Microbial colonization of Provox voice prosthesis in the Indian scenario.

INTRODUCTION: Tracheoesophageal speech using the voice prosthesis is considered to be the "gold standard" with success rates as high as 90%. Despite significant developments, majority eventually develop dysfunction due to microbial deterioration. We did a pilot study of 58 laryngectomy patients who developed prosthesis dysfunction. MATERIALS AND METHODS: A total of 58 laryngectomy patients who had their dysfunctional prosthesis removed were included in this study. Dysfunctional prostheses were removed and examined. Esophageal and tracheal flanges were examined separately. After obtaining pure fungal and bacterial cultures, the yeast strains were identified. Bacteria were identified with the light microscope and gram staining. We analyzed prosthesis lifespan and probable factors affecting it. RESULTS: Central leak was found in 43% cases while in 57% peri-prosthetic leakage was the most common reason for prosthesis replacement. Microbial analysis revealed a combination of yeast and bacteria in approximately 55% culture samples. Out of these, almost 90% had the presence of single yeast species with bacteria. Pure fungal culture was identified in rest of the 45% cultures while none detected pure bacterial forms. Candida tropicalis was the solitary yeast in 81% while Candida albicans was seen in 10% as the solitary yeast. Bacterial isolates revealed Klebsiella pneumonia in 19%, Escherichia coli in 8% while Staphylococcus aureus was grown in 1% cultures. The consumption of curd (P = 0.036, 95% confidence intervals [CI]: 2.292-64.285) to have a significant correlation of the mean prosthesis lifespan. Consumption of curd (P = 0.001, 95% CI: 0.564-2.008) and history of prior radiotherapy (P = 0.015, 95% CI: 0.104-0.909) had a significant bearing on the Provox prosthesis lifespan. CONCLUSIONS: Candida is the most common organism grown on voice prosthesis in Indian scenario. Consumption of curd and history of prior radiotherapy significantly affect Provox prosthesis lifespan.

Alarming prevalence of community-acquired multidrug-resistant organisms colonization in children with cancer and implications for therapy: A prospective study.

BACKGROUND: Infection or colonization with multidrug-resistant organisms (MDRO) is associated with high mortality and morbidity. Knowledge of MDRO colonization may help in planning empirical antibiotic approach in neutropenic patients, which is known to improve patient outcomes. While routine cultures are positive and may help direct antibiotic therapy in only up to 15% neutropenic patients, surveillance cultures are positive in more than 90% of cancer patients. AIMS: To assess the rate of MDRO carrier status at presentation and rate of conversion to MDRO during the treatment. MATERIALS AND METHODS: Rectal swabs of all the outpatients presenting to pediatric oncology unit were sent within 7 days from date of registration from January 2014 to December 2014. Furthermore, stool cultures/rectal swabs of all patients who got directly admitted to the pediatric ward at presentation were sent within 24 h. Repeat rectal swabs were sent again for patients from this cohort when they got readmitted to the ward at least 15 days after last discharge or when clinically indicated. RESULTS: Baseline surveillance rectal swabs were sent for 618 patients, which included 528 children with hematological malignancies and 90 children with solid tumors. Forty-five (7.3%) showed no growth. Of the remaining 573, 197 (34.4%) patients were colonized by two organisms and 30 (5.2%) by three organisms. Three hundred and thirty-four (58.4%) showed extended spectrum beta-lactamase (ESBL) Enterobacteriaceae, of which 165 (49.5%) were ESBL sensitive to beta-lactam with beta-lactamase inhibitors combinations and 169 (50.5%) were resistant to combinations. One hundred and sixteen (20.2%) were carbapenem-resistant Enterobacteriaceae (CRE) and 65 (11.4%) had vancomycin-resistant enterococci in baseline cultures. Only 63 (21%) patients were colonized by a sensitive organism in their baseline surveillance cultures. Morbidity (Intensive Care Unit stay) and mortality was higher in patients colonized by MDR organisms. There was a significant correlation between the place of residence and CRE colonization status with the highest rate (60%) of CRE colonization observed in children from East India. The repeat cultures showed the further conversion of sensitive isolates to MDRO in 80% of these children, of which 40% each converted from non-ESBL and non-CRE to ESBL and CRE, respectively. CONCLUSION: This is the first study illustrating the alarming high prevalence of community-acquired MDRO colonization, especially CRE, which has grave implications for therapy for children with cancer potentially compromising delivery of aggressive chemotherapy and affecting outcomes. This incidence further increases during the course of treatment. Knowing the baseline colonization also guides us for the planning of chemotherapy as well as antibiotic approach and infection control strategies. Local antibiotics stewardship including education of the healthcare workers as well as national level interventions to prevent antibiotic misuse in the community is critical to minimize this problem.

