Phenotypic expression of blast resistance gene Pi54 is not affected by its chromosomal position.

KEY MESSAGE: This is a novel report in which chromosomal position of the rice blast resistance gene Pi54 was not found to affect significantly the resistance phenotype or morphological traits. Blast disease caused by Magnaporthe oryzae is a serious constraint in rice production at global level. Pi54 gene imparts resistance against M. oryzae. Three different transgenic lines containing Pi54 and its orthologue Pi54rh were shown to be resistant to different races of M. oryzae. To determine the chromosomal location of Pi54 gene in transgenic lines, inverse PCR was performed. Our analysis showed that in two transgenic lines, Pi54 gene was integrated on chromosomes 6 and 10 at 12.94 and 22.30 Mb, respectively. Similarly, Pi54rh allele was integrated on chromosome 1 at 16.25 Mb. The Pi54 gene present on chromosome 6 was located in a non-coding region whereas in the other TP-Pi54 line, the gene was introgressed on chromosome 10 in between the coding region of SAP domain gene. The Pi54rh was also located in the non coding region flanked by the retrotransposon genes. These rice lines were evaluated for eight different traits related to seed and plant morphology and agronomic features for two consecutive years. The transgenic lines containing Pi54 gene have higher tiller number, grain weight, epicotyl length, and yield compared to the non-transgenic control. Multivariate correlation analysis shows that blast resistance was positively correlated with the number of tillers; thousand grain weight and epicotyl length. These results will facilitate precise utilization of Pi54 gene and its orthologue in breeding programs for the development of rice cultivars with broad spectrum and durable resistance to M. oryzae.

Polysaccharides and lipoproteins as reactants for the synthesis of pharmaceutically important scaffolds: A review.

The growing number of diseases in the past decade has once again highlighted the need for extensive research on the development of novel drugs. There has been a major expansion in the number of people suffering from malignant diseases and types of life-threatening microbial infections. The high mortality rates caused by such infections, their associated toxicity, and a growing number of microbes with acquired resistance necessitate the need to further explore and develop the synthesis of pharmaceutically important scaffolds. Chemical entities derived from biological macromolecules like carbohydrates and lipids have been explored and observed to be effective agents in the treatment of microbial infections and diseases. These biological macromolecules offer a variety of chemical properties that have been exploited for the synthesis of pharmaceutically relevant scaffolds. All biological macromolecules are long chains of similar atomic groups which are connected by covalent bonds. By altering the attached groups, the physical and chemical properties can be altered and molded as per the clinical applications and needs, this ring them potential candidates for drug synthesis. The present review establishes the role and significance of biological macromolecules by articulating various reactions and pathways reported in the literature.

Antifungal activity of novel azetidine tethered chitosan synthesized via multicomponent reaction approach.

The increasing incidences of fungal infections among Covid-19 infected patients is a global public concern and urgently demands novel antifungals. Biopolymers like chitosan hold unique structural properties and thus can be utilized in the synthesis of biologically important scaffolds. To address the current scenario, the author's synthesized novel chitosan-azetidine derivative by adopting one-pot multicomponent reaction approach. The influence of chemical modification on the structural characteristics was investigated by means of spectroscopic techniques viz. FT-IR and 1HNMR and elemental analysis. Additionally, the authors investigated the antifungal potential of chitosan-azetidine derivative against Aspergillus fumigatus 3007 and the results indicated higher antifungal effect with an antifungal inhibitory index of 26.19%. The SEM and confocal microscopy images also reflected a significant inhibitory effect on the morphology of fungal mycelia, thus reflecting the potential of synthesized chitosan-azetidine derivativeas a potential antifungal agent.

Long-term quality-of-life outcomes in children undergoing adenotonsillectomy for obstructive sleep apnoea: a longitudinal study.

OBJECTIVES: To assess a cohort of patients who underwent adenotonsillectomy for obstructive sleep apnoea, 4 years after surgery for evidence of continued and long-term improvement in quality of life. We also sought to compare our results to Child Health Questionnaire scores obtained from our previous study. We also compared our data with a healthy UK children population from normative data available. DESIGN: Longitudinal study. SETTINGS: University Hospital Tertiary Referral Centre. PARTICIPANTS: A 4-year follow-up study of 37 children who underwent adenotonsillectomy for obstructive sleep apnoea confirmed on polysomnography. There were 19 boys and 18 girls from our initial cohort. The primary caregiver completed the validated Child Health Questionnaire Parental Form version-28, 4 years after initial surgery. Our control group consist of 221 healthy children aged 6-18 that were included as 'normal' controls in a study looking at children with juvenile arthritis. The children were defined as healthy by a physician and/or after declaration by the parent. MAIN OUTCOME MEASURE: Child Health Questionnaire Parental Form version-28 scores. RESULTS: A total of 33 patients (89%) from our initial cohort were contacted. The mean age was 10.6 (median, 11; range, 5-16). When compared with results obtained 3 months postoperatively, the mean scores were higher in five domains and were statistically significant in three subscales (Role Limitations P < 0.00001; Bodily Pain P < 0.002; and Global Health P < 0.02). There was a significant deterioration in Behaviour subscale (P < 0.0007) in spite of surgery. Compared with controls, 4-year follow-up scores were higher in five domains with the Global Health domain (P < 0.0004) being statistically significant. When the 4-year follow-up scores were compared with preoperative values, these were higher in all 13 domains with statistically significant improvements in nine domains, indicating that improvements had persisted 4 years after surgery. At 4 years, however, the means scores in many domains remain lower when compared with controls. CONCLUSION: Quality-of-life data are an important measure when deciding on a specific clinical intervention. In the short term, quality-of-life measures have been shown to improve after adenotonsillectomy for obstructive sleep apnoea. Our study demonstrates that the benefits of surgery are still persistent and the children continue to improve in the long term.

