Application of a novel sorting system for equine mesenchymal stem cells (MSCs)     

Catherine L. Radtke  Rodolfo Nino-Fong  Blanca P. Esparza Gonzalez  Laurie A. McDuffee 《Canadian journal of veterinary research》2014,78(4):290-296

The objective of this study was to validate non-equilibrium gravitational field-flow fractionation (GrFFF), an immunotag-less method of sorting mesenchymal stem cells (MSCs) into subpopulations, for use with MSCs derived from equine muscle tissue, periosteal tissue, bone marrow, and adipose tissue. Cells were collected from 6 young, adult horses, postmortem. Cells were isolated from left semitendinosus muscle tissue, periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and left supragluteal subcutaneous adipose tissue. Aliquots of 800 × 10³ MSCs from each tissue source were separated and injected into a ribbon-like capillary device by continuous flow (GrFFF proprietary system). Cells were sorted into 6 fractions and absorbencies [optical density (OD)] were read. Six fractions from each of the 6 aliquots were then combined to provide pooled fractions that had adequate cell numbers to seed at equal concentrations into assays. Equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells were consistently sorted into 6 fractions that remained viable for use in further assays. Fraction 1 had more cuboidal morphology in culture when compared to the other fractions. Statistical analysis of the fraction absorbencies (OD) revealed a P-value of < 0.05 when fractions 2 and 3 were compared to fractions 1, 4, 5, and 6. It was concluded that non-equilibrium GrFFF is a valid method for sorting equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells into subpopulations that remain viable, thus securing its potential for use in equine stem cell applications and veterinary medicine.  相似文献   

Perfrin,a novel bacteriocin associated with netB positive Clostridium perfringens strains from broilers with necrotic enteritis     

Leen Timbermont  Lina De Smet  Filip Van Nieuwerburgh  Valeria R Parreira  Gonzalez Van Driessche  Freddy Haesebrouck  Richard Ducatelle  John Prescott  Dieter Deforce  Bart Devreese  Filip Van Immerseel 《Veterinary research》2014,45(1):40

Necrotic enteritis in broiler chickens is associated with netB positive Clostridium perfringens type A strains. It is known that C. perfringens strains isolated from outbreaks of necrotic enteritis are more capable of secreting factors inhibiting growth of other C. perfringens strains than strains isolated from the gut of healthy chickens. This characteristic could lead to extensive and selective presence of a strain that contains the genetic make-up enabling to secrete toxins that cause gut lesions. This report describes the discovery, purification, characterization and recombinant expression of a novel bacteriocin, referred to as perfrin, produced by a necrotic enteritis-associated netB-positive C. perfringens strain. Perfrin is a 11.5 kDa C-terminal fragment of a 22.9 kDa protein and showed no sequence homology to any currently known bacteriocin. The 11.5 kDa fragment can be cloned into Escherichia coli, and expression yielded an active peptide. PCR detection of the gene showed its presence in 10 netB-positive C. perfringens strains of broiler origin, and not in other C. perfringens strains tested (isolated from broilers, cattle, sheep, pigs, and humans). Perfrin and NetB are not located on the same genetic element since NetB is plasmid-encoded and perfrin is not. The bacteriocin has bactericidal activity over a wide pH-range but is thermolabile and sensitive to proteolytic digestion (trypsin, proteinase K). C. perfringens bacteriocins, such as perfrin, can be considered as an additional factor involved in the pathogenesis of necrotic enteritis in broilers.  相似文献   

Identification of Apoptotic Bodies in Equine Semen     

AB Caselles  A Miro‐Moran  A Morillo Rodriguez  JM Gallardo Bolaños  C Ortega‐Ferrusola  GM Salido  FJ Peña  JA Tapia  IM Aparicio 《Reproduction in domestic animals》2014,49(2):254-262

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.  相似文献   

Effect of Overnight Staining on the Quality of Flow Cytometric Sorted Stallion Sperm: Comparison with Tradtitional Protocols     

CM Balao da Silva  C Ortega Ferrusola  JM Gallardo Bolaños  M Plaza Dávila  P Martín‐Muñoz  JM Morrell  H Rodriguez Martínez  FJ Peña 《Reproduction in domestic animals》2014,49(6):1021-1027

Flow cytometry is considered the only reliable method for the separation of X and Y chromosome bearing spermatozoa in equines. The MoFlo SX DP sorter is highly efficient, allowing the production of foals of the desired sex. However, to achieve acceptable pregnancy rates the currently used protocol requires working with fresh semen obtained close to, or at, the sorting facility. An alternative protocol was tested during two consecutive breeding seasons. Fresh stallion semen was cooled for 20 h, during which staining with Hoechst 33342 took place. On the following day, this sample was flow sorted and compared with spermatozoa from the same ejaculate that had been sexed on the previous day. All sperm parameters evaluated remained unchanged when fresh sorted and refrigerated sorted semen were compared. Pre‐sorting storage at 5°C did not alter sperm velocities nor kinetics, viability or membrane permeability, production of reactive oxygen species, mitochondrial membrane potential or DNA fragmentation index of the sorted sample. The findings open for the possibility of using semen from stallions housed far from the sorting facilities. Processed and stained sperm could be shipped refrigerated on the previous day, sorted and inseminated on the next day.  相似文献   

