Spermatic and oxidative profile of domestic cat (Felis catus) epididymal sperm subjected to different cooling times (24, 48 and 72 hours)          下载免费PDF全文

DSR Angrimani  KK Nagai  BR Rui  LC Bicudo  JDA Losano  MM Brito  MCP Francischini  M Nichi 《Reproduction in domestic animals》2018,53(1):163-170

Cooling stored epididymal samples for several days allows facilities to transport and process genetic material post‐mortem. Improvements to this practice allow the preservation of sperm from domestic cats, which are the ideal study model for wild felids. However, the modifications in spermatic features and the oxidative profile are not fully understood in cats. This information is necessary for the development of biotechniques, such as new extenders for cryopreservation. Therefore, the purpose of this study was to evaluate the spermatic and oxidative profile in samples from the epididymal cauda of domestic cats cooled at 5°C for 24, 48 and 72 hr. Spermatozoa were collected from the epididymis cauda. Evaluations consisted of computer‐assisted sperm analysis (CASA), plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, sperm DNA integrity (toluidine blue), mitochondrial activity (3′3 diaminobenzidine), activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD), measurement of lipid peroxidation (TBARS) and protein oxidation. A decrease in sperm motility parameters was observed after 72 hr of cooling (i.e. total and progressive) with a higher percentage of minor (37.7 ± 6.3%) and total defects (53.4 ± 6.3%). Additionally, a decrease in high mitochondrial activity (Class I: 16.6 ± 2.2%) occurred after 72 hr. The decrease in motility rates after a long cooling time probably was caused by the increase in sperm abnormalities. A long cooling time causes cold shock and mitochondrial exhaustion, but there was no observed change with the oxidative stress condition. Therefore, cat epididymal sperm stored at 5°C appear to maintain a high quality for up to 48 hr of cooling time.  相似文献   

A fast,low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species          下载免费PDF全文

BR Rui  DSR Angrimani  LC Bicudo  JDA Losano  M Nichi  RJG Pereira 《Reproduction in domestic animals》2018,53(1):171-175

Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high‐precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For this purpose, we used six animals of each specie (n = 30), in reproductive age. Sperm was collected by different methods according to the particularities of each species, and such samples were divided into two aliquots: a sperm sample was kept at 5°C (considered as intact sperm DNA), and the remaining samples were submitted to the induction of DNA fragmentation by exposure to ultraviolet light for 4 hr. Samples were then mixed with the intact sample to obtain known and progressive proportions of sperm with fragmented DNA (0%, 25%, 50%, 75% and 100%). Semen smears were performed and subjected to staining with TB. Blue‐stained spermatozoa were considered to have DNA fragmentation. We observed high linear regression coefficients between the expected proportion of damaged DNA and the results of TB for dog, cat, ram, bull and stallion samples. In conclusion, TB stain was considered a fast and effective technique for the study of spermatozoa DNA in several species.  相似文献   

Effect of anticonvulsant dosages of potassium bromide on thyroid function and morphology in dogs     

Paull LC  Scott-Moncrieff JC  DeNicola DB  Glickman N  Refsal KR  Glickman LT 《Journal of the American Animal Hospital Association》2003,39(2):193-202

A placebo-controlled experiment was performed to evaluate the effect of potassium bromide on the canine thyroid gland. Basal total thyroxine, free thyroxine, and basal thyrotropin serum concentrations were evaluated over a 6-month period in potassium bromide-treated and control dogs. A thyrotropin-releasing hormone stimulation test was also performed in all dogs at the beginning and conclusion of the study. Thyroid histopathology was compared between treated and control dogs at the end of the study. No difference was detected in any parameter between the two groups at the end of the study. A decline in thyroid hormone concentrations over the course of the study did occur in both groups of dogs. Potassium bromide does not appear to have a significant effect on canine thyroid function or morphology.  相似文献   

Pharmacy and dispensing courses: a new era     

Paull N 《The Journal of small animal practice》2002,43(8):377

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Pineapple organic acid metabolism and accumulation during fruit development     

Parson Saradhuldhat  Robert E. Paull 《Scientia Horticulturae》2007

Developmental changes in pineapple (Ananas Comosus (L.) Merrill) fruit acidity was determined for a ‘Smooth Cayenne’ high acid clone PRI#36-21 and a low acid clone PRI#63-555. The high acid clone gradually increased in fruit acidity from 1.4 meq/100 ml 6 weeks from flowering, and peaked a week before harvest at ca 10 meq/100 ml. In contrast, the low acid clone increased in acidity 6 to 8 weeks after flowering, peaked 15 weeks after flowering at ca. 9 meq per/100 ml and then sharply declined in 2 weeks to 6 meq/100 ml. The increased in total soluble solids (TSS) of the low acid clone began 6 weeks after flowering and for the high acid clone at 12 weeks after flowering. The increase in titratable fruit acidity (TA) paralleled the changes in the citric acid content of both clones. Citric acid content increased from less than 1 mg/g at 6 weeks after flowering to 6 to 7 mg/g, 9 weeks later. The malic acid concentration in both clones varied between 3 and 5 mg/g and showed no marked changes just before harvest. The developmental changes in fruit potassium were significantly correlated with fruit acidity and fruit total soluble solids in both the high and low acid clones. Developmental changes in acid-related enzymatic activities showed an increase in citrate synthase (EC 4.1.3.7) activity that occurred a week before harvest, coincided with the peak in citric acid in the high acid clone. An increase in aconitase (ACO, EC 4.2.1.3) activity was observed just before harvest as the decline in acidity occurred in the low acid clone. The activities of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), malate dehydrogenase (MDH, EC 1.1.1.37) and malic enzyme (ME, EC 1.1.1.40) did not parallel any changes in fruit acidity. The results indicated that the change in pineapple fruit acidity during development was due to changes in citric acid content. The major difference in acid accumulation occurred in the low acid clone just before harvest when acidity declined by one-third. The activities of citrate synthase and aconitase possibly played a major role in pineapple fruit acidity changes.  相似文献   

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia     

C Fergani  JE Routly  DN Jones  LC Pickavance  RF Smith  H Dobson 《Reproduction in domestic animals》2014,49(3):433-440

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.  相似文献   

Genetics of tolerance to high concentrations of soil boron in peas (Pisum sativum L.)     

