Biodegradation of 4-nitrophenol by a Rhodococcus species and a preliminary insight into its toxicoproteome based on mass spectrometry analysis

Aim: The objective of this study was to understand the possible fate of 4-NP through the molecular mechanism and to identify potential enzymes involved in 4-NP biodegradation by Rhodococcus sp. strain BUPNP1 Methodology: Biodegradation of 4-NP was detected spectrophotometrically at 400 nm and also confirmed by TLC and HPLC. Comparative study of proteomes was performed by 2-D gel electrophoresis followed by peptide mass fingerprinting and bioinformatic analysis to identify and/ or predict the possible functions of over-expressed proteins in 4-NP treated cells of BUNP1. Results: Utilization of 4-nitrophenol and its hydrolysis intermediate 4-nitrocatechol (4-NC) and 1,2,4-Benzenetriol as sole carbon source indicated the presence of genomic information encoding the enzyme necessary for the operation of 4-nitrophenol degradation pathway in the strain BUPNP1. It could transform 4-NP into 4-NC by monooxygenase whose major activity was detected during initial stage of degradation. The 4-NC further depleted in the medium to release nitrite ions. In order to investigate the molecular changes occurring during degradation, a comparative study of proteome profiles was carried out where; 4-nitrophenol treated cells were compared against cells grown on glucose as control. The comparative study indicated expression of several protein spots under 4-nitrophenol treated condition. Interpretation: This study showed the potential of BUPNP1 strain belonging to genus Rhodococcus towards induced expression of some unique proteins which might have possible role in 4-NP biodegradation process

Knowledge on Care of Autistic Child among the Mother’s Attending Protibondhi Foundation, Dhaka.

Objective: Autism has a strong genetic basis, although the genetics of autism are complex and it is unclear. This descriptive type of cross sectional study was conducted to explore the knowledge on care of autistic children among the mother’s attending Protibondhi Foundation, Dhaka. Methods: A total of 385 samples were selected purposively, but due to time and budget constrict it was 150 finally. Data were collected by using an anonymous, pre-tested, semi structured and self-administered questionnaire. Results: The study revealed that more than 50% of the respondent’s were in the age group 16-30 years, majority of the respondent’s were Muslim. Almost 84% of the respondents were SSC-Graduate and majority of them (nearly 50%) were housewife .The monthly family income was ranged between Taka 25,001-50,000 and its percentage was 53.3%. The knowledge about type of autism is very low, the data showed that only two types of autism they knew and the respondent’s knowledge about the risk factor was also very low; only 24% of the respondents had knowledge about complications during pregnancy and delivery related risk factors. About 28% needed physiotherapy, 30% needed occupational therapy, 34% needed speech therapy & 44.7% needed psychotherapy. Here significant association was found between age, education and knowledge about autism (p<0.05). Conclusion: Overall knowledge on care of autistic child was not satisfactory among respondents.

Prevalence of multidrug-resistant tuberculosis among Category II pulmonary tuberculosis patients.

Background & objectives: Multidrug-resistant tuberculosis (MDR-TB) has emerged as a significant global health concern. The most important risk factor for the development of MDR-TB is previous anti-tuberculosis therapy. Category II pulmonary TB includes those patients who had failed previous TB treatment, relapsed after treatment, or defaulted during previous treatment. We carried out this study to ascertain the prevalence of MDR-TB among category II pulmonary TB patients. Methods: This was a cross-sectional, descriptive study involving category II pulmonary TB patients diagnosed between 2005 and 2008. All sputum-positive category II TB cases were subjected to mycobacterial culture and drug-susceptibility testing (DST). MDR-TB was defined as TB caused by bacilli showing resistance to at least isoniazid and rifampicin. Results: A total of 196 cases of sputum-positive category II pulmonary tuberculosis patients were included. Of these, 40 patients (20.4%) had MDR-TB. The mean age of MDR-TB patients was 33.25 ± 12.04 yr; 9 patients (22.5%) were female. Thirty six patients showed resistance to rifampicin and isoniazid; while 4 patients showed resistance to rifampicin, isoniazid and streptomycin. The prevalence of MDR-TB among category-II pulmonary tuberculosis patients was 20.4 per cent. Interpretation & conclusions : The prevalence of MDR-TB in category II TB patients was significant. However, nation-wide and State-wide representative data on prevalence of MDR-TB are lacking. We stress the importance of continuous monitoring of drug resistance trends, in order to assess the efficacy of current interventions and their impact on the TB epidemic.

