Genetic and molecular analysis of leaf blast resistance in Tetep derived line RIL4 and its relationship to genes at Pita/Pita2 locus.

The Vietnamese indica landrace 'Tetep' is known worldwide for its durable and broad spectrum-resistance to blast. We performed genetic and molecular analyses of leaf blast resistance in a Tetep derived recombinant inbred line 'RIL4' which is resistant to both leaf and neck blast. Phenotypic analysis of segregating F2 progenies suggested that leaf blast resistance in RIL4 was controlled by a dominant gene tentatively designated as Pi-l(t). The gene was mapped to a 2.4 cm region close to the centromere of chromosome 12. The search for the gene content in the equivalent genomic region of reference cv. Nipponbare revealed the presence of five NBS-LRR genes, two of which corresponded to the alleles of Pita and Pi67 genes previously identified from Tetep. The two other genes, LOC_Os12g17090, and LOC_Os12g17490 represented the homologs of stripe rust resistance gene Yr10. The allelic tests with Pita2 and Pi67 lines suggested that the leaf blast resistance gene in RIL4 is either allelic or tightly linked to these genes. The genomic position of the leaf blast resistance gene in RIL4 perfectly coincided with the genomic position of a neck blast resistance gene Pb2 previously identified from this line suggesting that the same gene confers resistance to leaf and neck blast. The present results were discussed in juxtaposition with past studies on the genes of Pita/Pita2 resistance gene complex.

Expression and significance of leukocyte membrane cofactor protein transcript in systemic lupus erythematosus.

Membrane cofactor protein (MCP) is a complement regulatory protein ubiquitously expressed on most nucleated cells. Since MCP protects autologous cells from complement-mediated injury, it is suggested to have a protective role against the self-tissue damage in inflammatory conditions such as systemic lupus erythematosus (SLE). However, the relevance of MCP in human SLE is not well explored. To assess the significance of MCP in SLE, we studied expression of leukocyte MCP transcript in 60 healthy individuals (controls) and 60 patients with SLE and correlated that with the levels of circulating immune complex (CIC), C3, C3d and SLEDAI scores. The levels of leukocyte MCP transcript were significantly higher (p < 0.001) in patients with SLE than the controls. Furthermore, MCP transcript levels exhibited significant positive correlations with SLEDAI scores and CIC level and a negative correlation with C3d level in patients. Twelve patients were followed-up until remission. The levels of MCP transcripts decreased significantly during remission as compared with the state of active disease. These findings suggest that in SLE, the expression of leukocyte MCP at the mRNA level is closely related to disease activity. A protective role of MCP in response to increased disease burden may be speculated. The follow-up study suggested MCP as a potential disease marker.

Postoperative nausea and vomiting in patients undergoing total abdominal hysterectomy under subarachnoid block: a randomized study of dexamethasone prophylaxis.

BACKGROUND: Postoperative nausea and vomiting is a common distressing problem in patients undergoing gynaecological surgery under anaesthesia including central neuraxial blockade, which requires frequent medical interventions. OBJECTIVES: We aimed to find out the antiemetic effect of prophylactic dexamethasone for prevention of postoperative nausea and vomiting in patients undergoing total abdominal hysterectomy under subarachnoid block. Influences of dexamethasone on patient satisfaction and postoperative analgesia were also observed as secondary objectives. METHODS: This was a prospective, randomized, double blind, placebo controlled study conducted in BPKIHS, from January 2009 to April 2009. This study involved 80 American Society of Anaesthesiologist Physical Status I and II patients undergoing total abdominal hysterectomy under subarachnoid block. Patients were divided into two groups of 40 each to receive either 4 mg of dexamethasone (group D) or normal saline (group N) in volume of 2 ml intravenously 1 hour prior to subarachnoid block. Surgery was allowed to start with block height of at least T8 dermatome. Intraoperative and postoperative nausea and vomiting was observed using nausea and vomiting scale every 4 hour for 24 hours. RESULTS: Seven (17.4%) patients in group D and 11 (27.5%) patients in group N had nausea and vomiting in the intraoperative period (P=0.284). Sixteen (40%) patients in group D experienced nausea and vomiting in the postoperative period as compared to 27 (67.5%) in group N (P =0.0136). Accordingly, the mean requirement of rescue antiemetic was less in group D compared to Group N (P=0.042). Further, only 15 (37.5%) patients in group D required postoperative supplemental analgesic as compared to 23 (57.5%) in group N (P=0.058). After 24 hrs of surgery, 26 (65%) patients expressed satisfaction in group D as compared to 16 (40.0%) in group N (P =0.025). CONCLUSIONS: Use of dexamethasone prior to subarachnoid block in patients undergoing total abdominal hysterectomy significantly reduces the incidence of nausea and vomiting and the requirement of antiemetic in the postoperative period, with better patient satisfaction.

