Analysis of antibiotic resistance and virulence patterns in Klebsiella pneumoniae isolated from human urinary tract infections in India.

In this study, 504 urine samples collected from patients suffering from urinary tract infection (UTIs) were screened for the presence of Klebsiella pneumoniae. The overall occurrence of Klebsiella spp. and K. pneumoniae was found to be 23·2% (117/504) and 16·8% (85/504) respectively. Antibiotic susceptibility testing of 85 K. pneumoniae isolates was carried out by disc diffusion which revealed alarming levels of antibiotic resistance (ABR). Antimicrobial resistance was prominently observed against cefpodoxime (76·47%) followed by ampicillin (70·59%), ceftriaxone (52·94%), cefoxitin (50·59%), amoxyclav (48·24%), ofloxacin (45·88%), cefotaxime (44·71%), cefepime (43·53%) and doxycycline hydrochloride (40%). A small percentage of strains also exhibited resistance to other antimicrobials in the range of 7-35%. Around 77·6% of the isolates were found to be resistant to three or more antibiotic classes and 66·7% of the isolates had multiple antibiotic resistance index values >0·2. Screening of virulence genes in 85 K. pneumoniae isolates revealed that uge gene was the most predominant (11/85, 12·9%), followed by rmpA (9/85, 10·5%), kfu (4/85, 4·7%) and aerobactin genes (2/85, 2·35%). Further, the overall percentage of biofilm producers were found to be 17·65% (15/85). This study warrants hospitals and health care centres to reduce misuse of antibiotics and manage UTI with appropriate treatment after performing antibiotic susceptibility testing.

Studies on the effect of antidiabetic drugs on collagen in rats.

To assess the effects of antidiabetic drugs on collagen, the study has been carried out. However, at present, no conclusive data available on the In vivo effect of antidiabetic drugs in collagen. It was therefore thought worthwhile to undertake a systemic study on the metabolic turnover of connective tissue proteins, particularly collagen which will help to understand the various changes taking place in conventional antidiabetic drug treated rats. The hydroxyproline content was determined by the method of Neumann and Logan. There was no significant difference in mean body weight of animals in treated groups when compared to control animals on 50th day. In drug treated group G5, urinary excretion of hydroxyproline was slightly increased in contrast to that of the rats in other groups and showed a significant difference (p < 0.001) between control and G5 group. The proportion of soluble collagen in treated group was somewhat higher than in the control but while taking the significance value there are no statistically significant changes. Liver showed the highest concentration of 0.98 g collagen/100 g tissue, whereas kidney and heart showed 0.57 and 0.72 g collagen/100 g tissue. Observations of the present study collectively indicate that the anabolic rate of collagen metabolism is not altered significantly by the administration of antidiabetic drugs in normal rats.

Assessment of Drug-Drug Interactions among Renal Failure Patients of Nephrology Ward in a South Indian Tertiary Care Hospital.

Polypharmacy is common in drug prescriptions of chronic kidney disease patients. A study of the prescription patterns of drugs with potential interactions would be of interest to prevent drug related adverse events. A prospective observational study of six months (Dec 2009-May 2010) was carried out among the chronic kidney disease patients admitted to the nephrology ward of a South Indian tertiary care hospital. The pattern and rates of drug-drug interactions seen in the prescriptions of these patients was studied. Among the 205 prescriptions included, a total of 474 interactions were reported, making 2.7 interactions per prescription with incidence rates of 76.09%. Around 19.62% of interactions were of major severity. Most common interactions were found between ascorbic acid and cyanocobalamine (12.45%), clonidine and metoprolol (3.80%) respectively. Hypo or hypertension (31.65%), decreased drug efficacy (29.11%) and hypo or hyperglycemia (14.14%), were the most commonly reported clinical outcomes of the drug interactions. Cardiovascular drugs (calcium channel blockers and beta blockers; 52%) constitute the major class of drugs involved in interactions. As most of the interactions had a delayed onset, long term follow-up is essential to predict the clinically significant outcomes of these interactions. Hence, drug interactions are commonly seen in the prescriptions of chronic kidney disease patients which can lead to serious adverse events if not detected early. Need for collaboration with a clinical pharmacist and electronic surveillance, which are absent in developing countries like India, is emphatic.

Collagen-chitosan polymeric scaffolds for the in vitro culture of human epidermoid carcinoma cells.

