Isolation, structure characterization, and immunomodulating activity of a hyperbranched polysaccharide from the fruiting bodies of Ganoderma sinense

A polysaccharide (GSP-6B) with a molecular mass of 1.86 × 10? Da was isolated from the fruiting bodies of Ganoderma sinense . Chemical composition analysis, methylation analysis, infrared spectroscopy, and nuclear magnetic resonance spectroscopy were conducted to elucidate its structure. GSP-6B contains a backbone of (1→6)-linked-β-D-glucopyranosyl residues, bearing branches at the O-3 position of every two sugar residues along the backbone. The side chains contain (1→4)-linked-β-D-glucopyranosyl residues, (1→3)-linked-β-D-glucopyranosyl residues, and nonreducing end β-D-glucopyranosyl residues. An in vitro immunomodulating activity assay revealed that GSP-6B could significantly induce the release of IL-1β and TNF-α in human peripheral blood mononuclear cell (PBMC) and showed no toxicity to either PBMC or a human macrophage cell line THP-1. GSP-6B could also activate dendritic cells (DC) by stimulating the secretion of IL-12 and IL-10 from DC.     

Purification and localization of nitric oxide synthases from hybrid tilapia (Nile tilapia x Mozambique tilapia)

The aims of this study were to purify and localize the nitric oxide synthases (NOSs) from hybrid tilapia (Nile tilapia Oreochromis niloticus x Mozambique tilapia O. mossambicus). The purification procedures involved affinity chromatography with a 2', 5'-ADP-agarose 4B column and ion exchange with a diethylaminoethanol Bio-Gel A column. The results from gel filtration assays showed that the molecular weights of neuronal NOS (nNOS) and inducible NOS (iNOS) were 178 and 120 kDa, respectively. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis results showed that there were three bands with molecular weights of 89, 47, and 29 kDa from the purified nNOS. However, only one band, with a molecular weight of 120 kDa, appeared on the gel from the purified iNOS. Hybrid tilapia nNOS was a dimer structure, while iNOS appeared to be a monomer structure. Moreover, our results revealed that the activities of nNOS and iNOS were significantly higher after the addition of Ca+2 or Mg+2 ions individually. However, when L-arginine and NADPH were present, the addition of 1 mM of either ion did not further increase the activity. The chemical L-N(G)-methyl-L-arginine could inhibit the activities of the purified NOSs with or without L-arginine. Western blot analyses showed only an 89-kDa immunoreactive band from the extracts of cerebrum; however, we did not find the specific bands in other tissues, such as gill, intestine, liver, spleen, and anterior kidney. We found another 120-kDa immunoreactive protein band with the rabbit antirat iNOS serum against iNOS from the extracts of anterior kidney and spleen. The results of immunohistochemistry with the rabbit antihuman nNOS serum indicated that the nNOS existed in the cerebellum, olfactory bulb, diencephalons, and nerve cell bodies and neuronal fibers of the spinal cord. Interestingly, only macrophages from anterior kidney and spleen showed positive reactions with the rabbit antirat iNOS serum. In the same way, the endothelial NOS (eNOS) located in the heart and epithelial cells of the blood vessels reacted positively with the rabbit antibovine eNOS serum.     

Reproducible infection model for Clostridium perfringens in broiler chickens

Experiments were carried out to establish an infection and disease model for Clostridium perfringens in broiler chickens. Previous experiments had failed to induce disease and only a transient colonization with challenge strains had been obtained. In the present study, two series of experiments were conducted, each involving four groups of chickens with each group kept in separate isolators. A coccidial vaccine given at 10 times the prescribed dosage was used to promote the development of necrotic enteritis. In the first experiment, cultures of C. perfringens were mixed with the feed at day 9, 10, 11, and 12, and the coccidial vaccine was given at day 10, whereas in the second experiment, C. perfringens cultures were mixed with the feed at day 17, 18, 19, and 20, and the coccidial vaccine was given at day 18. Chickens were examined at day 9, 11, 12, and 15 (Experiment 1), and at day 17, 18, 20, and 24 (Experiment 2). There was no mortality in any of the groups; however, chickens in the groups receiving both coccidial vaccine and C. peifringens developed the subclinical form of necrotic enteritis, demonstrated by focal necroses in the small intestine, whereas chickens in control groups or groups receiving only coccidial vaccine or only C. perfringens cultures developed no necroses. The results underline the importance of predisposing factors in the development of necrotic enteritis.     

Antibacterial carbazole alkaloids from Clausena harmandiana twigs

Three new carbazole alkaloids, harmandianamines A-C (1-3), together with fifteen known compounds (4-18) were isolated from the twigs of Clausena harmandiana. The structures were elucidated by spectroscopic methods, including UV, IR, NMR, and MS. The antibacterial activity against Escherichia coli TISTR 780, Salmonella typhimurium TISTR 292, Staphylococcus aureus TISTR 1466 and methicillin-resistant S. aureus (MRSA) SK1 of some isolated compounds was also evaluated. Compound 6 exhibited significant antibacterial activity against MRSA SK1 with an MIC value of 0.25 μg/mL which higher than that of standard drug, vancomycin (MIC value=1 μg/mL) whereas compounds 14 and 5 showed strong activity with MIC values of 4 and 8 μg/mL, respectively. Only compound 14 showed strong antibacterial activity against S. aureus TISTR 1466 with an MIC value of 4 μg/mL.     