Intestinal alkaline phosphatase preserves the normal homeostasis of gut microbiota.

BACKGROUND AND AIMS: The intestinal microbiota plays a critical role in maintaining human health; however, the mechanisms governing the normal homeostatic number and composition of these microbes are largely unknown. Previously it was shown that intestinal alkaline phosphatase (IAP), a small intestinal brush border enzyme, functions as a gut mucosal defence factor limiting the translocation of gut bacteria to mesenteric lymph nodes. In this study the role of IAP in the preservation of the normal homeostasis of the gut microbiota was investigated. METHODS: Bacterial culture was performed in aerobic and anaerobic conditions to quantify the number of bacteria in the stools of wild-type (WT) and IAP knockout (IAP-KO) C57BL/6 mice. Terminal restriction fragment length polymorphism, phylogenetic analyses and quantitative real-time PCR of subphylum-specific bacterial 16S rRNA genes were used to determine the compositional profiles of microbiotas. Oral supplementation of calf IAP (cIAP) was used to determine its effects on the recovery of commensal gut microbiota after antibiotic treatment and also on the colonisation of pathogenic bacteria. RESULTS: IAP-KO mice had dramatically fewer and also different types of aerobic and anaerobic microbes in their stools compared with WT mice. Oral supplementation of IAP favoured the growth of commensal bacteria, enhanced restoration of gut microbiota lost due to antibiotic treatment and inhibited the growth of a pathogenic bacterium (Salmonella typhimurium). CONCLUSIONS: IAP is involved in the maintenance of normal gut microbial homeostasis and may have therapeutic potential against dysbiosis and pathogenic infections.

Serogroup distribution and virulence characteristics of sorbitol-negative Escherichia coli from food and cattle stool.

AIMS: To (i) study the serogroup distribution and virulence characteristics of non-sorbitol-fermenting Escherichia coli isolates from foods of animal origin and cattle faeces and (ii) re-examine the true sorbitol and beta-D-glucuronidase (GUD) reactions of sorbitol-negative (Sor(-)) strains from MacConkey sorbitol agar (SMAC) to assess their phenotypic similarity with E. coli O157. METHODS AND RESULTS: One hundred and thirty Sor(-)E. coli were isolated from 556 food samples and 177 cattle stool samples using cefixime tellurite-supplemented SMAC (CT-SMAC) and chromogenic HiCrome MS.O157 agar respectively. Based on typing of somatic antigen, the isolates were classified into 38 serogroups. PCR results identified about 40% strains, belonging to O5, O8, O20, O28, O48, O60, O78, O82, O84, O101, O110, O123, O132, O156, O157, O-rough and OUT as Shiga toxigenic. Majority of O5, O84, O101, O105, O123, O157, O-rough and OUT strains were enterohaemolytic. Further, 39.2% and 63.1% of Sor(-) isolates from CT-SMAC fermented sorbitol in phenol red broth and hydrolysed 4-methylumbelliferyl-beta-D-glucuronide (MUG) respectively. Members of serogroups O5, O28, O32, O81, O82, O84, O101, O-rough lacked both the sorbitol fermentation (broth test) and GUD activity and might create confusion in phenotypic identification of E. coli O157. CONCLUSIONS: Sor(-)E. coli isolates from raw meat, milk, shrimp and cattle stool belonged to 38 serogroups, with E. coli O157 constituting only 14.6% of the isolates. Many of these nonclinical Sor(-) strains were potentially pathogenic. Nearly 39% of these Sor(-)E. coli from CT-SMAC fermented sorbitol in broth, indicating the need for confirmation of sorbitol reaction in broth. SIGNIFICANCE AND IMPACTS OF THE STUDY: Classical sorbitol utilization and GUD tests are not likely definitive tests for E. coli O157. Further improvement of differential media based on these phenotypic properties is necessary for detection of pathogenic serotypes from foods and environmental samples.

Outcomes of different subgroups of smear-positive retreatment patients under RNTCP in rural West Bengal, India.