Improving tracheostomy care: a prospective study of the multidisciplinary approach.

OBJECTIVES: Suboptimal standards in tracheostomy care have been highlighted as a growing concern in view of the increasing demands for intensive care services. Our objective is to assess the impact of our model for tracheostomy care on patients with short-term tracheostomies (<4 months in situ) following their discharge from the intensive care unit. The model has three components: The St Mary's tracheostomy care bundle checklist, a dedicated tracheostomy multidisciplinary team and an educational programme. DESIGN: A 38-month prospective cohort study. SETTING: A London Teaching Hospital. PARTICIPANTS: A total of 102 patients with tracheostomy within the 19-month pre-intervention cohort and 95 patients in the 19-month post-intervention cohort. MAIN OUTCOME MEASURES: The number of clinical incidents, mean time taken for decannulation, mean total tracheostomy time and total number of days spent in the intensive care unit were assessed before and after the intervention. RESULTS: Time to decannulation following intensive care unit discharge decreased from 21 to 11 days, as did the mean total tracheostomy time, from 34 to 25 days (both statistically significant with a P < 0.0001 Mann-Whitney U-test). The number of critical incidents, which included all patients prior to exclusion, substantially declined following the introduction of intervention from 58 to 7 in the second year after intervention. CONCLUSIONS: A multidisciplinary care model significantly expedited the decannulation process and reduced the overall time that a tracheostomy was in situ. The intervention was associated with a reduction in clinical incidents and shorter intensive care unit admissions, which can be associated with significant monetary savings.

Carbohydrate hitched imidazoles as agents for the disruption of fungal cell membrane.

The fungal diseases represent an increasing global health burden and have transformed from a rare curiosity to the leading cause of human mortality. The present manuscript reports the antifungal potential of two novel compounds possessing a carbohydrate and an imidazole moiety. Antifungal susceptibility test determined the growth inhibition potential of the synthesized compounds against Aspergillus niger 9689 and it was observed that compounds D and E gave an antifungal inhibitory index of 66.66 and 56.67% respectively. Further, ultra-structure analysis of the treated fungal mycelia through scanning electron microscope (SEM) and confocal microscopy indicated significant membrane permeability and disintegration of fungal cell membrane, thus highlighting the probable role of the synthesized compounds as inhibitors of fungal lanosterol 14α-demethylase. In silico studies corroborated with the in-vitro results, as the synthesized compounds interacted with the critical amino acids present at the active site of the fungal enzyme (lanosterol 14α-demethylase).

Acute cortical necrosis in Falciparum malaria: an unusual manifestation.

The spectrum of acute renal failure in Falciparum malaria varies from mild urinary abnormalities to acute renal failure. Acute tubular necrosis has been reported in 1% patients, and acute cortical necrosis has rarely been reported. We present a case of acute cortical necrosis in a young patient with Falciparum malaria who had a prolonged oligo-anuric course followed by partial recovery of renal function.

Driving standards in tracheostomy care: a preliminary communication of the St Mary's ENT-led multi disciplinary team approach.

OBJECTIVES: To assess tracheostomy care and improve standards following the introduction of an ENT-led multidisciplinary tracheostomy ward round service. DESIGN: Prospective third cycle audit. SETTING: Tertiary academic London hospital serving an inner city population of multi-ethnic background (St Mary's Hospital, Paddington, London). PARTICIPANTS: Patients with a tracheostomy discharged from ITU to general wards. IMPLEMENTED ACTIONS: Establishment of an ENT-led Tracheostomy Multidisciplinary Team (TMDT). Weekly TMDT ward round to manage patients with a tracheostomy. ENT-led educational and training sessions for allied healthcare professionals. MAIN OUTCOME MEASURES: Compliance with local tracheostomy care guidelines (St Mary's tracheostomy care bundle) and time to tracheostomy tube decannulation. RESULTS: Preliminary results of 10 patients show improved compliance with tracheostomy care guidelines, established in 2004, rising to 94%. Average time to decannulation was significantly reduced from 21 to 5 days (P-value = 0.0005, Mann Whitney Wilcoxon Test). The mean total tracheostomy time was reduced from 34 to 24 days although this was not statistically significant (P-value = 0.13, Mann Whitney Wilcoxon Test). CONCLUSIONS: The introduction of regular ENT-led multidisciplinary input for patients with a tracheostomy significantly improved compliance with nursing care standards. There was also a reduction in the total length of time tracheostomy tubes remain in situ, with time to decannulation significantly reduced.

