Clinical Use of Deslorelin for the Control of Reproduction in the Bitch

This study was conducted to evaluate clinical efficacy of deslorelin for inhibiting reproduction in the bitch. Ten adult healthy bitches or bitches with mammary neoplasia for which owners were requesting suppression of cyclicity without performing gonadectomy were administered a 4.7- or a 9.4-mg deslorelin implant subcutaneously. The first implant of deslorelin was administered in anoestrus (n = 5) or in dioestrus (n = 5). Treatment was repeated every 5 months for as long as necessary based on the clinical situation of the dog and owner's desires. Some of the bitches implanted in anoestrus came in heat within 4–15 days after treatment, while none of the bitches implanted in dioestrus showed heat during treatment. Suppression of reproductive cyclicity was successfully achieved in 6/10 bitches for 1–4 years. No behavioural and local/general side-effects were observed in any of the treated bitches. The 4.7-mg deslorelin implant may work well for suppression of cyclicity provided that it is administered in dioestrus and at intervals of 4.5 months. A 9.4-mg implant may be more suitable for this use although its efficacy may also be shorter than 12 months. Owner compliance is an important limiting factor.     

Measurements of the equation of state of deuterium at the fluid insulator-metal transition

A high-intensity laser was used to shock-compress liquid deuterium to pressures from 22 to 340 gigapascals. In this regime deuterium is predicted to transform from an insulating molecular fluid to an atomic metallic fluid. Shock densities and pressures, determined by radiography, revealed an increase in compressibility near 100 gigapascals indicative of such a transition. Velocity interferometry measurements, obtained by reflecting a laser probe directly off the shock front in flight, demonstrated that deuterium shocked above 55 gigapascals has an electrical conductivity characteristic of a liquid metal and independently confirmed the radiography.     

Oestradiol Induced Inhibition of Neuroendocrine Marker Expression in Leydig Cells of Adult Rats

The objectives of this work were to determine the changes in the expression of neuroendocrine markers in Leydig cell by oestradiol treatment, and to determine whether testosterone is able to recover partially the effects of hormonal suppression induced by oestradiol. Adult male rats were injected daily with either 50 microg of oestradiol or oestradiol plus testosterone propionate (25 mg every 3 days) for 15 days. The animals were sacrificed and testicles were dissected and processed by routine histological protocols. FSH and LH serum levels were determined by radioimmunoassay. The visualization of antigens was achieved by the streptavidin-peroxidase immunohistochemical method. Antibodies against chromogranin A (CrA), S-100 protein (S-100), P substance (PS), synaptofisin (SYN), neurofilament protein (NF), gliofibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were used. The mean LH and FSH serum concentrations were consistently suppressed with hormonal treatments. Intermediate filaments (NF and GFAP) showed no difference in their expression. The expression of S-100, NSE and SYN was significantly lower in both hormone-treated groups. In oestradiol-treated rats, the immunoreactivity of CrA and SP decreased significantly but was restored after testosterone supplementation. Although the nature and functions of many of these substances in Leydig cells remain unknown, these results are consistent with the hypothesis that the expression of some neuroendocrine markers is hormonally controlled.     

Characterization of the cytokine pattern of porcine bone marrow-derived cells treated with 1alpha,25(OH)D

The biologically active form of vitamine D(3) [1alpha,25(OH)(2)D(3)] has recently been described not only to influence bone metabolism but also to exert immunomodulating activities, which may have an impact on bone formation/resorption as well. In this study, we analysed the effects of 1alpha,25(OH)(2)D(3) on the cytokine pattern of porcine bone marrow-derived cells from piglets aged 1-3 weeks. After culture for 1 week, the number of osteoclasts was determined, with tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated cells being considered osteoclasts. Cultured bone marrow cell-derived mRNA was subjected to semiquantitative RT-PCR specific for a panel of porcine cytokines (IL-1alpha, IL-6, IL-8, IL-10, and TNF-alpha). In addition, an immunofluorescence analysis using anti-porcine mAbs specific for IL-1beta, IL-2, IL-4, IL-6, IL-12, TNF-alpha, and IFN-gamma was performed. In order to prove the existence of a porcine homologue of the receptor activator of NF-kappaB ligand (RANKL) bone marrow cell- as well as porcine white blood cell-derived mRNA was investigated by RT-PCR using primer pairs specific for murine RANKL. Cell culture supernatant was analysed for soluble RANKL by means of an ELISA designed for quantification of human RANKL. By means of RT-PCR, expression of IL-1alpha, IL-6, IL-8, IL-10 and TNF-alpha mRNA could be found in cells cultured with and without 1alpha,25(OH)(2)D(3). Immunofluorescence analysis revealed that IL-1, IL-6, and TNF-alpha were produced by both stromal cells and osteoclasts. Besides its known osteoclastogenic effects, 1alpha,25(OH)(2)D(3) tended to downregulate the respective cytokines, but significantly upregulated RANKL expression. The homology between the porcine RANKL-specific sequence and the corresponding human RANKL sequence was 79%. The data found support the idea that porcine bone marrow cell cultures may provide a suitable alternative to murine systems in human osteological research.     

Fungi isolated from bovine udders,and their possible sources

AIM: To identify fungi isolated from infections of the bovine mammary gland, and establish their possible sources.

