Effect of modified oligopeptides from the beetle Allomyrina dichotoma on Escherichia coli infection in mice

The novel peptides based on a putative active site of defensin, an anti-bacterial peptide from the beetle Allomyrina dichotoma, were synthesized. These synthetic oligopeptides exhibited strong anti-bacterial activity in vitro, even against antibiotic-resistant pathogenic bacteria. Then, anti-bacterial activity of two newly synthesized peptides, RLYLRIGRR-NH(2) (peptide A) and RLRLRIGRR-NH(2) (peptide B) was also examined by macroscopic and histopathologic assessment during the course of infection in mice inoculated with antibiotic-resistant pathogenic Escherichia coli (E. coli) in vivo. Peptide B decreased the mortality of mice inoculated with antibiotic-resistant pathogenic E. coli. The results of macroscopic and histopathologic examinations revealed that peptide B could protect the mice from infection. In contrast, peptide A failed to protect mice from infection with antibiotic-resistant pathogenic E. coli. Also, modified peptides A and B produced no toxicity or side effects in mice. These results suggest that peptide B is useful for developing novel antibiotics against antibiotic-resistant pathogenic bacteria.     

Comparative anatomical study on the relationships between the vestigial pelvic bones and the surrounding structures of finless porpoises (Neophocaena phocaenoides)

Morphology of the modern cetaceans represents the results of adaptation of the ancestral terrestrial mammals to aquatic life through their evolutional processes. Some of the primitive fossil cetaceans are known to have both fore and hind limbs, whereas the pelvic bones of modern cetaceans are, in general, a pair of slender rod-like structures within the abdominal wall muscles just anterior to the anus with no articulations to the axial skeleton in both sexes. It is interesting and important to consider the causes and processes of how the hind limbs were lost and how the pelvis was reduced during the process of adaptation. In the present study, we tried to evaluate the topography and function of rudimentary pelvic bones of the finless porpoise (Neophocaena phocaenoides), one of the members of the odontocete cetaceans, with special references to the structures around the pelvic bones. Some soft tissues such as M. ischiocavernosus relating to the pelvic bone are transformed following the drastic reduction of the pelvis. This transformation tells us that the cetaceans adapted to the aquatic life during evolutional processes chose the tail flukes driven by the powerful trunk muscles for locomotion, instead of modifying the hind limbs into hind flippers as seen in pinnipeds. On the other hand, it is evident that a function of the pelvic bones of the male finless porpoise was supporting the penis as those of terrestrial mammals. It is noteworthy that the morphological features of the ancestral terrestrial mammals can be traced when they are carefully compared with those of the finless porpoise.     

Usefulness of MRI compared with CT for diagnosis of mesenteric lymphoma in a dog

We evaluated the usefulness of MRI and compared it with CT for diagnosis of mesenteric lymphoma in a dog. The results in the plain CT, dynamic CT and plain MR (T1WI and T2WI) images suggested that the mass was a large single nodular lesion with abundant blood perfusion. On enhanced MRI(T1WI) , the mass was depicted as a tumor with adhesion to the gut wall. Exploratory laparotomy confirmed the mass was consistent with the findings on enhanced MRI. We think that MRI might be a useful imaging tool for diagnosis of canine mesenteric lymphoma.     

Immunohistochemical localization of steroidogenic enzymes in the testis of Hokkaido Sika deer (Cervus nippon yesoensis)

The testes from 15 adult male Hokkaido Sika deer (Cervus nippon yesoensis) were collected during the rutting season (October and November). We investigated the localization of 4 kinds of steroidogenic enzymes (P450scc, 3betaHSD, P450c17 and P450arom) immunohistochemically in these testicular samples. The specific immunoreactivities to these enzymes were detected only in the cytoplasm of Leydig cells. This differs to the enzyme distributions reported previously in Japanese black bear, Japanese raccoon dog, Hokkaido brown bear and American black bear, in which the same immunoreactivities were detected in Leydig cells, Sertoli cells and/or spermatogenic cells. The current study suggests that in the testes of the Hokkaido Sika deer, testosterone and estradiol-17beta may be synthesized in the Leydig cells only.     

Fate of cystic ovarian follicles and the subsequent fertility of early postpartum dairy cows

The fate of cystic ovarian follicles that developed spontaneously during the early postpartum period of 50 lactating dairy cows was traced by ultrasonography to characterise the follicular dynamics in relation to the fertility of the cows. The absence of postpartum ovulations caused by repeated waves of anovulatory large follicles was also characterised and evaluated. Fifteen of the 50 cows developed cystic follicles, and these follicles became follicular cysts in five of the 15 cows. Most of the cystic follicles emerged before the first postpartum ovulation of the cows. The transition from cystic follicles to follicular cysts delayed the cows' first ovulation, oestrus and insemination, but had less influence on their fertility after they had recovered spontaneously. In addition to the 15 cows that developed cystic follicles or follicular cysts, six of the cows had five to 13 waves of follicles before their first ovulation. These repeated waves of follicles caused a more severe delay in the early postpartum reproductive events but did not affect the cows' fertility.     

