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MicroRNAs (miRNA) are important regulators of oocyte maturation, playing a key role in modulating gene expression both in a temporal- and spatial-specific manner. These small non-coding RNAs are involved in important processes during oocyte maturation, acting as messengers between the oocyte and its surrounding cumulus cells. Despite its significance, the bidirectional communication mechanism is still unknown. To test miRNA communication between oocyte and surrounding cumulus cells through the gap junctions the gap junctions were either blocked with carbenoxolone or not. MiRNA sequencing of oocytes at 1, 6, and 22 h of in vitro maturation was then performed. Among the differentially expressed miRNAs, bta-miR-21-5p, a regulator of cumulus cell viability and oocyte maturation, was the only previously known miRNA. Furthermore, by labeling a bta-miR-21-5p mimic with FAM, crossing of this miRNA through the gap junctions within the cumulus-oocyte complex could be visualized and internalization in the oocyte was confirmed by RT-qPCR. In conclusion, this study provides, for the first time, evidence that miRNA communication within the bovine cumulus-oocyte complex is enabled through the gap junctional network. [ABSTRACT FROM AUTHOR]

Background: Within the follicular fluid, extracellular vesicles (EVs) guide oocyte growth through their cargo microRNAs (miRNAs). Here, we investigated the role of EVs and their cargo miRNAs by linking the miRNAs found in EVs, derived from the fluid of an individual follicle, to the ability of its oocyte to become a blastocyst (competent) or not (non-competent)., Methods: Bovine antral follicles were dissected, categorized as small (2-4 mm) or large (5-8 mm) and the corresponding oocytes were subjected to individual maturation, fertilization and embryo culture to the blastocyst stage. Follicular fluid was pooled in 4 groups (4 replicates) based on follicle size and competence of the corresponding oocyte to produce a blastocyst. Follicular fluid-derived EVs were isolated, characterized, and subjected to miRNA-sequencing (Illumina Miseq) to assess differential expression (DE) in the 4 groups. Functional validation of the effect of miR-34c on embryo development was performed by supplementation of mimics and inhibitors during in vitro maturation (IVM)., Results: We identified 16 DE miRNAs linked to oocyte competence when follicular size was not considered. Within the large and small follicles, 46 DE miRNAs were driving blastocyst formation in each group. Comparison of EVs from competent small and large follicles revealed 90 DE miRNAs. Cell regulation, cell differentiation, cell cycle, and metabolic process regulation were the most enriched pathways targeted by the DE miRNAs from competent oocytes. We identified bta-miR-34c as the most abundant in follicular fluid containing competent oocytes. Supplementation of miR-34c mimic and inhibitor during IVM did not affect embryo development. However, blastocyst quality, as evidenced by higher cell numbers, was significantly improved following oocyte IVM in the presence of miR-34c mimics, while miR-34c inhibitors resulted in the opposite effect., Conclusion: This study demonstrates the regulatory effect of miRNAs from follicular fluid-derived EVs on oocyte competence acquisition, providing a further basis for understanding the significance of miRNAs in oocyte maturation and embryonic development. Up-regulation of miR-34c in EVs from follicular fluid containing competent oocytes and the positive impact of miR-34c mimics added during IVM on the resulting blastocysts indicate its pivotal role in oocyte competence., (© 2024. The Author(s).)

Background: The southern African yellow dog tick, Haemaphysalis elliptica, occurs in eastern and southern Africa and adults infest domestic and wild carnivores. This tick species is also a vector of the highly virulent Babesia rossi pathogen, the causative agent of canine babesiosis in sub-Saharan Africa. Sustained high levels of efficacy of a parasiticide are not only important in protecting dogs against the direct effects of tick infestation, but also in reducing the risk of tick-borne diseases. Sarolaner (Simparica™ chewable tablets) has been reported to be effective against the major tick species infesting dogs in Europe and the USA, including representatives from the genera Amblyomma, Ixodes, Rhipicephalus and Dermacentor. Until now no efficacy evaluations have been reported against species of the genus Haemaphysalis. The objective of the study was to confirm the efficacy of a single 2 mg sarolaner/kg oral dose of Simparica™ against induced infestations with H. (R.) elliptica, an important parasite of dogs in southern Africa., Methods: This blinded, randomised, single centre, placebo controlled efficacy study followed a parallel group design and was conducted on two groups consisting of eight purpose-bred dogs each. Animals were treated orally, once on Day 0, with either a placebo compound (Group 1) or Simparica™ (Group 2). Simparica™ was administered orally at a dose rate of 2 mg sarolaner/kg body weight. The dogs were infested with ticks on Days - 7, - 2, 5, 12, 19, 26 and 33, with removal counts conducted on Days - 5, 2, 7, 14, 21, 28 and 35., Results: A single oral administration of Simparica™ (sarolaner) at a minimum dose of 2 mg/kg resulted in a 100% efficacy against existing infestations of H. (R.) elliptica on dogs and a 100% reduction in live ticks following weekly re-infestations for 35 days. Moreover, the immediate and persistent high levels of efficacy observed in this study for 35 days is consistent with those observed in previous studies against ticks in other genera., Conclusions: The efficacy of sarolaner (Simparica™), administered orally to dogs at the minimum label dose of 2.0 mg/kg, was demonstrated against existing and weekly re-infestations of H. (R.) elliptica for at least 5 weeks. Efficacy of 100% was achieved against existing infestations as well as weekly re-infestations.

