Altering palmitic acid and stearic acid ratios in the diet of early-lactation Holsteins under heat stress: Feed intake, digestibility, feeding behavior, milk yield and composition, and plasma metabolites.

The objective of this study was to evaluate the effects of varying the ratio of dietary palmitic (C16:0; PA) and stearic (C18:0; SA) acids on nutrient digestibility, production, and blood metabolites of early-lactation Holsteins under mild-to-moderate heat stress. Eight multiparous Holsteins (body weight = 589 ± 45 kg; days in milk = 51 ± 8 d; milk production = 38.5 ± 2.4 kg/d; mean ± standard deviation) were used in a duplicated 4 × 4 Latin square design (21-d periods inclusive of 7-d data collection). The PA (88.9%)- and SA (88.5%)-enriched fat supplements, either individually or in combination, were added to diets at 2% of dry matter (DM) to formulate the following treatments: (1) 100PA:0SA (100% PA + 0% SA), (2) 66PA:34SA (66% PA + 34% SA), (3) 34PA:66SA (34% PA + 66% SA), and (4) 0PA:100SA (0% PA + 100% SA). Diets offered, in the form of total mixed rations, were formulated to be isonitrogenous (crude protein = 17.2% of DM) and isocaloric (net energy for lactation = 1.69 Mcal/kg DM), with a forage-to-concentrate ratio of 40:60. Ambient temperature-humidity index averaged 72.9 throughout the experiment, suggesting that cows were under mild-to-moderate heat stress. No differences in DM intake across treatments were detected (mean 23.5 ± 0.64 kg/d). Increasing the dietary proportion of SA resulted in a linear decrease in total-tract digestibility of total fatty acids, but organic matter, DM, neutral detergent fiber, and crude protein digestibilities were not different across treatments. Decreasing dietary PA-to-SA had no effect on the time spent eating (340 min/d), rumination (460 min/d), and chewing (808 min/d). As dietary PA-to-SA decreased, milk fat concentration and yield decreased linearly, resulting in a linear decrease of 3.5% fat-corrected milk production and milk fat-to-protein ratio. Feed efficiency expressed as kg 3.5% fat-corrected milk/kg DM intake decreased linearly with decreasing the proportion of PA-to-SA in the diet. Treatments had no effect on milk protein and lactose content. A linear increase in de novo and preformed fatty acids was identified as the ratio of PA to SA decreased, while PA and SA concentrations of milk fat decreased and increased linearly, respectively. A linear reduction in blood nonesterified fatty acids and glucose was detected as the ratio of PA to SA decreased. Insulin concentration increased linearly from 10.3 in 100PA:0SA to 13.1 µIU/mL in 0PA:100SA, whereas blood ß-hydroxybutyric acid was not different across treatments. In conclusion, the heat-stressed Holsteins in early-lactation phase fed diets richer in PA versus SA produced greater fat-corrected milk and were more efficient in converting feed to fat-corrected milk.

Non-infectious meningitis and CNS demyelinating diseases: A conceptual review.

Many cases of aseptic meningitis or meningoencephalitis, unresponsive to antimicrobial treatments, have been reported recently in patients with established/new-onset central nervous system (CNS) inflammatory demyelinating diseases (CNSIDDs). Given the higher probability of infectious etiologies, CNSIDDs are rarely considered among the differentials in meningitis or meningoencephalitis cases. We gathered and tabulated cases of non-infectious, steroid-responsive meningitis or meningoencephalitis associated with neuromyelitis optica spectrum disorder (NMOSD) and myelin oligodendrocyte glycoprotein-associated disease (MOGAD). This conceptual review highlights the need to bolster routine infectious workups with immunological workups in cases of meningoencephalitis or meningitis where potential autoimmune etiologies can be suspected. Although differentiating CNSIDDs with meningeal involvement from infectious meningitis may not substantially affect acute treatment strategies, long-term management and follow-up of the two are entirely different. We also discuss future research directions and hypotheses on how CNSIDDs may be associated with meningitis-like presentations, e.g. overlapping glial fibrillary acidic protein astrocytopathy or autoimmune encephalitis, alterations in regulatory T-helper cells function, and undetected viral agents.

Prediction and Synthesis of Dysprosium Hydride Phases at High Pressure.

