The effects of a glyphosate-based herbicide on the bovine gametes during an in vitro embryo production model.

Roundup® (R), while it is the most used herbicide globally, and its residues are ubiquitous in urban and suburban areas, its impact on vertebrates' safety remains highly debated. Here, in three in vitro experiments, we investigated the effects of a very low dose (1 ppm) of R on the fertilization capacity and embryo development in cattle. In the first experiment, frozen-thawed bull semen exposed to R for 1 h exhibited reduced motility parameters but unaffected fertilization ability. However, after in vitro fertilization, the rates of embryo formation were significantly lower compared to the untreated controls. In the second experiment, oocytes exposed to R during in vitro maturation showed reduced cleavage rates, and the embryo yield on days 7, 8, and 9 of embryo culture was significantly lower than that of the controls. In the third experiment, oocytes were matured in the presence of R and in a medium containing both R and Zinc, chosen to offer antioxidant protection to the oocytes. Day-7 blastocysts were analyzed for the expression of genes associated with oxidative stress, apoptosis, and epigenetic reprogramming. Exposure to R markedly suppressed embryo formation rates compared to the controls. The combination of R with Zinc restored the blastocyst yield, which on days 8 and 9 was comparable to that of the controls and higher than the groups exposed only to R on all days. The gene expression analysis revealed that R promotes oxidative stress development, triggers apoptosis, and induces epigenetic changes in developing embryos, while zinc presence alleviates these adverse effects of R. These findings imply that even at very low doses, R could be highly toxic, leading to functional abnormalities in both gametes, potentially affecting fertility in both genders.

Chios ram testicular blood flow and echotexture changes depending on age, season and ejaculation process.

The present study aimed to investigate the impact of age, season and ejaculation on ram testicular blood flow and echotexture. The survey was conducted biweekly on 7 Chios rams for one year, including breeding and non-breeding periods. The rams were divided into 2 age groups: 3 rams 2-6 years old (mature) and 4 rams 9-13 years old (old). Hemodynamic indices [Pulsatility index (PI), Resistive index (RI), End-diastolic velocity (EDV), testicular artery Diameter (D), Time-averaged maximum velocity (TAVM), Blood flow volume (BFV)] and echotexture parameters [Mean value (MV), Contrast (Con), Gray value distribution (GVD), Run length distribution (RunLD), Long run emphasis (LRunEm), Entropy (Ent), Correlation (Cor), Standard deviation (StD), Gray variance (GV) and Gradient mean value (GMV)] were evaluated in each testis before and after ejaculation. Ejaculation did not affect testes blood flow or echotexture (p>0.05). PI and RI were higher in the breeding period compared to the non-breeding period, for both testes (p⟨0.001). Left testis GV and Cor before ejaculation were lower (p=0.01) and higher (p=0.03), respectively, in the breeding compared to the non-breeding period. Left testis D (p=0.005) and BFV (p⟨0.001) were higher in old compared to mature rams after ejaculation. Right testis Con (p=0.03) and Cor (p=0.05) before ejaculation were higher in old rams, whereas right testis Ent after ejaculation was higher in mature rams (p=0.05). In conclusion, testicular blood flow and echotexture are affected by season and ram age, but not by ejaculation.

Breed differences of bull frozen-thawed semen.

