The relationship between bovine blastocyst formation in vitro and follicular fluid amino acids.

Follicular fluid has been found as a possible source of metabolic predictors for oocyte competence, and it is conveniently accessible during ovum pick-up (OPU). We used the OPU procedure to recover oocytes from 41 Holstein heifers for in vitro embryo production in this study. Follicular fluid was collected during OPU in order to establish a link between follicular amino acids and blastocyst formation. Each heifer's oocytes were collected, matured in vitro for 24 h and fertilized separately. The heifers were then divided into two groups based on blastocyst formation: those that produced at least one blastocyst (the blastocyst group, n = 29) and those that did not (the failed group, n = 12). The blastocyst group had higher follicular glutamine concentrations and lower aspartate levels than the failed group. Furthermore, network and Spearman correlation analyses revealed a link between blastocyst formation and aspartate (r = -0.37, p = 0.02) or glutamine (r = 0.38, p = 0.02). The receiver operator characteristic curve revealed that glutamine (AUC = 0.75) was the greatest predictor of blastocyst formation. These findings revealed that follicular amino acid levels in bovines can be used to predict blastocyst development.

Draft Genomic Sequences of Pseudomonas moorei Strains Isolated from Wild Cranberry Bogs in Massachusetts.

Pseudomonas moorei has been used to detoxify recalcitrant environmental contaminants from the pharmaceutical industry. Two P. moorei strains were isolated from soil in the pristine wild cranberry bogs of the Cape Cod National Seashore that putatively encode genes for degradation of 4- and 5-chlorosalicylates, acetaminophen, and diclofenac.

Innovating Care in Multiple Sclerosis: Feasibility of Synchronous Internet-Based Teleconsultation for Longitudinal Clinical Monitoring.

The 'coronavirus disease of 2019' crisis has recently forced an expedited adoption of teleconsultation (TC) in most medical domains. Short-term digital interventions have generally been associated with feasibility, clinical benefits, user satisfaction, and cost-effectiveness in patients with multiple sclerosis (MS) but outcomes after repeated utilization over extended periods need to be further evaluated. In this feasibility study, 60 subjects with MS were 1:1 randomized to receive standard care augmented by four TCs using an audiovisual Internet platform (intervention) versus standard care alone (controls), over a period of 12 months. Effects on functional status, medical costs, and satisfaction were explored as secondary outcomes. Eighty-nine out of 108 scheduled TCs (82.4%) were completed, and 26 patients could complete at least one TC (86.7%), meeting our prespecified feasibility target of 80%. The intervention did not lead to significant differences in functional status (with the potential exception of fatigue) nor medical costs. Most interventional patients declared themselves to be (very) satisfied about the quality of care and technical aspects associated with the TCs. Our results demonstrate that longitudinal clinical monitoring using real-time audiovisual TC over the Internet is feasible and well-received by patients with MS. Such an approach can be a promising new care strategy.

Epidermal growth factor alleviates the negative impact of urea on frozen-thawed bovine sperm, but the subsequent developmental competence is compromised.

Upon insemination, sperm cells are exposed to components of the female reproductive tract (FRT) fluids, such as urea and epidermal growth factor (EGF). It has been shown that both urea and EGF use EGF receptor signaling and produce reactive oxygen species (ROS) that are required at certain levels for sperm capacitation and acrosome reaction. We therefore hypothesized that during bovine sperm capacitation, a high level of urea and EGF could interfere with sperm function through overproduction of ROS. High-level urea (40 mg/dl urea is equal to 18.8 mg/dl of blood urea nitrogen) significantly increased ROS production and TUNEL-positive sperm (sperm DNA fragmentation, sDF) percentage, but decreased HOS test score, progressive motility, acrosome reaction and capacitation. The EGF reversed the negative effects of urea on all sperm parameters, with the exception of ROS production and DNA fragmentation, which were higher in urea-EGF-incubated sperm than in control-sperm. The developmental competence of oocytes inseminated with urea-EGF-incubated sperm was significantly reduced compared to the control. A close association of ROS production or sDF with 0-pronuclear and sperm non-capacitation rates was found in the network analysis. In conclusion, EGF enhanced urea-reduced sperm motility; however, it failed to reduce urea-increased sperm ROS or sDF levels and to enhance subsequent oocyte competence. The data suggests that any study to improve sperm quality should be followed by a follow-up assessment of the fertilization outcome.