Crystal structure prediction (CSP) methods recently proposed a series of new rare-earth (RE) hydrides at high pressures with novel crystal structures, unusual stoichiometries, and intriguing features such as high-Tc superconductivity. RE trihydrides (REH3) generally undergo a phase transition from ambient P63/mmc or P3̅c1 to Fm3̅m at high pressure. This cubic REH3 (Fm3̅m) was considered to be a precursor to further synthesize RE polyhydrides such as YH4, YH6, YH9, and CeH9 with higher hydrogen contents at higher pressures. However, the structural stability and equation of state (EOS) of any of the REH3 have not been fully investigated at sufficiently high pressures. This work presents high-pressure X-ray diffraction (XRD) measurements in a laser-heated diamond anvil cell up to 100 GPa and ab initio evolutionary CSP of stable phases of DyH3 up to 220 GPa. Experiments observed the Fm3̅m phase of DyH3 to be stable at pressures from 17 to 100 GPa and temperatures up to ∼2000 K. After complete decompression, the P3̅c1 and Fm3̅m phases of DyH3 recovered under ambient conditions. Our calculations predicted a series of phases for DyH3 at high pressures with the structural phase transition sequence P3̅c1 → Imm2 → Fm3̅m → Pnma → P63/mmc at 11, 35, 135, and 194 GPa, respectively. The predicted P3̅c1 and Fm3̅m phases are consistent with experimental observations. Furthermore, electronic band structure calculations were carried out for the predicted phases of DyH3, including the 4f states, within the DFT+U approach. The inclusion of 4f states shows significant changes in electronic properties, as more Dy d states cross the Fermi level and overlap with H 1s states. The structural phase transition from P3̅c1 to Fm3̅m observed in DyH3 is systematically compared with other REH3 compounds at high pressures. The phase transition pressure in REH3 shows an inverse relation with the ionic radius of RE atoms.

The Protective Effects of Alpha Lipoic Acid on Human Sperm Function During Freezing-Thawing.

BACKGROUND: Sperm cryopreservation is presently used for conservation of male gametes in assisted reproduction technologies (ART). Despite the benefits of sperm banking, freeze-thawing process is injurious to sperm integrity due to induced oxidative stress by cold stress. Oxidative stress reduces sperm motility, viability and DNA integrity. OBJECTIVE: To investigate the effect of alpha lipoic acid (ALA) on human sperm function during the freeze-thawing process. MATERIALS AND METHODS: Thirty semen samples were collected and different concentrations (0, 0.05, 0.1, 0.2, 0.4, 0.8, and 8mM) of ALA were added to a sperm freeze medium and its effects on sperm motility, DNA damage, and lipid peroxidation of frozen-thawed spermatozoa were assessed. RESULTS: The addition of 0.2 mM ALA to the sperm freeze medium resulted in significant improvement in percentage of sperm motility, less DNA damage and decreased lipid peroxidation during freeze-thawing process (p<0.05). CONCLUSION: ALA improves the cryo-protective capacity of sperm freeze medium used for human semen by protecting the sperm from ROS attack induced by the freezing-thawing process. We suggest that sperm freeze medium supplemented with 0.2 mM ALA would be beneficial for the cryopreservation of male gametes in ART.

Unraveling the Hidden Martensitic Phase Transition in BaClF and PbClF under High Pressure Using an Ab Initio Evolutionary Approach.

We predict crystal structures of MClF (M = Ba and Pb) compounds by performing an ab initio evolutionary simulation at ambient as well as high pressure. We propose a structural transition sequence in MClF compounds as follows: P4/ nmm → Pmcn → P63/ mmc below 100 GPa. The predicted ambient and intermediate phases are consistent with X-ray and Raman spectroscopic measurements, while the newly proposed high pressure P63/ mmc phase is thermodynamically more favorable than the previously proposed monoclinic ( P21/ m) phase. It is found that the P4/ nmm → Pmcn transition is first order in nature, while the Pmcn → P63/ mmc transition is a martensitic phase transition, which is accompanied by a slight volume change and is of a displacive nature. The austenite and martensite phases coexist in a wide pressure range, especially for PbClF. The martensite phase transition is mainly driven by (1) tilting and transformation of distorted heptahedron to pentahedron environment of MCl6, which leads to negative area compressibility, and (2) cooperative displacive movement of F- ions to form a trigonal bypyramidal (MF5) structure around a metal cation. Overall, the metal cation coordination increases from 9 (MF4Cl5- P4/ nmm) to 10 (MF4Cl6- Pmcn) and, further, to 11 (MF5Cl6- P63/ mmc) under high pressure. The predicted ambient and high pressure phases are mechanically and dynamically stable under the studied pressure range. Electronic structure, bonding, and optical properties are calculated and discussed using new parametrization of Tran Blaha modified Becke Johnson potential. We find nearly isotropic optical properties (except for the ambient phase of PbClF), even though all the predicted ambient and high pressure phases are structurally anisotropic.