Epidemiology of blood stream infections in pediatric patients at a Tertiary Care Cancer Centre.

BACKGROUND: Blood stream infections (BSI) are among the most common causes of preventable deaths in children with cancer in a developing country. Knowledge of its etiology as well as antibiotic sensitivity is essential not only for planning antimicrobial policy, but also the larger infection prevention and control measures. AIMS: To describe the etiology and sensitivity of BSI in the pediatric oncology unit at a tertiary cancer center. MATERIALS AND METHODS: All the samples representative of BSI sent from pediatric oncology unit during the period of January to December, 2013 were included in the study, and analyzed for microbiological spectrum with their antibiotic sensitivity. RESULTS: A total of 4198 samples were representative of BSI. The overall cultures positivity rate was 6.97% with higher positivity rate (10.28%) from central lines. Of the positive cultures, 208 (70.9%) were Gram-negative bacilli (GNB), 71 (24.2%) were Gram-positive organisms, and 14 (4.7%) were Candida species. Lactose fermenting Enterobacteriaceae i.e., Escherichia coli (28.4%), Klebsiella pneumoniae (22.1%), and Enterobacter (4.8%) accounted for 55.3% of all GNB. Pseudomonas accounted for 53 (25.5%) and Acinetobacter 19 (9.1%) of GNB. Among Gram-positive isolates, staphylococci were the most frequent (47.8%), followed by Streptococcus pneumoniae 17 (23.9%), beta-hemolytic streptococci 11 (15.5%), and enterococci 9 (12.68%). Of GNB, 45.7% were pan-sensitive, 24% extended spectrum beta-lactamase (ESBL) producers, 27% were resistant to carbapenems, and 3.4% resistant to colistin. Pseudomonas was most sensitive, and Klebsiella was least sensitive of GNB. Of the staphylococcal isolates, 41.67% were methicillin-resistant Staphylococcus aureus (MRSA) and 10% of Coagulase Negative Stapylococci (CONS) were methicillin. CONCLUSION: A high degree of ESBL producers and carbapenem-resistant Enterobacteriaceae is concerning; with emerging resistance to colistin, raising the fear of a return to the preantibiotic era. An urgent intervention including creating awareness and establishment of robust infection control and antibiotic stewardship program is the most important need of the hour.

Frequency of bacterial isolates and pattern of antimicrobial resistance in patients with hematological malignancies: A snapshot from tertiary cancer center.

BACKGROUND: Infections are the most important cause of mortality in patients with high-risk febrile neutropenia. Emergence of multi-drug resistant organisms (MDROs) has become a major challenge for hemato-oncologists. Knowledge of the prevalent organisms and their antimicrobial sensitivity can help deciding the empirical therapy at individual centers and allows timely measures to reduce the risk of antimicrobial resistance. AIMS: To evaluate the frequency of bacterial isolates from all the samples and the pattern of bacterial bloodstream infections and incidence of MDROs. SETTINGS AND DESIGN: This is a retrospective analysis from a tertiary care cancer center. MATERIALS AND METHODS: From January to June 2014 information on all the samples received in Department of Microbiology was collected retrospectively. The data from samples collected from patients with hematological cancers were analyzed for types of bacterial isolates and antimicrobial sensitivity. RESULTS: A total of 739 isolates were identified with 67.9% of isolates being Gram-negative. The predominant Gram-negative organisms were Escherichia coli, Psuedomonas spp. and Klebsiella spp. Among the bacterial bloodstream infections, 66% were Gram-negative isolates. MDROs constituted 22% of all isolates in blood cultures. Incidence of resistant Gram-positive organisms was low in the present dataset (methicillin resistant Staphylococcus aureus and vancomycin-resistant enterococci-1.3%). CONCLUSIONS: The analysis reconfirms the Gram-negative organisms as the predominant pathogens in bacteremia seen in patients with hematological cancers. The high frequency of multi-drug resistance in the dataset calls for the need of emergency measures to curtail further development and propagation of resistant organisms.