Standing Laparoscopic Peritoneal Flap Hernioplasty of the Vaginal Rings does not Modify the Sperm Production and Motility Characteristics in Intact Male Horses     

LA Gracia‐Calvo  LJ Ezquerra  M Martín‐Cuervo  ME Durán  H Tapio  JM Gallardo  FJ Peña  C Ortega‐Ferrusola 《Reproduction in domestic animals》2014,49(6):1043-1048

Laparoscopic hernioplasty techniques have been developed in the recent years to avoid the recurrence of inguinal hernias and to spare the testicles for breeding purposes in stallions. However, there have been no previous comprehensive and systematic studies of the reproductive outcomes and prognoses for stallions after inguinal hernioplasty. Therefore, the objective of this study was to assess the possible effects of one of these techniques (standing laparoscopic peritoneal flap hernioplasty) on the sperm production and motility characteristics of six healthy stallions that received this procedure based on 1‐year follow‐ups. There were no significant differences in the measured sperm variables (assessments based on the DSO, MOT, PMOT, VSL, VCL and VAP) during 1‐year follow‐ups.  相似文献   

Diffuse Extreme-Ultraviolet Emission from the Coma Cluster: Evidence for Rapidly Cooling Gases at Submegakelvin Temperatures     

R Lieu  JPD Mittaz  S Bowyer  JO Breen  FJ Lockman  EM Murphy  C Hwang 《Science (New York, N.Y.)》1996,274(5291):1335-1338

The central region of the Coma cluster of galaxies was observed in the energy band from 0.065 to 0.245 kiloelectron volts by the Deep Survey telescope aboard the Extreme Ultraviolet Explorer. A diffuse emission halo of angular diameter approximately 30 arc minutes was detected. The extreme-ultraviolet (EUV) emission level exceeds that expected from the x-ray temperature gas in Coma. This halo suggests the presence of two more phases in the emitting gas, one at a temperature of approximately 2 x 10(6) kelvin and the other at approximately 8 x 10(5) kelvin. The latter phase cools rapidly and, in steady state, would have produced cold matter with a mass of approximately 10(14) solar masses within the EUV halo. Although a similar EUV enhancement was discovered in the Virgo cluster, this detection in Coma applies to a noncooling flow system.  相似文献   

Aurora hypotheses     

Tsurutani B  McPherron R  Gonzalez W 《Science (New York, N.Y.)》1988,239(4845):1228

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Chromium propionate improves performance and carcass traits in broilers     

Veerle Van Hoeck  Mahendra Sonawane  Antonio L. Gonzalez Sanchez  Iris Van Dosselaer  Chris Buyens  Dany Morisset 《动物营养（英文）》2020,6(4):480-487

There is evidence to suggest that poultry may have a dietary requirement for metabolically available chromium (Cr) that exceeds the amount provided through wheat soybean meal diets. The objective of the present study was to investigate the effects of dietary supplemental organic Cr from Cr propionate at different dose levels (control = 0 μg/kg, T1 = 200 μg/kg, T2 = 400 μg/kg) on the growth performance, carcass traits, and meat quality of broilers. Weight gain and feed intake of each treatment were recorded at the start and after 14, 28 and 35 d, and feed conversion ratios (FCR) were calculated accordingly. At 35 d of age, birds were randomly selected and euthanized for carcass evaluation. Results of the first trial indicate that both Cr propionate treatments increased final body weight (P < 0.05), feed efficiency (P < 0.05) and body weight gain (P < 0.0001). Furthermore, Cr propionate supplementation improved (P < 0.0001) all carcass characteristics. Interestingly, with increased Cr dosage, carcass yield, dressing percentage and breast meat yield increased linearly (P < 0.0001). The second study reveals that the feed intake in the control group was significantly higher compared to both Cr propionate supplemented groups (T1 & T2). Furthermore, the Cr propionate supplemented T2 group displayed a significantly lower FCR than the control and T1 group (P = 0.027). Finally, Cr propionate supplementation increased the dressing percentage compared to control birds (P < 0.0001). In the third experiment, Cr propionate supplementation (T1 & T2) increased final body weight and decreased FCR compared with the control treatment. These effects were highly significant (P < 0.0001) throughout all feeding phases of the trial. Cr propionate supplementation also increased (P < 0.0001) carcass yield, dressing percentage, breast meat yield, leg and thigh weights compared with the control treatment. In conclusion, growth performance, feed conversion, carcass yield, breast and leg meats of broiler birds can be significantly improved by dietary inclusion of Cr propionate. Cr propionate can be supplemented to broiler birds from 1 d old of age at a level that provides 200 or 400 μg/kg organic Cr and can increase the efficiency of broiler production.  相似文献   

NPAS2: a gas-responsive transcription factor     

Dioum EM  Rutter J  Tuckerman JR  Gonzalez G  Gilles-Gonzalez MA  McKnight SL 《Science (New York, N.Y.)》2002,298(5602):2385-2387

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Rate dependency hypothesis     

Gonzalez FA  Byrd LD 《Science (New York, N.Y.)》1977,198(4322):1183

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