Association between total globozoospermia and sperm chromatin defects.

Globozoospermia is a severe sperm morphological anomaly leading to primary infertility and low fertilisation following intracytoplasmic sperm injection (ICSI). This phenotype is observed in less than 0.1% of infertile men and is determined by small, round-headed spermatozoa with absence of an acrosomal cap, acrosome protease and also cytoskeletal proteins. Failure of oocyte activation is considered as the main cause of fertilisation failure in these individuals post-ICSI. Therefore, artificial oocyte activation (AOA) along with ICSI is commonly implemented. However, based on previous report, fertilisation rate remains low despite implementation of ICSI-AOA. Therefore, other mechanisms like sperm chromatin packaging and DNA fragmentation may account for low fertilisation and development post-ICSI-AOA. Therefore, this study aims to assess and compare the degree of sperm protamine deficiency and DNA fragmentation in large population of infertile men with total globozoospermia (30 globozoospermic men presenting with 100% round-headed spermatozoa) with 22 fertile individuals using chromomycin A3 and TUNEL assay respectively. Results clearly show that mean of sperm concentration and percentage of sperm motility were significantly lower, while percentage of sperm abnormal morphology, protamine-deficient and DNA-fragmented spermatozoa were significantly higher in infertile men with globozoospermia compared to fertile men. Therefore, increased sperm DNA damage in globozoospermia is likely related to defective DNA compaction and antioxidant therapy before ICSI-AOA could be recommended as an appropriate option before ICSI-AOA.

Cathepsin B inhibitor improves developmental competency and cryo-tolerance of in vitro ovine embryos.

BACKGROUND: Cathepsin B is a lysosomal cysteine protease involved in apoptosis and oocytes which have lower developmental competence show higher expression of Cathepsin B. Furthermore, expression of Cathepsin B show a decreasing trend from oocyte toward blastocyst stage. RESULTS: Present study assessed the effect of cathepsin B inhibitor, E-64, on developmental competency and cryo-survival of pre-implantation ovine IVF derived embryos. Cathepsin B inhibitor was added during day 3 to 8 of development. One µM E-64 was defined as the optimal concentration required for improving blastocyst rate. This concentration also reduced DNA fragmentation and BAX as apoptotic markers while increasing total cell number per blastocyst and improving anti-apoptotic marker, the BCL2. We further showed that addition of 1.0 µM of E-64 during day 3 to 8 of development improved re-expansion and hatching rates of blastocysts post vitrification. E-64 also reduced rate of DNA fragmentation and BAX expression and increased total cell number per blastocyst and BCL2 expression post vitrification. However, addition of E-64 post vitrification reduced the hatching rate. CONCLUSION: Therefore, it can be concluded that inhibition of cathepsin B in IVC, not only improves quality and quantity of blastocysts but also improves the cryo-survival of in vitro derived blastocysts.

Novel magnesium borides and their superconductivity.

With the motivation of searching for new superconductors in the Mg-B system, we performed ab initio evolutionary searches for all the stable compounds in this binary system in the pressure range of 0-200 GPa. We found previously unknown, yet thermodynamically stable, compositions MgB3 and Mg3B10. Experimentally known MgB2 is stable in the entire pressure range 0-200 GPa, while MgB7 and MgB12 are stable at pressures below 90 GPa and 35 GPa, respectively. We predict a reentrant behavior for MgB4, which becomes unstable against decomposition into MgB2 and MgB7 at 4 GPa and then becomes stable above 61 GPa. We find ubiquity of phases with boron sandwich structures analogous to the AlB2-type structure. However, with the exception of MgB2, all other magnesium borides have low electron-phonon coupling constants λ of 0.32-0.39 and are predicted to have Tc below 3 K.