Pattern of infection, therapy, outcome and risk stratification of patients with febrile neutropenia in a tertiary care oncology hospital in India.

CONTEXT: Indian febrile neutropenia (FN) data are limited, especially in adult solid tumor patients. AIMS: The aim was to study patterns of presentation, source of infection, management and outcome and to evaluate the factors which may correlate with outcome. MATERIALS AND METHODS: A retrospective analysis of prospective data of FN patients at a tertiary care oncology teaching hospital in India between 2007 and 2012. A standardized form was filled for each patient. Patient management was at the discretion of the treating physician. Multinational Association for Supportive Care in Cancer (MASCC) score was retrospectively calculated. Failure of therapy was defined as death, organ failure, shifting from outpatient to inpatient or requirement of intensive care support. SPSS version 16 was used for analysis. RESULTS: A total of 388 FN episodes were included: 256 in hematolymphoid and 132 in solid tumor patients. 156 episodes were high-risk by MASCC score. Focus of infection was clinical in 45% and radiologic in 16%. Blood cultures were positive in 18% cases, most commonly Gram-negative organisms (72%). 93% patients were treated with an antibiotic combination of third-generation cephalosporin/beta-lactamase inhibitor, with aminoglycoside or fluoroquinolone. Antibiotic sensitivity to ceftriaxone was low at 38% while sensitivity to cefoperazone/sulbactam and piperacillin/tazobactam ranged between 50% and 55% and for carbapenems 75%. Failure of therapy occurred in 156 episodes, most commonly due to the need for second line antibiotics. Mortality was 5.5%. On univariate analysis, MASCC score, age, type of malignancy, prophylactic growth factors, presence of focus of infection, hemoglobin and nadir platelet count correlated with FN complications. CONCLUSION: Gram-negative bacteremia continues to be the predominant cause of FN in our setup.

Comparison of isolates and antibiotic sensitivity pattern in pediatric and adult cancer patients; is it different?

BACKGROUND: Infection is a common cause of mortality and morbidity in cancer patients. Organisms are becoming resistant to antibiotics; age appears to be one of the factors responsible. We analyzed common organisms and their antibiotic sensitivity pattern in the correlation with age. METHODS: This is a single institutional, retrospective analysis of all culture positive adult and pediatric cancer patients from January 2007 to December 2007. For statistical analysis, Chi-square test for trend was used and P values were obtained. Of 1251 isolates, 262 were from children <12 years of age and 989 were from adolescents and adults (>12 years of age). Gram-negative organisms were predominant (64.95) while Gram-positive constituted 35.09% of isolates. RESULTS: The most common source in all age groups was peripheral-blood, accounting to 47.8% of all samples. The most common organisms in adults were Pseudomonas aeruginosa (15.3%) while in children it was coagulase negative Staphylococcus aureus (19.8%). Antibiotic sensitivity was different in both groups. In pediatric group higher sensitivity was seen for Cefoparazone-sulbactum, Cefipime, Amikacin, and Tobramycin. No resistance was found for Linezolid. CONCLUSIONS: The isolates in both children and adults were predominantly Gram-negative though children had proportionately higher Gram-positive organisms. High-dose cytarabine use, cotrimoxazole prophylaxis, and frequent use of central lines in children especially in hematological malignancies could explain this observation. Children harbor less antibiotic resistance than adults; Uncontrolled, cumulative exposure to antibiotics in our community with increasing age, age-related immune factors and variable bacterial flora in different wards might explain the higher antibiotic resistance in adults. Thus age is an important factor to be considered while deciding empirical antibiotic therapy.

Detection of bacterial growth in blood components using oxygen consumption as a surrogate marker in a tertiary oncology setup.

Microbiological contamination of blood and blood products is a well-recognised transfusion risk. This study was performed in the blood bank of our oncology centre, with an objective to detect bacterial contamination in our blood products using oxygen consumption as a surrogate marker [Pall Enhanced Bacterial Detection System (eBDS)]. Results revealed that the percentages of failed units were 1.16% for random donor platelets (RDP), 0.81% for single donor platelets (SDP) and 2.94% for packed red blood cells (PRBCs), of which one RDP and one SDP grew coagulase-negative staphylococcus, while one PRBC culture grew Gram-positive bacilli.