INTRODUCTION: This record-based study was undertaken at Bagula Tuberculosis Unit, Nadia, West Bengal, India to compare outcomes among sputum-positive TB retreatment patient groups (relapse, failure and treatment after default) at completion of therapy, under the Revised National Tuberculosis Control Program (RNTCP). METHODS: A total of 234 registered cases of TB retreatment (category II) between January 1999 and June 2005 were analysed and compared by Z-test for proportion. RESULTS: There was a uniform distribution in terms of age, grades of sputum positivity and sputum conversion at 2 and 3 months among the three groups. In spite of this, a favourable outcome was most likely for relapse cases, and cases with a low grade of sputum positivity in all three subgroups. Unfavourable outcome was most likely among the treatment failure subgroup and those with high grades of sputum positivity. CONCLUSION: The results are likely to be due to an increased incidence of multi-drug resistant TB in these patients. In rural areas of the developing world, as in India, there is a heavy burden of TB and resources are limited. Culture and drug sensitivity patterns prior to commencing therapy should be performed for failure and default patients who present with an initially high load of bacilli in their sputum.

Refractory Pseudomonas Bacteremia in a 2-Year-Old Sterilized by Bacteriophage Therapy.

Here, we report a complex case that involved a pediatric patient who experienced recalcitrant multidrug-resistant Pseudomonas aeruginosa infection complicated by bacteremia/sepsis; our antibacterial options were limited because of resistance, allergies, and suboptimal source control. A cocktail of 2 bacteriophages targeting the infectious organism introduced on 2 separate occasions sterilized the bacteremia.

A review on adverse event profiles of epidermal growth factor receptor-tyrosine kinase inhibitors in nonsmall cell lung cancer patients.

The epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of EGFR-mutant nonsmall cell lung cancer (NSCLC). These EGFR TKIs demonstrate a different adverse event (AE) profile as compared to conventional chemotherapy agents. They are more commonly associated with cutaneous AEs and diarrhea while hematological AEs occurred commonly with chemotherapy agents. These AEs are the extension of pharmacological effect and occur as a result of blockade of EGFR-regulated pathways in the skin and gastrointestinal tract. This review article sheds light on the safety profile of first-, second-, and third-generation EGFR TKIs based on data obtained from several clinical trials conducted in NSCLC patients and highlights trials comparing these agents with the conventional chemotherapy agents. The strategies to manage EGFR TKI-related AEs are also reviewed.

Critical evaluation of research articles in relation to informed consent.

The concept of research bioethics arose and developed over the last three decades mainly in response to moral crisis created by rapidly advancing medical science and technology. Accordingly, reinterpretation of some fundamental principles of ethics is needed. At the heart of the ethical principle of respect for person, lies the informed consent process. Nuremberg Code, Helsinki Declaration of World Medical Association (WMA), Council for International Organizations of Medical Sciences (CIOMS) Guidelines etc. demonstrated the importance of informed consent in biomedical research involving human subjects. This article was planned to evaluate the reflection of informed consent in the published research articles. Fifty-two articles published in national and international journals were studied among which informed consent was found to be taken only in 21(40.4%) occasions and the same was not taken in 31(59.6%) occasions. Study showed that out of 38 articles published in national journals, informed consent was taken from only in 12(31.6%) situations, while out of 14 articles published in international journals, informed consent was taken in 9(64.3%) situations. Out of 36 observational studies, informed consent (both written and verbal) was taken from only 27.8% cases, whereas out of 16 interventional experiments, informed consent was taken in 68.8% situations.

Percutaneous dilational tracheostomy: an initial experience in community based teaching hospital.

UNLABELLED: Percutaneous dilational tracheostomy (PDT) is frequently performed in the intensive care unit to prevent the long term complications associated with prolonged endotracheal intubation. OBJECTIVE: To report the analysis of our experience with percutaneous dilation tracheostomy. STUDY DESIGN: A prospective documentation of 40 patients who received percutaneous dilational tracheostomy in a multidisciplinary intensive care unit during a 12-month period. METHOD: The patients demographic, indications of intubation and PDT, time required to perform the procedure, complications and the outcome of these patients in the intensive care unit were noted. RESULT: Among 425 patients, 40 underwent percutaneous dilational tracheostomy that included 22 females and 18 males with the median age of 35 years. Prolonged ventilatory support was the most common indication for tracheostomy. The average duration of intubation before PDT was 5 days. Median procedure time was 20 minutes. Complications included minor bleeding in two (5%), subcutaneous emphysema with pneumothorax in two patients (5%), tracheal stenosis in three (7.5%), tracheo-esophageal fistula and glottic granuloma in one patient each (2.5%). Among forty patients, 28 (70%) were discharged to the ward, 8 died in intensive care unit and 4 left hospital against medical advice. CONCLUSION: Percutaneous dilational tracheostomy is a safe, quick and effective way for long term airway management in critically ill patients.

