Target silencing of porcine SPAG6 and PPP1CC by shRNA attenuated sperm motility.

The quality of sperm significantly influences the reproductive efficiency of pig herds. High-quality sperm is necessary for efficient fertilization and to maximize the litter numbers in commercial pig farming. However, the understanding of genes regulating porcine sperm motility and viability is limited. In this study, we validated porcine sperm/Sertoli-specific promoters through the luciferase reporter system and identified vital genes for sperm quality via loss-of-function means. Further, the shRNAs driven by the ACE and SP-10 promoters were used to knockdown the SPAG6 and PPP1CC genes which were provisionally important for sperm quality. We assessed the effects of SPAG6 and PPP1CC knockdown on sperm motility by using the sperm quality analyzer and flow cytometry. The results showed that the ACE promoter is active in both porcine Sertoli cells and sperms, whereas the SP-10 promoter is operating exclusively in sperm cells. Targeted interference with SPAG6 and PPP1CC expression in sperm cells decreases the motility and increases apoptosis rates in porcine sperms. These findings not only offer new genetic tools for targeting male germ cells but also highlight the crucial roles of SPAG6 and PPP1CC in porcine sperm function.

The Survival and Reproduction of Rhopalosiphum padi (Hemiptera: Aphididae) on Different Plants: Exploring the Possible Host Range for a Serious Wheat Pest.

Rhopalosiphum padi (L.) is one of the most economically important pests of wheat worldwide; however, the host ranges of R. padi remain unclear. Particularly, it is unknown which plants R. padi can survive and reproduce on after the harvest of crops. The results revealed that the survival, developmental times, longevity, and fecundity of the aphid varied among the 13 Gramineae weeds, with the life-history parameters significantly differing. The virginoparae could survive long-term and reproduce on 11 of the 13 weeds. Gramineae weeds can possibly play a significant role in the buildup of R. padi populations as reservoirs. The virginoparae could survive long term and reproduce on Iris lactea Pall. var. chinensis (Fisch.) Koidz (Liliflorae: Iridaceae), Iris tectorum Maxim. (Liliflorae: Iridaceae), Cyperus rotundus L. (Cyperales: Cyperaceae), and Brassica oleracea L. var. capitata (Rhoeadales: Cruciferae), but not on Fagopyrum esculentum Moench (Polygonales: Polygonaceae), F. tataricum (L.) Gaertn. (Polygonales: Polygonaceae), Chlorophytum comosum (Thunb.) Baker (Liliflorae: Liliaceae), and Ophiopogon japonicas (Thunb.) Ker-Gawl (Liliflorae: Liliaceae). Rhopalosiphum padi can survive and reproduce on non-Gramineae plants of different families. Detailed host range information would be helpful for more effective control of insect pests. The design and implementation of sustainable pest management strategies should consider the aphid population on weeds and other host plants.

Improvement of the homogeneity of protein-imprinted polymer films by orientated immobilization of the template.

A method for preparing homogeneous protein-imprinted polymer films with orientated immobilization of template is described. The template methyl parathion hydrolase (MPH) was modified with a peptide linker (Gly-Ser)(5)-Cys and was immobilized on a cover glass with a fixed orientation via the linker. The activity of the fusion enzyme (MPH-L) was evaluated by determining the product's absorbance at 405 nm (A(405)). Both the free and the immobilized MPH-L showed higher retention of the bioactivity than the wide type enzyme (MPH-W) as revealed by the A(405) values for MPH-L(free)/MPH-W(free) (1.159/1.111) and for MPH-L(immobilized)/MPH-W(immobilized) (0.348/0.118). The immobilized MPH-L also formed a more homogeneous template stamp compared to the immobilized MPH-W. The molecularly imprinted polymer films prepared with the immobilized MPH-L exhibited high homogeneity with low Std. Deviations of 80 and 200 from the CL intensity mean volumes which were observed for batch-prepared films and an individual film, respectively. MPH-L-imprinted polymer film also had a larger template binding capacity indicated by higher CL intensity mean volume of 3900 INT over 2500 INT for MPH-W-imprinted films. The imprinted film prepared with the orientated immobilization of template showed an imprinting factor of 1.7, while the controls did not show an imprinting effect.

Two generation reproduction and teratogenicity studies of feeding cyadox in Wistar rats.

