Plasma high-mobility group box 1 (HMGB1) in dogs with various diseases: comparison with C-reactive protein

High-mobility group box 1 (HMGB1), a nonhistone chromosomal protein, has recently been suggested as a late mediator of the inflammatory cascade. Blood HMGB1 levels are increased in a number of human diseases, and HMGB1 has been suggested to be a useful marker for disease severity and prognosis. The objective of this study was to assess the clinical usefulness of HMGB1 in dogs. Plasma HMGB1 levels, as well as C-reactive protein (CRP), a typical canine inflammatory marker, were measured in dogs with various diseases, especially systemic inflammatory response syndrome (SIRS), and dogs that had undergone surgery. HMGB1 gradually increased and attained a maximum level 72 hr after surgery, whereas CRP increased rapidly, peaking at 24 hr. Although both HMGB1 and CRP levels were significantly increased in dogs with various diseases compared with the control dogs, no correlation was found between the HMGB1 and CRP values. HMGB1 levels in the SIRS group were significantly elevated compared with those in the non-SIRS group. However, the increase in HMGB1 levels above the reference range was not indicative of SIRS. Instead, the presence of increased HMGB1 and CRP levels above the reference ranges significantly affects the poor outcome of SIRS. The present study indicates that HMGB1 is a novel canine inflammatory marker and is distinct from CRP. However, the additional clinical value of HMGB1 measurement remains unclear, and further studies are warranted.     

Epigenetic assessment of environmental chemicals detected in maternal peripheral and cord blood samples

Epigenetic alteration is an emerging paradigm underlying the long-term effects of chemicals on gene functions. Various chemicals, including organophosphate insecticides and heavy metals, have been detected in the human fetal environment. Epigenetics by DNA methylation and histone modifications, through dynamic chromatin remodeling, is a mechanism for genome stability and gene functions. To investigate whether such environmental chemicals may cause epigenetic alterations, we studied the effects of selected chemicals on morphological changes in heterochromatin and DNA methylation status in mouse ES cells (ESCs). Twenty-five chemicals, including organophosphate insecticides, heavy metals and their metabolites, were assessed for their effect on the epigenetic status of mouse ESCs by monitoring heterochromatin stained with 4￠,6-diamino-2-phenylindole (DAPI). The cells were surveyed after 48 or 96 h of exposure to the chemicals at the serum concentrations of cord blood. The candidates for epigenetic mutagens were examined for the effect on DNA methylation at genic regions. Of the 25 chemicals, five chemicals (diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se) and octachlorodipropyl ether (S-421)) caused alterations in nuclear staining, suggesting that they affected heterochromatin conditions. Hg and Se caused aberrant DNA methylation at gene loci. Furthermore, DEP at 0.1 ppb caused irreversible heterochromatin changes in ESCs, and DEP-, Hg- and S-421-exposed cells also exhibited impaired formation of the embryoid body (EB), which is an in vitro model for early embryos. We established a system for assessment of epigenetic mutagens. We identified environmental chemicals that could have effects on the human fetus epigenetic status.     

Glomerular lipidosis accompanied by renal tubular oxalosis in wild and laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)

Glomerular lipidosis is a disease characterized by lipid accumulation in mesangial cells but that has not been fully investigated in avian species. We examined four wild and two laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)--an endangered avian species--presenting vacuolar deposits in the glomeruli. All cases had vacuolar deposits in the glomeruli. In the wild cases, fewer than 30% of all glomeruli were affected, compared with more than 90% in the laboratory-reared cases. In the wild cases, most deposits were mild and restricted to the mesangial areas of glomeruli. In the laboratory-reared cases, nearly all of the deposits covered entire glomeruli. Electron microscopy of mild deposits revealed vacuoles in the cytoplasm of mesangial cells. These vacuoles were positive for Sudan III, Sudan black B, oil red O, Nile blue, periodic acid-Schiff, Schultz test, and digitonin stain and were negative for performaric acid-Schiff stains. Based on these results, we diagnosed the glomerular lesion as glomerular lipidosis caused by uptake of low-density lipoprotein in mesangial cells. Except for one wild case, all cases exhibited renal tubular oxalosis. The severity of tubular oxalosis tended to be related to the severity of glomerular lipidosis: In cases of mild glomerular lipidosis, tubular oxalosis was also mild or absent. We therefore diagnosed the primary lesion as glomerular lipidosis accompanied by tubular oxalosis. The four wild cases came from different zones and therefore had no opportunities to interbreed and no common relatives. We believe these data support the hypothesis that glomerular lipidosis is a disease of the general population ofJapanese rock ptarmigans. This is the first report of glomerular lipidosis accompanied by renal tubular oxalosis in an avian species.     

Multi-locus sequence typing of Salmonella enterica subsp. enterica serovar Enteritidis strains in Japan between 1973 and 2004

Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) was responsible for a worldwide pandemic during the 1980s and 1990s; however, changes in the dominant lineage before and after this event remain unknown. This study determined S. Enteritidis lineages before and after this pandemic event in Japan using multilocus sequence typing (MLST). Thirty S. Enteritidis strains were collected in Japan between 1973 and 2004, consisting of 27 human strains from individual episodes, a bovine strain, a liquid egg strain and an eggshell strain. Strains showed nine phage types and 17 pulsed-field profiles with pulsed-field gel electrophoresis. All strains had homologous type 11 sequences without any nucleotide differences in seven housekeeping genes. These MLST results suggest that S. Enteritidis with the diversities revealed by phage typing and pulsed-field profiling has a highly clonal population. Although type 11 S. Enteritidis may exhibit both pleiotropic surface structure and pulsed-field type variation, it is likely to be a stable lineage derived from an ancestor before the 1980s and/or 1990s pandemic in Japan.     

