Characterization of glycolipid biosurfactant from Pseudomonas aeruginosa PA 1 and its efficacy against Phytophtora infestans

Aim: The aim of the present study was to identify biosurfactant produced by Pseudomonas aeruginosa and to determine the effectiveness of biosurfactants (rhamnolipid) against P. infestans causing late blight of potato. Methodology: Pseudomonas aeruginosa strains were isolated from soil samples and the potential strain PA 1, selected because of its antagonistic ability, was used to optimize anti-metabolite production and its characterization using HPLC-Mass spectrometry. Rhamnolipid based formulation was developed and its efficacy was tested against late blight disease. Results: The results revealed that four rhamnolipids congeners were identified, among them three were mono-rhamno-di-lipidic congeners and one was di-rhamno-di-lipdic congeners, abundantly present in the crude biosurfactant obtained from P. aeruginosa PA1. The mass spectra of mono-rhamno-di-lipidic Rha-C12-C14 peak value (m/z 584), Rha-C12.1-C10.CH3 peak value (m/z 545), Rha-C12-C-12-CH3 peak value (m/z 575) and di-rhamno-di-lipidic. Rha-Rha-C10-C10 peak value (m/z 651) were also detected. Rhamnolipid-based formulation was developed and evaluated at different concentration ranging from 0.012 and 0.3% in detached leaf test. Significant reduction in lesion area was recorded at 0.2% concentration (lesion area 0.06 cm2 as against 9.8 cm2 on 5th day of inoculation). Interpretation: Microbial produced rhamnolipid based formulation at 0.2% concentration was found effective against late blight of potato in detached leaf test. Further, it could be used in field study as green chemical which would help in replacing application of chemicals in agriculture.

Influence of root-knot nematode infestation on antioxidant enzymes, chlorophyll content and growth in Pogostemon cablin (Blanco) Benth.

Plants adapt themselves to overcome adverse environmental conditions, and this involves a plethora of concurrent cellular activities. Physiological experiments or metabolic profiling can quantify this response. Among several diseases of Pogostemon cablin (Blanco) Benth. (Patchouli), root-knot nematode infection caused by Meloidogyne incognita (Kofoid and White) Chitwood causes severe damage to the plant and hence, the oil production. In the present study, we identified M. incognita morphologically and at molecular level using sequenced characterized amplified region marker (SCAR). M. incognita was artificially inoculated at different levels of second stage juveniles (J2) to examine the effect on Patchouli plant growth parameters. Peroxidase and polyphenol oxidase enzyme activity and changes in the total phenol and chlorophyll contents in M. incognita was also evaluated in response to infection. The results have demonstrated that nematode infestation leads to increased peroxidase activities in the leaves of the patchouli plants and thereby, increase in phenolic content as a means of defence against nematode infestation. Chlorophyll content was also found decreased but no changes in polyphenol oxidase enzyme activity.

An experience with 156 patients attending a newly organized pain and palliative care clinic in a tertiary hospital.

Context: Pain and palliative care clinic (PCC). Aims: The primary object of this study was to enumerate the demographic characteristics of patients attending a newly organized PCC. The secondary purpose was to detect symptom prevalence and frequency of different cancers in these patients. Settings and Design: Prospective cross-sectional descriptive study. Materials and Methods: A prospective cross-sectional descriptive study was done on patients referred to the PCC of a tertiary hospital in North India. Comprehensive details of all patients were recorded systematically on the first visit on a proforma specially prepared for the newly established palliative care clinic. Statistical Analysis Used: The descriptive statistics of palliative care data was presented in terms of frequencies and percentages (%) for categorical variables. Results: The data collected at our PCC showed that out of 156 patients, 87 were males and 69 were females. Patients of all ages varying from 6 to 85 years were seen. Most patients (82.1%) lived with their families, and 28 (17.1%) patients lived alone and had no financial support. The most common primary diagnoses were head and neck cancers (38.5%), carcinoma cervix (15.4%), breast cancer (10.3%), colorectal cancer (6.4%), and lung cancer (4.5%). Frequency of seven most common symptoms was pain (100%), insomnia (64.1%), loss of appetite (34.6%), nausea (32.7%), vomiting (32.1%), constipation (31.4%) and sore mouth (28.8%). Conclusions: Population-based studies determine the actual magnitude of sufferers and suffering and show that palliative care services should be included as an essential component in a tertiary care hospital. The objective should be to reach out to the patient and help in improving the patent's quality of life in every way possible.

A microbiological study of bottled mineral water marketed in Ludhiana.

