of these patients. Of the 100 culture positives, 75 were found to be positive by auramine-rhodamine and Ziehl-Neelsen staining technique. However, with the latter method the diagnosis was missed on first examination of eight smears. With periodic acid-Schiff staining mycobacteria could be visualised in 71 smears.

Resistotyping as a new epidemiological marker for Klebsiella pneumoniae.

Resistogram typing was established with the help of 30 randomly chosen clinical isolates of K. pneumoniae using sodium arsenate, malachite green, boric acid, potassium tellurite, mercuric chloride, antimony potassium tartarate and sodium arsenite. The resistance to these chemicals was designated as A, B, C, D, E, F and G respectively. The technique was then applied to 152 clinical isolates of K. pneumoniae. A total of 35 patterns were obtained. Common patterns were ABEFG and ABFG. There was no clustering of the strains in any of the resistogram patterns as even the commonest pattern had only 10.5 per cent of the strains. When combined with klebocin typing, it provided better discrimination of strains as strains belonging to seven klebocin types could be subdivided into 68 resistogram patterns. The reverse was also possible, i.e., the strains belonging to seven resistogram patterns could be subdivided into 38 klebocin types. The former procedure thus offered better discrimination of the strains.

Subconjunctival infection with Dirofilaria repens.

Cases of zoonotic dirofilariasis infection, caused by Dirofilaria repens, occur widely throughout European, African, Middle Eastern, and Asian countries. The reports of this infection in humans in India are limited, and we herein report the case of a 23-year-old man who presented with subconjunctival swelling in the right eye with no local symptoms. A large nematode was visualized on examination and the intact worm was surgically removed. The parasite was identified as a male D. repens.

Radio-active iodine uptake in vitiligo.

Vitiligo and thyroid disease are commonly associated disorders. Twenty-two clinically euthyroid vitiligo patients were studied for functional assessment of thyroid by radioactive iodine uptake assay. Half of them showed abnormal uptake values at 24 hours. Of these patients, 90% had lower values indicating a tendency towards developing hypothyroid state. Subclinical thyroid dysfunction in vitiligo appears to be an adaptive change.

Sensitivity pattern of Salmonella serotypes in Northern India.

BACKGROUND: Enteric fever continues to be a major public health problem, especially in the developing countries of the tropics. We determined the incidence of Salmonella bloodstream infections and their antimicrobial resistance patterns from May to August in the years 1997-2001 in Haryana, a large state of India. The minimum inhibitory concentration (MIC) was also determined for 60 isolates of S. typhi to various commonly used antimicrobial agents. MATERIAL AND METHODS: Blood cultures of 6,956 patients (PUO/septicemia) were processed by standard procedures and the Salmonella spp. isolates were identified with specific antisera and with standard biochemical tests. Antimicrobial susceptibilities were determined by Stokes disc diffusion method. The MIC of 60 randomly isolated strains of S. typhi was determined by agar dilution method using Mueller Hinton Agar medium. RESULTS: Isolation rates of Salmonella spp. increased in 2000 and 2001. Multidrug resistance (MDR) in S. typhi had increased while in S. paratyphi it had decreased markedly. Ninety per cent chloramphenicol sensitivity was seen in S. typhi by MIC method. There was a decrease in the susceptibility to ciprofloxacin of S. typhi with MIC showing an upward trend. All S. typhi tested were sensitive to third generation cephalosporins and aminoglycosides with MIC well below the breakpoint. DISCUSSION: Our study indicates that MDR in S. typhi is on the rise in our area. There is also re-emergence of chloramphenicol sensitivity. Rising MIC values of ciprofloxacin may lead to prolonged treatment, delayed recovery or pose treatment failure. Thus, sensitivity pattern of causative organism must be sought before instituting appropriate therapy to prevent further emergence of drug resistance.

Actinomycotic cholecystitis.

Actinomycotic cholecystitis is rare. We report a 65-year-old woman who was admitted with repeated attacks of pain in the abdomen and vomiting. Clinical and laboratory findings suggested the diagnosis of cholecystitis with cholelithiasis. At cholecystectomy the gall bladder was inflamed with a small perforation, and contained many calculi. Gram's staining of the gall bladder wall revealed Gram-positive actinomycotic colonies.

Enterobacter bacteremia.

