Prevention of biofilm formation with a coating of 2-methacryloyloxyethyl phosphorylcholine polymer

Device-associated infections are serious complications, and their prevention is an issue of considerable importance. Since biofilms are responsible for these refractory infections, effective methods to inhibit biofilm formation are required. In this investigation, stainless steel plates with and without 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, i.e., poly (MPC-co-n-butyl methacrylate) (PMB) coating, were incubated in a medium containing bacteria. In the course of incubation, half of the specimens received antibiotics. The specimens were stained for nucleic acid and polysaccharides, and then examined with a confocal laser scanning microscope. The numbers of bacteria on the specimen surfaces were evaluated by an ATP assay. On the surfaces of the specimens without PMB coating, the formation of a biofilm enveloping bacteria was confirmed. The addition of antibiotics did not effectively decrease the number of bacteria. On the other hand, on the surfaces of the specimens with PMB coating, no biofilm formation was observed, and the number of bacteria was significantly decreased. The addition of potent antibiotics further decreased the number of bacteria by 1/100 to 1/1000 times. The PMB coating combined with the validated use of antibiotics might provide a method for the simultaneous achievement of biocompatible surfaces of devices and the prevention of device-associated infections.     

Cell wall modification during development of mealy texture in the stony-hard peach “Odoroki” treated with propylene

The melting peach cultivar “Akatsuki” [Prunus persica L. Batsh] develops melting texture during ripening, but the stony-hard peach cultivar “Odoroki”, softens and develops a mealy texture when treated with propylene. Cell wall polysaccharides were fractionated as 1,2-diaminocyclohexanetetraacetic acid (CDTA)-, Na₂CO₃-, NaOH-soluble fractions, or the cellulosic residues that remained after solvent extraction. Size-exclusion chromatography of the CDTA-soluble fraction revealed a limited depolymerization of polyuronides in mealy “Odoroki” fruit, accompanying a lower level of endo-polygalacturonase (endo-PG) and a higher level of exo-PG activities compared to melting “Akatsuki” fruit. “Akatsuki” fruit showed increases in the arabinose, galactose and rhamnose contents in the CDTA-soluble fraction. These neutral sugars decreased in the Na₂CO₃-soluble and cellulosic residue fractions during ripening. Similar changes were observed for neutral sugars other than arabinose during softening in mealy “Odoroki” fruit. Arabinose content did not increase in the CDTA-soluble fraction of “Odoroki” fruit; however, arabinose content decreased in the Na₂CO₃-soluble fraction and in the cellulosic residue as observed for “Akatsuki” fruit. A significant increase in α-l-arabinofuranosidase activity, about 10 times higher than for “Akatsuki” fruit, was observed in propylene-treated mealy “Odoroki” fruit. Differences in ripening-associated metabolic changes in the cell walls of these peach cultivars seem to be related to their differences in texture.     

齐氏田中鳑鲏的早期发育研究

通过对齐氏田中鳑鲏(Tanakia chii)人工授精和培育(22℃恒温环境)阶段早期发育的观察,描述了其仔稚鱼发育形态特征。初孵仔鱼卵黄囊饱满,呈淡黄色,表皮布满微突;3日龄仔鱼卵黄囊两侧向上突起,尾部鳍褶延展变大;6日龄仔鱼眼囊发育;8日龄仔鱼眼囊有黑色素,晶状体形成;11日龄仔鱼口形成,脑开始发育,体上有黑色素出现;15日龄仔鱼鳃沟和肛门形成,背鳍和臀鳍的鳍基出现,尾鳍下叶鳍条形成;21日龄仔鱼头与躯干分界明显,鳔分化为前鳔和后鳔,背部和臀部鳍条数目分别为8和10枚,身体黑色素连接成条带状,虽然残存有卵黄囊,但已经开口并摄食。31-36日龄仔鱼胸鳍呈膜状鳍褶,腹鳍出现鳍条,尾鳍呈深叉形。61-66日龄稚鱼,鳞片出现,体中线后半部具一条黑色素带,稚鱼出现集群游动现象。仔鱼体长的生长呈阶段性:1-26日龄仔鱼生长较为平缓;26-81日龄生长较快。综合21日龄仔鱼卵黄囊残留很少并开口摄食的特征,我们推测在孵化后21日为自然条件下齐氏田中鳑鲏的出蚌时间。同时,仔鱼的感觉、摄食、呼吸和游泳器官在早期阶段快速发育。这些器官在仔鱼的早期快速发育,使出蚌后的仔鱼在最短的时间内获得与早期生存相关的各种能力,提高了出蚌后仔鱼的生存能力。     

Characterization of monoclonal antibodies against fowl adenovirus serotype 1 (FAV1) isolated from gizzard erosion

Six clones of monoclonal antibodies (Mabs) to fowl adenovirus (FAV) serotype 1 were produced. All Mabs reacted positively by enzyme-linked immunosorbent assay. Three Mabs recognized the putative 100-kD hexon protein and reacted to serotype 1 specifically by western blot analysis but did not react to other FAV serotypes (2, 3, 4, 5, 6, 7, and 8a). These Mabs will be useful for immunodiagnosis of FAV serotype 1 infection in chickens with gizzard erosion and in further research studies involving the genomes and proteins of FAV serotype 1.     

