Effect of intramammary injection of rboGM-CSF on milk levels of chemiluminescence activity, somatic cell count, and Staphylococcus aureus count in Holstein cows with S. aureus subclinical mastitis

The effect of intramammary injection of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF, 400 microg/10 mL) on quarter milk levels of chemiluminescence (CL) activity, and somatic cell count (SCC) and shedding pattern of Staphylococcus aureus was investigated. Ten Holstein cows, naturally infected with S. aureus were used, with either early-stage or late-stage subclinical mastitis. Injection of rboGM-CSF caused a remarkable increase in milk CL activity with a peak at 6 h after the cytokine injection in the early- and late-stage groups. In the early-stage group, milk SCC stayed around preinjection level at 6 h, rose significantly on days 1 and 2, and was followed by a smooth and significant decline to an under preinjection level (below 200 000 cells/mL) on day 7 postinjection. Alternatively, in the late-stage group, milk SCC rose significantly at 6 h after the cytokine injection and maintained high levels thereafter. The milk S. aureus count decreased drastically by the cytokine injection in the early-stage group. The bacterial count was moderately decreased in the late-stage group, but increased back to preinoculation levels on day 7 after the cytokine injection. The results suggest that the rboGM-CSF has a potential as a therapeutic agent for S. aureus infection causing subclinical mastitis of dairy cows, if the cytokine is applied at the initial stage of infection.     

Time-course comparison of pulmonary inflammation induced by intratracheal instillation of four different nickel oxide nanoparticles in male Fischer rats

Occupational exposure to nickel oxide (NiO) is an important cause of respiratory tract cancer. Toxicity is known to be associated with the dissociated component, i.e. nickel (II) ions. To address the relationship between physicochemical properties, including solubility in artificial lysosomal fluid, of NiO and time-course changes in the pulmonary response, we conducted an intratracheal instillation study in male Fischer rats using four different well-characterized NiO products, US3352 (NiO A), NovaWireNi01 (NiO B), I small particle (NiO C), and 637130 (NiO D). The NiOs were suspended in purified water and instilled once intratracheally into male F344 rats (12 weeks old) at 0 (vehicle control), 0.67, 2, and 6 mg/kg body weight. The animals were euthanized on days 3, 28, or 91 after instillation, and blood analysis, bronchoalveolar lavage fluid (BALF) testing, and histopathological examination were performed. The most soluble product, NiO B, caused the most severe systemic toxicity, leading to a high mortality rate, but the response was transient and surviving animals recovered. The second-most-soluble material, NiO D, and the third, NiO A, caused evident pulmonary inflammation, and the responses persisted for at least 91 days with collagen proliferation. In contrast, NiO C induced barely detectable inflammation in the BALF examination, and no marked changes were noted on histopathology. These results indicate that the early phase toxic potential of NiO products, but not the persistence of pulmonary inflammation, is associated with their solubility.     

Comparison of dendritic cell-mediated immune responses among canine malignant cells

Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials.     

Artificially induced tetraploid masu salmon have the ability to form primordial germ cells

Diploid gametes generated with tetraploid animals are a stepping stone to improving chromosome manipulation techniques. However, artificially induced tetraploid individuals generally die soon after hatching. Diploid gametes could be induced by in vivo cultures of tetraploid primordial germ cells (PGCs) through germ-line chimera. In the present study, characteristics of PGCs were studied in inviable tetraploid masu salmon, Oncorhynchus masou. Histological observation of tetraploid embryos revealed that the same or smaller numbers of PGCs were observed and they migrate into the genital ridges as did diploid PGCs during gonadogenesis. By whole-mount in situ hybridization using vasa messenger RNA (mRNA), 4–35 vasa-positive signals were detected in a pair of genital ridges of tetraploids. By cytological observation of genital ridge cell suspensions, several large round cells were observed, some of which extended pseudopodia. They also contained large nuclei and round granules in their cytoplasm, characteristics of PGCs. As the results suggest that inviable artificial tetraploids have PGCs, we expect to achieve diploid gamete production through surrogate propagation and tetraploid fish production.     

Behavior of phenolic substances in the decaying process of plants

The behavior of phenolic substances in the decaying process of rice straw, ladino clover, and fanen leaves of red oak under moist conditions, and also of rice straw under various conditions were compared in the laboratory. The amounts of phenolic substances, divided into either humic acid and fulvic acid fractions, or ether-extractable, butanol-extractable and organic solvent-unextractable fractions, and the amounts of individual phenolic acids were periodically determined during incubation for 150 days. The following results were obtained.

1) The amounts and behavior of phenolic substances in various fractions differed considerably among the plant materials. The total amount of phenolic substances was remarkably larger in red oak leaves than in the others during the whole period of incubation. The amounts of phenolics in the fulvic acid fraction changed to a larger extent than those in the humic acid fraction during the decaying process of plant materials. The changes in total amount of phenolics in decaying red oak leaves and ladino clover were mainly due to changes in the level of relatively hydrophilic phenolics in the fulvic acid fraction, but the changes in decaying rice straw were mainly due to changes in the level of relatively lipophilic phenolics.

