电泳引入CB-HRP在鸡端脑内的有效扩散范围的研究

将CB－HRP经电泳引入火鸡左侧古纹状体，动物分别存活2h、5h、12h、17．5h、24h、48h，灌杀。做脑干和大脑半球的冰冻连续切片，TMB成色，中性红复染，光镜下明、晴视野对照观察。结果发现：1．CB－HRP被引入古纹状体后，其酶液扩散范围于2h最大直径为1．689mm；5h达到2.675mm;8h亦达2.675mm，12h时为2．253mm；17．5h时为1．830mm；24h时为0.845mm;48h时为0.422mm.2．CB－HRP被引入12h时在耳蜗居核、顶盖灰质及三叉神经感觉主核出现小范围的顺行标记给本，及至48h时脑干内出现广泛的颁行标记物。经测算其顺行运输速率可达96mm／d。     

The solubilization of myofibrillar proteins of vertebrate skeletal muscle in water

Myofibrillar proteins of vertebrate skeletal muscles are insoluble in solutions of ionic strength that approximate physiological conditions. We established a method to solubilize more than 80% of chicken breast muscle myofibrillar proteins in water for the use of meat as a source of food protein. SDS‐polyacrylamide gel electrophoretic patterns of water‐soluble myofibrillar proteins demonstrated that all identified myofibrillar proteins except connectin/titin were soluble in water. A part of α‐actinin was released from myofibrils by repeated washing with 2.5 mmol/L NaCl and 5 mmol/L L‐histidine solution, and subsequent destruction of connectin/titin in washed myofibrils by ultrasonication resulted in solubilization of a large fraction of chicken breast muscle myofibrillar proteins in water. Myofibrillar proteins of chicken leg, pork loin, beef shoulder loin, and lamb were also solubilized in water using this procedure.     

Measles virus infection induces interleukin-8 release in human pulmonary epithelial cells

Measles virus (MV) infection primarily targets epithelial cells of the respiratory tract, which have the potential to synthesize a variety of cytokines. In this report, we studied the effect of MV infection on the production of interleukin (IL)-8 by the pulmonary epithelial cells. A549 cells, a lower airway epithelial cell line, produced IL-8 after MV inoculation in a dose- and time-dependent manner. The IL-8 production was little affected by UV-inactivation of MV and scarcely suppressed by cycloheximide treatment. These results indicated that MV particle binding to and/or incorporation into cells stimulated IL-8 expression in A549 cells.     

A novel method for estimating the adaptive ability of guide dogs using salivary sIgA

Salivary secretory immunoglobulin A (sIgA) concentrations of prospective guide dogs for the blind were determined to clarify whether salivary sIgA is useful in evaluating the potential suitability of guide dogs for the blind. Saliva was collected from 73 prospective guide dogs in the kennel on day 1 (the day of separation from volunteer puppy-raisers), 2, 3, 7 and 14 during the estimation period (at about 1 year old). We selected particularly suitable dogs (superior dogs) and unsuitable dogs (inferior dogs) on the basis of the trainers' estimation. All dogs were divided into two groups, those were acceptable dogs would become the guide dogs and rejected dogs could not become guide dogs. The sIgA concentrations in superior dogs gradually increased from day 1 to 14 and those in inferior dogs remained at low levels. Moreover, the sIgA concentrations on day 14 in the acceptable dogs were significantly higher than those in rejected dogs. The cut-off point of sIgA concentrations on day 14 using an ROC curve was 90 EU/ml, and the specificity of the estimation at this point (70.4%) was higher than that of trainers' estimation (50%). Moreover, parallel testing using both trainers' estimation and sIgA estimation, showed that specificity was further improved (79.5%). The present study showed that sIgA concentration was extremely useful in estimating the adaptive ability for guide dogs for the blind.     

Identification and characterization of lactic acid bacteria isolated from mixed pasture of timothy and orchardgrass,and its badly preserved silage

In order to understand the relationship between lactic acid bacteria (LAB) species and silage fermentation, a total of 65 LAB strains isolated from mixed pasture of timothy (Phleum pratense L.) and orchardgrass (Dactylis glomerata L.), and its badly preserved silages were subjected to phenotypic and genetic analysis. According to these analyses, the isolates were divided into 13 groups, including Enterococcus gallinarum, Lactobacillus acidipiscis, L. coryniformis subsp. coryniformis, L. coryniformis subsp. torquens, L. curvatus, L. paraplantarum, L. plantarum subsp. argentoratensis, L. plantarum subsp. plantarum, L. sakei subsp. carnosus, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Leuconostoc pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Weissella hellenica, Weissella paramesenteroides and Carnobacterium divergens. This is the first report to document that C. divergens, L. acidipiscis, L. sakei subsp. carnosus, L. garvieae, phenotypically novel L. lactis subsp. cremoris, E. gallinarum and W. hellenica are present in vegetative forage crops. L. plantarum group strains were most frequently isolated from the badly preserved silages. Some isolates showed a wide range of growth preferences for carbohydrate utilization, optimal growth pH and temperature in vitro, indicating that they have a high growth potential. These results are useful in understanding the diversity of LAB associated with decayed silage of timothy and orchardgrass.     

