Biotypes and virulence factors of Gardnerella vaginalis isolated from cases of bacterial vaginosis.

The present study was conducted to correlate the biotypes of Gardnerella vaginalis strains isolated from cases of bacterial vaginosis and their virulence factors. Thirty-two strains of G. vaginalis isolated from cases of bacterial vaginosis were biotyped. Adherence to vaginal epithelial cells, biofilm production, surface hydrophobicity, phospholipase C and protease activity were tested on these isolates. Biotype 1 was the most prevalent (8; 25%), followed by biotype 2 (7; 21.9%) and biotypes 5 and 8 (5; 15.6%). We did not find any statistical correlation between G. vaginalis biotypes and its virulence factors. Virulence factors expressed by G. vaginalis were not associated with a single biotype.

Significance of MRSA strains in community associated skin and soft tissue infections.

The study was conducted to determine the antibiotic susceptibility profile of community-associated methicillin resistant Staphylococcus aureus (CAMRSA) strains isolated from infections. S. aureus strains were isolated from clinical specimens using the standard procedures. CDC definition was used to classify CAMRSA. Antibiotic susceptibility test was done using Kirby-Bauer disk diffusion method. Double disk diffusion method (D-test) was used to detect inducible macrolide, lincosamide and streptogramin B resistance (inducible MLS B resistance ). A total of 83 CAMRSA were isolated from abscesses and other skin infections in persons without known risk factors for MRSA infection. All CAMRSA were susceptible to vancomycin. Out of 83 CAMRSA, 13 (15.65%) were D-test positive (inducible MLS B positive) and 6 (7.23%) were positive for constitutive MLS B resistance. Eight strains (9.63%) were resistant to tetracycline and 26 (31.32%) strains were resistant to erythromycin. Increased rate of inducible clindamycin resistance among CAMRSA indicates the importance of identification of such strains by D test to avoid treatment failure when clindamycin is used.

Effect of stress on production of heat labile enterotoxin by Escherichia coli.

Enterotoxigenic Escherichia coli (ETEC) is an important pathogen responsible for secretory diarrhoea. The production of heat labile enterotoxin (LT), by ETEC, is largely responsible for the pathogenesis of diarrhoea. In the present study we investigated the effect of stress factors such as temperature, pH, osmotic stress and nutritional limitation on the production of LT by ETEC using in-house GMI-ELISA. Four strains of E. coli consisting, one standard strain MTCC 723 and three clinical isolates were used in the study. Maximum amount of LT (OD 3.285) was produced at 37 0 C followed by 40 0 C (OD 3.305). Growth of E. coli in medium with pH 8.6 resulted in maximum amount of LT production (OD 3.489). LT was not detectable when bacteria were grown in medium with pH < or =7.2 and > or = 9.2. Sodium chloride concentration of 0.2 M stimulated maximum amount of LT production. Maximum amount of LT was produced when the bacteria were grown in medium containing 2.5 g/l of glucose. All the stress factors had a significant effect on the LT production by E. coli , though quantitative differences in the various strains were observed.

Factors affecting the nasal carriage of methicillin-resistant Staphylococcus aureus in human immunodeficiency virus-infected patients.

Human immunodeficiency virus-infected patients attending skin outpatient department were studied for nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) and associated factors affecting nasal colonization. Nasal swabs were used for isolation of S. aureus. MRSA were detected by agar screen and agar dilution methods. Careful examination for dermatoses was carried out. Forty-six of the 60 (76.67%) outpatients with HIV infection were colonized with S. aureus in the anterior nares. Significant number of S. aureus carriers were in the 31-40 year age group. Methicillin resistance was found in eight (17.39%) isolates. Of the 46 S. aureus strains, 29 (63%) were resistant to erythromycin, 69.5% to co-trimoxazole and 41.3% to ciprofloxacin. Co-trimoxazole use was found to be a risk factor for S. aureus carriage ( P = 0.0214) but not for methicillin resistance. Hospital stay for more than 10 days was a risk factor for methicillin resistance whereas stay for more than 25 days was found to be a highly significant risk factor. Dermatophytosis and herpes simplex virus infection were other risk factors for nasal carriage of S. aureus.

