SUBPOPULATION STRUCTURE AND CHANGES AFTER CRYOPRESERVATION OF SPERMS FROM HIGH AND LOW FERTILITY WATER BUFFALO / ESTRUCTURA DE LAS SUBPOBLACIONES Y CAMBIOS DESPUÉS DE LA CRIOPRESERVACIÓN ESPERMÁTICA EN BÚFALOS DE AGUA CON ALTA Y BAJA FERTILIDAD

The aim of this study was to identify the sperm subpopulation structure in buffalo bulls with high and low fertility and to determine how sperm subpopulations change after semen cryopreservation. Semen was obtained from four bulls with high fertility (HF) and four bulls with low fertility (LF) and was cryopreserved. A total of 64 ejaculates were assessed for their sperm kinematics using computer assisted sperm analyzer (CASA). Ward’s Hierarchical Dendogram and K-Means clustering method were used to identify the subpopulations. In experiment 1, two significantly different (P≤0.05) sperm subpopulations were observed: Subpopulation 1 (SP1): sperms travel longer distances most rapidly and progressively, and Subpopulation 2 (SP2): sperms travel shorter distances slower but highly progressive. A higher percentage of SP1 was found in HF bulls (47.27); whereas, a higher percentage of SP2 was found in LF bulls (54.89). A low negative relationship (r=-0.18) was observed for the fertility level and sperm subpopulation structure. This implies that sperms that travel longer distances most rapidly and progressively (SP1) are most likely associated to high fertility, while sperms that travel shorter distances slower but highly progressive (SP2) are associated with low fertility. In experiment 2, based on the change in SP1 after cryopreservation, significantly higher sperm survival was observed in samples from HF bulls (29.97) as compared to those from LF bulls (31.64). During post thaw, there were more SP1 sperms in samples from HF bulls (27.52) than in those from LF bulls (26.74). Thus, semen containing higher proportion of SP1 sperms are more resistant to cryopreservation and have greater chances of obtaining high fertility. Overall, the identification of sperm heterogeneity in water buffaloes can be associated to sperm survival after cryopreservation and fertility. RESUMENEl propósito de este estudio fue identificar la estructura de las subpoblaciones espermáticas en toros bufalinos con alta y baja fertilidad y determinar los cambios luego de la criopreservación. El semen se obtuvo de cuatro búfalos con alta fertilidad (HF) y cuatro con baja fertilidad (LF) y fue criopreservado. Un total de 64 eyaculados fueron evaluados para parámetros cinéticos usando un analizador espermático asistido por computadora (CASA). El método de dendograma jerárquico de Ward y el método K-means fueron utilizados para identificar las subpoblaciones. En el experimento 1, dos subpoblaciones espermáticas estadísticamente diferentes (p<0.05) fueron observadas: Subpoblación 1 (SP1): espermatozoides que viajan largas distancias más rápida y progresivamente, y la Subpoblación 2 (SP2): espermatozoides que viajan distancias cortas de forma lenta pero muy progresivamente. Un mayor porcentaje de SP1 fue encontrado en los búfalos HF (47,27); mientras que un mayor porcentaje de SP2 fue encontrado en los búfalos LF (54.89). Una relación baja pero negativa (r = -0,18) fue observada para el nivel de fertilidad y la estructura de la subpoblación espermática. Esto implica que los espermatozoides que viajan largas distancias más rápida y progresivamente (SP1) están más asociados a la alta fertilidad, mientras que los que viajan distancias cortas más lento y con alta progresividad (SP2) están asociados a una baja fertilidad. En el experimento 2, en base a los cambios en SP1 luego de la criopreservación, un mayor porcentaje de espermatozoides permaneció en esta subpoblación en los búfalos HF (27,52) que en los LF (26,74). Por lo tanto, semen con una alta proporción de espermatozoides dentro de SP1 son más resistentes a la criopreservación y tienen mayor probabilidad de obtener una mayor fertilidad. En general, la identificación de la heterogeneidad espermática en búfalos de agua puede ser asociada a la sobrevivencia luego de la criopreservación y a la fertilidad

Influence of Ovarian Follicle Sizes and Estrous Signs on Pregnancy Following Progesterone-Based Fixed Time Artificial Insemination in Water Buffaloes

