The lncRNAs at X Chromosome Inactivation Center: Not Just a Matter of Sex Dosage Compensation

Non‐coding RNAs (ncRNAs) constitute the majority of the transcriptome, as the result of pervasive transcription of the mammalian genome. Different RNA species, such as lncRNAs, miRNAs, circRNA, mRNAs, engage in regulatory networks based on their reciprocal interactions, often in a competitive manner, in a way denominated “competing endogenous RNA (ceRNA) networks” (“ceRNET”): miRNAs and other ncRNAs modulate each other, since miRNAs can regulate the expression of lncRNAs, which in turn regulate miRNAs, titrating their availability and thus competing with the binding to other RNA targets. The unbalancing of any network component can derail the entire regulatory circuit acting as a driving force for human diseases, thus assigning “new” functions to “old” molecules. This is the case of XIST, the lncRNA characterized in the early 1990s and well known as the essential molecule for X chromosome inactivation in mammalian females, thus preventing an imbalance of X‐linked gene expression between females and males. Currently, literature concerning XIST biology is becoming dominated by miRNA associations and they are also gaining prominence for other lncRNAs produced by the X‐inactivation center. This review discusses the available literature to explore possible novel functions related to ceRNA activity of lncRNAs produced by the X‐inactivation center, beyond their role in dosage compensation, with prospective implications for emerging gender‐biased functions and pathological mechanisms

Human MicroRNAs Interacting With SARS-CoV-2 RNA Sequences: Computational Analysis and Experimental Target Validation

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel RNA virus affecting humans, causing a form of acute pulmonary respiratory disorder named COVID-19, declared a pandemic by the World Health Organization. MicroRNAs (miRNA) play an emerging and important role in the interplay between viruses and host cells. Although the impact of host miRNAs on SARS-CoV-2 infection has been predicted, experimental data are still missing. This study started by a bioinformatics prediction of cellular miRNAs potentially targeting viral RNAs; then, a number of criteria also based on experimental evidence and virus biology were applied, giving rise to eight promising binding miRNAs. Their interaction with viral sequences was experimentally validated by transfecting luciferase-based reporter plasmids carrying viral target sequences or their inverted sequences into the lung A549 cell line. Transfection of the reporter plasmids resulted in a reduction of luciferase activity for five out of the eight potential binding sites, suggesting responsiveness to endogenously expressed miRNAs. Co-transfection of the reporter plasmids along with miRNA mimics led to a further and strong reduction of luciferase activity, validating the interaction between miR-219a-2-3p, miR-30c-5p, miR-378d, miR-29a-3p, miR-15b-5p, and viral sequences. miR-15b was also able to repress plasmid-driven Spike expression. Intriguingly, the viral target sequences are fully conserved in more recent variants such as United Kingdom variant B.1.1.7 and South Africa 501Y.V2. Overall, this study provides a first experimental evidence of the interaction between specific cellular miRNAs and SARS-CoV-2 sequences, thus contributing to understanding the molecular mechanisms underlying virus infection and pathogenesis to envisage innovative therapeutic interventions and diagnostic tools

Efficacy of eprinomectin pour-on against Dictyocaulus arnfieldi infection in donkeys (Equus asinus).

A trial to assess the efficacy of eprinomectin (EPM) against the lungworm Dictyocaulus arnfieldi was carried out on 15, naturally-infected donkeys. Ten animals were treated with a 'pour-on' EPM preparation (at a dose of 0.5mg/kg bodyweight), and five animals acted as controls. EPM was 100% effective in eliminating faecal larvae from day 7, until the end of study at day 28. No adverse drug-reactions or side-effects were observed in any of the treated donkeys

A novel ceRNA regulatory network involving the long non-coding antisense RNA SPACA6P-AS, miR-125a and its mRNA targets in hepatocarcinoma cells

MicroRNAs (miRNA), and more recently long non-coding RNAs (lncRNA), are emerging as a driving force for hepatocellular carcinoma (HCC), one of the leading causes of cancer-related death. In this work, we investigated a possible RNA regulatory network involving two oncosuppressive miRNAs, miR-125a and let-7e, and a long non-coding antisense RNA, SPACA6P-AS (SP-AS), all transcribed from the same locus, with SP-AS in the opposite direction and thus carrying complementary sequences to the miRNAs. In vitro experiments validated the binding of the miRNAs to SP-AS. Then, the boosting of either the miRNAs or SP-AS levels demonstrated their reciprocal inhibition. In addition, overexpression of SP-AS resulted in a reduced silencing activity of miR-125a and let-7e toward their key oncogenic targets, i.e., Lin28b, MMP11, SIRT7, Zbtb7a, Cyclin D1, CDC25B, HMGA2, that resulted significantly upregulated. Finally, the analysis of 374 HCC samples in comparison to 50 normal liver tissues showed an upregulation of SP-AS and a reverse expression of miR-125a, not observed for let-7e; consistently, miR-125a oncogenic targets were upregulated. Overall, the data depict a novel competing endogenous RNA (ceRNA) network, ceRNET, whereby miR-125a can regulate the expression of SP-AS, which in turn regulates the miRNA by competing with the binding to the mRNA targets. We speculate that the unbalancing of any network component may contribute to hepatocarcinogenesis

