Conjugal transfer of a virulence plasmid in the opportunistic intracellular actinomycete Rhodococcus equi.

Rhodococcus equi is a facultative intracellular, Gram-positive, soilborne actinomycete which can cause severe pyogranulomatous pneumonia with abscessation in young horses (foals) and in immunocompromised people, such as persons with AIDS. All strains of R. equi isolated from foals and approximately a third isolated from humans contain a large, ~81-kb plasmid which is essential for the intramacrophage growth of the organism and for virulence in foals and murine in vivo model systems. We found that the entire virulence plasmid could be transferred from plasmid-containing strains of R. equi (donor) to plasmid-free R. equi strains (recipient) at a high frequency and that plasmid transmission reestablished the capacity for intracellular growth in macrophages. Plasmid transfer required living cells and cell-to-cell contact and was unaffected by the presence of DNase, factors pointing to conjugation as the major means of genetic transfer. Deletion of a putative relaxase-encoding gene, traA, located in the proposed conjugative region of the plasmid, abolished plasmid transfer. Reversion of the traA mutation restored plasmid transmissibility. Finally, plasmid transmission to other Rhodococcus species and some additional related organisms was demonstrated. This is the first study showing a virulence plasmid transfer in R. equi, and it establishes a mechanism by which the virulence plasmid can move among bacteria in the soil.

Lot-to-lot consistency of a hexavalent DTwP-IPV-HB-PRP∼T vaccine and non-inferiority to separate DTwP-HB-PRP∼T and IPV antigen-matching vaccines at 6-8, 10-12, and 14-16 weeks of age co-administered with oral rotavirus vaccine in healthy infants in India: A multi-center, randomized, controlled study.

Background: Combination vaccines reduce the number of pediatric injections but must be as safe, immunogenic, and effective as each of the individual vaccines given separately. Additionally, consistency in manufacturing lots is essential for WHO prequalification. This study aimed to establish the lot-to-lot consistency of a fully liquid, hexavalent diphtheria (D)-tetanus (T)-whole-cell pertussis (wP)-inactivated poliovirus (IPV)-hepatitis B (HB)-Haemophilus influenzae b (PRP-T) (DTwP-IPV-HB-PRP∼T) vaccine and to demonstrate non-inferiority to licensed DTwP-HB-PRP∼T and IPV vaccines. Methods: A Phase III, randomized, active-controlled, and open-label study was conducted at multiple centers across India. Healthy infants who had received a birth dose of oral poliovirus vaccine and hepatitis B vaccine received one of three lots of DTwP-IPV-HB-PRP∼T or separate DTwP-HB-PRP∼T and IPV vaccines at 6-8, 10-12, and 14-16 weeks of age. Oral rotavirus vaccine was co-administered at 6-8 weeks of age and 10-12/14-16 weeks of age. DTwP-IPV-HB-PRP∼T lot-to-lot consistency and non-inferiority (pooled DTwP-IPV-HB-PRP∼T) versus DTwP-HB-PRP∼T and IPV post-third dose were assessed using seroprotection rates (anti-D, anti-T, anti-HBs, anti-PRP, anti-polio 1, 2, 3) and adjusted geometric mean concentrations (anti-PT, anti-FIM). Safety was assessed by parental reports. Results: Lot-to-lot consistency was demonstrated for DTwP-IPV-HB-PRP∼T and non-inferiority versus DTwP-HB-PRP∼T and IPV was confirmed with 95% CIs for seroprotection rate differences and adjusted geometric mean concentration ratios within pre-defined clinical margins. Pooled seroprotection rate was ≥ 99.7% for anti-D ≥ 0.01 IU/mL, anti-T ≥ 0.01 IU/mL, anti-HBs ≥ 10 mIU/mL, anti-PRP ≥ 0.15 µg/mL, and anti-polio 1, 2, and 3 ≥ 8 (1/dil) and vaccine response rate was 83.9% for anti-PT and 97.7% for anti-FIM. There were no safety concerns. Conclusions: Immunogenicity of three lots of the fully liquid DTwP-IPV-HB-PRP∼T vaccine was consistent and non-inferior to licensed comparators following vaccination at 6-8, 10-12, and 14-16 weeks of age. There were no safety concerns and no evidence of any effect of co-administration with rotavirus vaccine.

Ni2+-uptake in Pseudomonas putida strain S4: a possible role of Mg2+-uptake pump.

