Homeobox 27, a Homeodomain Transcription Factor, Confers Tolerances to CMV by Associating with Cucumber Mosaic Virus 2b Protein.

Transcription factors (TFs) play an important role in plant development; however, their role during viral infection largely remains unknown. The present study was designed to uncover the role transcription factors play in Cucumber mosaic virus (CMV) infection. During the screening of an Arabidopsis thaliana (Col-0) transcription factor library, using the CMV 2b protein as bait in the yeast two-hybrid system, the 2b protein interacted with Homeobox protein 27 (HB27). HB27 belongs to the zinc finger homeodomain family and is known to have a regulatory role in flower development, and responses to biotic and abiotic stress. The interaction between CMV 2b and HB27 proteins was further validated using in planta (bimolecular fluorescence complementation assay) and in vitro far-Western blotting (FWB) methods. In the bimolecular fluorescence complementation assay, these proteins reconstituted YFP fluorescence in the nucleus and the cytoplasmic region as small fluorescent dots. In FWB, positive interaction was detected using bait anti-MYC antibody on the target HB27-HA protein. During CMV infection, upregulation (~3-fold) of the HB27 transcript was observed at 14 days post-infection (dpi) in A. thaliana plants, and expression declined to the same as healthy plants at 21 dpi. To understand the role of the HB27 protein during CMV infection, virus accumulation was determined in HB27-overexpressing (HB27 OE) and knockout mutants. In HB27-overexpressing lines, infected plants developed mild symptoms, accumulating a lower virus titer at 21 dpi compared to wild-type plants. Additionally, knockout HB27 mutants had more severe symptoms and a higher viral accumulation than wild-type plants. These results indicate that HB27 plays an important role in the regulation of plant defense against plant virus infection.

Outbreak or pseudo-outbreak? Integrating SARS-CoV-2 sequencing to validate infection control practices in a dialysis facility.

BACKGROUND: The COVID-19 pandemic poses a particularly high risk for End Stage Renal Disease (ESRD) patients so rapid identification of case clusters in ESRD facilities is essential. Nevertheless, with high community prevalence, a series of ESRD patients may test positive contemporaneously for reasons unrelated to their shared ESRD facility. Here we describe a series of 5 cases detected within 11 days in November 2020 in a hospital-based 32-station ESRD facility in Southwest Wisconsin, the subsequent facility-wide testing, and the use of genetic sequence analysis to evaluate links between cases. METHODS: Four patient cases and one staff case were identified in symptomatic individuals by RT-PCR. Facility-wide screening was conducted using rapid SARS-CoV-2 antigen tests. SARS-CoV-2 genome sequences were obtained from residual diagnostic specimens. RESULTS: Facility-wide screening of 47 staff and 107 patients identified no additional cases. Residual specimens from 4 of 5 cases were available for genetic sequencing. Clear genetic differences proved that these contemporaneous cases were not linked. CONCLUSIONS: With high community prevalence, epidemiological data alone is insufficient to deem a case cluster an outbreak. Cluster evaluation with genomic data, when available with a short turn-around time, can play an important role in infection prevention and control response programs.

COVID-19-associated mucormycosis and treatments

In the current pandemic, COVID-19 patients with predisposing factors are at an increased risk of mucormycosis, an uncommon angioinvasive infection that is caused by fungi with Mucor genus which is mainly found in plants and soil. Mucormycosis development in COVID-19 patient is related to various factors, such as diabetes, immunocompromise and neutropenia. Excessive use of glucocorticoids for the treatment of critically ill COVID-19 patients also leads to opportunistic infections, such as pulmonary aspergillosis. COVID-19 patients with mucormycosis have a very high mortality rate. This review describes the pathogenesis and various treatment approaches for mucormycosis in COVID-19 patients, including medicinal plants, conventional therapies, adjunct and combination therapies.

Chilling Affects Phytohormone and Post-Embryonic Development Pathways during Bud Break and Fruit Set in Apple (Malus domestica Borkh.).