Effect of transient hypothyroidism during infancy on the postnatal ontogeny of luteinising hormone release in the agonadal male rhesus monkey (Macaca mulatta): implications for the timing of puberty in higher primates.

The present study examined whether a transient thyroid hormone (T(4)) deficit during infancy in male monkeys would compromise the arrest of luteinising hormone (LH) secretion during the infant-juvenile transition, and/or interfere with the pubertal resurgence of LH. Animals were orchidectomized and thyroidectomized (n = 3; Tx) or sham Tx (n = 3) within 5 days of birth. T(4) replacement was initiated in two Tx monkeys at age 19 weeks to reestablish a euthyroid condition. Blood samples were drawn weekly for hormone assay. Body weight, crown-rump length, and bone age were assessed throughout the study. Within a week of Tx, plasma T(4) declined to undetectable levels and, by 6-8 weeks of age, signs of hypothyroidism were evident. Transient hypothyroidism during infancy failed to prevent either arrest of LH secretion during the infant-juvenile transition or the pubertal resurgence of LH secretion, both of which occurred at similar ages to sham Tx animals. Although body weight exhibited complete catch-up with T(4) replacement, crown-rump length and bone age did not. Thus, bone age at the time of the pubertal LH resurgence in Tx animals was less advanced than that in shams. Although Tx did not influence qualitatively the pattern of gonadotrophin secretion, LH levels during infancy and after pubertal LH resurgence were elevated in Tx monkeys. This was not associated with changes in LH pulse frequency and amplitude, but half-life (53 versus 65 min) of the slow second phase of LH clearance was greater in Tx animals. These results indicate that hypothalamic mechanisms dictating the pattern of gonadotrophin-releasing hormone release from birth to puberty are not dependent on T(4) action during infancy, and fail to support the notion that onset of puberty is causally coupled to skeletal maturation. They also indicate that LH renal clearance mechanisms may be programmed in a T(4) dependent manner during infancy.

Effect of exposure to hydrogen peroxide on the virulence of Escherichia coli.

PURPOSE: To eliminate pathogenic bacteria, the host presents conditions that are stressful for bacteria. Oxidative stress arises when the concentration of pro-oxidants like hydrogen peroxide (H2O2 ) and superoxide anion increases to a level over the basal defence capacity of the cell. In the present study, we studied the effect of oxidative stress on the production of certain virulence factors by Escherichia coli . METHODS: E. coli was exposed to oxidative stress by growing in the presence of different concentrations of H2O2 . The effect of oxidative stress on the expression of surface hydrophobicity, adherence, haemolysin production, serum resistance and phagocytosis was studied. RESULTS: Oxidative stress caused a significant decrease in the expression of all the virulence factors of E. coli . CONCLUSIONS: Synthesis of virulence factors can be significantly altered by oxidative stress and such changes may affect the pathogenicity of E. coli.

Virulence factors and drug resistance in Escherichia coli isolated from extraintestinal infections.

PURPOSE: To determine the virulence factors produced by Escherichia coli isolated from extraintestinal infections, to study the drug resistance pattern in E. coli with special reference to extended spectrum beta -lactamase (ESBL) and to evaluate screening methods for ESBL. METHODS: A total of 152 isolates of E. coli from various extraintestinal infections were screened for virulence factors such as haemolysin, surface hydrophobicity, serum resistance and protease. All the isolates were also studied for antibiotic susceptibility pattern using modified Kirby Bauer disk diffusion method. ESBL production was screened by standard disk diffusion method and confirmed using phenotypic confirmatory method. RESULTS: Among 152 isolates, 36 (23.7%) were haemolytic, 42 (27.6%) were hydrophobic, 132 (86.8%) were serum resistant and only four were positive for protease. Multiple virulence factor were observed in 67 (44%) of isolates. Seventy-nine (51.4%) isolates produced ESBL. ESBL producing isolates showed multidrug resistance. There was a significant association ( P E. coli . CONCLUSIONS: The present study shows the expression of virulence factors and multidrug resistance in E. coli isolated from various extraintestinal infections. The study also shows that appropriate methods of detecting drug resistance and ESBL production are required for the judicious use of antibiotics in managing these infections.