How Robust are our Methods of Detecting Impaired Glomerular Filtration Rate in the Intensive Care Unit?

BACKGROUND: Serum creatinine is not a sensitive marker to assess early loss of renal function in acute kidney injury. Timed creatinine clearance and several formula used to predict glomreular filtration rate have not been validated. METHODS: In a prospective observational study in 50 adult patients admitted to the intensive care unit with apparent normal renal function, we assessed the glomerular filtration rate by the formula methods and timed creatinine clearance. RESULT: The mean serum creatinine was 0.77mg/dl, SD ± 0.15 (range 0.5-1.14 mg/dl). The mean measured creatinine clearance was 87.15 ml/min/1.73m(2), SD ± 20.5 (range 56.9-137 ml/min/1.73m(2)). In 25 (50%) patients, one hour urinary creatinine clearance was <80 ml/min/1.73m(2) and in two (4%) patients, the creatinine clearance was <60 ml/min/1.73m(2). Spearman correlation coefficient and regression analysis revealed a statistically significant correlation for the Cockcroft-Gault and predictive equations when compared with measured creatinine clearance. The differences between the predictive equations and creatinine clearance, as illustrated by the ±95% confidence interval in the Bland-Altman graphs was very significant [Cockcroft- Gault = -40.3 to 17.7 ml/min/ 1.73m(2), Modification of Diet in Renal Disease equation = -46.2 to 30.6 ml/min/1.73m(2) and the simplified Modification of Diet in Renal Disease equation = -72.8 to 24.8 ml/min/1.73m(2)]. CONCLUSION: Formula methods and creatinine clearance are more sensitive than serum creatinine in detecting early phase of acute kidney injury. However, there is no agreement between these methods of glomerular filtration rate estimation.

Deficient gene expression in protein kinase inhibitor alpha Null mutant mice.

Protein kinase inhibitor (PKI) is a potent endogenous inhibitor of the cyclic AMP (cAMP)-dependent protein kinase (PKA). It functions by binding the free catalytic (C) subunit with a high affinity and is also known to export nuclear C subunit to the cytoplasm. The significance of these actions with respect to PKI's physiological role is not well understood. To address this, we have generated by homologous recombination mutant mice that are deficient in PKIalpha, one of the three isoforms of PKI. The mice completely lack PKI activity in skeletal muscle and, surprisingly, show decreased basal and isoproterenol-induced gene expression in muscle. Further examination revealed reduced levels of the phosphorylated (active) form of the transcription factor CREB (cAMP response element binding protein) in the knockouts. This phenomenon stems, at least in part, from lower basal PKA activity levels in the mutants, arising from a compensatory increase in the level of the RIalpha subunit of PKA. The deficit in gene induction, however, is not easily explained by current models of PKI function and suggests that PKI may play an as yet undescribed role in PKA signaling.

Hepatitis C virus-related fibrosing cholestatic hepatitis in a renal transplant recipient.

Fibrosing cholestatic hepatitis (FCH) is a severe and progressive form of liver dysfunction seen in organ transplant recipients and immunosuppressed patients; it is usually associated with hepatitis B virus infection. We report 36-year-old man, a renal transplant recipient, also developed FCH with hepatitis C virus infection and succumbed to it.

Radiohalogenation of a monoclonal antibody using an N-succinimidyl 3-(tri-n-butylstannyl)benzoate intermediate.

N-Succinimidyl 3-(tri-n-butylstannyl)benzoate (ATE) was elevated for its utility in the radiohalogenation of monoclonal antibodies. The F(ab')2 fragment of monoclonal antibody OC 125 was labeled with 125I using the ATE reagent and with 131I using a conventional electrophilic iodination method (Iodogen). N-Succinimidyl 3-[125I]iodobenzoate was synthesized from ATE in greater than 90% yield and purified using a disposable silica gel cartridge. About 60-65% of the radioiodinated product was coupled to the F(ab')2 fragment after a 30-min reaction. Two procedures were investigated, one involving exposure of antibody to 35 nmol of ATE and the other to 240 nmol of ATE. Using Scatchard analyses, affinity constants for binding to CA 125 antigen for OC 125 F(ab')2 labeled using the low ATE, Iodogen, and high ATE procedures were determined to be (5.2 +/- 1.0) x 10(10), (2.5 +/- 0.9) x 10(10), and (4.2 +/- 2.4) x 10(9) M-1, respectively. Paired-label studies in athymic mice bearing OVCAR-3 tumors treated with injections of antibody labeled via both ATE and Iodogen demonstrated that use of the ATE method (a) reduced thyroid uptake to less than 0.1% of the injected dose, more than 100 times less than that observed with Iodogen; (b) resulted in more rapid clearance of activity from normal tissues; and (c) with the low ATE preparations, increased the uptake of radioactivity in tumor from 27 to 49%. At 96 h, tumor:tissue ratios were generally at least 4-fold higher when antibody was labeled via ATE. These results suggest that the ATE method may be a valuable approach for the radiohalogenation of antibodies.