Identification and characterization of miRNAs expressed in the bovine ovary

<p>Abstract</p> <p>Background</p> <p>MicroRNAs are the major class of gene-regulating molecules playing diverse roles through sequence complementarity to target mRNAs at post-transcriptional level. Tightly regulated expression and interaction of a multitude of genes for ovarian folliculogenesis could be regulated by these miRNAs. Identification of them is the first step towards understanding miRNA-guided gene regulation in different biological functions. Despite increasing efforts in miRNAs identification across various species and diverse tissue types, little is known about bovine ovarian miRNAs. Here, we report the identification and characterization of miRNAs expressed in the bovine ovary through cloning, expression analysis and target prediction.</p> <p>Results</p> <p>The miRNA library (5'-independent ligation cloning method), which was constructed from bovine ovary in this study, revealed cloning of 50 known and 24 novel miRNAs. Among all identified miRNAs, 38 were found to be new for bovine and were derived from 43 distinct loci showing characteristic secondary structure. While 22 miRNAs precursor loci were found to be well conserved in more than one species, 16 were found to be bovine specific. Most of the miRNAs were cloned multiple times, in which let-7a, let-7b, let-7c, miR-21, miR-23b, miR-24, miR-27a, miR-126 and miR-143 were cloned 10, 28, 13, 4, 11, 7, 6, 4 and 11 times, respectively. Expression analysis of all new and some annotated miRNAs in different intra-ovarian structures and in other multiple tissues showed that some were present ubiquitously while others were differentially expressed among different tissue types. Bta-miR-29a was localized in the follicular cells at different developmental stages in the cyclic ovary. Bio-informatics prediction, screening and Gene Ontology analysis of miRNAs targets identified several biological processes and pathways underlying the ovarian function.</p> <p>Conclusion</p> <p>Results of this study suggest the presence of miRNAs in the bovine ovary, thereby elucidate their potential role in regulating diverse molecular and physiological pathways underlying the ovarian functionality. This information will give insights into bovine ovarian miRNAs, which can be further characterized for their role in follicular development and female fertility as well.</p

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10&nbsp;years; 78.2% included were male with a median age of 37&nbsp;years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p&lt;0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p&lt;0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised

Exploitation of next generation sequencing technologies for unraveling metabolic pathways in medicinal plants: A concise review

Medicinal plants having high-value and unique active ingredients are attractive in different industries, including pharmaceutical, food and cosmetic sectors. The phytochemicals in these plants are synthesized through diverse biosynthetic pathways including those for mevalonate, shikimic, acetic acid and so forth. Several key enzymes catalyze different reactions in these pathways and convert precursors to final products. Identification of rate limiting enzymes in the biosynthetic pathways of the secondary metabolites and their encoding genes can reveal the bottlenecks of these pathways and ultimately lead us to develop an appropriate breeding strategy for selecting plants possessing desirable and high-quantity active ingredients. Next-generation sequencing methods, in the last two decades, have contributed, in many ways to a more detailed study of medicinal plants and a deeper exploration of these pathways. In this way, an NGS-based whole genome sequencing (WGS) approach and the data it provides, can lead to the development of new high-throughput molecular markers for more effective marker assisted selection (MAS), more accurate QTL and association mappings, easier genotyping by sequencing, a deeper study of phylogenetic relationships, a faster calculation of evolutionary rates of different genes, reconstruction of genetic diversity patterns between plant taxa, and ultimately domestication of wild high-value medicinal plants. In addition, RNA-Seq methods can have a similar application in the plants without a WGS as well as being used for representing the expression profile of the genes in plants with a great impact on functional genomics. NGS-based methods can greatly help medicinal plant researchers to identify genes involved in the biosynthetic pathways of important metabolites including phenolics, terpenoids, alkaloids, glycosides, etc. Subsequently, through metabolic engineering, it would be possible to increase the quantity and quality of sec-ondary metabolites and to develop new medicinal plant cultivars for their exploitation in an industrial scale

Induction of colitis in cftr-/- mice results in bile duct injury.