The objectives of the present study were to elucidate the importance of follicle sizes and estrous signs during Controlled Internal Drug Release-Synch-human Chorionic Gonadotropin (CIDR-Synch-hCG) protocol for Fixed Time Artificial Insemination (FTAI) and to evaluate their association with pregnancy in water buffaloes. Data from riverine buffaloes (n = 207) under the CIDR-Synch-hCG protocol were analyzed. Buffaloes were administered with Gonadotropin-Releasing Hormone (GnRH) with insertion of CIDR on Day 0. Prostaglandin (PGF2α) was given on Day 7 with the removal of CIDR. hCG was given on Day 9, and AI was performed on Day 10. Follicle measurements by ultrasonography were done on Days 0, 7, and 10, and follicle sizes on those days were categorized into I, II, and III. Estrus signs were taken on the day of AI. The pregnancy diagnosis was done on Day 30-35 post-AI. The average size of follicles in Category III is significantly higher than those of Categories I and II, regardless of the Days of the protocol. Pregnancy is significantly (P&lt;0.001) associated with Pre-Ovulatory Follicle (POF) size and uterine tonicity on the Day of AI but not with follicle sizes on Days 0 and 7, nor with mucus discharge discharge (P&gt;0.05). The overall pregnancy rate is 44.44% while performing AI with POF size ≥12.0 mm increased the probability of pregnancy rate to 56.25%. In conclusion, the present study demonstrated a follicle size-based CIDR-Synch-hCG protocol providing new fertility indicators to improve FTAI efficiency in buffaloes with huge application in other livestock species

Fourier transform infrared spectroscopy as a novel approach for analyzing the biochemical effects of anionic surfactants on a surfactant-degrading acrobacter butzleri strain

Cataloged from PDF version of article.Anionic surfactant-biodegrading capability of an Arcobacter butzleri strain was analyzed under aerobic conditions. The A. butzleri isolate displayed efficient surfactant-biodegrading capacity for sodium dodecyl sulfate (SDS) at concentrations of up to 100 mg/L in 6 days, corresponding to 99.0% removal efficiency. Fourier transform infrared spectroscopy was applied to observe the effects of varying concentrations of SDS on the biochemistry of bacterial cells. Results suggest that protein secondary structures were altered in bacterial cells at sufficiently high SDS concentrations, concurrent with SDS biodegradation

SLC25A22 is a novel gene for migrating partial seizures in infancy

Objective To identify a genetic cause for migrating partial seizures in infancy (MPSI). Methods We characterized a consanguineous pedigree with MPSI and obtained DNA from affected and unaffected family members. We analyzed single nucleotide polymorphism 500K data to identify regions with evidence of linkage. We performed whole exome sequencing and analyzed homozygous variants in regions of linkage to identify a candidate gene and performed functional studies of the candidate gene SLC25A22. Results In a consanguineous pedigree with 2 individuals with MPSI, we identified 2 regions of linkage, chromosome 4p16.1-p16.3 and chromosome 11p15.4-pter. Using whole exome sequencing, we identified 8 novel homozygous variants in genes in these regions. Only 1 variant, SLC25A22 c.G328C, results in a change of a highly conserved amino acid (p.G110R) and was not present in control samples. SLC25A22 encodes a glutamate transporter with strong expression in the developing brain. We show that the specific G110R mutation, located in a transmembrane domain of the protein, disrupts mitochondrial glutamate transport. Interpretation We have shown that MPSI can be inherited and have identified a novel homozygous mutation in SLC25A22 in the affected individuals. Our data strongly suggest that SLC25A22 is responsible for MPSI, a severe condition with few known etiologies. We have demonstrated that a combination of linkage analysis and whole exome sequencing can be used for disease gene discovery. Finally, as SLC25A22 had been implicated in the distinct syndrome of neonatal epilepsy with suppression bursts on electroencephalogram, we have expanded the phenotypic spectrum associated with SLC25A22. Ann Neurol 2013;74:873-882 © 2013 American Neurological Association

Psychiatric disorders among older prisoners: a systematic review and comparison study against older people in the community

Objectives: Despite emerging evidence that older prisoners experience poor mental health, literature in this area is still limited. In the present systematic review and meta-analysis, we report on the prevalence of psychiatric disorders among older prisoners and compare our findings against community studies on older people. Methods: We searched on Assia, PsycInfo, MedLine, Embase, Web of Science, Google and Gov.uk. We carried out bias assessments, rated studies for quality and ran a heterogeneity test. We meta-analysed prevalence rates of psychiatric disorders through an aggregate weighted mean and calculated Relative Risk and statistical significance against community studies. Sensitivity analyses were further performed. Results: We reviewed nine studies and obtained the following prevalence: “Any psychiatric disorder” 38.4%, depression 28.3%, schizophrenia/psychoses 5.5%, bipolar disorder 4.5%, dementia 3.3%, cognitive impairment 11.8%, personality disorder 22.9%, alcohol abuse 15.9%, anxiety disorders 14.2%, PTSD 6.2%. Older prisoners were found to have higher RR for every single psychiatric disorder against older people in the community, with the sole exception of alcohol abuse (RR=1) and dementia (RR=.75). The prevalence rates were statistically significantly higher (p<.05) among the prisoners for “Any psychiatric disorder”, depression and personality disorder. Overall, the sensitivity analyses confirmed our original results. Conclusion: Our findings point at a high prevalence of every single psychiatric disorder among older prisoners, who also experience rates of dementia and alcohol abuse comparable to those reported in the community. Our results have relevant implications for policy and practice in this area. Further research is crucial to confirm findings from this study

