RNA-seq analysis reveals an early defense response to tomato leaf curl New Delhi virus in potato cultivar Kufri Bahar.

Potatoes in India are very susceptible to apical leaf curl disease, which causes severe symptoms and greater yield losses. Because the majority of potato cultivars are susceptible to the virus, it is crucial to discover sources of resistance and investigate the mechanism of resistance/susceptibility in potato cultivars. In this study, the gene expression profile of two potato cultivars, Kufri Bahar (resistant) and Kufri Pukhraj (susceptible), varying in their level of resistance to ToLCNDV, was analyzed using RNA-Seq. The Ion ProtonTM system was used to sequence eight RiboMinus RNA libraries from inoculated and uninoculated potato plants at 15 and 20 days after inoculation (DAI). The findings indicated that the majority of differentially expressed genes (DEGs) were cultivar-or time-specific. These DEGs included genes for proteins that interact with viruses, genes linked with the cell cycle, genes for proteins involved in defense, transcription and translation initiation factors, and plant hormone signaling pathway genes. Interestingly, defense responses were generated early in Kufri Bahar, at 15 DAI, which may have impeded the replication and spread of ToLCNDV. This research provides a genome-wide transcriptional analysis of two potato cultivars with variable levels of ToLCNDV resistance. At an early stage, we observed suppression of genes that interact with viral proteins, induction of genes associated with restriction of cell division, genes encoding defense proteins, AP2/ERF transcription factors, and altered expression of zinc finger protein genes, HSPs, JA, and SA pathway-related genes. Our findings add to a greater comprehension of the molecular basis of potato resistance to ToLCNDV and may aid in the development of more effective disease management techniques.

Construction of a single nucleotide polymorphism linkage map and identification of quantitative trait loci controlling heat tolerance in cowpea, Vigna unguiculata (L.) Walp.

Plant tolerance to heat or high temperature is crucial to crop production, especially in the situation of elevated temperature resulting from global climate change. Cowpea, Vigna unguiculata (L.) Walp., is an internationally important legume food crop and an excellent pool of genes for numerous traits resilient to environmental extremes, particularly heat and drought. Here, we report a single nucleotide polymorphism (SNP) genetic map for cowpea and identification of the loci controlling the heat tolerance in the species. The SNP map consists of 531 bins containing 4,154 SNPs grouped into 11 linkage groups, and collectively spans 1,084.7 cM, thus having a density of one SNP in 0.26 cM or 149 kb. The 11 linkage groups of the map were aligned to the 11 cowpea chromosomes. Quantitative trait locus (QTL) mapping identified nine QTLs responsible for the cowpea heat tolerance on seven of the 11 chromosomes, with each QTL explaining 6.5-21.8% of heat tolerance phenotypic variation. Moreover, we aligned these nine QTLs to the cowpea genome. Each of the QTLs was positioned in a genomic region ranging from 209,000 bp to 12,590,450 bp, and the QTL with the largest effect (21.8%) on heat tolerance, qHT4-1, was located within an interval of only 234,195 bp. These results provide SNP markers useful for marker-assisted selection for heat tolerance and lay a foundation for cloning, characterization, and applications of the genes controlling the cowpea heat tolerance for heat tolerance genetic improvement in cowpea and related crops.

New-Onset Heart Failure From Viral Myocarditis and Inflammatory Cardiomyopathy: A Case Report.

An inflammatory cardiomyopathy is a form of nonischemic cardiomyopathy that results from myocarditis associated with cardiac dysfunction and ventricular remodeling. It can be caused by a wide array of pathogens and toxins. We present a case of a 69-year-old female with a history of asthma who was admitted to our facility with recurrent asthma exacerbations, likely triggered by viral upper respiratory tract infections. In 5 months, serial echocardiograms showed a significant decline in her left ventricular systolic and diastolic function. Cardiac catheterization showed no clinically significant coronary artery disease. Despite normal renal function, her troponin remained elevated. This is an interesting case of a viral upper respiratory tract infection that led to myocarditis and ultimately resulted in inflammatory cardiomyopathy.

Spraying of dsRNA molecules derived from Phytophthora infestans, along with nanoclay carriers as a proof of concept for developing novel protection strategy for potato late blight.