Resistance to cypermethrin, deltamethrin and chlorpyriphos in populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from small farms of the State of São Paulo, Brazil

This study was undertaken to determine the flea diversity on urban dogs and cats in Australia in 2009-2010. A total of 2530 fleas were recovered from 291 animals (151 dogs, 69 cats and 71 uncategorised dogs or cats) from veterinary clinics across five states of Australia. The majority of specimens were from coastal areas. The cat flea (Ctenocephalides felis felis) was the most frequent flea species identified (98.8%, 2500/2530). The only other flea species identified was the stickfast flea (Echidnophaga gallinacea) from Western Australia. Sequencing of the cytochrome oxidase subunit II mtDNA revealed a single haplotype across Australia within a subset of C. f. felis (n=19). Our study demonstrated dominance and haplotype homogeneity of C. f. felis on dogs and cats. Although Ctenocephalides canis was recovered from a feral fox, it was not identified from the sample of fleas analysed. This suggests that, under current conditions, it is unlikely that foxes are reservoirs of C. canis for domestic dogs or cats residing in coastal Australia, as previously speculated.     

Characterization of canine BCL6 cDNA and detection systems for its protein expression

BCL6 is known to be a key molecule in germinal center (GC) formation of lymph nodes, and its expression profiles have been implicated in the prognosis of diffuse large B-cell lymphoma in humans. The present study was carried out to characterize canine BCL6 cDNA and to indicate the technical methods for detection of the BCL6 protein in dog tissues. The deduced amino acid sequence of canine BCL6 showed close homology to that of human BCL6 (96.3%), especially in the zinc-finger motifs and POZ (poxvirus and zinc finger) domain with complete identity. Immunoblot analysis of a canine lymph node with an anti-human BCL6 monoclonal antibody revealed a band of 80 kDa. Immunohistochemical staining using the same antibody produced positive reactions in the cells exclusively localized in the GC of a canine lymph node. This study will be useful for the molecular classification of canine B-cell lymphomas with different prognoses.     

Effects of oral administration of colostrum whey in peripartum goat on antimicrobial peptides in postpartum milk

The present study was conducted to examine whether colostrum supplementation in peripartum goats increases the antimicrobial peptides in their milk. Goats were orally administered 2 ml of colostrum whey products (colostrum group) or water (control group) daily, from 2 weeks before until 2 weeks after kidding. Body weights of mothers and kids were measured. Blood, milk, and fecal samples were collected from the mothers, and blood samples were collected from the kids. Concentrations of milk antimicrobial peptides (beta‐defensin, cathelicidin, lactoferrin, S100A7, lactoperoxidase, and immunoglobulin A [IgA]) were determined. IgA and nutritional parameters (glucose, total cholesterol, triglyceride, ketone bodies, and non‐esterified fatty acids) were also determined in the blood of mothers and kids. Milk IgA and lactoferrin concentrations were higher in the colostrum group than in the control group. Conversely, lower milk concentrations of S100A7 were observed in the colostrum group than that in the control group. Plasma IgA concentrations were higher for kids from the colostrum group than for those from the control group. These results suggest that oral administration of colostrum in pregnant goats increases IgA concentration in postpartum milk, which can subsequently improve the health of their kids.     

Use of tween 20 as a substrate for assay of lipase activity in soils

Abstract

A method for assaying the soil lipase activity is described. It involves the titrimetric estimation of the amount of lauric acid released by the lipase activity when the soil is incubated with Tween 20 in the presence of toluene at 30°C for 18 h under agitation. The method is simple and precise and incubation without agitation is also possible. The method has been applied to six different kinds of soils. The lipase activity in the cultivated soils ranged from 22.5 to 75.5 mmol min^?1 g^?1 of dried soil. The K _m value for Tween 20 was 1.8 × 10^?4 m. The optimum pH was approximately 7.5. The hydrolysis of liveen 20 in soil was inhibited by glycerol which was the essential moiety of glyceride. The inhibition by glycerol was found to be competitive. These results indicate that Tween 20 is a potential substrate for the assay of the glyceride hydrolytic activity in soils.     

Changes in the microbial community structure in soils treated with a mixture of glucose and peptone with reference to the respiratory quinone profile

Based on the respiratory quinone profile, changes in the structure of microbial communities in the soil samples from Nagoya University Farm were monitored after the treatment with 1% of a mixture of glucose and peptone. Samples of two soils differing in the fertilization history were examined: CF-soil with the application of only chemical fertilizers and FYM-soil with the application of only farmyard manure at a high rate. In the CF-soil, the amount of water-soluble organic carbon (WOC), indicator of the mixture of glucose and peptone, decreased to the original level after 14 d. After 7 d, the soil pH reached the maximum level, then decreased gradually. Changes in the inorganic nitrogen levels in the water extract also reflected the 14-d period of mineralization. The amount of respiratory quinones reached maximum levels after 7 d and gradually decreased, reflecting the changes in the microbial biomass. The quinone composition significantly changed during the 14-d period and returned to a profile similar to the original one after 28 d. Diversity of quinones significantly decreased during the 14-d period due to the predominance of ubiquinone with 9 isoprenoid units. In the FYM-soil, the amount of WOC decreased to the original level after 1 d, and the pH and inorganic nitrogen levels in the water extract reflected the one-day mineralization period, and nitrification started after 3 d. Although the amount of quinones indicated an increase in the microbial biomass for 14 d, the quinone composition did not change. These findings suggested that long-term application of farmyard manure resulted in stable microbial communities in response to the incorporation of organic matter in soil.