CHMP1A encodes an essential regulator of BMI1-INK4A in cerebellar development

Charged multivesicular body protein 1A (CHMP1A; also known as chromatin-modifying protein 1A) is a member of the ESCRT-III (endosomal sorting complex required for transport-III) complex but is also suggested to localize to the nuclear matrix and regulate chromatin structure. Here, we show that loss-of-function mutations in human CHMP1A cause reduced cerebellar size (pontocerebellar hypoplasia) and reduced cerebral cortical size (microcephaly). CHMP1A-mutant cells show impaired proliferation, with increased expression of INK4A, a negative regulator of stem cell proliferation. Chromatin immunoprecipitation suggests loss of the normal INK4A repression by BMI in these cells. Morpholino-based knockdown of zebrafish chmp1a resulted in brain defects resembling those seen after bmi1a and bmi1b knockdown, which were partially rescued by INK4A ortholog knockdown, further supporting links between CHMP1A and BMI1-mediated regulation of INK4A. Our results suggest that CHMP1A serves as a critical link between cytoplasmic signals and BMI1-mediated chromatin modifications that regulate proliferation of central nervous system progenitor cells

Effects of isolation method and pre-treatment with ethylene glycol or raffinose before vitrification on in vitro viability of mouse preantral follicles

Effects of isolation and vitrification protocols on follicular survival after warming were examined. Mouse preantral follicles enzymatically or mechanically isolated from ovaries of 12-day-old mice were exposed either to 2 M ethylene glycol (EG) for 2 or 5 min, or to ascending concentrations (0.15 then 0.3 M) of raffinose for 2 or 5 min each (2-2 and 5-5 min). They were then exposed to a vitrification solution (VS) composed of 6 M EG and 0.3 M raffinose for 0.5, 1, or 2 min before vitrification. Mechanically isolated follicles showed higher survival than enzymatically isolated follicles, regardless of periods of exposure to EG or raffinose and subsequent exposure to VS. After 10 days of culture, follicular growth and maturational ability of oocytes derived from vitrified follicles exposed to 2 M EG for 5 min and to VS for 1 min were higher than those from follicles exposed to raffinose solutions for 2-2 min and to VS for 1 min. Histological evaluation revealed that exposure of preantral follicles to raffinose solutions caused cytoplasmic vacuolation in granulosa cells which could be due to cellular shrinkage during dehydration; whereas, exposure to 2 M EG induced morphological alterations in follicles only to a lesser extent

Effect of activation treatments of recipient oocytes on subsequent development of bovine nuclear transfer embryos

Effects of recipient oocyte activation methods on the development of nuclear transfer (NT) embryos were investigated. In Exp. 1, cell-cycle phase of serum-starved bovine cumulus cells was examined by flow cytometry. Majority (95.5%) of medium-sized (16-20 μm) cells that made up 56% of total cells was at the G_0/G_1 phase. NT embryos were constructed by electric fusion with the medium-sized serum-starved cumulus cells and bovine oocytes of 3 different preparations: enucleated oocytes treated with calcium ionophore A 23187 for 5 min and cycloheximide for 5 hr (A 23187/CHX), those treated with ethanol for 7 min and cycloheximide for 2 hr (ethanol CHX) and those without treatment. In Exp. 2 and 3, developmental competence of NT embryos constructed with A 23187/CHX- and ethanol/HX-treated oocytes was compared to that of NT embryos constructed with non-treated oocytes, respectively. Further, nuclear behavior in 3 different NT embryos was examined in Exp. 4. Within 1 hr after fusion, majority of the NT embryos constructed with non-treated oocytes showed condensed chromosome. Three hours after fusion, about 50% of NT embryos constructed with non-treated or ethanol/CHX-treated oocytes showed a single pronucleus-like structure. NT embryos constructed with ethanol/CHX-treated oocytes showed similar rates of fusion, cleavage and blastocyst formation to those of the non-treated oocytes. In contrast, NT embryos constructed with A 23187/CHX-treated oocytes did not show any pronucleus-like structure and showed lower cleavage rate and no development to blastocysts. The results indicate that ethanol/CHX-treated oocytes could support development of somatic cell NT embryos to the blastocyst stage at a similar rate to that of non-treated oocytes