BACKGROUND: Phytophthora infestans is a late blight-causing oomycetes pathogen. It rapidly evolves and adapts to the host background and new fungicide molecules within a few years of their release, most likely because of the predominance of transposable elements in its genome. Frequent applications of fungicides cause environmental concerns. Here, we developed target-specific RNA interference (RNAi)-based molecules, along with nanoclay carriers, that when sprayed on plants are capable of effectively reducing late blight infection. RESULTS: Targeted the genes unique to sporulation, early satge infection and the metabolism pathway stages based on in an our own microarray data. We used nanoclay as a carrier for sorbitol dehydrogenase, heat shock protein 90, translation elongation factor 1-α, phospholipase-D like 3 and glycosylphosphatidylinositol-anchored acidic serine-threonine-rich HAM34-like protein double-stranded (ds)RNAs, which were assessed by culture bioassay, detached leaf assay and spray methods, and revealed a reduction in growth, sporulation and symptom expression. Plants sprayed with multigene targeted dsRNA-nanoclay showed enhanced disease resistance (4% disease severity) and less sporulation (<1 × 103 ) compared with plants sprayed with dsRNA alone. CONCLUSION: The use of nanoclay with multigene targeted dsRNA was assumed to be involved in effective delivery, protection and boosting the action of RNAi as a spray-induced gene silencing approach (SIGS). A significant reduction in growth, sporulation, disease severity and decreased gene expression authenticates the effects of SIGS on late blight progression. This study demonstrated as a proof of concept the dsRNA-nanoclay SIGS approach, which could be used as an alternative to chemical fungicides and transgenic approaches to develop an environmentally friendly novel plant protection strategy for late blight. © 2022 Society of Chemical Industry.

Recurrent, Complicated Diverticulitis With Atypical Features.

Diverticular disease is a common condition responsible for significant costs to the healthcare system in the Western world. It ranges from asymptomatic diverticulosis to complicated diverticulitis. Here, we present a unique case of recurrent, complicated diverticulitis in a 62-year-old Caucasian male. Within a span of one year, he was hospitalized six times with diverticulitis before undergoing elective sigmoid colon resection. Imaging showed diverticulitis of distal descending and proximal sigmoid colon with sealed perforation, recurrent abscesses, and formation of colocutaneous fistulas. During each hospitalization, the patient was advised to follow up with general surgery and/or outpatient gastroenterology but chose not to do so. Eventually, he required an elective sigmoid colectomy with a takedown of the colocutaneous fistulas. In this case report, we discuss the atypical features and criteria for prophylactic colon resection in diverticulitis to highlight the importance of outpatient follow-up with general surgery and gastroenterology.

An Atypical Etiology of Acute Pericarditis: A Case Report.

Acute pericarditis is caused by inflammation of the pericardial sac and is characterized by sharp and pleuritic chest pain, which is accompanied by a pericardial friction rub and serial electrocardiogram (ECG) changes. Although there are many known etiologies of pericarditis, we present the first known case of a herbal supplement, the Tejocote root, causing acute pericarditis in a previously healthy 23-year-old female. Her ECG showed t-wave inversions that resolved with colchicine and non-steroidal anti-inflammatory drug therapy. Prior studies have demonstrated that it can cause adverse cardiovascular effects, but this is the first documented case of the Tejocote root causing acute pericarditis. This case report reinforces the importance of inquiring about supplements patients may be taking, especially when looking for the etiology of pericarditis.

Quantitative trait loci influencing days to flowering and plant height in cowpea, Vigna unguiculata (L.) Walp.

