Resumo Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.

Abstract To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

Ractopamine is a beta-adrenergic agonist that is approved for use in beef cattle, pigs and turkeys as a repartitioning agent to increase lean muscle deposition and decrease lipogenesis. Although the effects of dietary ractopamine on the proteome profile of post-mortem pork muscles have been examined, its influence on beef muscle proteome has not been studied. Therefore, the objective of this study was to examine the effect of ractopamine on the proteome profile of post-mortem beef longissimus lumborum (LL) muscle. LL muscle samples were obtained from the carcasses of six (n = 6) steers fed ractopamine (RAC; 400 mg ractopamine hydrochloride for 28 days) and six (n = 6) steers fed no ractopamine (CON). The muscle proteome was analysed using two-dimensional gel electrophoresis and tandem mass spectrometry. Five differentially abundant spots were identified, and all the spots were over-abundant in RAC. The identified proteins were involved in muscle structure development (F-actin-capping protein subunit beta-2; PDZ and LIM domain protein-3), chaperone activity (heat shock protein beta-1), oxygen transport (myoglobin), and glycolysis (L-lactate dehydrogenase A chain). These results suggested that dietary ractopamine could influence the abundance of enzymes associated with muscle development and muscle fibre type shift in beef LL muscle.

ABSTRACT The aim was to investigate the effect of glutamine (Gln) on broilers challenged with Salmonella Enteritidis. 240 1-day-old birds were divided into four groups in a completely randomized design, each of which included 6 replicates with 10 birds per replicate. Group I served as the unchallenged, untreated control (CON). All birds in groups II (SCC) - IV were challenged with 2.0 × 104 CFU/mL of S. Enteritidis. Birds in group III and IV were treated with 0.5% (Gln 1) and 1.0% (Gln 2), respectively, of Gln. The results indicated that S. Enteritidis infection led to a decrease in the average body weight at d 7, 14, and 21 (p<0.05). Chickens fed the Gln showed improved average body weights in comparison with the SCC group (p<0.05). At d 4, 7, 14, and 21, the Gln groups increased digestive enzyme (trypsin, lipase and amylase (except the amylase activity of jejunum at d 14 and d 21)) activities in the intestine (p<0.05), superoxide dismutase (SOD) (at d 14 jejunum; except at d 4, ileum) and catalase (CAT) (at d 4, and d 21, jejunum; d 4, ileum) activity in the serum (except at d 14) and intestinal mucosa (p<0.05), and the mRNA expression of SOD, CAT and nuclear respiratory factor 2 (Nrf2) of the intestinal mucosa compared with the SCC group (p<0.05). These results suggest that Gln as a feed additive could be effective for reducing the detrimental effects of S. Enteritidis infection of broilers.

ABSTRACT The objective of this study was to analyse the impact of dietary vitamin E supplementation on laying performance, serum reproductive hormones concentration and gene expression in ovary and follicles of anti-season breeding goose. A total of 210 anti-season breeding geese were divided into seven treatments with six replications. Each group was supplied with diets containing different vitamin E (DL-α-tocopherol acetate) contents (0, 10, 20, 40, 80, 160, 320mg/kg). We observed that the egg production and laying rate improved significantly at doses of 10 and 80 mg/kg, while the highest egg weight appeared in the 320 mg/kg group. Meantime, 80 mg/kg of vitamin E supplementation significantly improved the concentration of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) in serum (p<0.05). Dietary vitamin E supplementation significantly enhanced mRNA expression of FSHR, LHR and ESR1 at a dose of 80mg/kg, while PRLR increased at doses of 10 and 20mg/kg (p<0.05). It was found that the mRNA expression of Dicer increased at doses of 40 and 80 mg/kg of vitamin E supplementation in the ovary, SWF, LWF and SYF, respectively. Thence, Dietary vitamin E supplementation could improve egg laying performance, plasma reproductive hormones and the mRNA expression of reproductive hormone receptor genes in ovary, as well as the mRNA expression of Dicer in ovary, SWF, LWF and SYF. It was supposed that 80 mg/kg of vitamin E supplementation in dietary was appropriate to improve the fertility of anti-season breeding Xingguo grey goose.