Multistep analysis reveals the relationship between blood indices at the time of ovum pick-up and in vitro embryo production in heifers.

The inflammatory factors of complete blood count (CBC) are associated with a decrease in the in vitro embryo production (IVP) outcome in women. The relation between the blood indices and in vitro fertilization (IVF) outcomes in bovines remains to be elucidated. Using ovum pick-up (OPU), oocytes were retrieved from heifers (n = 60) and inseminated separately with sperm. The blastocyst formation was recorded on day 7 after insemination for each animal and the blood indices were evaluated at the time of OPU. Then, heifers were classified on the basis of (1) blastocyst formation, cleaved vs. failed, or (2) inflammation, low-grade inflammation (lymphocyte counts > 5.6 × 109/L) vs. no inflammation (lymphocyte counts < 5.6 × 109/L). Oocytes derived from heifers with higher lymphocytes, red blood cells (RBC), platelets, hematocrit, red cell distribution width (RDW-SD) and plateletcrit values and lower monocytes, eosinophils, mean corpuscular hemoglobin (MCH) and MCH concentration (MCHC) successfully developed to the blastocyst stage. Heifers with low-grade inflammation numerically had a higher percentage of blastocyst formation than normal heifers. The principle component analysis (PCA) showed that blastocyst formation had the strongest positive association with RDW-cv and RDW-SD, while having a strong negative association with mean corpuscular volume (MCV), hemoglobin, MCHC and MCH. The PCA determined that the number of grade A COCs and the percentage of COCs reached the cleavage stage had a negative association with white blood cells (WBC), lymphocytes, basophils and monocytes, and a positive correlation with platelet to lymphocyte ratio, platelet distribution width (PDW) and plateletcrit. Network mapping detected close similarities between BFR and RDW-SD, MPV, and lymphocytes. The area under the receiver operating characteristic curve (AUC) identified that, eosinophils (AUC 0.80), RDW-SD (AUC 0.76), monocytes (AUC 0.76) and lymphocytes (AUC 0.76) had a good predictive ability to detect heifers with high OPU-IVP outcome (≥60%). In conclusion, these findings suggest that CBC indices at the time of OPU were associated with the IVF outcome and may be incorporated into protocols for the identification of heifers with high potential for blastocyst formation.

A multilevel analysis identifies the different relationships between amino acids and the competence of oocytes matured individually or in groups.

High-protein diets contribute to an increase in urea follicular concentrations associated with decreased fertility. Urea has been shown to interfere with the epidermal growth factor (EGF)/EGFR system, which has been shown to have a beneficial effect during in vitro maturation (IVM) of oocytes. Of note, the number of cumulus-oocyte complexes (COCs) in the maturation medium can change the maturation and the developmental competence of COCs. Therefore, it was hypothesized that, the presence of urea and EGF may have a differential effect on the depletion/appearance of AAs and competence of COCs matured individually (I-IVM system) or in groups (G-IVM system). In the G-IVM system, COCs increased consumption (depletion) of AAs compared with other groups in the presence of high-level urea (40 mg/dl) + EGF (10 ng/ml). In the I-IVM system, the non-cleaved COCs depleted more AAs than the cleaved COCs, in particular in the presence of urea. The combination of urea and EGF increased the depletion of AAs in the G-IVM system. However, the EGF abrogated the urea-induced depletion of AAs by the I-IVM COCs. The use of N-acetyl-L-cysteine as an EGFR inhibitor canceled urea-induced depletion of AAs. This shows the inhibiting effect of urea over the EGF/EGFR system. In the presence of urea + EGF, COCs had a lower degree of developmental competence than control in both I- and G-IVM systems. Arginine had the best predictive power to identify highly competent COCs in the G-IVM system, while glutamine was the best predictor of the cleavage in the I-IVM system. In conclusion, this multi-level study shows that COCs matured individually or in groups may have different association with AAs metabolism. These findings provide new insights into the relationships between AA metabolism and the subsequent developmental competence of COCs.