Essential metal ion homeostasis is based on regulated uptake of metal ions, both during its scarcity and abundance. Pseudomonas putida strain S4, a multimetal resistant bacterium, was employed to investigate Ni(2+) entry into cells. It was observed that Mg(2+) regulates the entry of Ni(2+) and by this plays a protective role to minimize Ni(2+) toxicity in this strain. This protection was evident in both growth as well as viability. Intracellular accumulation of Ni(2+) varied in accordance with Mg(2+) concentrations in the medium. It was hypothesized that Ni(2+) enters the cell using a broad Mg(2+) pump, i.e. the CorA system, as the CorA inhibitor, i.e. Co(III) Hex, also inhibits Ni 2+ uptake. This led to the inference that Mg(2+)-based protection was basically due to competitive inhibition of Ni(2+) uptake. We also show that Zn(2+) can further regulate the entry of Ni(2+).

Oocyte retrieval and histological studies of follicular population in buffalo ovaries.

Ovaries (N = 250) from slaughtered buffaloes were collected to study follicular population and compare methods of oocyte retrieval. The number and size of surface follicles were recorded and grouped into different categories. Different sized follicles in relation to oocyte diameter were studied histologically. Yield of oocytes per ovary were less (P < 0.05) from ovaries bearing a corpus luteum (CL). Techniques used for oocyte recovery included slicing, follicle puncture and aspiration. The oocyte recovery rate was greatest (P < 0.05) using slicing. The average number of visible surface follicles was 5.20 +/- 0.97 with mean numbers of 2.5, 1.2, 0.82 and 0.62 per ovary for follicles sized 4, 8, 12 and 12mm respectively. Histological studies revealed large numbers of primordial follicles in prepubertal and atretic follicles in senile buffaloes. They also established a biphasic relationship of growth between oocyte diameter and follicular size.

Result of suprapubic cystostomy only as primary management of posterior urethral rupture following pelvic fracture.

Forty-five patients with posterior urethral injury following pelvic fractures were managed by suprapubic cystostomy alone as primary management. Simultaneous voiding cystourethrogram with retrograde urethrogram six weeks later revealed non-obliterative stricture in eight and total block in 36. Impotence was seen in 20 patients. Eight patients with non-obliterative stricture responded to optical internal urethrotomy. Out of 36 total block, 30 had long strictures in the posterior urethra and required transpubic urethroplasty. Impotence was not affected by transpubic urethroplasty.

Anterior fontanel size.

Anterior fontanel size was determined in a cross-sectional study of 445 infants ranging in age from newborn period to 2 years. The mean anterior fontanel size in neonates was 3.37 +/- 0.61 cm which decreased to 0.37 +/- 0.06 cm in 24 months age group. The age of closure of anterior fontanel was 12, 18 and 24 months in 40%, 70.4% and 91.3%, respectively.

Modified Levinson's test in rapid diagnosis of tuberculous meningitis.

Modified Levinson's precipitation test was done in 64 cases of TBM, 54 cases of TBM with inconsistent CSF finding, and 32 cases of pyogenic meningitis. The test was positive in 93.7% cases of TBM (sensitivity, 93.7%), 85.5% cases of TBM with doubtful diagnosis and in 9.4% cases of pyogenic meningitis (specificity 90.6%) compared to 79.7%, 72.2% and 18.8% in original Levinson's test, respectively. With CSF examination only 66% cases of TBM could be diagnosed while with modified Levinson's test and CSF analysis 89% cases could be diagnosed (p < 0.001). So modified Levinson's test for diagnosis of TBM is better than Levinson's test (p < 0.05) with an added advantage of time saving.

Prothrombin time in first week of life with special reference to vitamin K administration.

Prothrombin time was estimated in 100 neonates (80 full term and 20 preterm). Among the full term infants 50 were healthy and 30 sick. Prothrombin time was altered in neonates with birth hypoxia and prematurity (p less than 0.001). Vitamin K administration to anoxic babies resulted in improvement in prothrombin time after 48-72 hours (p less than 0.001). Four newborns has bleeding, 2 had anoxia and 2 were only in preterms who did not receive vitamin K after birth. It is concluded that vitamin K should be given to all preterms and those with difficult deliveries; term, healthy newborns do not need it.

Calf circumference as a predictor of low birth weight babies.