Estimation of buffalo cheese yield by using the chemical-physical parameters of the milk

The aim of this study was to estimate cheese yield by using the chemical- physical parameters of the milk. Analysis were performed on 325 milk samples with 80-219 days in milk interval. Furthermore, buffaloes which showed a ratio between theoretical cheese yield (calculated by Altiero formula) and real cheese yield at 28 hours higher (Group A) or lower (Group B) than 0.983, were compared taking into account 5 hypothetical analytical potentialities of laboratories: 1) Fat percentage; 2) Protein and fat percentages; 3) Protein and fat percentages, pH and SH; 4) Protein and fat percentages, pH, SH, urea, protein percentage corrected per urea, lactose, solids-not-fat (SNF) and SCC; 5) Protein and fat percentages, pH, SH, urea, protein percentage corrected per urea, lactose, SNF, SCC, TAMF, milk DM percentage, ash percentage and casein percentage. Correlation and regression analyses with stepwise method were performed for curd quantity in relation to the physic-chemical ad microbiological milk composition by using SPSS 15.0. As expected, R2 value was such high as the number of variables included in the calculation. A higher R2 value was observed in those samples characterized by a ThCY/28CY ratio < 0.983. ThCY calculated according to Altiero et al (1989), underestimated 28CY of +1.8 g/litre in all samples, whereas a difference between –2.2 (Laboratory 2) and +1.0 (Laboratory 3) g/litre was registered if the actual formula is utilized. According to Altiero formula, 28CY was overestimated of 9.6 g/litre in Group A, whereas it was underestimated of 1.8 g/litre in Group B. According to our study, the estimation of 28CY showed a difference between –9.3 (Laboratory 2) and 9 (Laboratory 1) g/litre in Group A and – 3.5 (Laboratory 1) e 0.0 (Laboratory 5) g/litre

embryonic mortality in buffalo naturally mated

The aim of this work was to evaluate the incidence of embryonic mortality in three different period of year in buffaloes naturally mated. The trial was carried out in a buffalo farm located in Caserta province between 2000-2006. In this period were registered natural insemination on 200 buffaloes. Pregnancy diagnosis was carried out on Day 30, confirmed on Day 45 and every 15th days until 90 days after natural mating. Buffaloes that were pregnant on Day 30 but not on Day 45 or Day 90 were considered to have undergone embryonic (EM) or fetal mortality (FM) respectively. EM and FM were 8.8% and 13.4% respectively throughout the experimental period. A high incidence (P<0.01) of FM was found in the transitional period (December-March) than in other months of the year. The incidence of embryonic mortality was significantly (P<0.01) higher between 28-60 days of gestation and lower after 71 day of gestation. The higher fetal mortality found in this study could be due the lower serum levels of progesterone normally found in transitional period in buffalo cows

milk protein and cheese yield in buffalo species

Buffalo milk samples differing significantly for cheese yield values were analysed by 2D electrophoresis in order to outline a protein profile, with specific regards to k-casein fractions. Four buffaloes, two of which showing high cheese yield and two with low cheese yield selected from a group of 135 subjects were chosen for the proteomic analyses. Six main spots in 2D gels were recognized as αs1-, αs2-, β- and k-casein, α-lactoalbumin, β-lactoglobulin. The main visible differences in the 2D gels between buffaloes with high vs. low cheese yield were found in the appearance of the four k-casein spots (spots numbers:20, 19, 16, 18) which differ in the number of phosphorilation and glycosilation. The area and the intensity of the four spots were calculated by using Melanie II (Bio-Rad) software. Samples with high cheese yield showed higher value of the by-products: area x intensity of spot 16, correspondent to k-casein with one phosphorilation site, and lower values of spots 19 and 20, of k-casein with more than one phosphorilation site and glycosilated

microRNA-377-3p downregulates the oncosuppressor T-cadherin in colorectal adenocarcinoma cells

