Genetic diversity and population structure of Fusarium fujikuroi causing Bakanae, an emerging disease of rice in India

45-52Bakanae caused by Fusarium fujikuroi (Nirenberg), is emerging as a serious threat for rice (Oryza sativa. L.) cultivation in India. In this study, 63 isolates of Fusarium fujikuroi isolated from symptomatic diseased plants were characterized for their morphology, pathogenicity and molecular variability using universal rice primers (URP). Of the 12 URPs used in the study, 6 primers could produce polymorphic fragments in all the isolates. The URP 17R primer was highly polymorphic (100%), whereas, the URP 1F primer produced 75% polymorphic fragments. A dendrogram obtained from the combined analysis of 6 URP primers categorized the isolates into four clusters, where most of the isolates from Punjab and Haryana were clustered separately. Mating type of the population was identified based on MAT-1 and MAT-2 region universal primers for Gibberella fujikuroi. Among the 63 isolates, 18 (28.57%) were identified as MAT-1 and 45 (71.42%) as MAT-2. The effective population number for mating type was 89% of the total population. Since the distribution frequencies of both mating types were not equal in the Indian population of F. fujikuroi, it could conclude that majority of the multiplication of isolates under field conditions was through asexual reproduction. However, the presence of both mating types in F. fujikuroi indicates that the population is also capable of sexual reproduction. Therefore, it is important to develop cultivars with inbuilt resistance to bakanae disease, taking into consideration the factors such as environmental conditions and variability of the pathogen in the area of intended cultivation

High Sensitivity Cardiac Troponin-T STAT in Type 2 Diabetes Mellitus Patients and Healthy Individuals: A Comparative Study

Introduction: Diabetes Mellitus (DM) is a metabolic disorder that shares the phenotype of hyperglycaemia, with several factors contributing to the disease, including decreased insulin secretion and glucose utilisation, as well as increased glucose production. There is a strong association between DM and Cardiovascular Disease (CVD). High-sensitivity cardiac troponin T (hs-cTnT), which is a marker of subclinical myocardial damage, is used in the risk stratification of asymptomatic individuals. Aim: To estimate and compare hs-cTnT Short Turn Around Time (STAT) levels in diabetic patients without Acute Myocardial Infarction (AMI) with age and sex matched controls and also to investigate the correlation between hs-cTnT STAT and Glycated Haemoglobin (HbA1c) levels. Materials and Methods: A comparative cross-sectional study was conducted in the Department of Biochemistry and Outpatient Clinic, Department of Medicine, Government Medical College, Kozhikode, Kerala, India, from April 2019 to April 2020. The study subjects were divided into two groups: Group 1 consisted of 58 patients with Type 2 Diabetes Mellitus (T2DM) without AMI, and Group 2 comprised 58 healthy individuals who were age and sex matched. No specific sampling technique was employed. After obtaining consent, T2DM patients who attended the outpatient clinic were evaluated with fasting blood glucose, HbA1c, Electrocardiogram (ECG), and hs-cTnT STAT estimation. Controls were selected and evaluated for the same from apparently healthy bystanders of other patients, medical and paramedical staff, and others willing to participate. Statistical analysis was performed using Statistical Package for the Social Sciences (SPSS) version 22.0 software. Results: The mean value of Fasting Blood Sugar (FBS) and HbA1c was higher in T2DM patients compared to healthy individuals. hs-cTnT showed a positive moderate correlation with HbA1c (rho=0.53), which was statistically significant (p-value<0.001). Simple linear regression analysis showed that in the case group, for a 1% increase in HbA1c levels, there was a 2.38 unit increase in hs-cTnT levels, which was statistically significant (p-value<0.001). Conclusion: hs-cTnT levels are significantly elevated in T2DM patients without overt CVD compared to age and sex matched healthy individuals. T2DM is a risk factor for increased levels of biomarkers for atherosclerotic CVD, and proper glycaemic control reduces the levels of hs-cTnT in T2DM patients

No excess of mitochondrial DNA deletions within muscle in progressive multiple sclerosis