A biodegradable polymer scaffold was developed using collagen and chitosan, in the form of interpenetrating polymeric network (IPN), for in vitro culture of human epidermoid carcinoma cells (HEp-2, Cincinnati). Glutaraldehyde was used as cross-linking agent for the development of scaffold. Various types of scaffolds were prepared using different proportionate mixtures of collagen and chitosan solutions in the ratio of 3:7, 4:6, 5:5, 6:4 and 7:3 (collagen:chitosan). These scaffolds were fully characterized by Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC) and Thermogravimetric analysis (TGA). Equilibrium swelling studies were carried out in phosphate buffer of physiological pH (7.4) to study its swelling characteristics at slightly alkaline pH. The scaffold that showed optimum swelling property was selected as the best scaffold for performing in vitro culture studies. In vitro culture studies were carried out using HEp-2 cells, over the selected scaffold and its growth morphology was determined through optical photographs taken at different magnifications at various days of culture. The results of the above studies suggest that the scaffolds prepared from collagen and chitosan can be utilized as a substrate to culture HEp-2 cells and can also be used as an in vitro model to test anticancerous drugs.

Studies on trypsin-modified bovine and human lens acylpeptide hydrolase.

Acylpeptide hydrolase removes the N -acetylated amino acids from the peptide substrates but not from intact proteins. Cleavage between amino acid residues 203--204 of the native acylpeptide hydrolase results in the formation of a 55 kDa truncated active enzyme in the bovine lens, in vivo. In this study we explored the hydrolytic properties of the truncated enzyme using lens beta- and gamma-crystallins as substrates. SDS--PAGE analysis indicated that the beta B2-crystallin was cleaved by truncated acylpeptide hydrolase into several protein fragments (10--26 kDa). No cleavage of the gamma-crystallins was observed under similar conditions. Both the acylpeptide hydrolase activity and the protease activity of the 55 kDa enzyme were completely inhibited by diisopropylfluorophosphate, p -chloromercuribenzoate and ebelactone, and moderately inhibited by N -tosyl phenylalanine chloromethyl ketone. SDS--PAGE analysis followed by fluorography of ((3)H) diisopropylfluorophosphate labeled human lens acylpeptide hydrolase preparation showed the presence of the 55 kDa truncated form of the enzyme, as observed in the bovine lens. The peptide (d)-AIKGDQFL-NH(2)--the amino acid sequence 200--207 of the native bovine acylpeptide hydrolase with an in vivo cleavage site of native protein--was hydrolysed by the lens protease(s) suggesting that the in vivo generation of the 55 kDa acylpeptide hydrolase may be mediated through a proteolytic processing. The protease(s) responsible for the cleavage of this peptide was inhibited by diisopropylfluorophosphate and p -chloromercuribenzoate.

A profile of lung cancer patients in Hyderabad.

One hundred and sixty cases of bronchogenic carcinoma were retrospectively analysed. Male to female ratio was 8.4:1, common age group being 40-60 years. The most common (38%) radiological presentation was mass lesion. The predominant histological cell type was squamous cell carcinoma (67%). Most of the patients presented in an advanced stage of the disease of inoperability. The study underlines the various diagnostic madalities in diagnosing lung cancer and emphasises the need for practising doctors to keep in mind the possibilities of bronchogenic carcinoma in all cases of unresolved pneumonia and mass lesion on chest x-ray especially in high risk patients.

Investigations on wound healing by using amphibian skin.

The present investigation is aimed to justify the validity of traditional practice of using the frog skin in wound healing. Studies were carried out on skin wounds from female albino rats of Wistar strain weighing 100-120 g. The wounds of standard size (5 cm2) were created in the experimental and control groups. The experimental group was dressed with dorsal skin of freshly sacrificed frogs (Rana tigrina) while the control group was dressed with cotton gauze. A faster healing was observed in the experimental group over the control group. Biochemical estimations of the wound granulation tissue were carried out every 2 days till the complete healing of the wound in both the groups. The collagen of frog skin was extracted and polyacrylamide gel electrophoresis was carried out to characterize and identity the molecular organization of collagen type.

Inflammatory fibrous histiocytoma of the posterior mediastinum.

A case of malignant fibrous histiocytoma (MFH) of inflammatory type from posterior mediastinum which is a rare site is reported here. Its clinico-pathological features are discussed.

Toxicity of dimethoate on urinary hydroxyproline in rats.