Feasibility of Real Time Internet-Based Teleconsultation in Patients With Multiple Sclerosis: Interventional Pilot Study.

BACKGROUND: Telemedicine (TM) is currently flourishing in rural and emergency settings, but its implementation in the routine management of chronic neurological disorders has developed with more hesitation. Limited access to specialized care facilities and expanding patient populations, combined with unprecedented mobility restrictions imposed by the coronavirus disease pandemic, are currently stressing the need for remote solutions in this field. Studies in patients with multiple sclerosis (MS) have been heterogeneous in objectives and methodology but generally support the concept that TM interventions produce clinical benefits, cost-effectiveness, and user satisfaction. Nonetheless, data on live interaction between patients and health care providers for MS teleconsultation purposes remain scarce. OBJECTIVE: The aim of this study is to demonstrate the feasibility of planned real time audiovisual teleconsultation over the internet for patients with MS. METHODS: A total of 20 patients with MS presenting at a specialized MS center in Belgium were recruited for this study. One teleconsultation was scheduled for each participant. Patients were provided a unique hyperlink by mail in advance, leading them automatically and directly to the virtual waiting room, where they could accept or decline our incoming call. All teleconsultations were performed by a trained medical student with the intention to keep the conversation similar to what is usually discussed during a classic face-to-face MS consultation; no remote physical exams were performed. The approach was considered feasible if at least 80% of the planned TM visits could be successfully completed at the foreseen moment. Patient satisfaction (technical quality, convenience, and overall quality of care) was evaluated at the end of each teleconsultation by means of 5-point Likert scales containing the categories very unsatisfied, unsatisfied, neutral, satisfied, and highly satisfied. RESULTS: Out of 20 consultations, 17 were successfully completed (85%). Failures were due to patients not responding (n=2) and technical issues (n=1). Out of the 17 consultations, 17 patients declared themselves satisfied or highly satisfied for technical quality, 15 patients for convenience, and 16 patients for overall quality of care. CONCLUSIONS: Planned real time audiovisual teleconsultation over the internet is feasible and highly appreciated in patients with MS. Incorporation of such services in routine clinical MS practice is expected to improve access to specialized care facilities for affected patients.

Preparation and characterization of WS2 decorated and immobilized on chitosan and polycaprolactone as biodegradable polymers nanofibers: Photocatalysis study and antibiotic-conjugated for antibacterial evaluation.

In the present work, WS2 nanoparticles and immobilized on chitosan and polycaprolactone as biodegradable polymers as photocatalyst were developed and studied for photocatalytic degradation of representative Neomycin as an aminoglycoside antibiotic. The WS2 nanoparticles were synthesized using the hydrothermal method. Further, the photocatalyst were characterized by different analytical instruments energy dispersive X-ray spectrometer (EDS), scanning electron microscopy (SEM), X-ray diffraction (XRD) analysis, and UV-Vis absorption spectroscopy, in order to understand their physical and optical properties. The mean crystallite sizes of WS2 nanoparticles, WS2/chitosan nanofibers and WS2/polycaprolactone nanofibers were 80.00, 70.14 and 68.71 nm, respectively. The optical absorption study revealed the presence of direct band-to-band transition with band-gap ranging from 2.0 to 1.8 eV for WS2, WS2/chitosan nanofibers and WS2/polycaprolactone nanofibers, respectively. The photocatalytic activity of the three photocatalyst was tested by UV-light-induced degradation of Neomycin antibiotic. The WS2/chitosan nanofibers and WS2/polycaprolactone nanofibers photocatalyst showed high amount of photodegradation in comparison to WS2 nanoparticles. The optimum degradation using WS2, WS2/chitosan nanofibers and WS2/polycaprolactone nanofibers occurred under UV light at pH:7 in 40 min. The bactericidal test was determined under light illumination (visible source light) and that the neomycin conjugated WS2/chitosan nanofibers and WS2/polycaprolactone nanofibers demonstrated good efficiency in antibacterial efficiency compared to pure WS2/chitosan nanofibers and WS2/polycaprolactone nanofibers.