To investigate the teratogenic potential and reproductive toxicity of cyadox, a growth promoting agent, Wistar rats (F(0)) were fed with diets containing cyadox (0, 50, 150 and 2500 mg/kg) or olaquindox (150 mg/kg), approximately equivalent to cyadox 5, 15, 250 or olaquindox 15 mg/kg b.w./day across two generations. Half of the pregnant rats (F(0), F(1b)) were subjected to caesarean section on gestational day 20 for teratogenic examination and the other half produced pups F(1a) and F(2a), respectively. At the 250 mg/kg b.w./day cyadox group, body weights of F(1b) pregnant rats and F(2a) on day 21 after birth decreased; fetal body lengths and tail lengths decreased; the number of fetal resorptions increased significantly; litter weights, number of viable fetuses decreased; number of embryo resorptions increased significantly; number of liveborn F(1a), F(1b) and F(2a) decreased. No macroscopic or microscopic change of any significance was found in the reproductive organs. Significant increases in the incidence of cervical ribs or lumbar ribs in F(2a) pups and significant increases of relative organ weight of testis and epididymis in F(1b) were observed at the 250 mg/kg b.w./day cyadox group. The NOAEL for reproduction/development of cyadox for rats was estimated to be 150 mg/kg diet, which was equivalent to approximately 15 mg/kg b.w./day.

Fluorescence detection techniques for protein kinase assay.

Traditional methods for protein kinase (PK) assay are mainly based on use of (32)P-labeled adenosine triphosphate (ATP); applications of such methods are, however, hampered by radioactive waste and short half-life of (32)P-labeled ATP. Therefore non-radioactive methods, such as fluorescence detection techniques are good alternative. In this review, we describe the principles of four fluorescence techniques (fluorescence intensity endpoint measurement, fluorescence resonance energy transfer (FRET), fluorescence polarization (FP), and fluorescence lifetime imaging) and provide an overview of applications of these fluorescence detection techniques in protein kinase assay, underlining their relative advantages and limitations. Research trends in this field are also highlighted. [figure: see text]

Experimental evaluation of cyadox phototoxicity to Balb/c mouse skin.

BACKGROUND: Cyadox is a veterinary drug mainly used as an effective antimicrobial promoter in animal husbandry. It was reported that the other quinoxaline-1,4-dioxide compounds had phototoxicity, but only few data are reported on phototoxicity of cyadox. This study was performed to evaluate the phototoxicity of cyadox on skin. METHODS: Eighty mice were equally divided into eight groups. Five groups with ultraviolet A (UVA) irradiation for 2 h (21 J/cm2) were administered at doses of 0, 10, 50, 200 mg/kg cyadox and 50 mg/kg olaquindox, respectively, and others as controls without irradiation administered at doses of 0, 200 mg/kg cyadox and 50 mg/kg olaquindox, respectively. Drugs were administered by gavage once daily with a suspension in 0.8% carboxymethyl-cellulose for consecutive 4 days. After administration, a recovery period of 7 days was arranged. Histopathological examination of auricular skin was performed on days 5 and 12. Measurement of auricular thickness, gross pathology and erythema score were conducted once daily. RESULTS: Cyadox groups (200, 50 and 10 mg/kg) with irradiation had erythema and oedema of auricular skin with dose-response relationship, which gradually convalesced after dosing and irradiation ceased. Severe erythema, oedema and necrosis of auricles were observed on olaquindox group with irradiation. CONCLUSION: The phototoxicity of cyadox was mild and reversible, which demonstrated a good safety profile of cyadox in terms of phototoxicity.

Subchronic oral toxicity study with cyadox in Wistar rats.

To investigate the potential subchronic toxicity of cyadox, groups of 15 male and 15 female Wistar rats were fed with diets containing cyadox (0, 50, 150 or 2500 mg/kg) or olaquindox (150 mg/kg), approximately equivalent to cyadox 5, 15, 250 or olaquindox 15 mg/kg b.w./day, for 13 weeks. Five rats/sex/group were sacrificed on days 30, 60 and 90. No test-material-related changes were seen in mortality, clinical signs, hematology, clinical chemistry, organ weight data and macroscopic examinations. Except that body weights of both sexes of the 2500 mg/kg cyadox group were significantly lower than controls beginning after the second week of treatment. Body weights of females of 150 mg/kg olaquindox group were significantly lower than those of the control group at weeks 3 and 4. Other groups were unaffected by treatments. Histopathological observations revealed that 2500 mg/kg cyadox or 150 mg/kg olaquindox induced swelling and fatty degeneration of the hepatocytes and proximal renal tubular epithelial cells. It was for the first time that changes were found in the liver and kidneys of rats fed 2500 mg/kg cyadox. The no-observed-adverse-effect level (NOAEL) of cyadox for rats was estimated to be 150 mg/kg dietary dose level.