The microbiological quality of bottled mineral water marketed in Ludhiana was examined, Twenty three brands were analyzed for presumptive coliform count by multiple tube tests, and E. coli count was confirmed by Eijkman test. Bacterial and fungal loads were tested by membrane filtration test. Out of 23 only one sample (4.4%) showed the presumptive coliform count to be 460 most probable number (MPN)l 1 00ml,and 1 was found to be positive when tested by Eijkman test for Ecoli. In the membrane filtration test three samples (13%) showed more than two types of bacteria. Different types of bacteria isolated included Bacillus sp (19/23). Pseudomonas spp (13123), Ecoli, Klebsiella sp and S.albus one each Fungi was isolated from five of twenty three. (22%) samples. Only one brand of mineral water was unfit for human consumption. The rest of the samples were contaminated with non pathogenic flora.

Coronary risk variables in young asymptomatic smokers.

BACKGROUND AND OBJECTIVES: Smoking plays a dominant role in premature atherosclerosis particularly among males in South Asian countries. It initiates and promotes atherosclerosis by altering cardiac haemodynamics, causing dyslipidaemia and producing oxidative damage. Not much information is available from our country. We therefore undertook this study to see the effect of smoking on electrocardiogram (ECG), blood pressure, lipids, apolipoprotein B level and free radical activity in young asymptomatic male smokers. METHODS: The study included 100 consecutive male subjects (50 smokers and 50 non smokers) aged 30-40 yr. Smoking profile, detailed cardiovascular assessment including ECG and lipid profile were evaluated in each subject. RESULTS: Of the 50 smokers, 22 (44%) had grade I hypertension as against 5 of 50 non smokers. Sinus tachycardia (10%) and P-pulmonale (8%) were the only notable ECG abnormalities. Dyslipidaemia was detected in 92 per cent smokers and 48 per cent non smokers (P<0.001). Total serum cholesterol, low density lipoprotein (LDL)-cholesterol, triglycerides and apolipoprotein B levels were significantly higher (P<0.001) in smokers compared to non smokers. LDL-cholesterol was > or =135 mg/dl in 94 per cent dyslipidaemic smokers. However, no significant difference was found in high density lipoprotein (HDL)-cholesterol. Smokers had significantly higher serum malondialdehyde levels (P<0.001) and low superoxide dismutase (P<0.001) compared to non smokers. INTERPRETATION AND CONCLUSION: Our data indicate that young asymptomatic male smokers tend to have hypertension, dyslipidaemia and increased production of free oxygen radicals, perhaps by attenuation of oxidative stress by cigarette smoking. This makes them prone for premature coronary artery disease. However, the findings need to be confirmed on a larger sample.

Y.enterocolitica gastroenteritis - A prospective study.

Two thousand faecal samples were collected from diarrhoeal patients of all age groups and processed by using MacConkey agar and Yersinia selective agar [Cefsulodin-Irgasan-Novobiocin (CIN) media]. One hundred and seventy faecal samples from non-diarrhoeal (healthy controls) were also processed. Samples were subjected to cold enrichment before processing. Three hundred and seventy-four (18.7%) faecal samples were positive for bacterial pathogens, of which 41 (2.05%) had Y.enterocolitica infection. Y.enterocolitica was more common in patients above 16 years of age 23/41 (56%). Higher rates of isolation were recorded during winter months. Of the typable isolates 21.9% belonged to 0:3 serotype followed by 01, 2a, 3 (17%) serotype. Twelve (29.2%) isolates were untypable. 51.2% isolates of Y.enterocolitica belonged to biotype IV whereas 24.4%, 19.5%, 4.9% belonged to type I, II and III respectively. Majority of the isolates were found sensitive to chloramphenicol, gentamicin, amikacin, ciprofloxacin, ampicillin, netromycin, cefoperazone, tobramycin and pefloxacin. All isolates were resistant to ofloxacin.

Clinical evaluation of sibutramine in obese type 2 diabetic patients refractory to dietary management.

MATERIAL AND METHODS: An open, non-comparative study was carried out to assess the efficacy and toleration profile of sibutramine, a new antiobesity drug, in promoting weight loss in obese type 2 diabetes mellitus subject who failed to reduce weight after strict dietary control. Twenty seven patients completed the study. Sibutramine was started as a single morning dose of 10 mg and was subsequently increased to 15 mg daily if weight loss was not satisfactory. The total duration of the study was twelve weeks with followup at every four weeks. Effect of drug was monitored in terms of weight reduction, changes in body mass index, waist circumference, hip circumference, waist/hip ratio and other metabolic parameters. A fixed dietary prescription and concomitant therapy with drugs, if required and not likely to interfere with the trial therapy, was permitted but was not changed during the study period. RESULTS: At the end of 12 week, mean weight reduction in study subjects was 4.16 kg (p < 0.001), the corresponding BMI decreased by 1.6 (p < 0.0001) and hip circumference by 3.68 (p < 0.001). However, there was no significant change in fasting blood glucose and Hb(A1c) values. CONCLUSION: The study indicates sibutramine to be an effective and well tolerated agent leading to significant reduction in parameter of obesity in obese type 2 diabetic subjects.