Urea influences amino acid turnover in bovine cumulus-oocyte complexes, cumulus cells and denuded oocytes, and affects in vitro fertilization outcome.

High-protein diets often lead to an increase in urea concentration in follicular fluid of dairy cows, which may reduce oocyte competence. In the present study, maturation media were supplemented with urea (0, 20, 40 mg/dl), and amino acids (AAs) turnover was evaluated in the 24-h spent media of specimens (cell types), bovine cumulus-oocyte complexes (COCs), cumulus cells (CCs), or denuded oocytes (DOs). The main effects of urea and cell type, and their interaction were significant on the individual turnover (expect threonine, glycine, and tyrosine) and total turnover, depletion, and appearance of AAs. The results showed a high level of urea and DOs increased the depletion of all AAs and that of essential and non-AAs, respectively. Sensitivity analysis revealed the highest sensitivity of isoleucine, lysine, and tryptophan to urea, especially in DOs. Principal component analysis (PCA) evaluated the strong correlations between the turnover of: (1) glutamine, aspartic acid or glycine, and developmental competence and fertilization of COCs; (2) serine, isoleucine, valine or glutamic acid, and cleavage rate of DOs; and (3) serine, glutamine, aspartic acid or alanine, and CCs viability. In conclusion, urea significantly changed the turnover of AAs by COCs, CCs and DOs, and reduced the subsequent developmental competence of bovine oocytes.

Urea changes oocyte competence and gene expression in resultant bovine embryo in vitro.

SummaryNutrition influences the microenvironment in the proximity of oocyte and affects early embryonic development. Elevated blood urea nitrogen, even in healthy dairy cows, is associated with reduced fertility and there is high correlation between blood urea levels and follicular fluid urea levels. Using a docking calculation (in silico), urea showed a favorable binding activity towards the ZP-N domain of ZP3, that of ZP2, and towards the predicted full-length sperm receptor ZP3. Supplementation of oocyte maturation medium with nutrition-related levels of urea (20 or 40 mg/dl as seen in healthy dairy cows fed on low or high dietary protein, respectively) dose-dependently increased: (i) the proportion of oocytes that remained uncleaved; and (ii) oocyte degeneration; and reduced cleavage, blastocyst and hatching rates. High levels of urea induced shrinkage in oocytes, visualised using scanning electron microscopy. Urea downregulated NANOG while dose-dependently upregulating OCT4, DNMT1, and BCL2 expression. Urea at 20 mg/dl induced BAX expression. Using mathematical modelling, the rate of oocyte degeneration was sensitive to urea levels; while cleavage, blastocyst and hatching rates exhibited negative sensitivity. The present data imply a novel role for urea in reducing oocyte competence and changing gene expression in the resultant embryos.

Synthesis of MnO2/cellulose fiber nanocomposites for rapid adsorption of insecticide compound and optimization by response surface methodology.

The MnO2/Cellulose fiber Nanocomposites have been prepared via the microwave-assisted hydrothermal method. The characteristic structure of MnO2/Cellulose fiber Nanocomposites was analyzed using X-ray diffraction, photoluminescence and UV-vis spectra, Transmission electron Microscopy, N2 adsorption-desorption and Scanning electron microscopy instrumental techniques. BET surface area and crystallite size values of MnO2/cellulose fiber nanocomposites have been found as 87.064m2/g and 70.0nm, respectively. Response Surface Methodology (RSM) has been used for adsorption of Insecticide compound such as Toxaphene by prepared adsorbent. MnO2/Cellulose fiber Nanocomposites shows maximum removal of 96.5% at initial Toxaphene concentration of 5.0mg/L, pH 3 and adsorbent dose of 5.0g/L. Kinetic and equilibrium data follow pseudo-second order and Langmuir isotherm model, respectively. Adsorption capacity of MnO2/Cellulose fiber Nanocomposites has been found to be 5.465mg/g.