The effect of age on the bacteria isolated and the antibiotic-sensitivity pattern in infections among cancer patients.

BACKGROUND: Patients with cancer are predisposed to infections. Antimicrobial patterns and antibiotic sensitivity change with increasing age, making choice of empirical therapy more complicated. MATERIALS AND METHODS: This single-center study aims to try and assess the influence of age on microbiology and antibiotic sensitivity of organisms causing infection in patients with malignant disease. RESULTS: The five most common bacterial pathogens isolated were Pseudomonas sp (245, 26.2%) > Enterocococcus sp (109, 11.66%) > Staphylococcus aureus (107, 11.44%) > Escherichia coli (106, 11.34%) > Klebsiella sp (99, 10.59%). There was no significant change in the distribution of Gram-positive and Gram-negative bacteria with age. However, there was an increase in the occurrence of the Enterobacteriacea group and a decrease in infections caused by nonlactose fermenters with increasing age. The ESBL production increased from 10.52% (12-19 years) to 24.88% (> 50 years) as did oxacillin resistance (from 14.3% to 28.1%) among S. aureus isolates. The activity of most antimicrobial agents decreased with increasing age. The decreasing trend of activity was statistically significant for meropenam (73.3-41.2%) against Pseudomonas sp. and for the activity of the aminoglycosides for Acinetobacter sp (61.1-17.4% for amikacin). CONCLUSIONS: This suggests that empirical antibiotic therapy needs to be changed on the basis of the age of the patient. It also appears that combination therapy is essential for the empirical treatment of infections in elderly patients with cancer.

Decontamination of laryngoscope blades: is our practice adequate?

BACKGROUND: The laryngoscope has been identified as a potential source of cross-infection, because of blood and bacterial contamination. In India, there are no guidelines for cleaning and disinfection of anesthesia-related equipment. Practices for decontamination of laryngoscopes vary widely and in most healthcare institutes, laryngoscope blades are re-used after cleaning with tap-water. MATERIALS AND METHODS: We prospectively compared two techniques for decontamination of laryngoscope blades - a) washing with tap-water and b) washing with tap-water followed by disinfection by immersing in 5% v/v (volume/volume, 1:20 dilution) aldehyde-free biguanide agent for 10 min. We calculated the cost-effectiveness of using 5% v/v aldehyde-free biguanide agent for disinfection of laryngoscopes. We also conducted a survey to assess the decontamination practices in other Indian hospitals. RESULTS: Overall bacterial growth was 58% (29 out of 50 blades) after tap-water cleaning (of which 60% were pathogenic organisms) versus 3.4% (one out of 29 blades) after tap-water cleaning followed by immersion in disinfectant (all of which were commensals). The cost of disinfection with biguanide was Indian Rupees 1.13 (20 US cents) per laryngoscope. Most hospitals in India do not have guidelines regarding laryngoscope decontamination between uses, and cleaning with tap water is a commonly used method. CONCLUSION: Cleaning of laryngoscope blades with tap-water is a commonly used but inadequate method for decontamination. Washing with tap-water followed by disinfection with 5% v/v aldehyde-free biguanide for at least 10 min is an effective and inexpensive alternative. National guidelines for the decontamination of anesthesia equipment are necessary.

Blood stream infections in cancer patients: a single center experience of isolates and sensitivity pattern.