The availability of sufficient chilling during bud dormancy plays an important role in the subsequent yield and quality of apple fruit, whereas, insufficient chilling availability negatively impacts the apple production. The transcriptome profiling during bud dormancy release and initial fruit set under low and high chill conditions was performed using RNA-seq. The comparative high number of differentially expressed genes during bud break and fruit set under high chill condition indicates that chilling availability was associated with transcriptional reorganization. The comparative analysis reveals the differential expression of genes involved in phytohormone metabolism, particularly for Abscisic acid, gibberellic acid, ethylene, auxin and cytokinin. The expression of Dormancy Associated MADS-box, Flowering Locus C-like, Flowering Locus T-like and Terminal Flower 1-like genes was found to be modulated under differential chilling. The co-expression network analysis indentified two high chill specific modules that were found to be enriched for "post-embryonic development" GO terms. The network analysis also identified hub genes including Early flowering 7, RAF10, ZEP4 and F-box, which may be involved in regulating chilling-mediated dormancy release and fruit set. The results of transcriptome and co-expression network analysis indicate that chilling availability majorly regulates phytohormone-related pathways and post-embryonic development during bud break.

Effect of Demographic Status and Lifestyle Habits on Glycaemic Levels in Apparently Healthy Subjects: A Cross-Sectional Study.

Aim. To identify the effects of sociodemographic status, family history, and lifestyle habits on fasting blood glucose (FBG) and fasting serum insulin (FSI) levels in apparently healthy subjects. Methods. Information was gathered using an interviewer-administered questionnaire from 227 apparently healthy nondiabetic subjects residing in a suburban area in Sri Lanka. Venous blood samples were collected after an overnight fast for FBG and FSI analysis. Correlations and differences were analyzed using SPSS (ver. 17) software. Results. The majority of the subjects were females, having secondary or tertiary education, monthly income ≥Rs. 25,000 (USD 175), and a positive family history of diabetes. Among the subjects, 10.1% were identified as prediabetics and the majority had familial diabetes with monthly income ≥Rs. 25,000 (USD 175). Subjects with high income had significantly higher mean FBG. In addition FBG had a significant correlation with age. Males and subjects with less than 6 hours/day sleeping duration at night had significantly higher FBG. Subjects with less vigorous physical activity and longer sitting duration had significantly higher FSI levels. Conclusions. Increasing age, higher income, positive familial history of diabetes, sedentary lifestyle, and short sleep at night have positive impact on glycaemic status in apparently healthy subjects.

Chilling-Mediated DNA Methylation Changes during Dormancy and Its Release Reveal the Importance of Epigenetic Regulation during Winter Dormancy in Apple (Malus x domestica Borkh.).

Winter dormancy is a well known mechanism adopted by temperate plants, to mitigate the chilling temperature of winters. However, acquisition of sufficient chilling during winter dormancy ensures the normal phenological traits in subsequent growing period. Thus, low temperature appears to play crucial roles in growth and development of temperate plants. Apple, being an important temperate fruit crop, also requires sufficient chilling to release winter dormancy and normal phenological traits, which are often associated with yield and quality of fruits. DNA cytosine methylation is one of the important epigenetic modifications which remarkably affect the gene expression during various developmental and adaptive processes. In present study, methylation sensitive amplified polymorphism was employed to assess the changes in cytosine methylation during dormancy, active growth and fruit set in apple, under differential chilling conditions. Under high chill conditions, total methylation was decreased from 27.2% in dormant bud to 21.0% in fruit set stage, while no significant reduction was found under low chill conditions. Moreover, the demethylation was found to be decreased, while methylation increased from dormant bud to fruit set stage under low chill as compared to high chill conditions. In addition, RNA-Seq analysis showed high expression of DNA methyltransferases and histone methyltransferases during dormancy and fruit set, and low expression of DNA glcosylases during active growth under low chill conditions, which was in accordance with changes in methylation patterns. The RNA-Seq data of 47 genes associated with MSAP fragments involved in cellular metabolism, stress response, antioxidant system and transcriptional regulation showed correlation between methylation and their expression. Similarly, bisulfite sequencing and qRT-PCR analysis of selected genes also showed correlation between gene body methylation and gene expression. Moreover, significant association between chilling and methylation changes was observed, which suggested that chilling acquisition during dormancy in apple is likely to affect the epigenetic regulation through DNA methylation.