A. Bagheri  J. G. Paull  A. J. Rathjen 《Euphytica》1996,87(1):69-75

Summary The inheritance of tolerance to high concentrations of soil boron in pea (Pisum sativum L.) was studied in five cross combinations and their reciprocals. Segregation patterns for boron response in F₂ populations and F₃ derived families were established by visual assessment of leaf damage. The segregation ratios were explained in terms of two major gene loci interacting in an additive manner with incomplete dominance at each locus. Evaluation of selected tolerant and susceptible families indicated that tolerant families contained a significantly lower concentration of boron in shoots than susceptible families.  相似文献   

Responses of wheat and barley genotypes to toxic concentrations of soil boron     

J. G. Paull  B. Cartwright  A. J. Rathjen 《Euphytica》1988,39(2):137-144

Summary The growth and yield of seven wheat and two barley cultivars or lines, previously found to show different degrees of boron tolerance under field conditions, were compared in a pot experiment at a range of soil boron treatments. Soil treatments ranged up to 150 mg/kg applied B. Extractable B in soils ranged up to 103 mg/kg.At the highest B treatment seedling emergence was delayed, but the percentage emergence was not reduced. The degree of boron toxicity symptom expression varied between the wheat cultivars and lines, with the two most tolerant, Halberd and (Wq^*KP)^*WmH)/6/12, displaying the least symptoms.The concentration of boron applied to the soil which produced a significant depression of growth and yield varied between cultivars. For example, the yield of (Wq^*KP)^*WmH)/6/12 was not affected at the 100 mg/kg applied boron treatment, while the grain yield for (Wl^*MMC)/W1/10 was significantly reduced at the 25 mg/kg treatment.There was a linear increase in boron concentration in tillers at the boot-stage with increasing concentration of boron in the soil. The most boron tolerant genotypes had the lowest tissue boron concentrations in each of the treatments. Halberd and (Wq^*KP)^*WmH)/6/12 had approximately half the boron concentrations of the more sensitive genotypes at the 25 and 50 mg/kg treatments. Differential tolerance of boron within the tissue was also observed. Both Stirling and (Wl^*MMC)/W1/10 had significantly reduced total dry matter and grain yields at the 25 mg/kg treatment, while the concentrations of boron in boot stage tillers at this treatment were 118 and 100 mg/kg, respectively. On the other hand, Halberd and (Wq^*KP)^*WmH)/6/12 had tissue boron concentrations of 144 and 131 mg/kg, respectively, at the 50 mg/kg treatment but yield was unaffected.The relative responses in the pot experiment, for wheat, were in close agreement with field results. Halberd and (Wq^*KP)^*WmH)/6/12 had the highest grain yields, with the lowest concentrations of boron in the grain when grown under high boron conditions in the field. In pots these two genotypes proved to be the most tolerant of boron. For barley the advantage in grain yield in the field, expressed by WI-2584 compared with Stirling, was not repeated in pots. WI-2584 was, however, more tolerant than Stirling on the basis of total dry matter production.The results show that useful variation in boron tolerance exists among wheat, and that breeding should be able to provide cultivars tolerant to high levels of boron.  相似文献   

Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex   总被引：2，自引：0，他引：2  

Lee JH  Paull TT 《Science (New York, N.Y.)》2004,304(5667):93-96

The complex containing the Mre11, Rad50, and Nbs1 proteins (MRN) is essential for the cellular response to DNA double-strand breaks, integrating DNA repair with the activation of checkpoint signaling through the protein kinase ATM (ataxia telangiectasia mutated). We demonstrate that MRN stimulates the kinase activity of ATM in vitro toward its substrates p53, Chk2, and histone H2AX. MRN makes multiple contacts with ATM and appears to stimulate ATM activity by facilitating the stable binding of substrates. Phosphorylation of Nbs1 is critical for MRN stimulation of ATM activity toward Chk2, but not p53. Kinase-deficient ATM inhibits wild-type ATM phosphorylation of Chk2, consistent with the dominant-negative effect of kinase-deficient ATM in vivo.  相似文献   

Biological communities at the Florida escarpment resemble hydrothermal vent taxa   总被引：4，自引：0，他引：4  

Paull CK  Hecker B  Commeau R  Freeman-Lynde RP  Neumann C  Corso WP  Golubic S  Hook JE  Sikes E  Curray J 《Science (New York, N.Y.)》1984,226(4677):965-967

Dense biological communities of large epifaunal taxa similar to those found along ridge crest vents at the East Pacific Rise were discovered in the abyssal Gulf of Mexico. These assemblages occur on a passive continental margin at the base of the Florida Escarpment, the interface between the relatively impermeable hemipelagic clays of the distal Mississippi Fan and the jointed Cretaceous limestone of the Florida Platform. The fauna apparently is nourished by sulfide rich hypersaline waters seeping out at near ambient temperatures onto the sea floor.  相似文献