BACKGROUND: Up to 10% of patients who develop a nosocomial blood stream infection (BSI) in the hospital have an underlying malignancy. The treatment of infections in patients with malignancy often relies on the use of established guidelines along with the consideration of the local microbiology and antibiotic sensitivity patterns of possible etiologic agents. AIMS: This study attempts to identify the likely etiologic agents and the antibiotic sensitivity profile of BSIs in cancer patients. SETTINGS AND DESIGN: This was a retrospective study. METHODS AND MATERIAL: The study was conducted at a tertiary care center for cancer patients, in which samples representing blood stream infections sent from the Medical Oncology services of the hospital during the year of 2007 were analysed. The microbiological profile and antibiotic sensitivity pattern of these isolates was studied. RESULTS: There were 484 isolates that represented BSIs. The most common bacterial isolates from patients with cancer were Pseudomonas spp. (30.37%), Staphylococcus aureus (12.6%) and Acinetobacter spp. (11.57%). Meropenem was the most effective antibiotic with 71.2% sensitivity to the bacterial isolates it was tested against. Oxacillin resistance was seen in 18% of S. aureus isolates. CONCLUSION: Gram-negative bacteria were more common as etiologic agents of BSIs in cancer patients. The poor activity of the primary empirical agents for infections in cancer namely ceftazidime and piperacillin-tazobactam is alarming.Strict regulation of vancomycin use should be considered in areas where there is a low prevalence of methicillin-resistant S. aureus (MRSA).

Comparison of in vitro activities of ceftazidime, piperacillin-tazobactam, and cefoperazone-sulbactam, and the implication on empirical therapy in patients with cancer.

BACKGROUND: Infection is a common cause of morbidity and mortality in cancer patients. In most of these cases empirical treatment is provided because the focus of infection is not identified. Empiric antibiotics provided to these patients are based on isolates, sensitivity, and on guidelines. Here we have compared three antibiotics recommended as empirical treatment by the Infectious Disease Society of America (IDSA). AIMS: To compare the three antibiotic sensitivities for gram negative isolates at our institute. OBJECTIVE: To choose the optimal antibiotic as the empirical treatment for cancer patients developing infections. MATERIALS AND METHODS: We collected the data on isolates and antibiotic sensitivity patterns of isolates for ceftazidime, piperacillin + tazobactum, and cefoperazone from the medical oncology department. We subsequently compared the sensitivity of these three antibiotics. STATISTICAL METHODS: The isolates were mapped using the WHONET 5.4 software. The analysis was conducted using SPSS 15.0 for Windows. McNemar Chi-square test was used to compare the sensitivity percentages between any two antibiotics. The agreement between the antibiotic and the gold standard was calculated using the Kappa statistic. Two tailed p values were reported. RESULTS: The results showed that there was a difference among sensitivities for these antibiotics. It appears that the sensitivity of ceftazidime was inferior to the two other antibiotics. Also cefoperazone has better sensitivity as compared to piperacillin + tazobactum. CONCLUSION: In spite of these three antibiotics being recommended by IDSA our data suggest that it should not be followed blindly and local sensitivity data is important for formulating institutional guidelines for using antibiotics.

Maternal nutrition and birth size among urban affluent and rural women in India.

BACKGROUND: Varying results of worldwide intervention programs to pregnant mothers necessitate the need to understand the relationship between maternal nutrition and birth size among well nourished and undernourished mothers. OBJECTIVE: To examine this relationship among urban affluent mothers and to compare the findings with those on rural Indian mothers. SUBJECTS: Data collected on urban affluent mothers (n = 236) was compared with rural mothers (n = 633). DESIGN: Mothers were contacted at 18 +/- 2 and 28 +/- 2 wk of gestation for anthropometry, dietary intakes [24-hr recall, Food Frequency Questionnaire (FFQ)] and after delivery for neonatal anthropometry. RESULTS: Despite large differences in nutritional status of urban and rural mothers ( pre-pregnant weight 55.9 +/- 9.2 Vs 41.5 +/- 5.2 kg, respectively) maternal fat intakes at 18 wk were associated with birth weight (p < 0.05), length (p < 0.01) and triceps skin fold thickness (p < 0.05) of the newborn in urban and rural mothers. Consumption of fruits was associated with birth length (p < 0.05) in urban (18wk) and with birth weight (p < 0.01) and length (p < 0.01) in rural (28wk) mothers, when their energy intakes were low. Maternal consumption of milk too, was associated with newborn's triceps (p < 0.01) in urban (28wk) while with birth weight (p < 0.05) and length (p < 0.05) in rural (18wk) mothers. The findings mainly underscore the importance of consumption of micronutrient rich foods, when energy intakes are limiting during pregnancy, for improving birth size. CONCLUSIONS: Creating nutritional awareness and motivating rural mothers for consuming micronutrient rich foods like green leafy vegetables and seasonal fruits that are easily available in rural areas, will be a much affordable solution for combating the problem of low birth weight rather than waiting for improvement in the existing nationwide programs for pregnant women.