Reversal of subclinical adrenal insufficiency through antituberculosis treatment in TB patients: a longitudinal follow up.

BACKGROUND & OBJECTIVE: Subclinical adrenal insufficiency has been shown to occur in patients with tuberculosis. Whether this insufficiency can be reversed with therapy and on long-term follow up, is not known. We studied the effect of antituberculosis treatment (ATT) with respect to reversal of the adrenal insufficiency, as assessed by response to standard dose adrenocorticotropin (ACTH) stimulation test in TB patients. METHODS: One hundred and five HIV-negative tuberculosis patients were studied. Of these, 72 patients had pulmonary and 33 had extrapulmonary forms of the disease. Baseline (pre-treatment) standard-dose ACTH stimulation test was done on all the subjects, following which, they were put on standard antituberculosis therapy, depending on the type of disease and were followed up for a period of 30 months. ACTH stimulation tests were performed at follow up, every 6 months. RESULTS: Baseline (pre-treatment) standard-dose ACTH stimulation test revealed an impaired response in 52 of 105 patients (49.5%). At 6 months, the percentage of responders had increased to 71 per cent with a gradual increasing trend noted thereafter. At 24 months, 31 of the 32 patients (97%) who were followed up demonstrated a normal response to ACTH stimulation. The percentage of responders was comparable in both pulmonary [21 of 22 patients (95%)] and extrapulmonary TB [10 of 10 patients (100%)] groups at follow up. INTERPRETATION & CONCLUSION: Our study shows that nearly half of patients with active tuberculosis had a subclinical adrenal insufficiency indicated by an impaired response to ACTH stimulation test. This insufficiency reverse with therapy in most patients on long-term follow up.

Contraceptive practices and awareness of emergency contraception in educated working women.

BACKGROUND: Regular contraceptive use and emergency contraception are tools to prevent pregnancies. AIMS: This study was designed to investigate knowledge and use of contraceptive methods and awareness of emergency contraception among women working in the hospital. SETTINGS: Educated workingwomen in a medical college hospital. DESIGN: Cross-sectional study. MATERIALS AND METHODS: The study was carried out among women belonging to three categories: staff nurses, ministerial staff and others. Married as well as unmarried employees in the reproductive age group were interviewed. A pretested mixed questionnaire containing open as well as closed ended questions was administered. The women were asked questions concerning knowledge and use of contraceptive methods and awareness of emergency or postcoital contraception. RESULTS: Of the 284 employees 258 women consented for the interview. All the subjects were literate and majority (97.2%) had an urban background. Of the 190 married women, 154 (81.1%) practiced contraception, among them (73.3%) were regular users. Eighty respondents underwent abortions of which 46 had spontaneous and 34 had induced abortions. Among the available contraceptive methods, condom was the most popular method in 89 (57.8%) followed by Copper T in 38 women (24.7%). The use of hormonal contraception was very low 2.6%. Print and electronic media were the common source of public awareness in 149 subjects (57.7%). Twenty-nine women (11.2%) were aware and only three women used emergency contraception. CONCLUSIONS: A high percentage of females in this literate workingwomen population used contraception; however, the awareness of emergency contraception was low.

Increasing HIV seropositivity among adult tuberculosis patients in Delhi.

We report the HIV seropositivity among adult TB patients from our hospital, a tertiary care hospital in north India between 2000-2002. Of the 555 patients with various forms of tuberculosis, 52 were found to be seropositive (9.4%). In 1994-1999, the HIV seropositivity in this hospital was only 0.4 per cent (2 of 500 patients). This communication describes a dramatic increase in seropositivity and highlights the importance of continued HIV serosurveillance in patients with TB.