Supplementation of vitamin E improves beef colour stability by minimizing lipid oxidation-induced myoglobin oxidation. Mitochondria affect myoglobin redox stability, and dietary vitamin E influences mitochondrial functionality in skeletal muscles. Nonetheless, the influence of vitamin E on the mitochondrial proteome of beef skeletal muscles has yet to be investigated. Therefore, the objective of this study was to examine the effect of dietary vitamin E on mitochondrial proteome of post-mortem beef longissimus lumborum (LL) muscle. Beef LL muscle samples (24 hours post-mortem) were obtained from the carcasses of nine (n = 9) vitamin E-fed (VITE) (1000 IU vitamin E for 89 days) and nine (n = 9) control (CONT) (diet without supplemental vitamin E) heifers. The mitochondrial proteome was analysed using two-dimensional gel electrophoresis and mass spectrometry, and nine differentially abundant spots were identified. All the differentially abundant spots were over-abundant in CONT, and the proteins were electron transport chain enzymes (NADH dehydrogenase iron-sulphur protein 8, NADH dehydrogenase flavoprotein 2, and cytochrome c oxidase subunit 5B), metabolic enzymes (succinate-CoA ligase (ADP-forming) subunit beta; dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex; pyruvate dehydrogenase protein X component), and enzymes involved in ATP regeneration (creatine kinase S-type). The low abundance of these proteins in VITE may decrease mitochondrial activity, resulting in low oxidative activity. These findings suggest that the strong antioxidant activity of vitamin E leads to less expression of mitochondrial oxidative enzymes in beef skeletal muscles.

Ractopamine is a beta-adrenergic agonist that increases leanness and carcass weight in finishing pigs. Our previous study observed that dietary ractopamine increased the abundance of several glycolytic enzymes in the sarcoplasmic proteome of post-rigor pork longissimus thoracis muscle. Pork semimembranosus is an economically important muscle and demonstrates differences in biochemistry compared with longissimus thoracis. Nonetheless, the effects of ractopamine on sarcoplasmic proteome of semimembranosus have not been evaluated yet. Therefore, this study examined the influence of ractopamine on sarcoplasmic proteome of post-rigor pork semimembranosus. Analyses of sarcoplasmic proteome of semimembranosus muscles from control (CON; diet without ractopamine) and ractopamine-fed (RAC; 7.4 mg/kg for 14 days followed by 10.0 mg/kg for 14 days) barrows revealed that haemoglobin subunit beta, alpha-crystallin B, and titin fragments were over-abundant in CON. In contrast, myosin light chain 1/3 and tripartite motif-containing protein 72 were over-abundant in RAC. The low abundance of haemoglobin subunit beta and alpha crystallin B in RAC could be attributed to fibre type shift (from oxidative to glycolytic) in response to ractopamine. The over-abundance of MLC 1/3 and tripartite motif-containing protein 72 in RAC could be due to the increased myofibrillar protein synthesis and muscle mass in ractopamine-fed pigs. Dietary ractopamine decreased the abundance of sarcoplasmic proteins involved in oxygen transport and chaperone activity, but increased the abundance of proteins involved in muscle contraction and plasma membrane repair in pork semimembranosus muscle.