Blood pressure among overweight adolescents from urban school children in Pune, India.

OBJECTIVES: To examine the magnitude of overweight and its association with blood pressure (BP) among adolescents. DESIGN: Cross-sectional study with all children in age range 9-16 years (n=1146 boys and 1077 girls) from two schools catering to urban affluent high socio-economic class (HSE), for anthropometric measurements by trained investigators and BP measurement by a pediatrician using sphygmomanometer. RESULTS: The prevalence of overweight based on conventional body mass index (BMI) cutoff was 27.5% for boys and 20.9% for girls but varied for different indicators. Prevalence of high systolic blood pressure (HSBP) was 12.0% in boys and 9.7% in girls and increased with increasing levels of BMI, weight, triceps skin fold thickness (TSFT) and percent body fat. Mean level of SBP among overweight children was significantly (P<0.001) higher by about 12 mm Hg, whereas that for diastolic blood pressure was higher by 8 mm Hg (P<0.001) as compared to their non-overweight (age, sex-matched) counterparts. This was true in both sexes and for all indicators used for assessing overweight. Prevalence of HSBP increased suddenly beyond BMI value of 20 kg/m2 in boys and 21.5 kg/m2 in girls, beyond TSFT value of 12 mm for boys and 14 mm for girls whereas such cutoffs for body fat were above 25% in both sexes. These cutoffs appear much lower than the conventional ones and therefore indicate the need for validation of conventional cutoffs in different populations. CONCLUSIONS: Our findings highlight that BP measurement needs to be a routine part of physical examination in school children, and the use of cutoffs anchored to metabolic risks may be essential for assessment of obesity.

Comparison of 5% povidone-iodine solution against 1% povidone-iodine solution in preoperative cataract surgery antisepsis: a prospective randomised double blind study.

BACKGROUND/AIM: Povidone-iodine (PI, Betadine) is routinely used as a preoperative topical antiseptic in cataract surgery as it has been shown to reduce the incidence of postoperative endophthalmitis. However, the concentration used clinically is variable. In vitro studies have shown that PI is paradoxically more effective at lower concentration. This study was undertaken to determine if this effect was reproducible in vivo. METHODS: A prospective randomised double blind study was carried out in the ophthalmic theatre in a district general hospital. 105 patients attending for routine cataract surgery were randomly allocated to have their conjunctival fornices irrigated preoperatively with either PI 1% (group A) or PI 5% (group B). Conjunctival swabs were taken, in identical fashion, both before and 1 minute after irrigation. The number and species of bacterial colonies cultured from each swab was counted. The difference in the median number of bacterial colonies from pre-irrigation to post-irrigation cultures was then compared between the groups. RESULTS: Bacterial cultures were gained from 100 patients (33 male, 67 female, mean age 74 years, range 30-95 years). Group B (5% PI) showed a decrease in median colony forming units (CFU) pre-irrigation from 100 to 40 CFU post-irrigation (a drop of 60%). This was greater than in group A (1% PI) where the reduction was 120 CFU pre-irrigation to 100 CFU post-irrigation (a drop of 16.7%) (Mann-Whitney test, p<0.05). At higher initial bacterial loads (CFU pre-irrigation >1000), the difference in median between the two groups became larger as the number of pre-irrigation bacteria increased. In group B pre-irrigation CFU reduced from 3340 to 110 post-irrigation (a drop of 96.7%) compared with group A: 5000 CFU pre-irrigation to 3000 post-irrigation (a drop of 40%) (Mann-Whitney test, p=0.0014). CONCLUSION: Despite in vitro evidence of higher bactericidal efficacy of PI at more dilute concentrations, 5% PI is more effective than 1% PI in decreasing the human conjunctival bacterial flora in vivo, particularly in the presence of heavier initial bacterial load.

Fas and Fas ligand protein and mRNA in normal and atretic mouse ovarian follicles.