The effect of the administration of three different concentrations (2.25, 1.13 and 0.56 mg/100 g body weight) of dimethoate on the urinary excretion of total, non dialysable and free hydroxyproline were studied in female albino rats. Compared to controls, the urine contents were significantly lower in higher concentration (2.25 mg/100 g body weight) of dimethoate treated rats. The results clearly indicate that the urine contents were not much affected in lower concentration (0.56 mg/100 g body weight) of dimethoate treated rats compared to the other concentrations (1.13 and 0.56 mg/100 g body weight) of dimethoate treated rats.

Toxic effects of different concentrations of dimethoate on lysosomal enzymes of female albino rats.

The effect of three different concentrations of dimethoate on the activity of certain lysosomal enzymes, viz. beta-glucuronidase, beta-N-acetylglucosaminidase, cathepsin B and cathepsin D in serum, skin, liver, kidney and spleen and the stability of liver and kidney lysosomes was studied in female albino rats. The activity of beta-glucuronidase, beta-N-acetylglucosaminidase, cathepsin D was found to increase in serum and tissues in higher concentration (2.25 mg/100 g body weight) of dimethoate treated rats. A significant increase in the rate of release of beta-glucuronidase was found in the liver and kidney of higher concentration of dimethoate treated rats compared to controls. The results demonstrate that the activity of lysosomal enzymes increased in higher concentration of dimethoate treated rats than the lower concentration (0.56 mg/100 g body weight) of dimethoate treated rats.

Effects of dimethoate on collagen metabolism in rats.

The effects of three different doses of dimethoate on the collagen metabolism in the tissues of female albino rats were studied by measuring the specific and total activities of 3H-hydroxyproline in the dermal, gingival and uteral collagen fractions and in the urine. Compared to controls, the total activity of 3H-hydroxyproline in the soluble collagen and in the urine at 12 h after the administration of 3H-proline was significantly lower by 44.45 and 58.12 per cent in the higher dose (2.25 mg/100 g body weight) of dimethoate treated groups respectively. The urinary excretion of hydroxyproline and the total activity of urinary 3H-hydroxyproline measured after 28 days of injection of labelled proline were decreased by 45.56 and 32.68 per cent in higher doses of dimethoate treated animals respectively but the excretions of urinary 3H-hydroxyproline were decreased by 6.36 and 2.88 per cent in lower doses of dimethoate (0.56 mg/100 g body weight) treated animals. The results of the present investigation clearly indicate that the synthesis of collagen is decreased in the higher doses of dimethoate treated animals compared to lower doses of dimethoate treated animals. In addition, the rates of catabolism of both soluble and insoluble collagens were decreased in higher doses of dimethoate treated rats. In concludes that the lower doses of dimethoate (0.56 mg) treated rats were less affected than the higher doses of dimethoate (2.25 mg) treated rats.

Toxic effect of dimethoate on the crosslinking of gingival and uteral collagen in female albino rats fed with normal and high protein diets.

The effect of dimethoate on gingival and uteral collagen crosslinking was studied in normal and high protein diets fed female albino rats. The gingival and uteral samples were collected from all the groups of rats and percent reversibility of neutral salt soluble collagen gel and the solubility of insoluble collagen in KCNS or urea were determined. The analysis of gingival and uteral samples showed alpha 1 and alpha 2 sub units of neutral salt soluble collagen appreciably increased beta chains and aldehyde content significantly decreased in dimethoate treated animals compared to controls. The results indicate that due to the effect of dimethoate, the crosslinking and maturation of collagen are impaired.

Toxicological effects of an organophosphorus pesticide (dimethoate) on urinary collagen metabolites in normal and high protein diets fed female albino rats.

The effect of an organophosphorus pesticide (dimethoate) on the urinary excretion of hydroxyproline (total, nondialysable, dialysable and free fractions) and hydroxylysylglycosides, glucosylgalactosyl hydroxylysine and galactosehydroxylysine was investigated in two groups of female albino rats fed with normal and high protein diets. In comparison to controls, dimethoate treated animals were found to excrete significantly decreased amounts of urinary hydroxyproline fractions from 7th day onwards. The excretion of total hydroxylysylglycoside in urine parallels the excretion of hydroxyproline. The urinary output of both glu-gal-hyl and gal-hyl was also appreciably lower from dimethoate treated animals. The normal ratio of glu-gal-hyl and gal-hyl found in the urine of dimethoate treated animals was discussed in light of decreased turn over of collagen in both bone and skin. The effect of dimethoate in rats fed with high protein diet was comparatively less than those fed with normal diet.