PIP: To identify an inexpensive, reliable predictor of low birth weight capable of being utilized by untrained personnel, anthropometric measures were performed on 1600 newborns in Kanpur, India. This analysis indicated that, for a birth weight of 2500 g, the critical limit of calf circumference was 10.8 cm or less. Critical limits for crown-heel length, head circumference, chest circumference, midarm circumference, and thigh circumference were equal to or less than 45.2 cm, 32.2 cm, 30.1 cm, 9.0 cm, and 15.4 cm. All these measurements were significantly correlated with birth weight. This correlation was highest for calf circumference (r = 0.98), thigh circumference (r = 0.93), and chest circumference (r = 0.86). The highest sensitivity of detecting low-birth-weight infants was found for calf circumference (98.4%), followed by thigh circumference (91.6%) and head circumference (76.7%); specificity was 98.2% for thigh circumference and 90.0% for calf circumference. Traditional birth attendants can be trained easily to screen out high-risk newborns by measuring calf circumference.^ieng

Poliomyelitis with special reference to immunization status.

One hundred ninety one children below 5 years of age suffering from poliomyelitis were analyzed to find out the immunization status and its correlation with the incidence of poliomyelitis. Effects of age, sex, immunization status and seasonal variation on the morbidity and mortality status were studied. The maximum number of cases were admitted during the months of July (23.6%) and August (23.1%). Of 191 cases, 143 (74.9%) had no immunization and 48 (25.1%) were partially immunized. A total of 155 (81.2%) cases had spinal polio, 23 (12.01%) bulbo-spinal polio, and 13 had (6.8%) bulbar polio. Serious illness (bulbospinal and bulbar type) was more in partially immunized children (25%) as compared to unimmunized children (16.8%). The mortality rate was more than two times higher in the partially immunized (29.6%) as compared to unimmunized children (11.2%). The possible explanation for high mortality in partially immunized children could be due to the adverse effect of OPV which has not been studied so far.

Field Trial of Relative Efficacy of Abate and Bacillus thuringiensis Against Simulium himalayense Larvae (Diptera simulidae).

Simulium himalayense (Diptera simulidae) are known pests which cause severe biting nuisance in North-East region and breed in its fast flowing rivers and rivulets. A field trial was undertaken to control this nuisance with the help of larvicides and to help ascertain relative efficacy of Abate and B thuringiensis against Simulidae. Abate proved to be an effective larvicide for river treatment wherein 0.1 PPM resulted in 100% larval reduction while in small streams, the treatment with Teknar (Bacillus thuringiensis) with 10 PPM for 10 minutes resulted in 80% larval reduction. The results of the present study showed that Abate was more effective in rivers while B thuringiensis proved to be useful in controlling the breeding in the rivulets. An integrated approach using chemical and biological agents is recommended and considered ecologically sound as it does not affect the fish and potability of water.

Occurrence of polymelia in a female child.

We report a rare case of polymelia in a 6-month-old female child who presented with developed lower limbs and an additional underdeveloped left lower limb.

Role of hypertonic saline and mannitol in the management of raised intracranial pressure in children: A randomized comparative study.

OBJECTIVE: To compare the efficacy and side effects of 3% hypertonic saline and mannitol in the management of raised intracranial pressure in children. DESIGN: Prospective randomized study. SETTING: Pediatric intensive care unit (PICU) in a tertiary care hospital. SUBJECT: 200 patients with raised intracranial pressure. MATERIALS AND METHODS: Patients were randomized into two statistically comparable groups; Group A (n = 98) was treated with mannitol while Group B (n = 100) was treated with 3% hypertonic saline. Group C (n = 2) included those members of Group A in whom serum osmolality ≥320 mosmol/kg and were then treated with 3% hypertonic saline. Both Drugs were given at a loading dose of 5 ml/kg stat followed by 2 ml/kg in every 6 h(both have same osmolarity) for two days in their respective groups. Besides monitoring, blood pressure (NIBP), mean arterial pressure (pre and post 30 min of drug), serum sodium, chloride and osmolality were measured. Intracranial pressure was assessed indirectly by measuring mean arterial ressure "MAP". Student paired 't' test was applied. RESULTS: Decrease in MAP was highly significant (P<0.001) at 0 h in males 0,6 h in females, and moderately significant at 12,36 h in females and significant(P<0.05) at 6,24,42 h in males of Group B. Decrease in coma hours was a highly significant finding (P<0.001) in Group B. In Group B, serum sodium and chloride increased significantly but remained within acceptable limits. There was no difference in osmolality and mortality (fisher Z). CONCLUSION: Mannitol has several side effects, 3% hypertonic saline is a safe and effective alternative in managing cerebral edema.