Pattern of animal bites and post exposure prophylaxis in rabies: A five year study in a tertiary care unit in Sri Lanka.

BACKGROUND: Rabies is a global problem which occurs in more than 150 countries and territories including Sri Lanka, where human deaths from rabies are in decline whilst resources incurred for prevention of rabies are in sharp incline over the years. In this backdrop, we aim to audit the post-exposure treatment (PET) in rabies and the pattern of animal bites in a tertiary care hospital in Sri Lanka. METHODS: This study was carried out at Teaching Hospital Peradeniya (THP), in the Central Province of Sri Lanka from 2007-2012 where a registry of all PET has been maintained. The data from registries were extracted after obtaining permission from the hospital authority for analysis. RESULTS: There were 19 661 cases of animal exposure presented to the THP over the study period of 5 ears. Of them, the majority-17431(88.66 %) were definitive animal bites whilst scratches accounted for 2147(10.92 %) and 83(0.42 %) were miscellaneous exposures. According to the severity grading of injuries, 7 362(37 %) were major bites and 12 226(62 %) were minor bites. The domestic unvaccinated dogs and cats were responsible for 10,662 (54 %) and 3,982 (20 %) of exposures respectively. The total cost incurred for both anti-rabies vaccine and anti rabies serum during the study period is 24,795,888.00 Sri Lankan rupees (190,737.60US$). CONCLUSIONS: The pattern of animal bite shows high dominance of domestic dogs and cats exposures. The age of victims ranged from infancy to old-age with higher incidence among children. Even though PET is costly, continued surveillance and rabies control is still necessary along with public education and vaccination of domestic pets.

Factors associated with neonatal deaths in Chitwan district of Nepal.

BACKGROUND: Neonatal mortality has remained unchanged since 2006 in Nepal. Reducing neonatal mortality is indispensable to reduce child mortality. The objective of this study was to investigate the factors associated with neonatal mortality. This study assesses socio-demographic factors, maternal health care and newborn care practices contributing to neonatal deaths in Chitwan district of Central Nepal. METHODS: A case-control study was conducted during April-July 2012. The study used a mixed-method approach, in which records of neonatal deaths were obtained from the District Public Health Office and a comparison group, survivors, was obtained from the same community. A total of 198 mothers (of 99 neonatal deaths and 99 survivor neonates) were included in the survey. Focus group discussions, in-depth interviews and case studies were also conducted. Maternal characteristics were analyzed using descriptive statistics, Mc Nemar's Chi square test and multivariable backward conditional logistic regression analysis. Qualitative data were analyzed by narrative analysis method. RESULTS: More than four-fifth of mothers (86%) had antenatal check-up (ANC) and the proportion of four or more ANC was 64%. Similarly, the percentage of mothers having institutional delivery was 62%, and postnatal check-up was received by 65% of mothers. In multivariable analysis, low birth weight [adjusted odds ratio: 8.49, 95% CI (3.21-22.47)], applying nothing on cord [adjusted odds ratio: 5.72, 95% CI (1.01-32.30)], not wrapping of newborn [adjusted odds ratio: 9.54, 95% CI (2.03-44.73)], and no schooling of mother [adjusted odds ratio: 2.09, 95% CI (1.07-4.11)] were significantly associated with an increased likelihood of neonatal mortality after adjusting for other confounding variables. Qualitative findings suggested that bathing newborns after 24 h and wrapping in clean clothes were common newborn care practices. The mothers only attended postnatal care services if health problems appeared either in the mother or in the child. CONCLUSION: Results of this study suggest that the current community based newborn survival intervention should provide an even greater focus to essential newborn care practices, low birth weight newborns, and female education.

Femoral artery pseudoaneurysm as a complication of angioplasty. How can it be prevented?

Femoral pseudoaneurysm is a common complication of repeated femoral puncture during cardiac catheterisation. We describe here the development of femoral pseudoaneurysms in a patient with Takayasu's arteritis, which healed in response to conservative treatment, and review the literature on the prevention and treatment of femoral pseudoaneurysm.