Selection of physiological spermatozoa during intracytoplasmic sperm injection.

Sperm genomic integrity has a significant effect on intracytoplasmic sperm injection (ICSI) outcomes, especially post-implantation. Spermatozoa selected based on motility and morphology do not guarantee the genomic integrity of spermatozoa. Nearly fifty percentage of spermatozoa in infertile men with normal morphology present different degrees of DNA fragmentation. However, capacitated or hyperactivated spermatozoa show lower degrees of DNA fragmentation. Therefore, selection of hyperactivated spermatozoa may improve ICSI outcome. Routinely, for ICSI, fast-moving spermatozoa with A or B motility pattern are mainly selected for injection. The result of this study shows that in processed semen samples, hyperactivated spermatozoa are mainly observed in B motility pattern while, in viscous medium like polyvinylpyrrolidone (PVP), hyperactivated spermatozoa are mainly present in spermatozoa with C pattern of motility (nonprogressive). Therefore, we propose spermatozoa with C motility pattern which contains the main population of physiological or hyperactivated spermatozoa should be selected for ICSI.

Effects of Tempol and Quercetin on Human Sperm Function after Cryopreservation.

　　BACKGROUND: Quercetin is a flavonoid with high reactive oxygen species (ROS) scavenging and ion chelating activity. It also enhances the activity of antioxidant enzymes and reduces enzymatic activity such as NADPH oxidase and NADH-dependent oxido-reductase. Tempol, as a superoxide dismutase mimetic agent, converts superoxide to less toxic hydrogen peroxide (H2O2), but cannot reduce highly toxic hydroxyl radicals in Fenton or Haber-Weiss reactions mediated with free iron or cupper. OBJECTIVE: The study was to compare the effect of Quercetin and Tempol in an optimized commercial cryo-protective media on ROS induced cryoinjury for the first time. MATERIALS AND METHODS: Following administration of these compounds during freezing process, sperm motility, viability, ROS production and DNA integrity were assessed before and after freezing/thawing process. RESULTS: Data showed that 10 µM Quercetin and 5 µM Tempol significantly improved sperm motility and viability, but they together had no additive effect. Supplementation with Quercetin alone or combination of Quercetin with Tempol decrease the ROS concentration, but the reduction was not significant for Tempol alone compared to control group. Quercetin and Tempol significantly decrease DNA fragmentation. CONCLUSION: The supplementation of Quercetin or Tempol, but not their combination improves the quality of cryopreserved human semen.

Soil heavy metals are associated with the distribution of multiple sclerosis in Isfahan, Iran.

BACKGROUND: Few epidemiological studies have explored the effects of soil pollution on multiple sclerosis (MS) risk in Asia. METHOD: Based on catchment areas, Isfahan province is divided into five regions (Central, North, East, West, and South), and Soil sampling performed in catchment area with the highest range of MS prevalence and incidence. Samples were analyzed for cobalt (Co), lead (Pb), cadmium (Cd), copper (Cu), zinc (Zn), and absorbable forms of Pb, Cd, and Co. Linear regression is used to examine the association of soil heavy metals with prevalence of MS in central part of Isfahan province. RESULT: Multiple sclerosis prevalence ranged from 5.62 to 156.65 in different townships. Based on regression analysis, in case of considering fixed amounts for the rest of elements, a one (SD) increase of absorbable Pb and absorbable Cd are associated with 0.385 (SD) (P < 0.0001) increase and 0.209 (SD) (P < 0.007) decrease in MS prevalence, respectively. CONCLUSION: This study documented a significant association between exposure to absorbable Pb and Cd in soil with prevalence of MS in Isfahan. Further work is warranted to confirm this association and if validated, to understand the mechanisms behind the association.

Novel superhard B-C-O phases predicted from first principles.

