Immunohistochemical and ultrastructural analyses of cytoplasmic blood plasma inclusions of rat hepatocytes

In the present study, we investigated the histological, immunohistochemical and ultrastructural characteristics of cytoplasmic blood plasma inclusions that spontaneously occurred in a rat liver. Histologically, a number of cytoplasmic inclusions were observed in the liver of an 8-week-old female SD rat. These inclusions were strongly positive for PAS staining and resistant to diastase digestion. Immunohistochemical analyses revealed that these inclusions were positive for albumin and IgG; however, most of them were negative for LAMP-1 and LAMP-2. Ultrastructurally, the inclusions were surrounded by limiting membranes and composed of moderately electron dense, homogenous materials. These characteristics described here represent valuable information for pathological examination in toxicity studies.     

Suitability of Japanese codling as a raw material for surimi-based products revealed by primary sequence analysis of myosin heavy chain and thermal gel properties

Fisheries Science - Complementary DNA encoding a part of myosin heavy chain was cloned from fast skeletal muscle of Japanese codling Physiculus japonicus. Japanese codling, a frequent catch off the...     

Occurrence of glucosamine in higher plants

Occurrence of the unknown basic ninhydrin positive substance in young plant tissue supplied with a high level of ammonium nitrogen has been reported in the previous paper (1). This substance disappeared when the supply of ammonium nitrogen was stopped or the source of nitrogen was changed from ammonium nitrogen to nitrace nitrogen, and this substance was verg scarcely detected when the level of ammonium nitrogen in culture solution was as low as 3 ppm or the foren of nitrogen source was nitrace nitrogen. It is very interesting to identify this substance, as the occurrence of this substance is a distinctive character of the plant which han been supplied with a higher level of ammonium nitrogen and has developed the symptoms of ammonia toxicity.     

The survival of multidrug‐resistant bacteria in thermophilic and mesophilic anaerobic co‐digestion of dairy manure and waste milk

Anaerobic digestion is considered as a promising method to manage animal waste with antibiotic‐resistant bacteria. Current research was conducted to investigate the survival of multidrug‐resistant bacteria (MDRB) resistant to three groups of antibiotics: (i) cefazolin, neomycin, vancomycin, kanamycin (group 1); (ii) penicillin, oxytetracycline, ampicillin, streptomycin (group 2); and (iii) cefazolin, neomycin, vancomycin, kanamycin, penicillin, oxytetracycline, ampicillin, streptomycin (group 3), in anaerobic digestion of dairy manure and co‐digestion of dairy manure and waste milk at 37°C and 55°C for 22 days, respectively. The population densities of three groups of MDRB on peptone, tryptone, yeast and glucose agar plates incubated at 30°C for 7 days before and after digestion showed 100% destruction in both digestates at thermophilic temperature. Overall reduction of more than 90% of three groups of MDRB was observed in mesophilic digestion with no significant differences (P > 0.05) between manure and milk mixture. Co‐digestion of dairy manure and waste milk always produced significantly (P < 0.05) higher total gas and methane gas than digestion of manure alone at both temperatures. Gas production in each case was significantly (P < 0.05) higher in thermophilic digestion than in mesophilic digestion. The results demonstrate that thermophilic co‐digestion of dairy manure and waste milk offers more benefits in terms of the environment and economy.     

Preliminary study of histological comparison on the growth patterns of long-bone cortex in young calf, pig, and sheep

Some young large farm animals show a laminar bone formation in the long-bone cortex. Such a laminar bone is gradually replaced by Haversian bone with osteons during their growth periods. In this preliminary study, we observed the transverse ground samples of tibia cortex in young calves, pigs, and sheep by backscattered electron imaging. The cortex bones of all the newborn (NB) animals were basically formed with laminar bone structures. The NB and 1-month-old (1-M) calves had a typical concentric structure of laminar bone, whereas the NB and 1-M pigs showed a wire-netting bone with laminar-bone units. The NB sheep was similar to the calf rather than the pig. In the growth rate of bone volume, sheep was similar to calf up to 6 months after birth (6-M). Such calf and sheep showed a more rapid ratio of bone volume than pig. A few osteons had initially appeared in the innermost layer of the 6-M calf. A 1-year-old (1-Y) calf showed scattered osteons in the bone cortex, but many laminar-bone units were still retained in the outer layer. A 6-M pig had many osteons in the entire cortex but only a few osteons in the outermost layer. In the 6-M sheep, no osteons were observed, whereas a 1-Y sheep showed a relatively small number of osteons mainly in the middle layer but a higher osteon-volume than the 1-Y calf. In the 1-Y sheep, the more widely absorbed areas by bone-remodeling with osteons were observed as compared with the 1-Y calf, and the bone volume was decreased from the 6-M into the 1-Y sheep because of the remarkable bone-absorption. Thus, calf kept on possessing many laminar-bone units for a longer time in the growth period than sheep, while pig showed the earliest bone-remodeling with osteons. These results may be caused by their different body size and withers height in calf and sheep after growing and the difference of the dependence upon mother's body during juvenile period between pig and calf with sheep. The initial region of osteon formation may be distinguishable among their animals, respectively. However, further detailed investigations of their young animals at successive stages will be necessary.     