METHODS: From a herd of 420 cows, milk samples were collected from all quarters at calving and cultured to detect causative organisms. Quarters identified as infected with fungi were further sampled during early lactation. Samples from feedstuffs, the feed pad and ends of teats were also collected and analysed for the presence of fungi.

RESULTS: Eleven of 420 cows were diagnosed with intramammary infections (IMI) caused by yeasts (nine cows, 10 quarters) and moulds (two cows, three quarters). Six of the yeast species had previously been reported as being responsible for mastitis. Elevated somatic cell counts (SCC) were observed in many quarters, but most infections were eliminated spontaneously. Two of the fungi isolated from milk samples were also isolated from feedstuffs and teat swabs, and seven other fungi isolated from milk samples were not isolated from feed, the feed pad or cows' teats.

CONCLUSIONS: Isolation of fungi from the udder is rarely reported in dairy cows in New Zealand. In this herd, contamination of the end of the teat originating from feedstuffs and possibly exacerbated by the use of a feed pad may have led to the establishment of IMI caused by fungi.

CLINICAL RELEVENCE: Fungi are infrequently if ever reported in mastitis trial data or surveys in New Zealand and are probably of little clinical significance.     

Assessment of Different Functional Parameters of Frozen–Thawed Buffalo Spermatozoa by Using Cytofluorimetric Determinations

Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen–thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR‐14/propidium iodide staining; mitochondrial function by JC‐1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC‐PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Δψ^high), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %‐DFI, ‐DFI, SD‐DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome‐reacted live (ARL) and acrosome‐reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respectively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca²⁺ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen‐thawed semen fertilizing potential.     

Heat Shock Protein 70 and Sex Steroid Receptors in the Follicular Structures of Induced Ovarian Cysts

The purpose of this study was to estimate the expression and relative amounts of estrogen (ER) and progesterone receptors (PR) and their isoforms as well as heat shock protein 70 (HSP70) in ovaries of rats with induced cystic ovarian disease (COD). Primary, secondary, tertiary, atretic and cystic follicles were evaluated by immunohistochemistry and total ovarian proteins were analyzed by Western blot. In the granulosa layer, growing and cystic follicles in the treated group have a higher expression of ERα than growing follicles of control individuals. In the theca interna layer, tertiary follicles presented a significantly higher expression of ERα in the treated group. An increase in total ERα protein was detected in the treated group. Granulosa cells of all growing, atretic and cystic follicles show a lower expression of ERβ in animals with COD, and the total protein expression of ERβ was lower in this group. The expression of PR was lower in the granulosa cell layer of tertiary and cystic follicles in treated animals, and theca interna layer had less intense immunostaining in this group. Although there were no differences in the expression of PR-B by Western blotting, the expression of PR-A was higher and the expression of PR-C was smaller in the treated group. An intense HSP70 immunostaining was observed in the cells of cystic follicles. By Western blotting, higher protein expression of HSP70 was detected in the ovarian samples of the control group than those of the treated ones. Ovaries of animals with COD exhibited an altered steroid receptor expression and subtype balance as compared with control animals, and an increase in HSP70 immunoexpression.     

CAN SONOGRAPHIC FINDINGS PREDICT THE RESULTS OF LIVER ASPIRATES IN DOGS WITH SUSPECTED LIVER DISEASE?

While abdominal ultrasound and ultrasound-guided fine-needle aspiration cytology are often combined to help determine the type of liver disease in dogs, little is known about the relationship that may exist between the results of these tests. We hypothesized that specific sonographic findings, or combinations of findings, may predict results of liver ultrasound-guided fine-needle aspiration cytology. Hepatic and extrahepatic sonographic findings were recorded prospectively using a standardized form in 70 dogs with clinically suspected liver disease and in which liver ultrasound-guided fine-needle aspiration cytology was performed. The predictive value of sonographic findings in regard to the category of cytology results was assessed with stepwise logistic regression analysis. Sonographic detection of a hepatic mass (≥3 cm; risk ratio [RR] 3.83, 95% Wald confidence intervals [95% CI] 2.42–3.93, P =0.0036), ascites (RR 3.82, 95% CI 1.94–4.28, P =0.0044), abnormal hepatic lymph node(s) (RR 3.01, 95% CI 1.22–4.88, P =0.0262), and abnormal spleen (RR 3.26, 95% CI 1.20–3.85, P =0.0274) were the most predictive of liver neoplasia on cytology. Conversely, sonographic detection of hepatic nodules (<3 cm; RR 1.97, 95% CI 0.95–2.96, P =0.0666) was most predictive of vacuolar hepatopathy on cytology. In dogs with suspected liver disease, several sonographic findings, alone or combined, are thus predictive of liver ultrasound-guided fine-needle aspiration cytology results. In the light of the fact that ultrasound-guided fine-needle aspiration cytology of the liver has limitations, these predictabilities could influence the selection of diagnostic tests to reach a reliable diagnosis.     

黔北地区主要竹种竹材气干密度与力学性质的关系研究

对黔北地区几种主要竹种的气干密度与力学性质的关系研究表明:各竹种的气千密度与力学性质除部分竹种外均存在一定的曲线或直线关系,其中在气干密度与抗剪强度关系中,除慈竹、花吊丝、毛脚龙竹、金佛山方竹等竹种外,其他竹种均可利用回归方程T12=ap12b,T12=ap12+b进行估测.在气干密度与顺纹抗拉强度关系中除硬头黄、花吊...