The objective of this study was to investigate the quality of frozen-thawed semen from different bull breeds. Commercial frozen-thawed bull semen samples (26 per breed, 130 totally) of five breeds (Holstein [Η], Brown Swiss [BS], Limousin [L], Belgian Blue [BB], Blonde d' Aquitaine [BA]) were used. After thawing, each semen sample was subjected to thermal resistance test (TR) for 0.5 and 1 hr at 38°C and hypo-osmotic swelling test (HOST) for 1 hr at 150 mOsm at 37°C. Additionally, all samples were evaluated at times 0 hr (thawing), 0.5 hr (TR), 1 hr (TR) for kinetics by CASA [progressive, immotile, rapid, medium, slow moving spermatozoa, curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (VSL), linearity (LIN), straightness (STR), beat cross-frequency (BCF), amplitude of lateral head displacement (ALH), wobble (WOB)]. Moreover, directly after thawing, all semen samples were evaluated for morphometry, morphology, viability and DNA fragmentation. Statistical analysis was conducted using a mixed model for repeated measures. The results showed (a) higher VCL after thawing in H, L breeds compared to BB and BA, (b) higher VAP after thawing in L compared to BB, BA, (c) higher values of progressive spermatozoa after TR in H, BS compared to BB, BA, (d) higher values of rapid spermatozoa after thawing and 0.5 hr of TR in H, BS, L compared to BB, BA, (e) lower viability in BA after thawing compared to H, BS, BB, (f) lower morphological abnormalities in H compared to L, BB, (g) higher head length in Η compared to BB. No significant differences were observed in the results from HOST and DNA fragmentation between breeds. In conclusion, quality characteristics of frozen-thawed bull semen are dependent on the breed. Frozen semen from BB and BA breeds should be handled more carefully after thawing, as it is more sensitive to stress.

Glomerular filtration rate estimation in patients with type 2 diabetes: creatinine- or cystatin C-based equations?

AIMS/HYPOTHESIS: It is argued that GFR estimation (eGFR) using cystatin C-based equations (eGFRcys) is superior to that using creatinine-based equations (eGFRcre). We investigated whether eGFRcys are superior to eGFRcre in patients with type 2 diabetes. METHODS: GFR was measured in 448 type 2 diabetic patients using (51)Cr-EDTA-measured GFR (mGFR) as the reference standard. Bias, precision and accuracy of eGFRcys and eGFRcre were compared. RESULTS: The most accurate eGFRcre equation (Chronic Kidney Disease Epidemiology Collaboration [CKD-EPI]), which produced the highest proportion of estimates that were within 30% and 10% of the reference standard (80.7% and 38.0% of samples, respectively) had a bias of 7.1 and precision of 12.0 ml min(-1) 1.73 m(-2). The calibrated eGFRcys with the highest accuracy (Tan-C), which produced the highest proportion of estimates that were within 30% (78.8%) and within 10% (39.0%) of the reference standard had a bias of -3.5 and precision of 18.0 ml min(-1) 1.73 m(-2). Moreover, the areas under the receiver operating curve were higher with eGFRcre (CKD-EPI and Modification of Diet in Renal Disease [MDRD]) than with eGFRcys for the diagnosis of mild (mGFR <90 ml min(-1) 1.73 m(-2)) and moderate (mGFR <60 ml min(-1) 1.73 m(-2)) chronic kidney disease. In patients with mGFR ≥90 ml min(-1) 1.73 m(-2), CKD-EPI was the least biased, the most precise and the most accurate equation. CONCLUSIONS/INTERPRETATION: In patients with type 2 diabetes, eGFRcys do not currently provide better eGFR than eGFRcre. At present, compared with eGFRcys, eGFRcre are better at predicting the stage of chronic kidney disease. In addition, CKD-EPI seems to be the best equation for eGFR in patients with normal renal function.

Network-centric Analysis of Genetic Predisposition in Diabetic Nephropathy.

Diabetic nephropathy is a serious, long-term complication of diabetes and the leading cause of end-stage renal disease throughout the world. Although this disease is progressively imposing a heavier burden on the health care system, in many aspects it remains poorly understood. In addition to environmental influences, there is abundant evidence in support of genetic susceptibility to microvascular complications of nephropathy in diabetic patients. Familial clustering of phenotypes such as end-stage renal disease, albuminuria and kidney disease have been reported in large scale population studies throughout the world demonstrating strong contribution of inherited factors. Recent genome-wide linkage scans identified several chromosomal regions that are likely to contain diabetic nephropathy susceptibility genes, and association analyses have evaluated positional candidate genes under linkage peaks. In this review we have extracted from the literature the most promising candidate genes thought to confer susceptibility to diabetic nephropathy and mapped them to affected pathways by using network-centric analysis. Several of the top susceptibility genes have been identified as network hubs and bottlenecks suggesting that they might be important agents in the onset of diabetic nephropathy.