OBJECTIVES: To study the incidence, antimicrobial susceptibility pattern, age and sex distribution, mortality and prognostic factors in cases of Enterobacter bacteremia. METHODS: A total of 18,745 indoor patients with suspected bacteremia were included in study. Enterobacter spp were identified and speciated using standard protocols. Antimicrobial susceptibility testing was done using Stoke's disc diffusion method and minimum inhibitory concentration (MIC) was determined by agar dilution method as recommended by NCCLS. RESULTS: Enterobacter spp ranking sixth was responsible for 4.6% of total group of significant bacteremias. The species most commonly causing bacteremia was E. cloacae (61.8%). The bacteremia was unimicrobial in 85.1% and part of a polymicrobial bacteremia in 14.8%. Portal of entry in decreasing order of frequency were unknown, respiratory tract, urinary tract and surgical wound. The most common clinical finding was fever (97.2%). Of Enterobacter isolates 72.9% were multiresistant to three or more antimicrobial agents. Overall mortality was 51.1%. CONCLUSION: Enterobacter spp are becoming increasingly important nosocomial pathogens. To prevent further emergence of multidrug resistance it may beprudent to avoid third generation cephalosporins and instead combination therapy may be used.

Bacteraemia in children.

OBJECTIVES: (i)To know the etiology of bacteraemia in children, (ii) To learn the antibiotic sensitivity pattern of the isolates. METHOD: Over the period of thirteen months 4,368 blood samples (for blood culture) were collected from children in the age group of 0 day-14 years, suspected of having fever and sepsis. Blood samples were collected for blood culture from each case. Organisms were isolated and identified by conventional methods. Antibiotic susceptibility for each isolate was determined by using modified Stokes method. RESULT: 1,001 cases (22.9%) were culture positive. Incidence of bacteraemia in neonates was 521(33.94%). Gram negative organisms were the most predominant isolates (88.8%). Commonest was Klebsiella 471 (47.1%) followed by Salmonella sp. 162 (16.2%) and Pseudomonas 80 (8%) whereas in gram positive, Staphylococcus aureus 76 (7.6%) was the most common. Maximum sensitivity was seen by sulbactum/cefaperazone combination-969 (98.2%) by all isolates. Linezolid 97 (99.0%) was the most sensitive drug for gram positive isolates. CONCLUSION: Gram negative multidrug resistant organisms were the main cause of septicemia in all the age groups. Therefore great caution is required in selection of antibiotic therapy.

A study of bacterial pathogens in acute pelvic inflammatory disease.

The study was carried out on two groups--the study group consisting of 50 women with acute pelvic inflammatory disease and the control group consisting of 20 women admitted for tubal ligation. Pouch of Douglas aspirates were collected under all aseptic precautions with 18 gauge medicut needle. The samples were transferred to thioglycollate broth. A direct Gram staining was done followed by plating on media for aerobic and anaerobic incubation. All cultures were sterile in the control group. There were both monomicrobial (56.7%) and polymicrobial (43.2%) isolates in the study group. The predominant aerobic isolates were coagulase negative Staphylococci, E. coli and Staphylococcus aureus. Amongst anaerobic isolates gram negative bacilli were the most common at 46.6% followed by gram positive cocci at 40%.

Sensitivity to the bactericidal effect of normal human serum on Klebsiella strains from different sources.

A total 270 Klebsiella strains isolated from clinical infections, faeces of healthy food handlers and environmental sources were studied. Of the strains tested 177 (65.6%) were serum sensitivity, 71 (26.3%) were serum resistant and the remaining 22 (8.1%) showed intermediate sensitivity to pooled normal human serum (PNHS). Faecal coliform (FC) reaction was positive in 59 per cent and 6 per cent for K. pneumoniae and K. oxytoca respectively.

Association of R plasmids of Klebsiella pneumoniae with resistance to heavy metals, klebocin and beta lactamase production and lactose fermentation.

One hundred and fifty clinical isolates of Klebsiella pneumoniae were studied for beta lactamase production, Klebocin production and resistance to antibiotics and metals. 97.3 and 82.7 per cent strains produced beta-lactamase and klebocin respectively. Resistance to arsenite, antimony, tellurite, mercury and silver was observed in 91,66,6, 47.7, 44.8 and 33.3 per cent respectively and to ampicillin, tetracycline and nalidixic acid in 93.3,60, 41.3, 34.4, 28.2, and 1.3 per cent respectively. Forty eight antibiotic and metal resistant strains were then conjugated with Escherichia coli 711 F-Lac-Nxr. Transferable drug resistance, metal ion resistance, beta lactamase production, klebocin production and lactose fermentation was seen in 30, 21, 10, 11 and 5 isolates respectively.

T and B lymphocytes in patients with enteric fever.

Estimation of T and B lymphocytes was done in 50 patients of enteric fever, 50 duration matched non enteric fever patients and 50 normal healthy individuals. The difference in both early and late rosette forming T lymphocytes was found to be statistically significant in enteric versus non-enteric patients. Significant difference was also observed in enteric versus normal individuals in case of late rosette forming T lymphocytes.

Evaluation of germ tube test in various media.