BACKGROUND: Mitochondrial dysfunction is an established feature of multiple sclerosis (MS). We recently described high levels of mitochondrial DNA (mtDNA) deletions within respiratory enzyme-deficient (lacking mitochondrial respiratory chain complex IV with intact complex II) neurons and choroid plexus epithelial cells in progressive MS. OBJECTIVES: The objective of this paper is to determine whether respiratory enzyme deficiency and mtDNA deletions in MS were in excess of age-related changes within muscle, which, like neurons, are post-mitotic cells that frequently harbour mtDNA deletions with ageing and in disease. METHODS: In progressive MS cases (n=17), known to harbour an excess of mtDNA deletions in the central nervous system (CNS), and controls (n=15), we studied muscle (paraspinal) and explored mitochondria in single fibres. Histochemistry, immunohistochemistry, laser microdissection, real-time polymerase chain reaction (PCR), long-range PCR and sequencing were used to resolve the single muscle fibres. RESULTS: The percentage of respiratory enzyme-deficient muscle fibres, mtDNA deletion level and percentage of muscle fibres harbouring high levels of mtDNA deletions were not significantly different in MS compared with controls. CONCLUSION: Our findings do not provide support to the existence of a diffuse mitochondrial abnormality involving multiple systems in MS. Understanding the cause(s) of the CNS mitochondrial dysfunction in progressive MS remains a research priority

TOM40 Mediates Mitochondrial Dysfunction Induced by α-Synuclein Accumulation in Parkinson's Disease.

Alpha-synuclein (α-Syn) accumulation/aggregation and mitochondrial dysfunction play prominent roles in the pathology of Parkinson's disease. We have previously shown that postmortem human dopaminergic neurons from PD brains accumulate high levels of mitochondrial DNA (mtDNA) deletions. We now addressed the question, whether alterations in a component of the mitochondrial import machinery -TOM40- might contribute to the mitochondrial dysfunction and damage in PD. For this purpose, we studied levels of TOM40, mtDNA deletions, oxidative damage, energy production, and complexes of the respiratory chain in brain homogenates as well as in single neurons, using laser-capture-microdissection in transgenic mice overexpressing human wildtype α-Syn. Additionally, we used lentivirus-mediated stereotactic delivery of a component of this import machinery into mouse brain as a novel therapeutic strategy. We report here that TOM40 is significantly reduced in the brain of PD patients and in α-Syn transgenic mice. TOM40 deficits were associated with increased mtDNA deletions and oxidative DNA damage, and with decreased energy production and altered levels of complex I proteins in α-Syn transgenic mice. Lentiviral-mediated overexpression of Tom40 in α-Syn-transgenic mice brains ameliorated energy deficits as well as oxidative burden. Our results suggest that alterations in the mitochondrial protein transport machinery might contribute to mitochondrial impairment in α-Synucleinopathies

Screening of DUB activity and specificity by MALDI-TOF mass spectrometry

Deubiquitylases (DUBs) are key regulators of the ubiquitin system which cleave ubiquitin moieties from proteins and polyubiquitin chains. Several DUBs have been implicated in various diseases and are attractive drug targets. We have developed a sensitive and fast assay to quantify in vitro DUB enzyme activity using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Unlike other current assays, this method uses unmodified substrates, such as diubiquitin topoisomers. By analyzing 42 human DUBs against all diubiquitin topoisomers we provide an extensive characterization of DUB activity and specificity. Our results confirm the high specificity of many members of the OTU and JAMM DUB families and highlight that all USPs tested display low linkage selectivity. We also demonstrate that this assay can be deployed to assess the potency and specificity of DUB inhibitors by profiling 11 compounds against a panel of 32 DUBs

Genetic diversity and population structure of Fusarium fujikuroi causing Bakanae, an emerging disease of rice in India

Bakanae caused by Fusarium fujikuroi (Nirenberg), is emerging as a serious threat for rice (Oryza sativa. L.) cultivation in India. In this study, 63 isolates of Fusarium fujikuroi isolated from symptomatic diseased plants were characterized for their morphology, pathogenicity and molecular variability using universal rice primers (URP). Of the 12 URPs used in the study, 6 primers could produce polymorphic fragments in all the isolates. The URP 17R primer was highly polymorphic (100%), whereas, the URP 1F primer produced 75% polymorphic fragments. A dendrogram obtained from the combined analysis of 6 URP primers categorized the isolates into four clusters, where most of the isolates from Punjab and Haryana were clustered separately. Mating type of the population was identified based on MAT-1 and MAT-2 region universal primers for Gibberella fujikuroi. Among the 63 isolates, 18 (28.57%) were identified as MAT-1 and 45 (71.42%) as MAT-2. The effective population number for mating type was 89% of the total population. Since the distribution frequencies of both mating types were not equal in the Indian population of F. fujikuroi, it could conclude that majority of the multiplication of isolates under field conditions was through asexual reproduction. However, the presence of both mating types in F. fujikuroi indicates that the population is also capable of sexual reproduction. Therefore, it is important to develop cultivars with inbuilt resistance to bakanae disease, taking into consideration the factors such as environmental conditions and variability of the pathogen in the area of intended cultivation