香港土壤研究 Ⅲ.土壤中多环芳烃的含量及其来源初探

通过对香港地区53个土壤样品多环芳烃(PAHs)含量的分析,并利用GIS空间数据管理功能,揭示了香港土壤中PAHs的含量和空间分布特征,同时,运用比值法定性地判断了土壤中PAHs的来源。研究结果表明:香港土壤中可以检测出16种美国环保署规定的优控PAHs中的15种(二苯并(a,h)蒽未被检测到),郊野土壤中PAHs的平均含量为34.2±16.0μg kg-1,而城区土壤中PAHs的平均含量为169±123μg kg-1,港岛动植物公园土壤中的苯并(a)芘的含量最高达到了47.2μg kg-1。在PAHs的来源上,前者的PAHs可能主要来自山火焚烧,而后者可能主要与城区的汽车尾气排放有密切的联系。     

Osteogenic proliferation and differentiation of canine bone marrow and adipose tissue derived mesenchymal stromal cells and the influence of hypoxia

The aim of this study was to compare the osteogenic and proliferative potential of canine mesenchymal stromal cells (cMSCs) derived from bone marrow (BM-cMSCs) and adipose tissue (AT-cMSCs). Proliferation potential was determined under varying oxygen tensions (1%, 5%, and 21% O(2)). Effects of reduced oxygen levels on the osteogenic differentiation of AT-cMSCs were also investigated. AT-cMSCs proliferated at a significantly faster rate than BM-cMSCs, although both cell types showed robust osteogenic differentiation. Culture in 5% and 1% O(2) impaired proliferation in cMSC from both sources and osteogenic differentiation in AT-cMSCs. Our data suggests that AT-cMSCs might be more suitable for use in a clinical situation, where large cell numbers are required for bone repair, due to their rapid proliferation combined with robust osteogenic potential. Our data also suggests that the inhibitory effects of hypoxia on both cell proliferation and differentiation should be considered when using MSCs in a potentially hypoxic environment such as a fracture site.     

Cloning of a novel phytase from an anaerobic rumen bacterium,Mitsuokella jalaludinii,and its expression in Escherichia coli

The full length phytase gene of Mitsuokella jalaludinii was successfully cloned and was found to be 1 047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, p ET32 a, for expression in Escherichia coli strain Rosetta-gami. Expression of the recombinant phytase gene was optimised and characterised. The recombinant phytase was estimated to be approximately 55 k Da by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, p H 4.5 and showed good p H stability from p H 3.5 to 5.5(78% relative activity). Metal ions such as Ca2+, Mg2+, and K+ were found to exert significant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+ greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced p H stability and metal ion effects should be taken into consideration during feed enzyme formulation.     

Neutrophil Gelatinase–Associated Lipocalin in Dogs with Naturally Occurring Renal Diseases

Background

Neutrophil gelatinase–associated lipocalin (NGAL) is released from renal tubular cells after injury and serves in humans as a real‐time indicator of active kidney damage, including acute kidney injury (AKI) and chronic kidney disease (CKD). However, NGAL concentrations in dogs with naturally occurring AKI or CKD rarely have been explored in detail.

Hypothesis/Objectives

The goal of this study was to evaluate whether NGAL can serve as a useful biomarker in dogs with naturally occurring renal disease.

Animals

Client‐owned dogs with renal disease (57) and control dogs without any disease (12) were examined.

Methods

Serum NGAL (sNGAL) and urine NGAL (uNGAL) concentrations were measured in each animal by a newly developed ELISA system. Demographic, hematologic, and serum biochemical data were recorded. Survival attributable to AKI and CKD was evaluated at 30 days and 90 days, respectively.

Results

Serum and urine NGAL concentrations in azotemic dogs were significantly higher than in nonazotemic dogs and were highly correlated with serum creatinine concentration (P < .05). Among CKD dogs, death was associated with significantly higher sNGAL and uNGAL concentrations compared with survivors. Receiver‐operating characteristic curve (ROC) analysis showed that sNGAL was better than serum creatinine concentration when predicting clinical outcomes for CKD dogs (P < .05). The best cutoff point for sNGAL was 50.6 ng/mL, which gave a sensitivity and a specificity of 76.9 and 100%, respectively. Furthermore, dogs that had higher concentrations of sNGAL survived for a significantly shorter time.

Conclusion

sNGAL is a useful prognostic marker when evaluating dogs with CKD.     

Sensitivity and specificity of the indirect-fluorescent antibody test and two enzyme-linked immunosorbent assays in canine dirofilariasis

Ninety dogs naturally infected with Dirofilaria immitis and 103 noninfected dogs, as determined by necropsy, were used to compare the sensitivity and specificity of the cuticular and somatic reactions of the indirect fluorescent antibody test (IFA-C and IFA-S, respectively) and 2 enzyme-linked immunosorbent assays (ELISA). In microfilaremic and amicrofilaremic heartworm-infected dogs, negative results were common for all serotests. In dogs without adult heartworms at necropsy, 32% to 49% were positive, using 1 ELISA, 27% to 29% were positive with the other ELISA, 15% to 36% were positive with the IFA-S, and 0% to 1% were positive using the IFA-C, depending on the classification of borderline reactions. The prevalence of false positive serotests was probably not due to the detection of precardial stages of D immitis in dogs obtained from areas of low endemicity. Until the causes of the false-positive tests are resolved, the use of currently available serotests for routine diagnostic screening or as criteria for instituting treatment is not recommended.