Phenotypic switching and its influence on expression of virulence factors by Candida albicans causing candidiasis in human immunodeficiency virus-infected patients.

PURPOSE: The purpose of the present study was to determine the degree of expression of virulence factors such as adherence, cell surface hydrophobicity (CSH) and production of proteinase by different morphological forms of Candida albicans causing oral candidiasis in human immunodeficiency virus (HIV)-infected individuals. METHODS: C. albicans 3153A and two strains isolated from oral thrush in HIV infected individuals were induced to undergo phenotypic switching by exposure to UV light and the degree of expression of virulence factors by the different morphological forms was studied. RESULTS: Three different morphological forms of C. albicans were obtained namely, star (S), wrinkled (W) and ring (R) types from the original smooth (O) variety. It was found that proteinase production was greatest with the W type followed by the R type and O type. The S type produced the least proteinase. Expression of cell surface hydrophobicity and adherence was greatest in the O type followed by the R and then the W type and finally the S type. CONCLUSIONS: The differential expression of virulence factors occurs with different phenotypic forms of C. albicans and this may provide a particular morphological type with a distinct advantage over other types in causing candidiasis.

Induction of a hypothyroid state during juvenile development delays pubertal reactivation of the neuroendocrine system governing luteinising hormone secretion in the male rhesus monkey (Macaca mulatta).

The present study aimed to determine the influence of thyroid status on the timing of the pubertal resurgence in gonadotrophin-releasing hormone pulse generator activity [tracked by circulating luteinising hormone (LH) levels] in male rhesus monkeys. Six juvenile monkeys were orchidectomised and then treated with the antithyroid drug, methimazole, from 15-19 months until 36 months of age, at which time thyroxine (T(4)) replacement was initiated. Four additional agonadal monkeys served as controls. Blood samples were drawn weekly for hormonal assessments. Body weight, crown-rump length and bone age were monitored at regular intervals. By 8 weeks of methimazole treatment, plasma T(4) had fallen sharply, and the decline was associated with a plasma thyroid-stimulating hormone increase. In controls, plasma LH levels remained undetectable until the pubertal rise occurred at 29.3 +/- 0.2 months of age. This developmental event occurred in only half of the methimazole-treated animals before 36 months of age when T(4) replacement was initiated. The hypothyroid state was associated with a profound arrest of growth and bone maturation, but increased body mass indices and plasma leptin levels. T(4) replacement in methimazole-treated monkeys was associated with the pubertal rise in LH in the remaining three animals and accelerated somatic development in all six animals. Although pubertal resurgence in LH secretion occurred at a later chronological age in methimazole-treated animals compared to controls, bone age, crown-rump length and body weight at that time did not differ between groups. There were no long-term differences in plasma prolactin between groups. We conclude that juvenile hypothyroidism in male primates causes a marked delay in the pubertal resurgence of LH secretion, probably occasioned at the hypothalamic level. Whether this effect is meditated by an action of thyroid hormone directly on the hypothalamus or indirectly as a result of the concomitant deficit in somatic development remains to be determined.

Bacterial adherence and humoral immune response in women with symptomatic and asymptomatic urinary tract infection.