Interaction of cytochrome c with tRNA and other polynucleotides.

Interaction between tRNA and other polynucleotides with cytochrome c was studied by visible spectroscopy, fluorescence spectroscopy and gel mobility shift assay in view of the recently reported important regulatory role of tRNA in cytochrome c mediated apoptotic pathway. Visible spectroscopy showed perturbation in the heme binding environment in cytochrome c with tRNA binding. Fluorescence titrations indicated that cytochrome c binds to different polynucleotides with differing affinities. A weak binding was observed with single stranded polyribonucleotides and polydeoxyribonuleotides and strong binding with tRNA and double stranded DNA as indicated by extent of fluorescence quenching and binding constants. Calculation of thermodynamic binding parameters from fluorescence titrations indicated that three molecules of cytochrome c bound with one tRNA molecule with binding constant of 1.9 × 10(6) M(-1). The perturbation of cytochrome c structure caused by the binding of tRNA could be affecting its role in mediating apoptosis.

Isolation of two strong poly (U) binding proteins from moderate halophile Halomonas eurihalina and their identification as cold shock proteins.

Cold shock proteins (Csp) are known to be expressed in response to sudden decrease in temperature. They are thought to be involved in a number of cellular processes viz., RNA chaperone activity, translation, transcription, nucleoid condensation. During our studies on ribosomal protein S1 in moderate halophile Halomonas eurihalina, we observed the presence of two strong poly (U) binding proteins in abundance in cell extracts from cells grown under normal growth conditions. The proteins can be isolated in a single step using Poly (U) cellulose chromatography. The proteins were identified as major cold shock proteins belonging to Csp A family by MALDI-TOF and bioinformatic analysis. Csp 12 kDa was found in both exponential and stationary phases whereas Csp 8 kDa is found only in exponential phase.

Outpatient High-Dose Urokinase Infusion Improves Dialysis Catheter Longevity: A Prospective Observational Study.

Catheter thrombosis is a major limiting factor affecting catheter survival in hemodialysis (HD). A previous retrospective study highlighted the utility of outpatient intradialytic high-dose urokinase (HDU) infusion in the treatment of HD catheter thrombosis. The present study was designed to prospectively evaluate the effectiveness of 250 000 IU urokinase given as a 3-hour infusion during HD. The infusion would follow detection of a drop in blood flow thought to be secondary to thrombosis [a "thrombotic event" (TE)] after exclusion of nonthrombotic causes of drop in blood flow [a "nonthrombotic event" (NTE)], such as systemic hypotension or suboptimal position of the catheter on x ray. An "event" was defined as a drop in pump blood flow by greater than or equal to 20% of the prescribed blood flow. Twenty patients (mean age 57.7 years; 13 males) with documented or potentially long-term catheters were recruited (total catheters used, 30) from an outpatient HD unit. A variety of catheter types were used. Catheter position was confirmed on x ray. Hemodialysis sessions were monitored for 6 months or until catheter removal (whichever was earlier). Thrombotic and NTEs (mostly related to drop in blood flow) were noted. The use of intradialytic HDU infusions was monitored. Improvement was defined as reestablishment of blood flow to within 10% of prescription. There were 24 TEs in 11 patients and 15 NTEs in 7 patients. Twenty-one of 22 (95%) TEs responded to HDU, with complete restoration of blood flow; in 1/22 HDU-treated instances there was a partial response. For 2/24 TEs the patients underwent catheter stripping with good results. Twelve of 15 NTEs occurred in the first 2 weeks of catheter placement versus 6/24 TEs (p = 0.001, chi-square). Catheter survival was longer than 24 weeks in 12/30 catheters. Nine catheters were lost to NTEs. No catheter was lost secondary to a TE. It is therefore concluded that intradialytic outpatient HDU is useful for the treatment of HD catheter thrombosis. It may obviate/reduce the need for catheter stripping or replacement. Randomized controlled trials with this approach are needed. A higher proportion of NTEs tended to occur earlier after catheter placement compared to TEs.