One of the most valuable tests for the rapid presumptive identification of Candida albicans is germ tube production. Previously researchers have concluded superior results with serum in germ tube test. However, handling of pooled human serum has danger of acquisition of hepatitis and HIV infections. Therefore, in the present study we compared various media--pooled human serum, sterile horse serum, peptone water and tryptic soy broth. It was observed that human serum remains to be the best with 100% positivity for identification of C. albicans followed by horse serum (76.3%), peptone water (61.8%) and tryptic soy broth (61.8%).

Sex chromatin in female breast tumour.

The incidence of sex chromatin in tumour cell nuclei was estimated in twenty five cases each of carcinoma and fibroadenoma breast using haematoxylin and eosin, thionine, aceto-orcein and Papanicolaou's stains on imprint-smears. The sex chromatin incidence in fibroadenoma was found similar to that of normal breast tissue but the incidence markedly decreased in carcinoma breast. Undifferentiated tumours with histologic grade III, nuclear grade I, no lymphocytic infiltrate and involvement of overlying skin, nipple and areola were more often associated with low sex chromatin incidence. No relationship was observed between sex chromatin incidence and tumour size or axillary lymph node metastasis.

Carcinoembryonic antigen and human chorionic gonadotrophin in breast carcinoma and prostatic specific acid phosphatase in prostate carcinoma.

Tumour markers viz carcinoembryonic antigen (CEA), human chorionic gonadotrophin (HCG) in 30 cases of carcinoma breast and prostatic specific acid phosphatase (PSAP) in 30 cases of carcinoma prostate were studied by peroxidase antiperoxidase technique in paraffin blocks of tissue. Twenty three (76.7%) and 20 (66.7%) cases were positive for CEA and HCG respectively. No correlation was observed between CEA and HCG status, and histological differentiation of the tumours. All the 29 cases (100%) of adenocarcinoma prostate were PSAP positive while a single case, negative for PSAP, was of transitional cell carcinoma.

Incidence of mycoses in bronchopulmonary disorders.

A total of 274 samples were collected--180 sputum samples, 82 bronchial secretions and 12 pleural aspirates. Main fungus was Candida albicans from sputum (45.5 percent), from bronchial secretions (14.6 percent). Rest were Aspergillus, Alternaria and Helminthosporium. All the pleural aspirates were negative for fungus.

Detection of anti-Proteus antibodies in sera of patients with rheumatoid arthritis.

This study was carried out to determine increased level of antibodies to certain bacteria in rheumatoid arthritis (RA) patients. Fifty RA patients and 25 each of those suffering from osteoarthritis (OA) and healthy controls were tested for antibodies to somatic ('O) and flagellar ('H') antigens of Proteus mirabilis, Escherichia coli, and for antibodies to 'O' antigen of Klebsiella pneumoniae by standard tube agglutination method. Anti-Proteus antibodies against 'O' and 'H' antigens could be demonstrated in 70% and 64% RA cases respectively, and were statistically significant. Antibodies could be demonstrated against E. coli 'O' and 'H' antigens in 86% and 92% RA cases respectively, and were statistically significant. Against K. pneumoniae 'O' antigen, no antibody could be demonstrated in any of RA and OA cases, and healthy controls. Of the 50 RA patients, 30 had raised CRP levels. Antibodies against P. mirabilis and E. coli antigens appeared to be independent from CRP. Thus a specific elevation in the immune response to P. mirabilis and E. coli has been demonstrated in patients with RA from India.

Prevalence of Trichomoniasis, Vaginal Candidiasis, Genital Herpes, Chlamydiasis, and Actinomycosis among Urban and Rural Women of Haryana, India.

Despite being curable reproductive tract infections (RTIs) including sexually transmitted infections continue to be a major health problem in developing countries. The present study was undertaken to know the prevalence of trichomoniasis, vaginal candidiasis, genital herpes, chlamydiasis, and actinomycosis in rural and urban women of Haryana by using wet mount, PAP smear, and fluorescent microscopic examination. Patients suspected of suffering from bacterial vaginosis were given treatment and were not included in the study. RTIs were seen in 16.6% of urban and 28.7% of rural women. The highest prevalence seen was that of trichomoniasis in both rural (24.2%) and urban (15.7%) women, followed by candidiasis (4.2% in rural and 0.6% in urban women), genital herpes (0.3% in rural and 0.2% in urban women), and chlamydiasis (0.02% in rural and 0.05% in urban women). Pelvic actinomycosis was seen in 1.4% of rural and 0.06% of urban women using intrauterine contraceptive devices. Mixed infection of Trichomonas vaginalis with Candida spp. was seen in 6.3% of rural women only. It is desirable to have a baseline profile of the prevalence of various agents causing RTIs in a particular geographic area and population which will help in better syndromic management of the patients.