It is unknown why some patients with inflammatory bowel disease develop primary sclerosing cholangitis. We have recently shown that patients with primary sclerosing cholangitis have an increased prevalence of mutations in the gene responsible for cystic fibrosis (CFTR) compared with individuals with inflammatory bowel disease alone. Our aim was to examine whether induction of colitis by oral dextran leads to bile duct injury in mice heterozygous or homozygous for mutations in CFTR. The effect of oral administration of docosahexaenoic acid to correct a fatty acid imbalance associated with cystic fibrosis was also examined to determine whether this would prevent bile duct inflammation. Wild-type mice and mice heterozygous and homozygous for CFTR mutations were given dextran orally for 14 days to induce colitis. Bile duct injury was quantitated by blinded histological scoring and measurement of serum alkaline phosphatase activity. The effect of pretreatment with docosahexaenoic acid for 7 days was examined. Treatment of mice with 100 mg dextran/day for 9 days followed by 85 mg/day for 5 days resulted in a significant increase in bile duct injury as determined by histological scoring in homozygous cystic fibrosis mice compared with wild-type mice (P = 0.005). The bile duct injury seen in cystic fibrosis mice was reflected in a threefold increase in serum alkaline phosphatase (P = 0.0006). Pretreatment with oral docosahexaenoic acid decreased both histological evidence of bile duct injury and serum alkaline phosphatase levels. In the setting of colitis, loss of CFTR function leads to bile duct injury

InteliRank: A Four-Pronged Agent for the Intelligent Ranking of Cloud Services Based on End-Users’ Feedback

Cloud Computing (CC) provides a combination of technologies that allows the user to use the most resources in the least amount of time and with the least amount of money. CC semantics play a critical role in ranking heterogeneous data by using the properties of different cloud services and then achieving the optimal cloud service. Regardless of the efforts made to enable simple access to this CC innovation, in the presence of various organizations delivering comparative services at varying cost and execution levels, it is far more difficult to identify the ideal cloud service based on the user’s requirements. In this research, we propose a Cloud-Services-Ranking Agent (CSRA) for analyzing cloud services using end-users’ feedback, including Platform as a Service (PaaS), Infrastructure as a Service (IaaS), and Software as a Service (SaaS), based on ontology mapping and selecting the optimal service. The proposed CSRA possesses Machine-Learning (ML) techniques for ranking cloud services using parameters such as availability, security, reliability, and cost. Here, the Quality of Web Service (QWS) dataset is used, which has seven major cloud services categories, ranked from 0–6, to extract the required persuasive features through Sequential Minimal Optimization Regression (SMOreg). The classification outcomes through SMOreg are capable and demonstrate a general accuracy of around 98.71% in identifying optimum cloud services through the identified parameters. The main advantage of SMOreg is that the amount of memory required for SMO is linear. The findings show that our improved model in terms of precision outperforms prevailing techniques such as Multilayer Perceptron (MLP) and Linear Regression (LR)

Synthesis and Characterization of Antibiotic&ndash;Loaded Biodegradable Citrate Functionalized Mesoporous Hydroxyapatite Nanocarriers as an Alternative Treatment for Bone Infections

The continuing growth of bacterial resistance makes the top challenge for the healthcare system especially in bone-infections treatment. Current estimates reveal that in 2050 the death ratio caused by bacterial infections can be higher than cancer. The aim of this study is to provide an alternative to currently available bone-infection treatments. Here we designed mesoporous hydroxyapatite nanocarriers functionalized with citrate (Ctr&ndash;mpHANCs). Amoxicillin (AMX) is used as a model drug to load in Ctr&ndash;mpHANCs, and the drug loading was more than 90% due to the porous nature of nanocarriers. Scanning electron microscopy shows the roughly spherical morphology of nanocarriers, and the DLS study showed the approximate size of 92 nm. The Brunauer&ndash;Emmett&ndash;Teller (BET) specific surface area and pore diameter was found to be about 182.35 m2/g and 4.2 nm, respectively. We noticed that almost 100% of the drug is released from the AMX loaded Ctr&ndash;mpHANCs (AMX@Ctr&ndash;mpHANCs) in a pH-dependent manner within 3 d and 5 d at pH 2.0 and 4.5, respectively. The sustained drug release behaviour was observed for 15 d at pH 7.4 and no RBCs hemolysis by AMX@Ctr&ndash;mpHANCs. The broth dilution and colony forming unit (CFU) assays were used to determine the antimicrobial potential of AMX@Ctr&ndash;mpHANCs. It was observed in both studies that AMX@Ctr&ndash;mpHANCs showed a significant reduction in the bacterial growth of S. aureus, E. coli, and P. aeruginosa as compared to Ctr&ndash;mpHANCs with no bacteria-killing. Thus, we proposed that Ctr&ndash;mpHANCs can be used as a drug carrier and a treatment option for bone infections caused by bacteria