2) Rice straw and ladino clover, especially the former, contained large amounts of p-coumaric and ferulic acids, but these decreased rapidly in the early stage of the decaying process. The amounts in red oak leaves were small, but did not decrease markedly during incubation.

3) The changes in amounts of phenolics in both humic acid and fulvic acid fractions in the decaying process of rice straw were largely influenced by temperature, moisture, and pH, but not to a large extent by C/N ratio and the presence of soil. At higher temperatures under moist conditions, phenolic substances disappeared rapidly. Also, acidification of the system inhibited the degradation process.     

Study of the Structure–Activity Relationship of an Anti-Dormant Mycobacterial Substance 3-(Phenethylamino)Demethyl(oxy)aaptamine to Create a Probe Molecule for Detecting Its Target Protein

The current tuberculosis treatment regimen is long and complex, and its failure leads to relapse and emergence of drug resistance. One of the major reasons underlying the extended chemotherapeutic regimen is the ability of Mycobacterium tuberculosis to attain a dormant state. Therefore, the identification of new lead compounds with chemical structures different from those of conventional anti-tuberculosis drugs is essential. The compound 3-(phenethylamino)demethyl(oxy)aaptamine (PDOA, 1), isolated from marine sponge of Aaptos sp., is known as an anti-dormant mycobacterial substance, and has been reported to be effective against the drug resistant strains of M. tuberculosis. However, its target protein still remains unclear. This study aims to clarify the structure–activity relationship of 1 using 15 synthetic analogues, in order to prepare a probe molecule for detecting the target protein of 1. We succeeded in creating the compound 15 with a photoaffinity group that retained antimicrobial activity, which proved to be a suitable probe molecule for identifying the target protein of 1.     

我国三种野生泥鳅遗传多样性的微卫星分析

本研究采用5个微卫星分子标记技术对我国黑龙江泥鳅(M.mohoity)、北方泥鳅(M.bipartitus)、泥鳅(M.anguillicaudatus)进行了遗传多样性分析。结果表明,三种泥鳅的5个微卫星基因座共检测到等位基因数27个,微卫星基因座的等位基因数范围为3~7个。5个微卫星基因座在三种泥鳅的多态信息含量都达到了高度多态的水平,平均PIC值在0.526~0.722之间,说明三种泥鳅具有丰富的遗传多样性。黑龙江泥鳅、北方泥鳅和泥鳅的观测杂合度Ho分别为0.978,0.889和0.742,期望杂合度He分别为0.683、0.781、0.721。三种泥鳅的遗传距离为0.189~0.559,泥鳅和北方泥鳅群体间遗传距离最小为0.189,亲缘关系较近,而泥鳅和黑龙江泥鳅群体间遗传距离最大为0.559,亲缘关系较远。Hardy-Weinberg平衡检验P值。得出所有个体在5个微卫星位点上都无偏离现象。聚类结果显示黑龙江泥鳅、北方泥鳅、泥鳅群体分为二支,泥鳅和北方泥鳅聚为一类,之后两者再与黑龙江泥鳅聚为一类。     

Oral administration of β-alanine modifies carnosine concentrations in the muscles and brains of chickens

Carnosine, and its derivative anserine, are known to function as anti‐oxidants and putative neurotransmitters. They are especially rich in the breast muscle (Musculus pectoralis superficialis, MPS) of chickens. To clarify whether the concentrations of carnosine and anserine are altered by dietary management, the effect of oral administration of their constituent, β‐alanine (β‐Ala), was determined in the MPS and brains of chickens. Birds were orally administered β‐Ala (22 mmol/kg) twice a day for five consecutive days (from 2 to 6 days old). In the MPS, carnosine was increased by β‐Ala, whereas anserine and taurine were decreased. The concentrations of other free amino acids in the MPS were also modified by β‐Ala. In the brain, the oral administration of β‐Ala increased anserine and carnosine and decreased taurine, but caused no changes to other free amino acid concentrations. These results suggest that orally administered β‐Ala increases carnosine concentrations in both the MPS and brains of chickens. However, the effects of β‐Ala on other concentrations differ depending on the tissues.     

Changes in hyaluronidase, acrosin, and N-acetylhexosaminidase activities of dog sperm after incubation

Hyaluronidase, acrosin and N-acetylhexosaminidase activities were examined in sperm collected from 12 beagle dogs and in culture medium after 0.5 hr and 7 hr of sperm incubation. The activities of the three enzymes were significantly higher at 7 hr than at 0.5 hr (P < 0.05, 0.01), and the increases were associated with sperm capacitation. It was considered that the three enzymes in the dog sperm are related to fertilization by reason of the findings of the release of these enzymes from the sperm into the medium after 7 hr of incubation.