Cystic echinococcosis (CE) is an anthropozoonotic disease with worldwide distribution and is caused by the cestode Echinococcus granulosus. Anaphylactic shock induced by CE rupture is a serious complication especially in patients with hydatid infections, as the resulting leakage of fluid contains highly toxic endogenous antigen. We aimed to isolate and identify the antigens of specific IgE and IgG1 (sIgE and sIgG1) in E. granulosus cyst fluid (EgCF). Crude antigen for EgCF was prepared from E. granulosus-infected sheep liver. Antigens were separated and identified by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D SDS-PAGE), two-dimensional gel electrophoresis (2-DE), and immunoblotting. Results of 1D SDS-PAGE and immunoblotting showed that 40.5 kDa protein was the major antigen of sIgE, and 35.5 kDa protein was the major antigen of sIgG1 in EgCF. Results of 2-DE and immunoblotting showed that main antigens of sIgE in EgCF were four proteins with pI values ranging from 6.5 to 9.0 and a molecular weight of 40.5 kDa. Main antigens of sIgG1 in EgCF were five proteins with pI values ranging from 6.5 to 9.0 and a molecular weight of 35.5 kDa. The antigens identified for sIgE and sIgG1 can provide critical insights into cellular and molecular mechanisms underlying anaphylactic shock induced by CE.

Since it was first identified in 1817, selenium has received considerable interest. Native selenium and a few selenium minerals were discovered several decades later. With the increasing number of selenium minerals, the occurrence of selenium minerals became the focus of much research. A great number of selenium deposits were reported all over the world, although few independent selenium deposits were discovered. Selenium is obtained mainly as a byproduct of other metals, and is produced primarily from the anode mud of copper refineries. This paper presents a comprehensive review of selenium minerals, as well as the treatment of copper refinery anode slimes for the recovery of selenium. Our focus is on the selenium minerals, including their discovery and occurrence, and the distribution of selenium resources. In addition, the main methods of recovering selenium from copper anode slimes are summarized.

The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period.

ABSTRACT Background: The aim of parapelvic cyst treatment is to have complete drainage of cyst fluid and prevent its further compression of the kidney and collective system. This study explores the efficacy and safety of surgical approaches with holmium laser ureteroscopic internal drainage in the treatment of parapelvic cyst. Methods: The compression effect of parapelvic cyst on the renal collective system was observed by rigid ureteroscope using retrograde ureteroscopic technology. With direct vision, the cyst wall on the obvious parapelvic compression site was cut. The diameter of the cyst wall cut was about 1 cm. The internal drainage was obtained by double-J tubes. When the cyst was in the inferior pole of kidney or where the rigid ureteroscope could not reach, a flexible ureteroscope was used. Results: In 28 cases of operation, 27 cases were successful. The cyst treatment time was eight to 40 minutes (average 26 minutes). During the operation, no massive haemorrhage, damage of nearby organ and ureter, or other complications happened. Time of follow-up was 10–72 months (average 39 months). The results of follow-up showed that in 22 cases, the cyst disappeared; the diameter of the cyst in four cases was reduced by more than half, and one case recurred. Conclusion: The treatment of parapelvic cyst by internal drainage operation using holmium laser and ureteroscopy was effective. The operation was safe with few complications.

RESUMEN Antecedentes: El objetivo del tratamiento de quiste parapélvico es lograr el drenaje completo del líquido del quiste y prevenir su compresión ulterior del riñón y el sistema colectivo. Este estudio explora la eficacia y seguridad de los abordajes quirúrgicos cn drenaje interno mediante ureteroscopia con láser de holmio en el tratamiento de quiste parapélvico. Métodos: El efecto de la compresión del quiste parapélvico del sistema colectivo renal fue observado mediante ureteroscopio rígido utilizando la tecnología ureteroscópica retrógrada. Con visión directa, la pared del quiste fue cortada en el sitio obvio de la compresión parapélvica. El diámetro de la pared del quiste cortada era alrededor de 1 cm. El drenaje interno se obtuvo con catéteres doble J. Cuando el quiste estaba en el polo inferior del riñón o donde no podía llegar el ureteroscopio rígido, se utilizaba un ureteroscopio flexible. Resultados: De 28 casos de operación, 27 casos tuvieron éxito. El tiempo de tratamiento del quiste fue de ocho a 40 minutos (un promedio de 26 minutos). Durante las operaciones, no hubo hemorragias masivas, daños de órganos y uréteres, u otras complicaciones. El tiempo de seguimiento fue de 10 a 72 meses (un promedio de 39 meses). Los resultados del seguimiento mostraron que en 22 casos, el quiste desapareció. En cuatro casos, el diámetro del quiste se redujo en más de la mitad, y reapareció en uno de los casos. Conclusión: El tratamiento de quiste parapélvico por operación de drenaje interno con láser de holmio y ureteroscopia fue eficaz. La operación fue segura con pocas complicaciones.