Anaerobic co-digestion of dairy manure and Japanese knotweed (Fallopia japonica) under thermophilic condition: Optimal ratio for biochemical methane production

Anaerobic co-digestion of animal manure and lignocellulosic biomass is a potent approach for sustainable biomethane production. Co-digestion of dairy manure (DM) and Japanese knotweed (JK), which was collected from a riverbank, was investigated at five different DM-to-JK mixing ratios (100:0, 90:10, 80:20, 60:40, and 0:100; wet weight basis) under thermophilic condition. The results showed that the methane yields obtain from the co-digestion of DM and JK were much higher than that obtained from JK alone (104 ml/gVS), which indicates the synergistic effect and the benefits of co-digesting JK with DM. The highest methane yield (232 ml/gVS) was obtained from the DM-to-JK ratio of 90:10, which was 14.9% and 123.1% higher than that from DM and JK alone, respectively. It also showed the highest synergistic effect (61 ml/gVS). However, further increase in JK ratios led to the decrease in methane yield and synergistic effect. Therefore, applying the co-digestion of DM and JK at a ratio of 90:10 is recommended for biomethane production.     

Atomic-level observation of disclination dipoles in mechanically milled, nanocrystalline Fe

Plastic deformation of materials occurs by the motion of defects known as dislocations and disclinations. High-resolution transmission electron microscopy was used to directly reveal the individual dislocations that constitute partial disclination dipoles in nanocrystalline, body-centered cubic iron that had undergone severe plastic deformation by mechanical milling. The mechanisms by which the formation and migration of such partial disclination dipoles during deformation allow crystalline solids to fragment and rotate at the nanometer level are described. Such rearrangements are important basic phenomena that occur during material deformation, and hence, they may be critical in the formation of nanocrystalline metals by mechanical milling and other deformation processes.     

Distinctive expression of a zinc-binding protein in rice callus grown in medium with high zinc concentration

We investigated the growth, contents of water-soluble protein and free amino acids of the callus of japonica rice (Oryza sativa L. cv. Nipponbare) cultured in liquid N6 medium containing a high concentration of zinc. Furthermore, we determined the N-terminal amino acid sequence of a Zn-binding protein expressed in large quantities in the callus. The addition of Zn stimulated the growth of the callus and increased the Zn concentration. The callus subjected to the high-Zn treatment (hereafter referred to as CHZn) contained a larger amount of soluble proteins and a smaller amount of free amino acids than the control callus. Zinc-binding proteins were separated by affinity chromatography. The SDS-PAGE pattern of these proteins showed a distinctive protein band of about 29 kDa. Especially, CHZn contained larger quantities of 29 kDa protein than the control callus. Twenty-seven N-terminal amino acids of the protein were sequenced as DYAPMTLTIVNNCPYPVWPGIQANSGH. Results of homology search to the amino acid sequences from the nr-aa database and the dbEST database suggested that this 29 kDa protein may be a novel zinc-binding protein and that the protein may regulate the concentration of free zinc in the cytoplasm of callus cells through its binding to zinc ions.     

Development of a canine trypsin-like immunoreactivity assay system using monoclonal antibodies

The radioimmunoassay (RIA) for trypsin-like immunoreactivity (TLI) is one of the most sensitive and specific tests for detecting exocrine pancreatic insufficiency (EPI). An abnormally low serum TLI concentration (<2.5 ng/ml) indicates end-stage EPI. Although RIA methods can be used to detect canine serum TLI, these procedures are beyond the capabilities of most veterinary clinics and general laboratories. Using monoclonal antibodies (mAbs), we developed an enzyme-linked immunosorbent assay (ELISA) for canine TLI and incorporated it into an immunochromatographic test (ICT) for the diagnosis of EPI. The ELISA was linear over TLI concentrations of 1-100 ng/ml. Levels of intra-assay coefficients of variance (CVs) were 1.8-6.1%, inter-assay CVs were 5.1-9.8%, and the recovery of TLI added to two samples of canine serum ranged from 89 to 111 and 93 to 108%, respectively. Good correlation (correlation coefficient, 0.974) occurred between the TLI values obtained by the ELISA method and those by RIA from 56 clinical samples. Serum TLI values in clinically healthy dogs ranged from 7.8 to 29.2 ng/ml by ELISA, and those from dogs with EPI were 0.0-0.6 ng/ml. The values were 0.0-287.4 ng/ml for dogs with pancreatitis, and those from dogs with gastrointestinal disease were 5.5-58.9 ng/ml. The only statistically significant difference (P<0.01) occurred between the TLI level of healthy dogs and those with EPI. The ICT kit showed high reproducibility, and the TLI values yielding negative results differed significantly (P<0.01) from those returning positive results. The ICT kit yielded negative results (indicating EPI) from clinical serum samples with TLI concentrations of 0.0-4.1 ng/ml by ELISA. Both the ELISA and ICT kit are useful tools in the diagnosis of canine EPI.     

One-step immunochromatography assay for detection of high-level canine serum trypsin-like immunoreactivity

We developed a one-step immunochromatography assay kit to measure high levels of canine trypsin-like immunoreactivity (cTLI) for bedside estimation of canine pancreatitis. The serum cTLI level can be determined within 10 min by visual comparison of color strengths in the test and reference zones. The serum cTLI levels determined by this method correlate well with canine TLI-ELISA and can be classified into 3 categories: cTLI levels higher than 60 ng/ml were considered positive; 20-60 ng/ml, weakly positive; and less than 20 ng/ml, negative. Twelve dogs suspected of pancreatitis were examined using this method; 4 dogs were positive, 2 were weakly positive, and 6 were negative. This test can detect a high level of serum cTLI and a positive result in the TLIH test will provide critical information for evaluation of pancreatitis in dogs.