A prospective study of sensitivity and specificity of adenosine deaminase estimation in the diagnosis of tuberculosis pleural effusion.

We prospectively evaluated the usefulness of adenosine deaminase [ADA] estimation in the diagnosis of tuberculosis [TB] pleural effusion. Seventy five subjects with pleural effusion were studied. Forty eight of them had TB pleural effusion [M:F: 37:11; mean age 33 +/- 14.4 years range 17-76] and the remaining 27 had pleural effusion due to causes other than TB [non-TB group] [M:F: 19:8; mean age 47.3 +/- 16.5 years; range 17-75]. Pleural fluid [PF] ADA levels were significantly higher in TB (n=48; mean 95.8 +/- 57.5 IU/L) compared with non-TB group (n=27; mean 30.7 +/- 27.2 IU/L) [p<0.001]. Serum ADA [S-ADA] levels were also significantly higher in TB (n=45; mean 39.6 +/- 18.3 IU/L) compared with non-TB group (n=26; mean 18.0 +/- 13.7 IU/L) [p<0.001]. PF-ADA levels were higher compared to S-SDA in TB (p <0.001) and non-TB groups [p<0.01]. Using a cut off of 35 IU/L, the sensitivity and specificity of PF-ADA in the diagnosis of TB was computed to be 83.3% and 66.6% respectively. At a cut-off level of 100 IU/L, PF-ADA was found to have a sensitivity 40% and specificity 100%. From this study it is concluded that, using 100 IU/L as the cut-off, it is possible to avoid pleural biopsy to ascertain the diagnosis of TB in as much as 40% of the patients.

HIV seropositivity among adult tuberculosis patients in Delhi.

We report HIV seropositivity among tuberculosis patients from our hospital between 1994-99. Of the 500 patients with various forms of tuberculosis, two were found to be seropositive (0.4%). This report contrasts with the HIV seropositivity reports from other parts of India where increasing HIV seropositivity has been reported. As the HIV infection is making rapid in-roads in India, it is suggested that continuous HIV sero-surveillance should be done in patients with tuberculosis.

Adrenocortical reserve and morphology in tuberculosis.

Ninety seven patients (63 males, mean age 31.8 years, SD 2.3) with various forms of tuberculosis were studied. All of them were HIV negative. Thirty normal control subjects (16 males, mean age 36.4 years, SD 1.8) were also studied. Fifty-eight of the 97 patients (59.8%) were malnourished (BMI < 18 kg/m2). The mean basal serum cortisol was lower in the TB group (n = 91) (351 nmol/1; SD 150) as compared to the normal control group (n = 8) (402 nmol/1; SD 93) but this difference did not attain statistical significance. Following administration of synthetic ACTH (cosyntropin), the 30 and 60 minutes mean serum cortisol values in the TB group were significantly lower as compared to the normal control group (p < 0.05). Forty five of the 91 patients (49.5%) who underwent the ACTH stimulation test had compromised adrenal reserve. Fourteen of the 86 patients (16.3%) in whom adrenal morphology was studied revealed adrenal gland enlargement on abdominal CT scan. ACTH stimulation was done in 12 of these 14 patients and eight of them had compromised adrenal reserve. Repeat ACTH stimulation done six months to one year after treatment in 13 patients revealed significantly increased 30 minutes (p < 0.05) and 60 minutes (p < 0.05) serum cortisol values. While nine of these 13 patients were negative responders before treatment, only three of them had evidence of compromised adrenal reserve after one year of antituberculosis treatment, (p < 0.05). Serum cortisol values in patients with drug-sensitive and drug-resistant tuberculosis did not differ significantly. Patients with drug-resistant tuberculosis had a higher prevalence of adrenal gland enlargement (7 of the 30) as compared to those with drug-sensitive tuberculosis (7 of the 56) (p = NS). Subclinical adrenal insufficiency is prevalent in a significant number of patients with both drug-sensitive and drug-resistant tuberculosis, and in some of these it is associated with adrenal gland enlargement. The compromised adrenal reserve and enlargement seem to reverse with therapy.