Colorectal cancer (CRC) is the second leading cause of death of malignant tumors worldwide. Recent studies point to a role for the adiponectin-receptor axis in colorectal carcinogenesis, and in particular to the oncosuppressive properties of the T-cadherin receptor. In addition, the loss of T-cadherin expression in tumor tissues has been linked to cancer progression and attributed to aberrant methylation of its promoter. Recognizing the pivotal role of microRNAs in CRC, this study explores their possible contribution to the downregulation of T-cadherin. A systematic bioinformatics analysis, restricted by microRNA expression data in the colon or in cultured colorectal cell lines, predicted twelve top-ranking target miRNA sites within the 3ʹ UTR of T-cadherin. Experimental validation analyses based on luciferase reporter constructs and miRNA mimic or miRNA inhibitor transfections toward colorectal adenocarcinoma cell lines indicated that miR-377-3p was able to directly bind to the T-cadherin sequence, and thus downregulating&nbsp;its expression. Given the oncogenic activity of miR-377 and the oncosuppressive activity of T-cadherin in CRC, the regulatory circuit highlighted in this study may add new insights into molecular mechanisms driving colorectal carcinogenesis, and perspectively it could be exploited to identify novel biomarkers and therapeutic targets

Relationship of body condition score and blood urea and ammonia to pregnancy in Italian Mediterranean buffaloes

The relationship of body condition score ( BCS) and blood urea and ammonia to pregnancy outcome was examined in Italian Mediterranean Buffalo cows mated by AI. The study was conducted on 150 buffaloes at 145 +/- 83 days in milk that were fed a diet comprising 14.8% crude protein, 0.9 milk forage units . kg(-1) dry matter and a non- structural carbohydrate/ crude protein ratio of 2.14. The stage of the oestrous cycle was synchronised by the Ovsynch- TAI programme and blood urea and ammonia levels were assessed on the day of AI. Energy corrected milk ( ECM) production and BCS were recorded bi- weekly. The pregnancy risk was 46.7% and was slightly lower in buffaloes with BCS 7.5. There were no significant differences in ECM, urea and ammonia between pregnant and non- pregnant buffaloes. However, pregnancy outcome was higher ( P = 0.02) in buffaloes with blood urea < 6.83 mmol . L-1. The likelihood of pregnancy for buffaloes with low urea blood level was 2.6 greater than for high urea level and exposure to a high urea level lowered the probability of pregnancy by about 0.25. The findings indicate that buffaloes are similar to cattle and increased blood levels of urea are associated with reduced fertility when animals are mated by AI

Effect of lifestyle changes on erectile dysfunction in obese men: a randomized controlled trial

Context: Healthy lifestyle factors are associated with maintenance of erectile function in men. Objective: To determine the effect of weight loss and increased physical activity on erectile and endothelial functions in obese men. Design, Setting, and Patients: Randomized, single-blind trial of 110 obese men (body mass index ≥30) aged 35 to 55 years, without diabetes, hypertension, or hyperlipidemia, who had erectile dysfunction that was determined by having a score of 21 or less on the International Index of Erectile Function (IIEF). The study was conducted from October 2000 to October 2003 at a university hospital in Italy. Interventions: The 55 men randomly assigned to the intervention group received detailed advice about how to achieve a loss of 10% or more in their total body weight by reducing caloric intake and increasing their level of physical activity. Men in the control group (n=55) were given general information about healthy food choices and exercise. Main Outcomes Measures: Erectile function score, levels of cholesterol and tryglycerides, circulating levels of interleukin 6, interleukin 8, and C-reactive protein, and endothelial function as assessed by vascular responses to L-arginine. Results: After 2 years, body mass index decreased more in the intervention group (from a mean [SD] of 36.9 [2.5] to 31.2 [2.1]) than in the control group (from 36.4 [2.3] to 35.7 [2.5]) (P<.001), as did serum concentrations of interleukin 6 (P=.03), and C-reactive protein (P=.02). The mean (SD) level of physical activity increased more in the intervention group (from 48 [10] to 195 [36] min/wk; P<.001) than in the control group (from 51 [9] to 84 [28] min/wk; P<.001). The mean (SD) IIEF score improved in the intervention group (from 13.9 [4.0] to 17 [5]; P<.001), but remained stable in the control group (from 13.5 [4.0] to 13.6 [4.1]; P=.89). Seventeen men in the intervention group and 3 in the control group (P=.001) reported an IIEF score of 22 or higher. In multivariate analyses, changes in body mass index (P=.02), physical activity (P=.02), and C-reactive protein (P=.03) were independently associated with changes in IIEF score. Conclusion: Lifestyle changes are associated with improvement in sexual function in about one third of obese men with erectile dysfunction at baseline