Cowpea (Vigna unguiculate (L.) Walp.) is a worldwide important multifunctional legume crop for food grain, vegetable, fodder, and cover crop. Nevertheless, only limited research has been conducted on agronomic traits. Here, we report quantitative trait locus (QTL) analysis of the days to flowering (DTF) and plant height (PH) using a dense SNP linkage map recently developed from a recombinant inbred line (RIL) population derived from a cross between Golden Eye Cream and IT98K-476-8. The population was phenotyped for DTF and PH through field and greenhouse trials under two environments. The QTLs controlling these traits were mapped using multiple-environment combined and individual trial phenotypic data. The combined data analysis identified one major QTL (qDTF9.1) for DTF, and one major QTL (qPH9.1) and a minor QTL (qPH4.1) for PH. qDTF9.1 and qPH9.1 were adjacent to each other on Chromosome 9 and each explained 29.3% and 29.5% of the phenotypic variation (PVE), respectively. The individual trial data analysis identified a minor QTL (qDTF2.1) on Chromosome 2 for DTF and two minor QTLs (qPH4.1 and qPH4.2) on Chromosome 4 for PH, while the major QTLs, qDTF9.1 and qPH9.1, were consistently identified in all trials conducted. Epistasis analysis revealed that qDTF9.1 interacted with one locus on Chromosome 4, contributed 50% of the PVE, and qPH9.1 interacted with one locus on each of Chromosomes 4 and 6, contributing 30% and 23% of the PVE, respectively, suggesting that epistasis plays an important role in the trait performance. These results, therefore, provide a deeper understanding of the genetic architecture of plant DTF and PH, and molecular tools necessary for cloning the genes and for enhanced cowpea breeding.

An insight into the downstream analysis of RB gene in F1 RB potato lines imparting field resistance to late blight.

Earlier studies have shown that level of late blight resistance conferred by the classical R gene (RB Rpi-blb1) is dependent on genetic background of the recipient genotype. This was revealed in the analysis of late blight response that belonged to a group of F1 progeny obtained from the cross between Kufri Jyoti and SP951, which showed wide variation in late blight resistance response in spite of possessing the same RB gene. The global gene expression pattern in the RB potato lines was studied in response to late blight infection using cDNA microarray analysis to reveal the background effect. Leaf samples were collected at 0, 24, 72 and 120h post inoculation (hpi) with Phytophthora infestans for gene expression analysis using 61031 gene sequences. Significantly upregulated (1477) and downregulated (4245) genes common in the RB-transgenic F1 lines at 24 and 72 hpi were classified into several categories based on GO identifiers and majority of genes were assigned putative biological functions. Highest expression of an NBS-LRR along with protease, pectin esterase inhibitors, chaperones and reactive oxygen species genes were observed which affirmed a significant role of these categories in the defence response of RB-KJ lines. Results suggest that the immune priming of plant receptors are likely to be involved in stability and functionality of RB to induce resistance against P. infestans. This study is important for effective deployment of RB gene in the host background and contributes immensely to scientific understanding of R gene interaction with host protein complexes to regulate defence system in plants.

Genotypic background of the recipient plant is crucial for conferring RB gene mediated late blight resistance in potato.

BACKGROUND: Late blight, caused by oomycetes pathogen Phytophthora infestans (Mont.) de Bary, is the most devastating potato disease in the world. RB gene from Solanum bulbocastanum has been shown to impart broad spectrum resistance against P. infestans races. In this study Katahdin transgenic event SP951 was used as male parent to cross with the popular Indian potato cultivars viz., Kufri Bahar (KB) and Kufri Jyoti (KJ) to enhance the late blight resistance. RESULTS: Populations of 271 F1seedlings from the crosses KB × SP951 (87) and KJ × SP951 (184) were screened for inheritance of RB transgene through PCR and bioassay. Disease response based on AUDPC of different hybrid lines varied from immunity to complete susceptibility. High degree of resistance (<25% infection) was observed in KJ × SP951 derived seedlings (85.2%), whereas level of resistance in KB × SP951 (36.4% infection) derived seedlings was of low order. CONCLUSION: This study provides valuable genetic materials for development of potentially durable late blight resistant potato varieties. Besides, it also corroborates the fact that efficacy of R gene is not solely dependent on its presence in the variety but largely depends on the genetic background of the recipient genotype.

An insight into differentially regulated genes in resistant and susceptible genotypes of potato in response to tomato leaf curl New Delhi virus-[potato] infection.