We explored the B-C-O system at pressures in the range 0-50 GPa by ab initio variable-composition evolutionary simulations in the hope of discovering new stable superhard materials. A new tetragonal thermodynamically stable phase B4CO4, space group I4[combining macron], and two low-enthalpy metastable compounds (B6C2O5, B2CO2) have been discovered. Computed phonons and elastic constants show that these structures are dynamically and mechanically stable both at high pressure and zero pressure. B4CO4 is thermodynamically stable at pressures above 23 GPa, but should remain metastable under ambient conditions. Its computed hardness is about 38-41 GPa, which suggests that B4CO4 is potentially superhard.

Long-term and efficient expression of human ß-globin gene in a hematopoietic cell line using a new site-specific integrating non-viral system.

Targeted integration of a therapeutic gene at specific loci in safe genomic regions by a non-viral vector can restore the function of the damaged gene. This approach also minimizes the potential genotoxic effects of transferred DNA. In this study, we have developed a non-viral vector that functions according to site-specific recombination (SSR). The vector contained a bacterial backbone and puromycin resistance gene (pur(r)), a ß-globin expressing cassette and an attB recombination site. We used phiC31 integrase to insert a copy of the vector into specific genomic locations of a human hematopoietic cell line. Site-specific integration of the vector with one or two copies in the transcriptionally active regions of the genome was confirmed. After genomic integration, we used Cre recombinase to remove the bacterial backbone and pur(r). This removal was verified by negative selection and genomic PCR screening. Following deletion of these sequences, the stable ß-chain expression was continued for several months in the absence of selective pressure. Consequently, this vector may potentially be a powerful tool for ex vivo correction of ß-globinopathies such as ß-thalassemia through successful genomic integration of a functional copy of the globin gene into the patient's target cells.

Glial cell line-derived neurotrophic factor in combination with insulin-like growth factor 1 and basic fibroblast growth factor promote in vitro culture of goat spermatogonial stem cells.

Growth factors are increasingly considered as important regulators of spermatogonial stem cells (SSCs). This study investigated the effects of various growth factors (GDNF, IGF1, bFGF, EGF and GFRalpha-1) on purification and colonization of undifferentiated goat SSCs under in vitro and in vivo conditions. Irrespective of the culture condition used, the first signs of developing colonies were observed from day 4 of culture onwards. The number of colonies developed in GDNF + IGF1 + bFGF culture condition was significantly higher than the other groups (p < 0.05). In contrast, the size of colonies developed in GDNF + EGF + LIF culture condition was significantly higher than the other groups (p < 0.05). Immunocytochemical stationing for specific biomarkers of somatic cells (vimentin, alpha-inhibin and α-SMA) and spermatogonial cells (PLZF, THY 1, VASA, alpha-1 integrin, bet-1 integrin and DBA) revealed that both cell types existed in developing colonies, irrespective of the culture condition used. Even though, the relative abundance of VASA, FGFR3, OCT4, PLZF, BCL6B and THY1 transcription factors in GDNF + IGF1 + bFGF treatment group was significantly higher than the other groups (p < 0.05). Additionally, goat SSCs developed in the latter culture condition could colonize within the seminiferous tubules of the germ-cell depleted recipient mice following xenotransplantation. Obtained results demonstrated that combination of GDNF with IGF1 and bFGF promote in vitro culture of goat SSCs while precludes uncontrolled proliferation of somatic cells.

A physiological, rather than a superovulated, post-implantation environment can attenuate the compromising effect of assisted reproductive techniques on gene expression in developing mice embryos.

Assisted reproductive techniques (ARTs) may perturb the pre-/peri-conception microenvironments, which subsequently threaten the health of offspring. This study aimed to investigate the effects of superovulation, vitrification, in vitro culture, and embryo transfer on the expression of epigenetic modulators, imprinted genes, and pluripotency markers in expanded blastocysts and Day-9.5 (D9.5) concepti. Results revealed that 53.4% (8/15) and 86.7% (13/15) of genes in the fetus and placenta, respectively, have similar patterns of transcription in all D9.5 concepti, despite the perturbed mRNA expression observed at the blastocyst stage for each embryo-production technique. These observations indicate a counterbalancing of the abnormal expression pattern analyzed at the blastocyst stage during post-implantation development, particularly when the uterus of a naturally synchronized foster mother is employed. Superovulation resulted in the most abnormal expression patterns compared to other treatment groups, although these same blastocysts were able to develop in a synchronized uterus. Thus, superovulation creates a hormonal environment that negatively affected gene expression and impairs fetal growth more adversely during post-implantation development than other ART protocols, such as in vitro culture, vitrification, or embryo transfer-although each did contribute negatively to the implantation and development process. Together, these results may have implications for treating infertility in humans.