PURPOSE: To determine the role of humoral immune response and bacterial adherence in the pathogenesis of symptomatic and asymptomatic urinary tract infection in women. METHODS: The study population consisted of 30 women with symptomatic UTI, 30 women with asymptomatic UTI and 30 healthy women as controls. Bacterial adherence to vaginal epithelial cells was studied and the concentration of serum and urine antibodies to mixed coliform antigen and clinical isolate was determined by ELISA. Surface hydrophobicity of the urine isolates was determined. Student's unpaired t test and Pearson's correlation coefficient test were used in the statistical analysis. RESULTS: Compared to asymptomatic UTI, significantly more number of bacteria adhered to the epithelial cells of women with symptomatic UTI (P< 0.001). All cases of UTI had significantly high concentration of urinary IgG antibody to mixed coliform antigens. Asymptomatic UTI cases had higher concentrations of urinary IgG, IgM and IgA antibodies to clinical isolate. Concentration of sIgA level was more in symptomatic UTI. Significant correlation was observed between urinary IgG and adherence of clinical isolate in case of asymptomatic UTI. CONCLUSIONS: The present study showed that greater receptivity of epithelial cells to bacteria may increase the susceptibility to UTI. Humoral immune response and local immunity may modify the pathogenesis of UTI.

Sterilisation of extracted human teeth for educational use.

Sixty intact, non-carious and unrestored teeth extracted due to periodontal disease were used to determine the most effective method of sterilisation. The teeth were divided into six groups, each containing 10 teeth. Group 1 teeth were immersed in 10% formalin for seven days, group 2 teeth were immersed in 3% hydrogen peroxide for seven days, group 3 teeth were immersed in 2.6% sodium hypochlorite for seven days, group 4 teeth were boiled in water at 100 degrees C for 20 minutes, group 5 teeth were autoclaved at 121 degrees C at 15 lbs psi for 30 minutes, and group 6 teeth were immersed in normal saline for seven days. After the treatment, the teeth were individually inoculated into trypticase soy broth and incubated for 48 hours. A questionnaire survey was also conducted to determine the awareness of dental students regarding infection due to extracted human teeth and the common disinfection/sterilisation methods used. Autoclaving at 121 degrees C, 15 lbs psi for 30 minutes and immersion in 10% formalin for seven days were effective in disinfecting/sterilising extracted human teeth. Chemicals such as 2.6% sodium hypochlorite, 3% hydrogen peroxide and boiling in water were not effective. The results indicate that autoclaving for 30 minutes or immersion in 10% formalin for seven days could be effectively used for disinfection/sterilisation of extracted human teeth.

Simulated conditions of microgravity suppress progesterone production by luteal cells of the pregnant rat.

The purpose of this study was to assess whether simulated conditions of microgravity induce changes in the production of progesterone by luteal cells of the pregnant rat ovary using an in vitro model system. The microgravity environment was simulated using either a high aspect ratio vessel (HARV) bioreactor with free fall or a clinostat without free fall of cells. A mixed population of luteal cells isolated from the corpora lutea of day 8 pregnant rats was attached to cytodex microcarrier beads (cytodex 3). These anchorage dependent cells were placed in equal numbers in the HARV or a spinner flask control vessel in culture conditions. It was found that HARV significantly reduced the daily production of progesterone from day 1 through day 8 compared to controls. Scanning electron microscopy showed that cells attached to the microcarrier beads throughout the duration of the experiment in both types of culture vessels. Cells cultured in chamber slide flasks and placed in a clinostat yielded similar results when compared to those in the HARV. Also, when they were stained by Oil Red-O for lipid droplets, the clinostat flasks showed a larger number of stained cells compared to control flasks at 48 h. Further, the relative amount of Oil Red-O staining per milligram of protein was found to be higher in the clinostat than in the control cells at 48 h. It is speculated that the increase in the level of lipid content in cells subjected to simulated conditions of microgravity may be due to a disruption in cholesterol transport and/or lesions in the steroidogenic pathway leading to a fall in the synthesis of progesterone. Additionally, the fall in progesterone in simulated conditions of microgravity could be due to apoptosis of luteal cells.