Apical leaf curl disease, caused by tomato leaf curl New Delhi virus-[potato] (ToLCNDV-[potato]) is one of the most important viral diseases of potato in India. Genetic resistance source for ToLCNDV in potato is not identified so far. However, the cultivar Kufri Bahar is known to show lowest seed degeneration even under high vector levels. Hence, microarray analysis was performed to identify differentially regulated genes during ToLCNDV-[potato] infection in a resistant (Kufri Bahar) and a susceptible cultivar (Kufri Pukhraj). Under artificial inoculation conditions, in Kufri Pukhraj, symptom expressions started at 15days after inoculation (DAI) and then progressed to severe symptoms, whereas no or only very mild symptoms were observed in Kufri Bahar up to 35 DAI. Correspondingly, qPCR assay indicated a high viral load in Kufri Pukhraj and a very low viral load in Kufri Bahar. Microarray analysis showed that a total of 1111 genes and 2588 genes were differentially regulated (|log2 (Fold Change)|>2) in Kufri Bahar and Kufri Pukhraj, respectively, following ToLCNDV-[potato] infection. Gene ontology and mapman analyses revealed that these altered transcripts were involved in various biological & metabolic processes. Several genes with unknown functions were 5 to 100 fold expressed after virus infection and further experiments are necessary to ascertain their role in disease resistance or susceptibility. This study gives an insight into differentially regulated genes in response to ToLCNDV-[potato] infection in resistant and susceptible cultivars and could serve as the basis for the development of new strategies for disease management.

Endoscopic management of pancreatic pseudocysts and walled-off pancreatic necrosis: A two-decade experience.

AIM: To determine long-term outcome of endoscopic management of pancreatic pseudocyst/walled-off pancreatic necrosis (WOPN) without necrosectomy. METHODS: One-hundred and sixty-five pancreatic pseudocysts/WOPN managed endoscopically over a period of 22 years were analyzed retrospectively for technical success, complications, and recurrence. RESULTS: Symptomatic 118 males and 47 females with mean age of 35.8 years were included. Alcohol was the most common etiology (41.2%). Transmural endoscopic drainage was done in 144 patients, while 21 patients underwent transpapillary drainage. All the patients were subjected to contrast computed tomography (CT) abdomen or routine/Doppler ultrasound. Endoscopic ultrasound was done in last 11 patients. One or two double pigtail 7 Fr stents were placed when clear watery fluid came out from cyst (130 patients, 78.8%), and nasocystic drainage (NCD) tubes were placed in addition to two 7 Fr stents when there were frank pus, thick dark fluid, or solid components inside the cyst (35 patients). All these patients settled on this treatment. Thirty-three of 35 patients of WOPN could be managed endoscopically without necrosectomy. Complications occurred in 9.2% of pseudocysts and 40% of WOPN. Thirty-five patients were followed up for more than 5 years (3 patients more than 10 years), and 130 patients were followed up for up to 5 years. Recurrence occurred in 8.1% of pseudocysts and 5.7% of WOPN. CONCLUSION: Majority of pancreatic pseudocysts/WOPN can be managed with endoscopic drainage without necrosectomy with high success, low complication, and recurrence rates.

Comparative Evaluation of Intra-Operative Peritoneal Lavage with Super Oxidized Solution and Normal Saline in Peritonitis Cases; Randomized Controlled Trial.

AIM: To investigate the efficacy of super-oxidized solution (SOS) over normal saline in peritonitis cases. Our objectives are to present the potential clinical impact of intraperitoneal lavage with solutions for early recovery of the patient by reducing the infection rate. MATERIAL AND METHODS: A double-blind random clinical trial was performed in 240 cases diagnosed as peritonitis at MM Institute of Medical Sciences and Research, Mullana, Ambala, India, from December 2013 to November 2015. Subjects were categorized into 2 groups, i.e. 120 cases in the group I and 120 cases in the group II. Both the groups underwent peritoneal lavage; the group I underwent normal saline lavage and group II received SOS. The effectiveness of both the solutions was compared. RESULTS: In both the groups majority of the patients (50.0%) were in the 21 - 40 years of age. The mean age in our study was 45.28 years, median was 45 and the standard deviation was 14.07. The bacterial load was higher in the group II as compared to the group I after the lavage with SOS solution. The superficial wound infection rate was higher in the group II (48 cases) compared to the group I (32). These data was significant with p value (0.040). Wound pain was found in 59 cases in group II as compared to 42 in group I. This difference was found to be statistically significant with p value 0.036. Post-operatively the morbidity and mortality was less in the group I as compared to group II. CONCLUSION: SOS significantly reduced the wound pain and infection hence resulting in early recovery. SOS is a high level disinfectant, non-flammable and no special training is required to handle it.

