Relationship between endogenous protein disulfide isomerase family proteins and glutenin macropolymer

The effects of endogenous protein disulfide isomerase (PDI) family proteins on the properties of gluten proteins in dough during breadmaking were determined using bacitracin, an inhibitor of PDI. Bread loaf volume in the presence of bacitracin was increased to 118% of that in the absence of bacitracin. The addition of bacitracin caused a decrease in the extension tolerance of the dough. The amount of sodium dodecyl sulfate (SDS)-insoluble glutenin macropolymer (GMP) in dough decreased to approximately 70% of that in flour during the 20 min of mixing for doughmaking. The addition of bacitracin to dough caused a dramatic GMP decrease, corresponding to ～20-30% of that in flour during the 20 min of mixing. The decrease in GMP was compensated by an increase in SDS-soluble glutenin polymer. Taken together, these results suggest that the endogenous PDI family proteins in flour suppress the depolymerization of GMP during dough mixing.     

Molecular characterization of recent Korean porcine reproductive and respiratory syndrome (PRRS) viruses and comparison to other Asian PRRS viruses

Twenty-eight PRRS viruses (PRRSVs) isolated from various pig farms in Korea between 2002 and 2003 were sequenced for open-reading frame (ORF) 5 and/or full-length genome and compared with numerous PRRSVs reported from North America, Europe and Asia. All Korean isolates examined were genetically of the North American genotype. The ORF5 sequence of one isolate was identical to Ingelvac PRRS MLV vaccine virus. ORF5 nucleotide sequence divergence of the remaining 27 Korean PRRSVs from VR-2332, the prototype of the North American PRRSV and parental strain of the MLV vaccine virus, ranged from 1.3% to 12.9%, which corresponded to 2.0% to 14.9% divergence at the amino acid level, raising a concern on the efficacy of the MLV vaccine. Phylogenetic analyses of ORF5 and/or full-length sequences revealed that the Korean PRRSVs formed a clade distinct from PRRSVs reported from other Asian countries (China, Taiwan, Japan, and Thailand). Our study demonstrated that PRRSVs of the North American genotype were introduced to the Korean swine population some time ago and have evolved independently from PRRSV in other Asian countries, suggesting that geographic separation might influence the molecular evolution of PRRSV. This should be taken into consideration when a national PRRS prevention and control policy for international trade is established.     

Decrease of pituitary adenylate cyclase activating polypeptide and its type I receptor mRNAs in rat testes by ethanol exposure

The present study was designed to evaluate the effect of ethanol on pituitary adenylate cyclase activating polypeptide (PACAP) and its typ I (PAC1) receptor expression in adult rat testes. Ethanol (3 g/kg i.p., 15% v/v in saline) was administrated to adult male rats for 10 days. Using northern blot analysis, the present study showed the reduction of PACAP mRNA levels in rat testes by ethanol administration. Also, ethanol decreased the expression level of PAC1 receptor in testes. In particular, in situ hybridization clearly showed the decrease of PAC1 receptor mRNA expression in Leydig cells, which produce testosterone. Furthermore, the serum level of testosterone was significantly decreased in the ethanol-treated group. In conclusion, our findings suggest that the decrease of PACAP and PAC1 receptor expression in rat testes by ethanol exposure may partly contribute to the suppression of male reproductive activity.     

The effects of growth and disease in serum keratan sulfate concentration in dogs

The purpose of this study is to investigate keratan sulfate (KS) concentration in the serum of puppies and the effects of age, body weight, breed and diseases. Serum samples from six neonatal dogs (4 Beagles, 2 Labrador Retrievers), and from 127 adult dogs with various diseases were collected at a Teaching Animal Hospital. Canine serum KS concentration was measured by inhibition enzyme-linked immunosorbent assay (ELISA). Samples from puppies were evaluated for growth-related changes, and samples from patients were evaluated for age, body weight, breed and disease-related changes. Serum KS concentration was high in puppies from birth to 4 months of age. KS values started to decrease from 4 months to 9 months of age, and then gradually reached to the plateau. Though in the small sample, mean KS concentration in a Labrador Retriever was higher than in Beagles during the first 10 months. The values of serum KS showed body weight-related increase within retrievers among teaching hospital population and there was significant increase in body weight-related change. Cartilage metabolism is high in canine immature joint and that activity continues for 5 months, and that higher in Labrador Retrievers rather than in Beagles. There was no effect from other factors, including age, body weight, breed and disease in all patients. Serum KS concentration of Retrievers is higher than Beagles, and that value increased with gain of body weight. We suggest that Retriever have higher cartilage metabolism with growth or ageing.     

Effects of Alendronate on A Disintegrin and Metalloproteinase with Thrombospondin Motifs Expression in the Developing Epiphyseal Cartilage in Rats

A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) have been reported to play a role in the degradation of aggrecan, a major component of cartilage. This study was performed to examine the effects of alendronate on the expression of ADAMTS in developing femoral epiphyseal cartilage. Primary cultured chondrocytes from this cartilage were treated with alendronate in vitro and postnatal day 1 rats were injected subcutaneously with alendronate (1 mg/kg) every second day in vivo . The number of cultured chondrocytes and their aggrecan mRNA levels were unaffected by the alendronate treatment at 10⁻⁶ to 10⁻⁴  m concentrations. The mRNA levels of ADAMTS-1, -2 and -9 in chondrocytes were also unaffected. However, the levels of ADAMTS-5 and -4 were reduced significantly by the same treatment. The thickness of the proliferating chondrocyte layers and the aggrecan mRNA levels in the epiphysis were unaffected by the alendronate treatment in vivo . However, the hypertrophied chondrocyte layers became significantly thicker, and the size of the secondary ossification centre was reduced significantly by the same treatment ( P  < 0.05). Both ADAMTS-4 and -5 mRNA expressions were also reduced significantly in vivo . The immunoreactivity against ADAMTS-4 was seen in hypertrophied chondrocytes and reduced significantly by the alendronate treatment. These results suggested that alendronate can inhibit the degradation of aggrecan in the articular cartilage by downregulating the expression of matrix enzymes such as ADAMTS-4 and -5.     