Paired box 7 inhibits differentiation in 3T3‐L1 preadipocytes

Myogenesis is precisely proceeded by myogenic regulatory factors. Myogenic stem cells are activated, proliferated and fused into a multinuclear myofiber. Pax7, paired box 7, one of the earliest markers during myogenesis. It has been reported that Pax7 regulates the muscle marker genes, Myf5 and MyoD toward differentiation. The possible roles of Pax7 in myogenic cells have been well researched. However, it has not yet been clarified if Pax7 itself is able to induce myogenic fate in nonmyogenic lineage cells. In this study, we performed experiments using stably expressed Pax7 in 3T3‐L1 preadipocytes to elucidate if Pax7 inhibits adipogenesis. We found that Pax7 represses adipogenic markers and prevents differentiation. These cells showed decreased expression of PDGFRα, PPARγ and Fabp4 and inhibited forming lipid droplets.     

Influence of Temperature on Growth and Production of Pectenotoxin-2 by a Monoclonal Culture of Dinophysis caudata

The effects of temperature on growth and production of Lipophilic Toxins (LT) by a monoclonal culture of Dinophysis caudata was studied. The cell density of D. caudata increased significantly with increasing temperature, and was the highest under 27, 30, and 32.5 °C. Temperature affected the average specific growth rate (µ) during the exponential growth phase (EG), which increased from 15 °C to 30 °C, and then decreased at 32.5 °C. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed that this strain of D. caudata produced only pectenotoxin-2 (PTX-2) whose concentration increased significantly with incubation period, except at 32.5 °C. It was significantly different between temperatures ≤18 °C, ≥21 °C, and 32.5 °C. The cellular toxin production (CTP, pg·cell⁻¹·day⁻¹) showed variation with growth phase and temperature, except at 32.5 °C. The average net toxin production (R_tox) was not affected by temperature. During EG, the average specific toxin production rate (µ_tox) increased significantly with increase in temperature, reaching a peak of 0.66 ± 0.01 day⁻¹ at 30 °C, and then decreased. Over the entire growth span, µ_tox was significantly correlated to µ, and this correlation was most significant at 27 and 30 °C. During EG, µ_tox was affected by both temperature and growth. This study shows that temperature affects growth and toxin production of this strain of D. caudata during EG. In addition, a positive correlation was found between toxin production and growth.     

Distribution and stem growth patterns of mangrove species along the Nakara River in Iriomote Island, Southwestern Japan

We examined variations in stand structure and tree growth of a mangrove forest along two gradients: from the river mouth to upper stream and from the riverside to inland, along the Nakara River, Iriomote Island, Japan. Bruguiera gymnorrhiza (L.) Lamk. occurred throughout the intertidal area, though Rhizophora stylosa Griff. and Kandelia candel (L) Druce did not occur upstream. Basal area and maximum tree height of B. gymnorrhiza decreased downstream. The growth rate in stem diameter of B. gymnorrhiza decreased downstream in relation with the soil salinity. The basal area and the maximum tree height of B. gymnorrhiza increased with the distance from the riverside. R. stylosa increased in stem diameter and growth rate toward the riverside.     

Rice phenolics efflux transporter 2 (PEZ2) plays an important role in solubilizing apoplasmic iron

Iron (Fe) is an essential micronutrient for plants; however, despite its abundance in soils it is not readily available in neutral to alkaline soils. Plants secrete phenolics, such as protocatechuic acid (PCA) and caffeic acid (CA), to absorb and utilize precipitated apoplasmic Fe from root surfaces. However, the synthesis and secretion of phenolics have not been well characterized in plants. We have identified and characterized a rice (Oryza sativa L.) mutant with reduced amounts of PCA and CA in xylem sap and root exudates; hence we named it phenolics efflux zero 2 (pez2). PEZ2 localized to the plasma membrane in onion (Allium cepa L.) epidermal cells and transported PCA when expressed in Xenopus laevis oocytes. PEZ2 expression was observed in whole root near root tips. Similarly, strong expression was observed in leaves. In line with reduced amounts of PCA and CA in xylem sap, the xylem Fe concentration was also low in pez2 plants. These results suggest that PEZ2 is involved in solubilization of apoplasmic Fe in rice.     

Nonulcerative keratouveitis as a manifestation of Leptospiral infection in a horse

A 2-year-old Thoroughbred filly presented with ocular pain and epiphora of the left eye. The pupil was miotic and the cornea edematous near the ventro-temporal limbus, but did not retain any fluorescein. The topical antibiotics and atropine and diclofenac, and systemic flunixin meglumine and antibiotic therapy did not resolve the condition. A pink and fleshy infiltrate developed near the limbus indicating nonulcerative keratouveitis. The anterior uveitis deteriorated as manifested by the presence of dyscoria, hypopyon, and organized fibrin in the anterior chamber. Ocular signs were improved by topical and subconjunctival corticosteroids, but repeatedly deteriorated as the frequency of medication was reduced. The horse was seropositive to three serovars of Leptospira interrogans. The animal was diagnosed as blind on day 91 by the absence of pupillary light and menace reflexes, and donated for histopathologic diagnosis. The corneal opacity was histologically fibrotic and infiltrated predominantly by lymphocytes with Descemet's membrane partially disrupted by macrophages. The choroid was infiltrated by lymphocytes, eosinophils and basophils, and was positive to IgG and C3. There were filamentous or spiral structures positive to Warthin-Starry stain in the renal cortex. There was also polymerase chain reaction amplification of the leptospiral gene in the kidney. From these findings nonulcerative keratouveitis was believed to be caused by systemic infection with Leptospira.