Cryopreservation of Rh negative blood for improved storage & utilisation by means of indigenous freezing bags & solutions & manual deglycerolisation.

BACKGROUND & OBJECTIVES: Cryopreservation allows donor blood to be stored for years, rather than weeks as in liquid storage. This is an established procedure in countries with a developed blood transfusion service. Cryopreservation has not been introduced in India, possibly because of the presumed high cost and complexity of the procedure. An attempt is made in this study, to cryopreserve Rh negative blood in a mechanical freezer using indigenous bags and solutions and manual deglycerolisation to reduce the cost of preservation. METHODS: RBCs to be frozen were weighed and transferred by sterile welding to an 800 ml freezing bag. Based on RBC weight, 400-500 ml of glycerol (5.7 MpH 6.8) was added as cryopreservative. After extracellular and intracellular equilibration, excess glycerol was removed by centrifugation. Glycerolised RBCs were frozen at -80 degrees C in a mechanical freezer. Cryopreserved units were thawed at 37 degrees C and deglycerolised. Twenty one patients were transfused with the frozen deglycerolised RBCs after the usual pre-transfusion tests. RESULTS: Haematocrit (Hct) values of the frozen deglycerolised units, complied with required standards, before addition of glycerol and post glycerolisation. RBC recovery was more than 80 per cent and Hct 83.3 per cent after deglycerolisation. Twenty one patients transfused with the frozen deglycerolised blood experienced no adverse effects and showed a 24 h mean post transfusion Hb increment of 0.73 g/dl. INTERPRETATION & CONCLUSION: Cryopreservation, unlike liquid storage, allows Rh negative blood of the required ABO groups to be stocked. Cryopreservation also improves its utilisation, reduces wastage and helps supplement the liquid stocks of Rh negative blood. The use of indigenous freezing bags and manual deglycerolisation has resulted in an inexpensive procedure, which can be integrated, with minor inputs, into any blood bank that is already preparing blood components.

Reappearance of Vibrio cholerae O139 during March-August, 1998 in Ludhiana (Punjab), India.

An outbreak of V. cholerae O139 was reported from Ludhiana in 1993. Strict bacteriological vigilance between 1994 and 1997 showed no isolates of O139, while 01 strains continued to be isolated. In 1998, 30 strains of V. cholerae O139 and 15 strains of V. cholerae 01 were isolated. Both strains appear to be endemic in Punjab.

Typing and antibiotic susceptibility patterns of Vibrio cholerae during six consecutive cholera seasons in north India.

A total of 10,427 diarrhoeal stool specimens were cultured for Vibrio cholerae between 1992 and 1997. The isolation rates were 2%, 2.6%, 6.7%, 7.08%, 0.9% and 2.6% in the years from 1992 to 1997 respectively. Till 1992, Vibrio cholerae 01 ogawa was the predominant strain. In 1993, 81.3% of the isolates were of 0139 Bengal strain and the rest were V. cholerae 01. From 1994 to 1997, V. cholerae 01 ogawa was the predominant strain and there were no isolation of 0139 strain. The predominant phage type in 1992 and 1993 were T2 and T27 thereafter. Most Vibrio cholerae strains were sensitive to tetracycline, gentamycin, netromycin, norfloxacin and furazolidine. Strains were resistant to cotrimaxozole till 1996, but were 100% sensitive in 1997. Strains were sensitive to chloramphenicol till 1993 but acquired resistance thereafter.

Prevalence of multi-drug resistant Salmonella typhi in Ludhiana Punjab.

A total of 945 strains of S. typhi isolated from blood cultures during 1989 to 1994 were studied. Their antibiotic susceptibility showed 580 (61.4%) of strains to be multidrug resistant. The 464 strains tested for their susceptibility to ciprofloxacin were all sensitive to the drug. Twenty three (17.9%) of the 128 strains of S. paratyphi A were resistant to chloramphenicol. The sole isolate of S. paratyphi B was sensitive to all antibiotics tested.

Outbreak of gastroenteritis due to a new strain of non O group 1 Vibrio cholerae, in Ludhiana in May-August 1993.

The isolation of the new serotype 0139 of non 01 V. cholerae from an outbreak of gastroenteritis is reported. The study of 35 such isolates revealed their similarity with the El tor vibrios biochemically and by other characters. All were strongly haemolytic and 97.1 per cent of the strains showed a positive haemagglutination.

Evaluation of peritoneovenous shunt in refractory cirrhotic ascites in Indian patients.

Ten patients with tense and refractory cirrhotic ascites underwent a modified peritoneovenous shunt procedure. Nine patients responded with complete resolution of ascites, mean weight gain of 5 (range 0-8) Kg, and rise in serum albumin level of 0.6 g/dL. Six patients improved from Child's class B to A. The longest survival period was 2.3 years. The main constraint in a developing country is the high cost of the shunt.