Effects of Persimmon Peel on Laying Performance,Nitrogen Availability,and Egg Quality in Laying Hens Provided with Shrimp Meal Diets

To determine whether persimmon peel (PP) showing high chitinase activity could alleviate the detrimental dietary effects of chitin-rich shrimp meal (SM), we assessed the laying performance, nitrogen (N) balance, and egg quality of laying hens provided with SM diets containing PP. We also examined the color and antioxidant properties of egg yolk, as we anticipated these would be improved by providing SM and PP. Seventy-two laying hens (45 weeks of age) were allotted to one of the nine dietary treatments (eight hens each), namely three levels of SM (0%, 10%, and 15%)×three levels of PP (0%, 6%, and 8%), and fed with the experimental diets over a period of 6 weeks. Hen-day egg production, feed intake, egg mass, feed conversion ratio, and N balance reduced with increasing levels of SM, whereas the reductions were recovered in a dose-dependent manner in response to increasing levels of PP; however, the SM0% treatment showed that PP exerted little effects. Notably, reductions in the Haugh unit and albumen height of eggs with increasing SM levels, and recovery by provision of increasing levels of dietary PP, were observed. Yolk color was improved by SM, although PP exerted little effect, whereas the antioxidant properties of yolk were enhanced by the inclusion of both SM and PP in diets. Furthermore, eggshell strength, weight, and thickness were enhanced with increasing levels of SM, whereas dietary PP had little effect on these parameters. Thus, we suggest that PP can alleviate the negative effects of dietary SM and improve egg quality, without causing a reduction in laying performance, provided that the level of supplementary PP in diets is less than 8%. These findings accordingly indicate that PP is a promising feed constituent for laying hens fed with SM diets.     

Development of two alternative Loop‐mediated isothermal amplification tools for detecting pathogenic pine wood nematodes

Loop‐mediated isothermal amplification (LAMP) detection tools have great potential for diagnosing the causal agents of infectious diseases in clinics and in agriculture. In this work, we developed two alternative LAMP protocols for detecting the pathogenic nematode Bursaphelenchus xylophilus, causal agent of pine wilt disease. We first identified a pectate lyase 3 gene as a biomarker for developing a LAMP Detection Kit, as there was no homologue in non‐pathogenic nematodes that live in pine timber or bark and show structural similarities to B. xylophilus. The first LAMP protocol used the Genie II equipment and an isothermal master mix containing Geobacillus sp. M 2.0 large fragment DNA polymerase showed approximately 10 times greater sensitivity with a shorter incubation period compared with that of the second LAMP protocol, which utilized a fluorescence metal indicator, calcein and an engineered Bacillus stearothermophilus DNA polymerase I, large fragment (Bst 2.0 DNA polymerase). However, the LAMP reactions with calcein and Bst 2.0 polymerase were the cost‐effective method because the reaction could be performed using a simple isothermal block and relatively inexpensive calcein as a fluorescence indicator visible to the naked eye.     

籼稻品种味度及淀粉RVA谱特性配合力分析

选用8个籼稻品种,采用Griffing双列杂交方法Ⅳ,对稻米味度及淀粉黏滞性谱（RVA谱）特性配合力进行分析。结果表明,(1) 在稻米味度、最高黏度、最低黏度、最终黏度、下降黏度值、黏滞峰消减值、回冷黏滞性恢复值等特性中,亲本基因的加性效应对杂种一代起主导作用,而糊化开始温度的遗传变异主要来自基因的非加性效应;(2)     

Toxicity of spray and fumigant products containing cassia oil to Dermatophagoides farinae and Dermatophagoides pteronyssinus (Acari: Pyroglyphidae)

The toxicity of formulations of oil of cassia, Cinnamomum cassia Blume, (20 and 50 g L(-1) sprays and 100% oil-based fumigant) to adult Dermatophagoides farinae Hughes and D. pteronyssinus Trouessart was examined using contact and vapour-phase toxicity bioassays. Results were compared with the lethal activity of three commercial acaricides: benzyl benzoate, dibutyl phthalate and diethyl-m-toluamide (deet). The contact toxicity of cassia oil to both dust mite species was comparable with that of benzyl benzoate but was higher than that of the other two acaricides. Sprays containing 20 and 50 g L(-1) cassia oil were effective against both mite species when applied to fabric, glass, paper, plastic, tin or wood substrates. Applications of the 50 g L(-1) spray to different space volumes and surface areas determined that 50-60 mg of cassia oil was needed to control dust mites in 3.4 m(3) or in 1 m(2). In tests with fumigant devices, toxicity varied according to the thickness of non-woven fabric covering the device, the exposure time, the number of fumigant devices used and the volume of the space sprayed. Fumigant toxicity to adult D. pteronyssinus was more pronounced with devices enclosed in thinner (40 microm) versus thicker (45 or 50 microm) non-woven fabric covers. A single fumigant device with a 40 microm thick non-woven fabric cover resulted in substantial control in a space of 0.05 m(3) but exhibited only moderate to weak control in spaces >or= 0.097 m(3) at 4 days after application. Two fumigant devices gave 88% mortality in a space of 1.73 m(3). Cassia oil applied as sprays or in fumigant devices appears to provide effective protection of humans from house dust mites.