Evaluation and comparison of postoperative levels of serum bilirubin, serum transaminases and alkaline phosphatase in laparoscopic cholecystectomy versus open cholecystectomy.

BACKGROUND: Laparoscopic cholecystectomy (LC) requires the creation of a pneumoperitoneum via insufflations of carbon dioxide; resulting in increased partial pressure of carbon dioxide (CO2) and intraperitoneal pressure which leads to the changes in pulmonary function and hemodynamic measurements. Hypercarbia leads to visceral organ ischemia including liver and venous stasis/thromboembolism or both due to impaired flow. The present study has been undertaken to see the changes in liver function tests (LFTs) after laparoscopic/open cholecystectomy (OC), the incidences of such change, their relation to age, sex, duration of surgery and to know the clinical significances of such disturbances. AIMS AND OBJECTIVES: To compare and correlate the serum level of bilirubin, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) in patients who underwent LC to those who underwent OC. MATERIALS AND METHODS: The present study was conducted in the Department of Surgery at MMIMSR, MM University, Mullana, Ambala. A total number of 200 patients diagnosed as cholelithiasis were included in the study from May 2012 to May 2014. These cases were randomly divided into two groups (A and B) consisting of 100 cases each. LC was performed in group A patients and OC was done in group B patients. Three blood samples were taken: (I) pre-operatively; (II) after 24 hours of surgery; and (III) after 72 hours of surgery for comparison of the enzyme level alterations. RESULTS: In LC patients, there were rise in the levels of serum bilirubin, AST and ALT after 24 hrs of surgery from the preoperative value and then again fall was noted (near to normal value) after 72 hrs of surgery except in that of ALP. ALP levels showed slight fall after 24 hrs of surgery and then slight rise after 72 hrs which was within the normal limit. Whereas in OC patients, there were slight variations in the liver enzymes (which were within the normal range). CONCLUSIONS: Transient elevation of serum bilirubin, AST and ALT occurs after LC or after OC. The alteration in the liver enzymes can be attributed to CO2 pneumoperitoneum, surgical manipulations, diathermy, patient position, and arterial injury may also other factors. These changes return to normal in 3-4 days after procedure and they have no clinical consequences in patients with normal hepatic function but they may still cause worry to the surgeon regarding the integrity of biliary tree.

Expression profiling of potato cultivars with contrasting tuberization at elevated temperature using microarray analysis.

Temperature is one of the most significant factors affecting potato yield. Night temperature beyond 18-22 °C drastically reduces tuber formation, constraining potato cultivation in tropics and subtropics. Identification of genes and pathways affected by high temperature is crucial for developing thermo tolerant cultivars for these regions. In the present study, two cultivars with contrasting tuberization behavior at night temperatures (24 °C) were selected for gene expression analysis using a customized microarray chip representing 39,031 potato genes. A total of 2500 genes were differentially expressed on 21 d and 4096 genes on 14 d after stress. Gene ontology and pathway analysis provided insights into the probable biological processes and pathways governing tuberization at elevated temperature. Pathway maps were constructed to graphically represent the gene expression patterns. Genes associated with photosynthesis, hormonal activity, sugar transporters and transcription factors were differentially expressed. The results are presented and discussed in terms of tuberization at high temperature. The effect of high temperature on expression of genes controlling tuberization was also analyzed. This study provided useful information on potato tuberization at elevated temperature and make available a framework for further investigations into heat stress in potato.

Storage behaviour and cooking quality of Indian potato varieties.