Apoptosis is the underlying mechanism of follicular atresia in the mammalian ovary. However, the apoptotic pathways governing this ovarian process are not completely elucidated. In the present study, expression of Fas and Fas ligand, the proximal members of the death receptor pathway, was evaluated in mouse ovarian follicles using immunofluorescence and in situ hybridization. Normal or atretic follicles were obtained from immature female Swiss mice after administration of 10 iu equine chorionic gonadotrophin for 48 or 72 h, respectively. Expression of both Fas and Fas ligand mRNA and protein was observed in granulosa cells of normal and atretic follicles. Although the oocytes of normal follicles failed to show any staining, those of atretic follicles stained intensely for Fas, indicating that the presence of Fas in the oocyte determines the fate of the follicle.

Expression of Fas and Fas ligand protein and mRNA in mouse oocytes and embryos.

During mammalian embryonic development, abnormal eggs and embryos are eliminated by apoptosis; however, the precise apoptotic pathways remain as yet unidentified. In the present study, expression of Fas and Fas ligand - the proximal members of the death receptor pathway, was evaluated in mouse preimplantation embryos by immunofluorescence and in situ hybridization techniques. Ovulated oocytes were collected from oviducts of cyclic mice on the day of oestrus (day 0), and one-cell, two-cell embryos and eight-cell morulae were collected from oviducts of mated animals on days 1, 2 and 3 of pregnancy, respectively. Blastocysts were flushed from the uterine horns on day 4. Expression of Fas and Fas L mRNAs and proteins was absent from embryos at days 0, 1 and 2. A marked increase in Fas and Fas L mRNA and protein expression was detected in all morphologically normal embryos on day 3 and day 4. In addition, embryos on days 3 and 4 were positive for terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining; however, absence of caspase 8 and 3 and intense localization of proliferating cell nuclear antigen confirmed the proliferative status of these embryos. Furthermore, TUNEL staining was absent in postimplantation embryonic sections obtained on day 6. The results of the present studies thus indicate an equilibrium between proliferation and apoptosis in the preimplantation embryo.

Invitro comparative evaluation of aminoglycosides at a cancer centre.

A comparative study of in vitro activity of amikacin, gentamicin, netilmicin, tobramycin and isepamicin was done. A total of 200 clinical isolates of Gram negative organisms from various clinical sources were tested. E.Coli was the most frequently isolated organism followed by Pseudomonas spp., Klebsiella spp., Proteus spp., Acinetobacter spp. and Enterobacter spp. Of the 5 aminoglycosides tested, isepamicin showed the highest numbers of susceptible isolates followed by amikacin, netilmicin, gentamicin and tobramycin. MIC 90 value of isepamicin was lower as compared to amikacin.

Mycobacterial pulmonary infection post allogeneic bone marrow transplantation.

Allogeneic bone marrow transplant recipients are prone to pulmonary infections caused by a wide spectrum of organisms. Since the first bone marrow transplatation (BMT) done in 1983 at the Tata Memorial Hospital, we have recently seen the first case of Mycobacterium Fortuitum Chelonae complex among 117 BMT (including 90 allogeneic and 27 autologous) patients. The patient was on immunosuppressants for chronic GVHD post allogeneic BMT done for CML-CP. He developed pulmonary mycobacterial infection 13 months post BMT. Diagnosis was difficult because of the atypical presentation, negative culture reports, and the presence of multiple pathogens due to immunosuppression. In our case the diagnosis was eventually established after examination of material obtained by bronchoscopy. Patient has shown response to antituberculosis drugs after 2 months. This shows the need to consider atypical mycobacterial infection in the differential diagnosis of pulmonary illness in the post allogeneic BMT setting.

Potential researchable areas in ARTs--oocyte maturation and embryo development.

Infertility is a commonly encountered situation occurring equally in both sexes. In vitro fertilization and embryo transfer (IVF-ET) and other assisted reproductive technologies (ARTs) have enhanced the possibilities for successful treatment to tackle infertility. However, ARTs currently face limitations due to the fact that although success rate is high for the initial stages such as ovulation induction and fertilization, it dwindles progressively so that the success rate of a take home baby is as low as 15-20%. Research centred around various stages in an IVF programme is therefore necessary to devise protocols that ensure a higher success rate. This review takes a look at the potential areas currently under research in the field of ARTs, such as, in vitro oocyte maturation, oocyte/embryo cryopreservation, embryo culture, preimplantation genetic diagnosis. Their applications, in clinical conditions such as cancer, have been discussed.