Clonal analysis of non-toxigenic Vibrio cholerae O1 associated with an outbreak of cholera.

We examined the clonal relationships among eight clinical isolates of non-toxigenic (NT) V. cholerae O1 associated with a cluster of cases of cholera in Warangal, Andhra Pradesh in south India and compared their relatedness to toxigenic O1 strains of classical and E1Tor biotypes and with O139 Bengal strains of V. cholerae by pulsed-field gel electrophoresis (PFGE). Phylogentic analysis of the NotI restriction fragment length polymorphism showed that all the NT. V. cholerae O1 strains formed a tight cluster with more than 80 per cent similarity. Interestingly, the NT V. cholerae O1 cluster was more closely related to V. cholerae O139 than to classical and E1Tor biotypes of V. cholerae O1 indicating closer genetic relationships between NT V. cholerae 01 and O139 Bengal strains that were isolated during the same time-frame.

Comparative analysis of factors promoting optimal production of cholera toxin by Vibrio cholerae O1 (classical & E1Tor biotypes) & O139.

Various culture media [AKI, Brain heart infusion broth (BHI), Casamino acid-yeast extract broth (CAYE), Casamino acid-yeast extract broth supplemented with 90 micrograms/ml of lincomycin (CAYE-L), Tryptic soy broth (TSB) and Yeast extract peptone (YEP)], cultural conditions (stationary and shaking) and incubation temperatures (30 degrees C and 37 degrees C) were evaluated to determine optimal conditions for production of cholera toxin (CT) by different biotypes (classical and E1Tor) and serogroups (O1 and O139) of V. cholerae. It was found that V. cholerae O1 E1Tor grown in CAYE-L and incubated at 30 degrees C with constant shaking was optimal for production of CT, while for the classical biotype and for the O139 serogroup, CT was maximally produced when grown in YEP and incubated at 30 degrees C in a shaker. Temperature appeared to be a prominent factor affecting the production of CT by the O1 E1Tor biotype when the media used were AKI, CAYE-L and YEP and also for the classical biotype when the media used were the AKI, BHI, CAYE and YEP. In the case of the O1 E1Tor biotype, CAYE-L was the best medium for CT production whereas for the classical biotype, CAYE-L was a poor medium as far as CT production was concerned. Irrespective of the media used, 30 degrees C shake culture condition seemed to be more favourable for supporting CT production except in CAYE medium for the O1 E1Tor biotype where incubation at 37 degrees C in a shaker was as good as incubation at 30 degrees C.

Effect of additional oral ofloxacin administration in the treatment of multidrug-resistant tuberculosis.

The effect of additional oral ofloxacin administration was evaluated in 19 HIV-negative patients with multidrug-resistant tuberculosis (MDR-TB). Their mean duration of illness was 3.6 years. Based on chest roentgenograms, 13 patients had advanced disease and 6 had moderate disease. In addition to 'second-line drugs', patients were treated with oral ofloxacin; high dose 800 mg per day (12 patients), low dose 400 mg per day (2 patients) and intermittent twice a week regimen of 800 mg a day (5 patients). With treatment, there was a significant clinical improvement. Sputum smear conversion occurred in 18 patients (mean duration 15 weeks) and this sputum smear negativity persisted for a mean duration of 7 months. Chest roentgenograms revealed regression of lesions in 18 patients. Reversible side effects were encountered in four patients. Results of present study suggest that additional administration of oral ofloxacin is both effective and safe for the treatment of MDR-TB.

Extensive CT scan abnormality in Wilson's disease.

A case of Wilson's disease with extensive white matter hypodensity, including in the basal ganglia, on CT scan is presented. Such extensive CT scan abnormality has not been described in Wilson's disease.

Acute necrotizing myelopathy with complete myelographic block.

Acute necrotizing myelopathy is a rare disorder. A case showing complete myelographic block extending from D3 to D6 level is described. The suspected aetiological factors for acute necrotizing myelopathy are discussed.