Experimental inoculation of beagle dogs with Ehrlichia species detected from Ixodes ovatus

Three beagle dogs were inoculated with mice spleen/liver homogenate infected with Ehrlichia species detected from Ixodes ovatus (EIO) and one dog was used as a control. All three infected dogs did not show clinical signs of disease except for mild pyrexia throughout the 41-day study period. Splenomegaly was observed from Day 7 post-inoculation (p.i.) in two of the dogs. Hematological and biochemical abnormalities included mild thrombocytopenia, hypoproteinaemia, hypoalbuminaemia and increased C-reactive protein values. One of the dogs' splenic aspirate sample was PCR-positive for Ehrlichia Day 7 p.i. and another dogs' blood and bone marrow aspirate sample was PCR-positive Day 41 p.i. Sequence analysis of the PCR products showed 100% homology with the 16SrRNA partial gene sequence of Ehrlichia sp. HF565. Antibody titers to EIO were observed in all three experimentally infected dogs starting from the first week p.i. and cross-reactivity with Ehrlichia canis was detectable in one of the dogs starting Day 7 p.i. These data suggest that infection of dogs with EIO is possible, though is probably of low pathogenic importance. Cross-reactivity of EIO infected dog serum with E. canis raises the likelihood of false E. canis seropositive dogs.     

Expression of canine Kdap in normal, hyperplastic and neoplastic epidermis

Keratinocyte differentiation-associated protein, Kdap, is a recently identified small secretory protein that may act as a soluble regulator for the cornification and/or desquamation of keratinocytes. To clarify the role of Kdap in the terminal differentiation of keratinocytes, detailed in situ localisation of Kdap was studied using canine skin with normal, hyperplastic and neoplastic epidermis. In normal canine trunk skin, Kdap was expressed by granular keratinocytes, with polarity to the apical side of the cells, suggesting that canine Kdap is present in lamellar granules, as in humans. Expression of Kdap was widespread in the spinous layers in hyperplastic epidermis, but was undetectable in squamous cell carcinomas. These findings suggest that Kdap is closely related to the delay of terminal differentiation and/or release of cells in hyperplastic epidermis.     

Lung toxicity of a vapor-grown carbon fiber in comparison with a multi-walled carbon nanotube in F344 rats

Carbon fibers have excellent physicochemical and electrical properties. Vapor-grown carbon fibers are a type of carbon fibers that have a multi-walled carbon tube structure with a high aspect ratio. The representative vapor-grown carbon fiber, VGCF^TM-H, is extremely strong and stable and has superior thermal and electrical conductivity. Because some high-aspect-ratio multi-walled carbon nanotubes (MWCNTs) have been reported to have toxic and carcinogenic effects in the lungs of rodents, we performed a 13-week lung toxicity study using VGCF^TM-H in comparison with one of MWCNTs, MWNT-7, in rats. Male and female F344 rats were intratracheally administered VGCF^TM-H at doses of 0.2, 0.4, and 0.8 mg/kg bw or MWNT-7 at doses of 0.4 and 0.8 mg/kg bw once a week for 8 weeks and then up to week 13 without treatment. The lung burden was equivalent in the VGCF^TM-H and MWNT-7 groups; however, the lung weight had increased and the inflammatory and biochemical parameters in the broncho-alveolar lavage fluid and histopathological parameters, including inflammatory cell infiltration, alveolar type II cells proliferation, alveolar fibrosis, pleural fibrosis, lung mesothelium proliferation, and diaphragm fibrosis, were milder in the VGCF^TM-H group than in the MWNT-7 group. In addition, the proliferating cell nuclear antigen (PCNA)-positive index in the visceral and pleural mesothelium was significantly higher in the MWNT-7 group than in the controls, but not in the VGCF^TM-H group. Thus, the results of this study indicate that the lung and pleural toxicities of VGCF^TM-H were less than those of MWNT-7.     

Involvement of neurokinin receptors in the control of pulsatile luteinizing hormone secretion in rats

It has recently been shown that neurokinin B, a tachykinin, is associated with GnRH pulse generation in sheep and goats. The aim of the present study was to clarify the role of tachykinin receptors in the control of LH secretion in rats. To this end, we evaluated the effect of CS-003, an antagonist for all three neurokinin receptors (NK1, NK2 and NK3 receptors), on pulsatile LH secretion in both sexes of rats with different routes of administration. Both oral and third ventricular administration of CS-003 suppressed LH secretion in both sexes of gonadectomized animals. Furthermore, intact male rats with oral administration of CS-003 showed decreased serum testosterone levels, which might be due to suppressed LH secretion. None of the three subtype-specific neurokinin receptor antagonists showed a significant effect on LH secretion in ovariectomized rats when each antagonist was singly administered. The present results suggest that neurokinins play a role in the control of pulsatile GnRH/LH secretion via multiple neurokinin receptors in both male and female rats.     

Reconstruction of stratum corneum in organotypically cultured canine keratinocyte-derived CPEK cells

The stratum corneum of epidermis is an essential barrier against the external environment and water loss. This study aimed to develop an organotypic culture model that targets the reconstruction of the stratum corneum using canine keratinocyte-derived CPEK cells. The CPEK cells cultured at the air-liquid interface became stratified and formed a stratum corneum-like layer on stratum spinosum- and stratum granulosum-like layers. The CPEK cells in the stratum granulosum-like layer expressed the cornified cell envelope (CCE)-related proteins loricrin and keratinocyte differentiation-associated protein. Organotypically cultured CPEK cells were considered to form a CCE at the stratum granulosum-like layer, allowing the formation of a stratum corneum-like layer. The organotypic culture of CPEK cells could be useful for studying the barrier function of canine stratum corneum.