Background: Canine babesiosis is a clinically significant emerging vector-borne disease caused among others by the protozoan Babesia canis. The efficacy of sarolaner (Simparica®; Zoetis; at the minimum recommended label dose of 2.0 mg per kg bodyweight) in the prevention of babesiosis was evaluated in twenty-four dogs randomly allocated to either a placebo-treated group or one of two sarolaner-treated groups. At 21 or 28 days after treatment administration, dogs were infested with 50 ± 4 Dermacentor reticulatus ticks of which 25% were confirmed to be infected with Babesia canis. Blood samples were collected from each dog prior to tick infestation and weekly thereafter until 49 days after infestation. The blood was assayed for B. canis antibodies using an indirect immunofluorescence test (IFAT) and for B. canis DNA by PCR assay. A dog was a priori defined as B. canis-positive if it tested positive by both IFAT and PCR at any time during the study., Results: No treatment-related adverse reactions were recorded during the study. All placebo-treated animals displayed clinical signs due to babesiosis and tested positive on both IFAT and PCR. None of the sarolaner-treated animals displayed any clinical symptoms or tested positive on both IFAT and PCR, resulting in a 100% efficacy in the prevention of canine babesiosis (P = 0.0002)., Conclusion: When given 21 or 28 days before tick infestation, a single treatment with sarolaner at the minimum recommended label dose of 2.0 mg per kg body weight prevented the transmission of B. canis by D. reticulatus to dogs.

Objective: To evaluate the effectiveness and safety of cefovecin sodium in the treatment of cats with naturally occurring skin infections (abscesses and infected wounds)., Design: Multicenter (26 sites), randomized, double-blind, controlled clinical trial., Animals: Client-owned cats of any breed with naturally occurring skin infections with associated clinical signs and confirmatory bacteriologic culture results., Procedures: Cats with clinical signs of skin and soft tissue infection were randomly allocated to receive a single dose of cefovecin (8 mg/kg [3.6 mg/lb], SC) followed by placebo drops administered orally once daily for 14 days or 1 SC placebo injection followed by cefadroxil (22 mg/kg [10 mg/lb], PO, once daily for 14 days). Only one 14-day treatment course was permitted., Results: Effectiveness of cefovecin in the treatment of cats with abscesses and infected wounds was similar to that of cefadroxil. At the final assessment on day 28, 97% (86/89) of cefovecin-treated cats and 91% (80/88) of cefadroxil-treated cats were considered treatment successes. There were no serious adverse events or deaths related to treatment., Conclusions and Clinical Relevance: 1 SC injection of 8 mg of cefovecin/kg for the treatment of cats with naturally occurring skin infections (wounds and abscesses) was safe and as effective as cefadroxil administered orally at 22 mg/kg, once daily for 14 days.

Objective: To evaluate the efficacy and safety of administration of cefovecin, compared with cefadroxil, for treatment of naturally occurring secondary superficial pyoderma, abscesses, and infected wounds in dogs., Design: Multicenter, randomized, positive-controlled clinical trial., Animals: 235 client-owned dogs., Procedures: Dogs with clinical signs of skin infection confirmed via bacteriologic culture were randomly allocated to receive a single SC injection of cefovecin (8 mg/kg [3.6 mg/lb]) followed by placebo administered PO twice daily for 14 days or cefadroxil (22 mg/kg [10 mg/lb]) administered PO twice daily for 14 days following a placebo injection. Two 14-day treatment courses were permitted. Treatment success was defined as reduction of clinical signs to mild or absent at the final assessment., Results: Clinical efficacy achieved with cefovecin in dogs was equivalent to that observed with cefadroxil. At the final assessment, 14 days following the completion of treatment (on day 28 or 42), 92.4% (109/118) of the cefovecin group and 92.3% (108/117) of the cefadroxil group were treatment successes. There were no serious adverse events or deaths related to treatment., Conclusions and Clinical Relevance: A single cefovecin injection (8 mg/kg) administered SC, which could be repeated once after 14 days, was safe and effective against naturally occurring skin infections in dogs and as effective as cefadroxil administered PO twice daily for 14 or 28 days.