A new species of Physomerus Burmeister (Hemiptera: Heteroptera: Coreidae: Coreinae), with a key to the species of India.

A new species from India, Physomerus centralis sp. nov. (Hemiptera: Heteroptera: Coreidae) is described and illustrated with both male and female genitalia. Morphological measurements and their ratios were taken as additional diagnostic characters. A key to the Indian species of the genus Physomerus Burmeister is provided.

Primitive neuroectodermal tumors of kidney: Our experience in a tertiary care center.

OBJECTIVES: Primitive neuroectodermal tumors (PNET) are rare highly aggressive neoplasms. The diagnosis is made by histopathology with the support of immunohistochemistry (IHC) and cytogenetics. The aggressive multimodality treatment is recommended for the management of these tumors. The purpose of our study is to review our experiences in the diagnoses and treatment of PNET of the kidneys. MATERIALS AND METHODS: We retrospectively reviewed the data of all the patients, who were treated for the PNET of the kidneys at our institute between April and March 2011 and compared with the available literature. RESULTS: A total of eight patients were treated for PNET of the kidney. Out of the eight patients, four were males and four females. Nearly 50% of our patients had inferior vena caval thrombus at the time of presentation. The diagnosis was made on histopathology supported by IHC. Out of the eight patients, one patient had intraoperative death due to massive pulmonary thromboembolism and another died on the 7th post-operative day due to disseminated intravascular coagulation and multiorgan failure. Rest six patients were treated with post-operative chemotherapy or a combination of chemotherapy and radiotherapy. For these six patients, overall median survival was 45 months with a 3 year disease-free survival of 66% and 5 year survival of 44%. CONCLUSIONS: PNET of the kidneys are rare peripheral neuroectodermal tumors with an aggressive clinical course. These tumors carry a very poor prognosis. An aggressive treatment approach using a combination of surgery, chemotherapy and radiotherapy is recommended for a reasonable survival in these tumors.

A new species of Stenolemus Signoret (Hemiptera: Heteroptera: Reduviidae: Emesinae) from India.

A new species of Stenolemus Signoret, 1858, Stenolemus annulatus sp. nov. (Reduviidae: Emesinae) is described from India. This new species is closely related to S. susainathani Wygodzinsky from Nilgiri Hills, South India and S. larat Wygodzinsky from Moluccas, Larat, by sharing common characteristics such as the ratio of distance between eyes and their width, long antennal first segment, slightly elevated postocular region of head and sharply divided pronotum into three parts, but can be easily separated by the following diagnostic characteristics: pronotum with petiole about as long as forelobe and 1.3 times shorter than hindlobe, first antennal segment with five annulations, rostrum creamy white and luteous at apex, basal and lateral areas of second segment; hind lobe of pronotum with 1+1 submedian conical rounded projections; forecoxa shorter than hind lobe of pronotum, foretibia with one subbasal and one submedian annulation and hind femora reaching apex of forewings. A key to Indian species of the genus Stenolemus is given. 

Uropathogenic Escherichia coli (UPEC) induced antimicrobial gene expression in the male reproductive tract of rat: evaluation of the potential of Defensin 21 to limit infection.

Escherichia coli (E. coli) is a common pathogen in epididymitis, which represents a prevalent entity in male reproductive tract infections (RTI). Although current treatment regimens using antibiotics are satisfactory, development of antimicrobial resistance by the pathogen represents a challenge in the management of RTI. Hence, identification of antimicrobial peptides as alternatives to antibiotics has gained importance. We demonstrate that in a rat epididymo-orchitis model induced with uropathogenic E. coli (UPEC) strain MTCC 729, the expression of defensins and defensin-like Spag11 genes are induced in the epididymis and testes. The induction of antimicrobial gene expression is paralleled by phosphorylation of the NF-kB subunit p65 and the inhibitor of NFkB (IkB-alpha), decreased levels of histone deacetylase 1 and increased methylation of Histone 3, indicating the role of classical Toll-like receptor mediated signaling and epigenetic regulation. Recombinant Defensin 21, when administered to UPEC-infected rats, substantially reduced the bacterial load in the epididymis and testis and proved to be more effective than gentamycin. The ability of Defensin 21 to limit RTI provides support that antibacterial proteins of the male reproductive tract may be used as potential alternatives to antibiotics in treatment of this disease.