In a greenhouse experiment, Medicago sativa was grown in iron-rich soil inoculated with arbuscular mycorrhizal fungi (Funelliformis mosseae, FM) and/or iron (Fe) -reducing bacteria (IRB, Klebsiella pneumoniae strain L17) with three exogenous labile C input forms (no C source, as a single dose, and as a pulse) at rates of P0 and P5 (0 and 5 mg P2O5 kg-1, KH2PO4), which is to understand the role of IRB in enhancing the Fe-P utilization with exogenous labile C in subtropical soil. The results showed when inoculated with IRB, labile C as a pulsed input significantly improved the mycorrhiza colonization and hyphal length density, and increased the shoot P content, microbial biomass C and glomalin content at P5 rate; meanwhile, the labile C input also promoted the Fe (II) production, when inoculated with IRB, labile C as a pulsed input increased the C/A extractable Fe (II), oxalate-extractable P and microbial biomass P content. These facts suggested that application of labile C as a pulsed input enhanced the ability of IRB in improving the Fe reduction and Fe-P utilization in subtropical soil.

In this study, poultry manure compost and organic fertilizer were added to soils derived from basalt, granite, and river alluvial deposits for use as soil parent materials. Following incubation periods of 30 or 60 days at 25 °C, the physical and chemical properties and adsorption characteristics of the soils were studied. The results indicated that the application of poultry manure compost or organic fertilizer increased the available phosphorus (P) and organic matter content in all soils; however, the quantity of P and organic matter decreased with an increase in incubation time. The organic materials added via the treatments reduced the strength of P adsorption by the soil derived from basalt. For soils derived from granite and river alluvial deposits, the strength of P adsorption declined after poultry manure compost was added, but increased after the application of organic fertilizer. All soil treatments derived from basalt fitted well with the three adsorption isotherm equations. For soils derived from granite and alluvial deposit as parent materials, the data conformed to the three adsorption models in the order: Langmuir > Freundlich > Temkin. The maximum phosphate buffering capacity (MPBC) and standard P retention (SPR) of soil derived from basalt decreased following the application of poultry manure or organic fertilizer, whereas changes in MPBC and SPR for soils derived from granite and alluvial deposits were dependent on the organic content and incubation time.

This paper investigates defect detection methodologies for rolling element bearings through vibration analysis. Specifically, the utility of a new signal processing scheme combining the High Frequency Resonance Technique (HFRT) and Adaptive Line Enhancer (ALE) is investigated. The accelerometer is used to acquire data for this analysis, and experimental results have been obtained for outer race defects. Results show the potential effectiveness of the signal processing technique to determine both the severity and location of a defect. The HFRT utilizes the fact that much of the energy resulting from a defect impact manifests itself in the higher resonant frequencies of a system. Demodulation of these frequency bands through use of the envelope technique is then employed to gain further insight into the nature of the defect while further increasing the signal to noise ratio. If periodic, the defect frequency is then present in the spectra of the enveloped signal. The ALE is used to enhance the envelope spectrum by reducing the broadband noise. It provides an enhanced envelope spectrum with clear peaks at the harmonics of a characteristic defect frequency. It is implemented by using a delayed version of the signal and the signal itself to decorrelate the wideband noise. This noise is then rejected by the adaptive filter that is based upon the periodic information in the signal. Results have been obtained for outer race defects. They show the effectiveness of the methodology to determine both the severity and location of a defect. In two instances, a linear relationship between signal characteristics and defect size is indicated.