The effect of guanfacine (2 mg once daily) on ambulatory blood pressure was studied with the Remler M 2000 recorder in 16 elderly hypertensive patients during a randomized, double-blind, placebo-controlled, balanced, cross-over study. Guanfacine significantly reduced heart rate and systolic and diastolic ambulatory blood pressure. The antihypertensive effect was maintained over the whole recording period. Systolic and diastolic blood pressure variability was not changed by guanfacine, neither when defined as standard deviation or variation coefficient of the mean, nor when defined as the range between the highest and lowest ambulatory blood pressure, suggesting that blood pressure variability is unrelated to sympathetic nervous system activity.

Chronic administration of nadolol has been reported to reduce blood pressure either without or with a concomitant fall of renal blood flow. We therefore studied the effects of nadolol 80 mg once daily on ambulatory blood pressure, renal and systemic haemodynamics in patients with mild to moderate essential hypertension. Ten patients took part in this randomized, double-blind, placebo-controlled, crossover study, each phase of which lasted 4 weeks. Nadolol significantly reduced ambulatory blood pressure and heart rate, but had no effect on blood pressure variability. Cardiac output was significantly reduced by nadolol and total peripheral resistance increased but without reaching statistical significance. Despite the fall in blood pressure and cardiac output, renal blood flow and glomerular filtration rate remained unchanged. The fraction of cardiac output reaching the kidneys rose significantly and renal vascular resistance was significantly reduced. Body weight, urinary sodium excretion and urine flow rate remained unchanged. We conclude that nadolol 80 mg once daily lowers ambulatory blood pressure in patients with mild to moderate hypertension without impairment of renal blood flow, indicating a redistribution of cardiac output to the kidneys. The mechanism of the renal vasodilator effect of nadolol remains to be determined.

A randomized, double-blind, placebo-controlled study was set up to study the effects of acute and chronic administration of carvedilol, a vasodilatory beta-blocker in essential hypertension. Acute administration of a single dose of 50 mg of carvedilol reduced systolic and diastolic blood pressure, without inducing reflex tachycardia. Renal blood flow was preserved; accordingly renal vascular resistance was significantly reduced. A significant reduction of glomerular filtration rate and filtration fraction was observed. Plasma renin activity (PRA) and plasma aldosterone were not changed. Chronic carvedilol treatment produced a significant fall in systolic and diastolic office and ambulatory blood pressure, heart rate, cardiac output, PRA and plasma aldosterone. Blood pressure variability was not changed. Renal blood flow, glomerular filtration rate and filtration fraction also remained unchanged; renal vascular resistance decreased significantly. It is concluded that carvedilol possesses definite antihypertensive and renal vasodilating properties, both acutely and after chronic treatment.

The effects of 4 weeks of placebo on clinic and on ambulatory blood pressure, measured non-invasively using the Remler M 2000 portometer, were studied in 46 hypertensive patients who were included in three consecutive double-blind randomized placebo-controlled trials with antihypertensive drugs. Placebo significantly reduced clinic blood pressure, but had no significant effect on ambulatory blood pressure.

The effect of the converting enzyme inhibitor lisinopril on renal and cardiac hemodynamics was studied in patients with moderate to severe primary hypertension, in a multicenter, double-blind, randomized clinical trial comparing the antihypertensive effect of lisinopril (LIS) and nifedipine (NIF). After a 2 week placebo run-in period, 15 patients were randomized in a 2:1 ratio to receive either LIS (20-80 mg q.d., n = 10) or NIF (20-40 mg b.i.d., n = 5). LIS significantly reduced blood pressure (BP) without changing heart rate or cardiac output. LIS significantly increased renal blood flow; glomerular filtration rate (GFR) was not changed. It can be concluded that LIS is an effective antihypertensive agent with a favorable renal hemodynamic profile.