Tubers of forty four indigenous potato varieties were assessed for storage behaviour at room temperature, tuber dry matter content and cooking quality during 2010, 2011 and 2012. The maximum, minimum temperatures and relative humidity during storage period ranged between 26 to 40 °C, 17-28 °C and 18 to 82 %, respectively. The lowest total weight loss was recorded in variety Kufri Pushkar (7.7 %) followed by Kufri Lalima (7.9 %), Kufri Surya (8.3 %), Kufri Red (9.2 %), Kufri Dewa, Kufri Sheetman (9.3 %), Kufri Chandramukhi, Kufri Jyoti (9.5 %), Kufri Sindhuri (9.6 %), Kufri Kuber (9.7 %), Kufri Chipsona-1 (9.8 %), Kufri Kundan (9.9 %) and Kufri Chamatkar (10.0 %). Highest tuber dry matter content (%) was observed in Variety Kufri Kundan (24.2) followed by Kufri Himsona (23.7), Kufri Frysona (23.6), Kufri Kuber (22.7), Kufri Chipsona-2 (22.3). Kufri Khasigaro (22.0), Kufri Sheetman (21.9), Kufri Chipsona-3 (21.7) and lowest in Variety Kufri Khyati and Kufri Pukhraj (16.1 %). Of the total varieties, 14 were adjudged as floury, 15 mealy, 14 waxy and one (Kufri Ashoka) as soggy. The total weight loss had highly significant and positive correlation with sprout weight/Kg tubers (r = 0.76**), physiological weight loss (r = 0.97**). Based on the results potato varieties namely, Kufri Chamatkar, Kufri Chipsona-1, Kufri Chandramukhi, Kufri Dewa, Kufri Jyoti, Kufri Kuber, Kufri Kundan, Kufri Lalima, Kufri Lauvkar, Kufri Pushkar, Kufri Red, Kufri Safed, Kufri Sheetman, Kufri Sindhuri possessed excellent keeping quality with medium to long tuber dormancy, low storage losses, medium to high tuber dry matter and good flavour. The information generated in this study can be utilized in the breeding programme. This can also help the farmer to choose and cultivate the potato varieties as per demand of the consumers.

Microarray analysis of gene expression patterns in the leaf during potato tuberization in the potato somatic hybrid Solanum tuberosum and Solanum etuberosum.

Genes involved in photoassimilate partitioning and changes in hormonal balance are important for potato tuberization. In the present study, we investigated gene expression patterns in the tuber-bearing potato somatic hybrid (E1-3) and control non-tuberous wild species Solanum etuberosum (Etb) by microarray. Plants were grown under controlled conditions and leaves were collected at eight tuber developmental stages for microarray analysis. A t-test analysis identified a total of 468 genes (94 up-regulated and 374 down-regulated) that were statistically significant (p ≤ 0.05) and differentially expressed in E1-3 and Etb. Gene Ontology (GO) characterization of the 468 genes revealed that 145 were annotated and 323 were of unknown function. Further, these 145 genes were grouped based on GO biological processes followed by molecular function and (or) PGSC description into 15 gene sets, namely (1) transport, (2) metabolic process, (3) biological process, (4) photosynthesis, (5) oxidation-reduction, (6) transcription, (7) translation, (8) binding, (9) protein phosphorylation, (10) protein folding, (11) ubiquitin-dependent protein catabolic process, (12) RNA processing, (13) negative regulation of protein, (14) methylation, and (15) mitosis. RT-PCR analysis of 10 selected highly significant genes (p ≤ 0.01) confirmed the microarray results. Overall, we show that candidate genes induced in leaves of E1-3 were implicated in tuberization processes such as transport, carbohydrate metabolism, phytohormones, and transcription/translation/binding functions. Hence, our results provide an insight into the candidate genes induced in leaf tissues during tuberization in E1-3.

Counselling services in prevention of mother-to-child transmission (PMTCT) in Delhi, India: an assessment through a modified version of UNICEF-PPTCT tool.

The study aims to assess the counselling services provided to prevent mother to child transmission of HIV (PMTCT) under the Indian programme of prevention of parent-to-child transmission of HIV (PPTCT). Five hospitals in Delhi providing PMTCT services were randomly selected. A total of 201 post-test counselled women were interviewed using a modified version of the UNICEF-PPTCT evaluation tool. Knowledge about HIV transmission from mother-to-child was low. Post-test counselling mainly helped in increasing the knowledge of HIV transmission; yet 20%-30% of the clients missed this opportunity. Discussion on window period, other sexually transmitted diseases and danger signs of pregnancy were grossly neglected. The PMTCT services during the antenatal period are feasible and agreeable to be provided; however, certain aspects, like lack of privacy, confidentiality of HIV status of the client, counsellor's 'hurried' attitude, communication skills and discriminant behaviour towards HIV-positive clients, and disinterest of clients in the counselling, remain as gaps. These issues may be addressed through refresher training to counsellors with an emphasis on social and behaviour change communication strategies. Addressing attitudinal aspects of the counsellors towards HIV positives is crucial to improve the quality of the services to prevent mother-to-child transmission of HIV.