A novel phase of beryllium fluoride at high pressure.

A previously unknown thermodynamically stable high-pressure phase of BeF2 has been predicted using the evolutionary algorithm USPEX. This phase occurs in the pressure range 18-27 GPa. Its structure has C2/c space group symmetry and contains 18 atoms in the primitive unit cell. Given the analogy between BeF2 and SiO2, silica phases have been investigated as well, but the new phase has not been observed to be thermodynamically stable for this system. However, it is found to be metastable and to have comparable energy to the known metastable phases of SiO2, suggesting a possibility of its synthesis.

Effect of epigenetic modification with trichostatin A and S-adenosylhomocysteine on developmental competence and POU5F1-EGFP expression of interspecies cloned embryos in dog.

Adult canine fibroblasts stably transfected with either cytomegalovirus (CMV) or POU5F1 promoter-driven enhanced green fluorescent protein (EGFP) were used to investigate if pre-treatment of these donor cells with two epigenetic drugs [trichostatin A (TSA), or S-adenosylhomocysteine (SAH)] can improve the efficiency of interspecies somatic cell nuclear transfer (iSCNT). Fluorescence-activated cell sorting (FACS), analyses revealed that TSA, but not SAH, treatment of both transgenic and non-transgenic fibroblasts significantly increased acetylation levels compared with untreated relatives. The expression levels of Bcl2 and P53 were significantly affected in TSA-treated cells compared with untreated cells, whereas SAH treatment had no significant effect on cell apoptosis. Irrespective of epigenetic modification, dog/bovine iSCNT embryos had overall similar rates of cleavage and development to 8-16-cell and morula stages in non-transgenic groups. For transgenic reconstructed embryos, however, TSA and SAH could significantly improve development to 8-16-cell and morula stages compared with control. Even though, irrespective of cell transgenesis and epigenetic modification, none of the iSCNT embryos developed to the blastocyst stage. The iSCNT embryos carrying CMV-EGFP expressed EGFP at all developmental stages (2-cell, 4-cell, 8-16-cell, and morula) without mosaicism, while no POU5F1-EGFP signal was observed in any stage of developing iSCNT embryos irrespective of TSA/SAH epigenetic modifications. These results indicated that bovine oocytes partially remodel canine fibroblasts and that TSA and SAH have marginal beneficial effects on this process.

Effects of heat shock during the early stage of oocyte maturation on the meiotic progression, subsequent embryonic development and gene expression in ovine.

Heat shock may affect different aspects of oocyte maturation and its subsequent development to the blastocyst stage. A series of in vitro experiments was performed to determine whether physiologically heat shock (41°C) disrupts the progression of the ovine oocytes through meiosis, activation and blastocyst formation. Cumulus-oocyte complexes (COCs) were aspirated from 2-6-mm follicles and cultured at 38.5°C (control) or 41°C (heat shock) for the first 12 h of maturation. The oocytes were incubated at 38.5°C during the last 10 h of maturation and 8 days after activation. Results showed that most of the oocytes matured under heat-shock conditions remained at the germinal vesicle breakdown (GVBD) stage and they showed an aberrant chromatin configuration. After heat shock, oocyte diameter and time spent for zona pellucida dissolution increased (P < 0.05). The heat-shocked group had a higher percentage of oocytes with incomplete migration of cortical granules (P < 0.05). The heat-shock condition decreased (P < 0.05) cleavage rates (56.19 versus 89.28%) and morula formation (26.85 versus 37.81%). However, there was no significant difference in blastocyst formation and percentage of hatched blastocysts. At 12 h, heat shock had an adverse effect on embryo quality and reduced inner cell mass number (P < 0.05). Quantitative gene expression analysis showed greater transcripts (P < 0.05) for Na/K-ATPase mRNA in heat-shocked oocytes. To sum up, heat shock has disruptive effects on ovine oocyte maturation and can impair cellular and molecular factors that are important for embryo development.