Expression of the Arabidopsis thaliana 2S albumin gene 3 in Saccharomyces cerevisiae.

The Arabidopsis thaliana (At) 2S albumin gene 3 (At2S3) has been cloned in YEp13 as a 3.5-kb genomic fragment. To study its expression in Saccharomyces cerevisiae, the accumulation in saturated cultures reached about 0.032% of the yeast total protein, and the product was localized in vacuolar bodies within the cell. The 13-kDa protein was processed to 9- and 4-kDa proteins, as obtained in transgenic tobacco plants.

Isolation, characterization and sequencing of a novel repetitive DNA from the mung bean Vigna radiata.

A family of highly reiterated, small (approx. 300 bp) sequences has been identified in DNA of the mung bean Vigna radiata. The members are extensively interspersed throughout the chromosomes with some clustering. They also occur extrachromosomally. There is no tissue-specificity to the repeat family but it is highly species-specific. The repetitive DNA hybridizes to total RNA as well as to polyadenylated RNA isolated from germinated mung beans. It has analogy with the human AluI family in the mode of isolation, size, genomic distribution, copy number and transcribability though they do not share any sequence homology. A repetitive DNA clone was selected from a shotgun genomic library of mung bean DNA in pBR322. The average copy number of the cloned repeat is estimated to be 8 x 10(4) per haploid genome, and thus constitutes approx. 5% of the total mung bean genome. The genomic organization and transcription of the cloned repeat is reported. Sequencing of the cloned repetitive DNA reveals the presence of the number of direct and inverted repeats and some short palindromic sequences.

Characterisation of a repetitive DNA family from Entamoeba histolytica containing Saccharomyces cerevisiae ARS consensus sequences.

Several repetitive DNA families were identified in Entamoeba histolytica DNA digested with Sau3AI. Characterisation of one of these repetitive DNA families showed the presence of multiple copies of Saccharomyces cerevisiae autonomously replicating sequence (ARS) core consensus sequences. The E. histolytica ARS consensus sequences allowed a yeast-integrating plasmid, YIP5, to replicate autonomously in S. cerevisiae. A 'bent DNA' fragment was located in one member of this E. histolytica repetitive DNA family.

Interaction of colchicine analogues with purified tubulin.

Binding of two colchicine analogues, desacetamidocolchicine and 2-methoxy-5-(2',3',4'-trimethoxyphenyl) tropone to purified tubulin have been studied. Both analogues bind to tubulin with a significant increase in fluorescence polarization of the drugs in solutions containing tubulin. The Kd for tubulin-drug complexes were found to be 1.25 X 10(-6) M and 1.08 X 10(-6) M for desacetamidocolchicine and 2-methoxy-5-(2',3',4'-trimethoxyphenyl) tropone, respectively. Scatchard analysis of the fluorescence titration curve of drug tubulin interaction also gives the values of stoichiometry and affinity constant. These were 0.8 and 1.6 X 10(6) M-1 for desacetamidocolchicine, and 0.9 and 0.58 X 10(6) M-1 for 2-methoxy-5-(2',3',4'-trimethoxyphenyl) tropone.

Inositol(1,3,4,5) tetrakisphosphate plays an important role in calcium mobilization from Entamoeba histolytica.

Calcium release from internal stores of Entamoeba histolytica, a parasitic protozoan, was observed by measuring fluorescence of Fura-2. Emptying of inositol(1,4,5)trisphosphate (Ins(1,4,5)P3)-sensitive calcium pools in permeabilized E. histolytica did not significantly affect subsequent calcium release by inositol(1,3,4,5)tetrakis-phosphate (Ins(1,3,4,5)P4). Similarly, prior depletion of Ins(1,3,4,5)P4-sensitive stores did not have any influence on subsequent calcium release by Ins(1,4,5)P3. The EC50 for calcium release was 0.15 microM with Ins(1,4,5)P3 and 0.68 microM with Ins(1,3,4,5)P4. In conclusion, the Ins(1,3,4,5)P4-sensitive calcium store in E. histolytica is separate and independent from the Ins(1,4,5)P3-sensitive pool.