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.     

Anticancer activities of thelephantin O and vialinin A isolated from Thelephora aurantiotincta

Thelephora aurantiotincta is an edible mushroom belonging to the genus Thelephora; it grows in symbiosis with pine trees. Recently, phytochemical investigations have revealed that the genus Thelephora is an abundant source of p-terphenyl derivatives. However, their bioactivity has not yet been well characterized. In screening for natural materials with anticancer activity, a T. aurantiotincta ethanol extract (TAE) was found to decrease cell viability in human hepatocellular carcinoma cells (HepG2). In this study, a new p-terphenyl derivative, thelephantin O, and a known compound, vialinin A, were isolated as the principal bioactive components of TAE. These compounds decreased cell viability in HepG2 and human colonic carcinoma cells (Caco2), but not in noncancerous human hepatocytes. This is the first report of the isolation from T. aurantiotincta of selective cytotoxic agents against cancer cells.     

Phylogenetic groups and cephalosporin resistance genes of Escherichia coli from diseased food-producing animals in Japan

A total of 318 Escherichia coli isolates obtained from different food-producing animals affected with colibacillosis between 2001 and 2006 were subjected to phylogenetic analysis: 72 bovine isolates, 89 poultry isolates and 157 porcine isolates. Overall, the phylogenetic group A was predominant in isolates from cattle (36/72, 50%) and pigs (101/157, 64.3%) whereas groups A (44/89, 49.4%) and D (40/89, 44.9%) were predominant in isolates from poultry. In addition, group B2 was not found among diseased food-producing animals except for a poultry isolate. Thus, the phylogenetic group distribution of E. coli from diseased animals was different by animal species. Among the 318 isolates, cefazolin resistance (minimum inhibitory concentrations: ≥32 μg/ml) was found in six bovine isolates, 29 poultry isolates and three porcine isolates. Of them, 11 isolates (nine from poultry and two from cattle) produced extended spectrum β-lactamase (ESBL). The two bovine isolates produced bla_CTX-M-2, while the nine poultry isolates produced bla_CTX-M-25 (4), bla_SHV-2 (3), bla_CTX-M-15 (1) and bla_CTX-M-2 (1). Thus, our results showed that several types of ESBL were identified and three types of β-lactamase (SHV-2, CTX-M-25 and CTX-M-15) were observed for the first time in E. coli from diseased animals in Japan.     

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.     

Characterization of ACE Inhibitory Peptides Prepared from Pyropia pseudolinearis Protein

More than 7000 red algae species have been classified. Although most of them are underused, they are a protein-rich marine resource. The hydrolysates of red algal proteins are good candidates for the inhibition of the angiotensin-I-converting enzyme (ACE). The ACE is one of the key factors for cardiovascular disease, and the inhibition of ACE activity is related to the prevention of high blood pressure. To better understand the relationship between the hydrolysates of red algal proteins and the inhibition of ACE activity, we attempted to identify novel ACE inhibitory peptides from Pyropia pseudolinearis. We prepared water soluble proteins (WSP) containing phycoerythrin, phycocyanin, allophycocyanin, and ribulose 1,5-bisphosphate carboxylase/oxygenase. In vitro analysis showed that the thermolysin hydrolysate of the WSP had high ACE inhibitory activity compared to that of WSP. We then identified 42 peptides in the hydrolysate by high-performance liquid chromatography and mass spectrometry. Among 42 peptides, 23 peptides were found in chloroplast proteins. We then synthesized the uncharacterized peptides ARY, YLR, and LRM and measured the ACE inhibitory activity. LRM showed a low IC₅₀ value (0.15 μmol) compared to ARY and YLR (1.3 and 5.8 μmol). In silico analysis revealed that the LRM sequence was conserved in cpcA from Bangiales and Florideophyceae, indicating that the novel ACE inhibitory peptide LRM was highly conserved in red algae.     

Rediscovery of Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) MK.) an ancient crop of the Indian subcontinent

Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) Mac Key, synonym: T. sphaerococcum Perc.) is endemic to southern Pakistan and northwestern India. It was one of the main winter crops grown by ancient Indian cultures. However, it disappeared from the record during the early twentieth century, especially after the Green Revolution brought modern wheat varieties into India and Pakistan. Whether or not Indian dwarf wheat is presently cultivated has been unclear. Here we report on the rediscovery of the cultivation of this wheat in northern Karnataka and southern Maharashtra in India. Molecular genetic analysis of the chloroplast DNA of the two specimens collected at location 3 revealed that both samples have a unique haplotype that is specific to Indian dwarf wheat. We found this wheat at three locations in 2010, but at only one of the three locations in 2011. Therefore, the future survival of this subspecies is uncertain. Further ethnobotanical research is urgently needed to conserve this unique genetic resource for the future.     

Soil aggregate as a favorable habitat for Bradyrhizobium japonicum strains

Abstract

Rhizobial cells are present in soils as saprophytes after the decay of host plant nodules, and must survive in the soil until the next encounter with the infection sites of the host plant root. Biotic and abiotic environmental factors affect the population size of these rhizobia in the soil (Vincent 1977). Precise estimation of the population size of the native and the introduced rhizobia in the soil is necessary to study the conditions for the successful nodule formation by introduced strains.