Application of response surface methodology to optimize the adsorption performance of a magnetic graphene oxide nanocomposite adsorbent for removal of methadone from the environment.

Magnetic graphene oxide (MGO) nanocomposites have been synthesized by the co-precipitation route. The surface structure of MGO nanocomposites were analyzed by XRD, TEM, SEM, VSM and nitrogen adsorption-desorption instrumental technique. Response surface methodology was used to optimize the removal of methadone by MGO nanocomposites in aqueous solution. Experiments were conducted based on Box-Behnken design (BBD). The influence of three parameters on the removal of methadone was examined using the response surface methodological approach. The significance of independent variables and their interactions were tested by analysis of variance (ANOVA). The optimum pH, adsorbent dose and temperature were found to be 6.2, 0.0098g and 295.7K, respectively. Under these conditions, removal efficiency of methadone was found to be 87.20mg/g.

Adsorption of toxic carbamate pesticide oxamyl from liquid phase by newly synthesized and characterized graphene quantum dots nanomaterials.

Graphene quantum dots have been synthesized using the microwave-assisted hydrothermal route. The surface textural and morphological structure of synthesized adsorbent i.e. graphene quantum dots was analyzed using various analytical techniques such as X-ray diffraction, Transmission electron Microscopy, Atomic Force Microscopy and N2 adsorption-desorption instrumental techniques. The application of graphene quantum dots as an adsorbent for the removal of noxious pesticide compound i.e. oxamyl from aqueous solutions was well investigated and elucidated. The impact of several effective parameters such as effect of agitation speed, pH, adsorbent dose, contact time, temperature and initial concentration on sorption efficiency was studied and optimized using batch adsorption experiments. The optimized pH for maximum oxamyl adsorption was found to be 8.0 and for the maximum adsorption rates the adsorbent dose of 0.6g was found to be optimum to carry out the adsorption with in less than 25min of contact time. From the results obtained, it is clear that for all contact times, an increase in oxamyl concentration resulted in increase in the percent oxamyl removal. The adsorption equilibrium and kinetic data were well fitted and found to be in good agreement with the Langmuir isotherm and pseudo-second-order kinetic model.

Electrografting of thionine diazonium cation onto the graphene edges and decorating with Au nano-dendrites or glucose oxidase: Characterization and electrocatalytic applications.

Thionine (Th) diazonium cation is covalently attached onto the glassy carbon (GC) electrode via graphene nanosheets (GNs) (GC-GNs-Th). The GC-GNs-Th electrode is subjected to further modifications to fabricate (i) glucose and (ii) nitrite sensors. Further modifications include: (i) direct immobilization of glucose oxidase (GOx) and (ii) electrodeposition of gold dendrite-like nanostructures (DGNs) on the GC-GNs-Th surface, constructing GC-GNs-Th-GOx and GC-GNs-Th-DGNs modified electrodes, respectively. The GC-GNs-Th-GOx biosensor exhibited a linear response range to glucose, from 0.5 to 6.0mM, with a limit of detection (LOD) of 9.6 µM and high sensitivity of 43.2 µAcm(-2)mM(-1). Also, the GC-GNs-Th-DGNs sensor showed a wide dynamic response range for NO2(-) ion with two linear parts, from 0.05 µM to 1.0 µM and 30.0 µM to 1.0mM, a sensitivity of 263.2 µAmM(-1) and a LOD of 0.01 µM. Applicability of the modified electrodes was successfully tested by determination of glucose in human blood serum and nitrite in water based on addition/recovery tests.