Intramolecular Cohesion of Coils Mediated by Phenylalanine–Glycine Motifs in the Natively Unfolded Domain of a Nucleoporin

The nuclear pore complex (NPC) provides the sole aqueous conduit for macromolecular exchange between the nucleus and the cytoplasm of cells. Its diffusion conduit contains a size-selective gate formed by a family of NPC proteins that feature large, natively unfolded domains with phenylalanine–glycine repeats (FG domains). These domains of nucleoporins play key roles in establishing the NPC permeability barrier, but little is known about their dynamic structure. Here we used molecular modeling and biophysical techniques to characterize the dynamic ensemble of structures of a representative FG domain from the yeast nucleoporin Nup116. The results showed that its FG motifs function as intramolecular cohesion elements that impart order to the FG domain and compact its ensemble of structures into native premolten globular configurations. At the NPC, the FG motifs of nucleoporins may exert this cohesive effect intermolecularly as well as intramolecularly to form a malleable yet cohesive quaternary structure composed of highly flexible polypeptide chains. Dynamic shifts in the equilibrium or competition between intra- and intermolecular FG motif interactions could facilitate the rapid and reversible structural transitions at the NPC conduit needed to accommodate passing karyopherin–cargo complexes of various shapes and sizes while simultaneously maintaining a size-selective gate against protein diffusion

Association of dialysis facility-level hemoglobin measurement and erythropoiesis-stimulating agent dose adjustment frequencies with dialysis facility-level hemoglobin variation: a retrospective analysis

<p>Abstract</p> <p>Background</p> <p>A key goal of anemia management in dialysis patients is to maintain patients' hemoglobin (Hb) levels consistently within a target range. Our aim in this study was to assess the association of facility-level practice patterns representing Hb measurement and erythropoiesis-stimulating agent (ESA) dose adjustment frequencies with facility-level Hb variation.</p> <p>Methods</p> <p>This was a retrospective observational database analysis of patients in dialysis facilities affiliated with large dialysis organizations as of July 01, 2006, covering a follow-up period from July 01, 2006 to June 30, 2009. A total of 2,763 facilities representing 436,442 unique patients were included. The predictors evaluated were facility-level Hb measurement and ESA dose adjustment frequencies, and the outcome measured was facility-level Hb variation.</p> <p>Results</p> <p>First to 99th percentile ranges for facility-level Hb measurement and ESA dose adjustment frequencies were approximately once per month to once per week and approximately once per 3 months to once per 3 weeks, respectively. Facility-level Hb measurement and ESA dose adjustment frequencies were inversely associated with Hb variation. Modeling results suggested that a more frequent Hb measurement (once per week rather than once per month) was associated with approximately 7% to 9% and 6% to 8% gains in the proportion of patients with Hb levels within a ±1 and ±2 g/dL range around the mean, respectively. Similarly, more frequent ESA dose adjustment (once per 2 weeks rather than once per 3 months) was associated with approximately 6% to 9% and 5% to 7% gains in the proportion of patients in these respective Hb ranges.</p> <p>Conclusions</p> <p>Frequent Hb measurements and timely ESA dose adjustments in dialysis patients are associated with lower facility-level Hb variation and an increase in proportion of patients within ±1 and ±2 g/dL ranges around the facility-level Hb mean.</p

Direct damage controlled seismic design of plane steel degrading frames

A new method for seismic design of plane steel moment resisting framed structures is developed. This method is able to control damage at all levels of performance in a direct manner. More specifically, the method: (a) can determine damage in any member or the whole of a designed structure under any given seismic load, (b) can dimension a structure for a given seismic load and desired level of damage and (c) can determine the maximum seismic load a designed structure can sustain in order to exhibit a desired level of damage. In order to accomplish these things, an appropriate seismic damage index is used that takes into account the interaction between axial force and bending moment at a section, strength and stiffness degradation as well as low cycle fatigue. Then, damage scales are constructed on the basis of extensive parametric studies involving a large number of frames exhibiting cyclic strength and stiffness degradation and a large number of seismic motions and using the above damage index for damage determination. Some numerical examples are presented to illustrate the proposed method and demonstrate its advantages against other methods of seismic design. © 2014, Springer Science+Business Media Dordrecht