Identification and cornification-related gene expression of canine keratinocyte differentiation-associated protein, Kdap

The outermost layer of skin, the epidermis, is cornified epithelial tissue composed of keratinocytes. To maintain the structure and function of the epidermis, the regulation of proliferation, differentiation, and cornification of keratinocytes is crucial, and various soluble factors secreted by keratinocytes are involved in these regulations. Previously, work has shown that keratinocytes secreted the protein Kdap (keratinocyte differentiation-associated protein) associated with the formation of cornified cell envelopes, a specialized protective barrier structure on the periphery of terminally differentiating keratinocytes. In the present report, the canine counterpart of human Kdap is identified and an attempt has been made to define its physiological role in canine keratinization. Canine Kdap (cKdap) showed structural features commonly observed in other counterparts and is secreted from transfected cells. The expression profile of cKdap mRNA, which was restrictively expressed in cornified epithelial tissues besides skin has also been determined. These findings indicate that there is a strong association between cKdap expression and cornification, which supports previous observations that Kdap is involved in the synthesis and/or degradation of cornified cell envelopes in humans and mice.     

Dietary apigenin suppresses IgE and inflammatory cytokines production in C57BL/6N mice

Flavonoids ubiquitously exist in plants, vegetables, fruits, and teas. We evaluated the effect of dietary apigenin, one of the well-known flavonoids, on the immune system in C57BL/6N mice. Mice were fed experimental diets containing apigenin for 2 weeks. After the experimental period, there was no significant difference in body and organ weights between the control and the apigenin group. The total immunoglobulin (Ig) E levels in mice fed apigenin were significantly suppressed, whereas levels of IgG, IgM, and IgA were not affected. We also examined the effect of the apigenin diet on cytokine expression in mice sera using a cytokine array. The production of regulated upon activation normal T cell expressed and secreted (RANTES) and soluble tumor necrosis factor receptor I (sTNFRI) in mice sera was down-regulated by the apigenin diet. These results suggest that a diet containing apigenin can reduce serum IgE and inflammatory cytokines such as RANTES and sTNFRI in mice.     

Serotonin derivatives, major safflower (Carthamus tinctorius L.) seed antioxidants, inhibit low-density lipoprotein (LDL) oxidation and atherosclerosis in apolipoprotein E-deficient mice

The effects of defatted safflower seed extract and its phenolic constituents, serotonin derivatives, on atherosclerosis were studied. Ethanol-ethyl acetate extract of safflower seeds (SSE) inhibited low-density lipoprotein (LDL) oxidation induced in vitro by an azo-containing free-radical initiator V70 or copper ions. Two serotonin derivatives [N-(p-coumaroyl)serotonin, CS; N-feruloylserotonin, FS] and their glucosides were identified as the major phenolic constituents of the extract. The study with chemically synthesized materials revealed that a majority of the antioxidative activity of SSE was attributable to the aglycones of these two serotonin derivatives. Orally administered CS and FS suppressed CuSO(4)-induced plasma oxidation ex vivo. Long-term (15 week) dietary supplementation of SSE (1.0 wt %/wt) and synthetic serotonin derivatives (0.2-0.4%) significantly reduced the atherosclerotic lesion area in the aortic sinus of apolipoprotein E-deficient mice (29.2-79.7% reduction). The plasma level of both lipid peroxides and anti-oxidized LDL autoantibody titers decreased concomitantly with the reduction of lesion formation. Serotonin derivatives were detected as both intact and conjugated metabolites in the plasma of C57BL/6J mice fed on 1.0% SSE diet. These findings demonstrate that serotonin derivatives of SSE are absorbed into circulation and attenuate atherosclerotic lesion development possibly because of the inhibition of oxidized LDL formation through their strong antioxidative activity.     

ATM engages autodegradation of the E3 ubiquitin ligase COP1 after DNA damage

The ataxia telangiectasia mutated (ATM) protein kinase is a critical component of a DNA-damage response network configured to maintain genomic integrity. The abundance of an essential downstream effecter of this pathway, the tumor suppressor protein p53, is tightly regulated by controlled degradation through COP1 and other E3 ubiquitin ligases, such as MDM2 and Pirh2; however, the signal transduction pathway that regulates the COP1-p53 axis following DNA damage remains enigmatic. We observed that in response to DNA damage, ATM phosphorylated COP1 on Ser(387) and stimulated a rapid autodegradation mechanism. Ionizing radiation triggered an ATM-dependent movement of COP1 from the nucleus to the cytoplasm, and ATM-dependent phosphorylation of COP1 on Ser(387) was both necessary and sufficient to disrupt the COP1-p53 complex and subsequently to abrogate the ubiquitination and degradation of p53. Furthermore, phosphorylation of COP1 on Ser(387) was required to permit p53 to become stabilized and to exert its tumor suppressor properties in response to DNA damage.     

Loss of virulence of the phytopathogen Ralstonia solanacearum through infection by φRSM filamentous phages

φRSM1 and φRSM3 (φRSM phages) are filamentous phages (inoviruses) that infect Ralstonia solanacearum, the causative agent of bacterial wilt. Infection by φRSM phages causes several cultural and physiological changes to host cells, especially loss of virulence. In this study, we characterized changes related to the virulence in φRSM3-infected cells, including (i) reduced twitching motility and reduced amounts of type IV pili (Tfp), (ii) lower levels of β-1,4-endoglucanase (Egl) activity and extracellular polysaccharides (EPS) production, and (iii) reduced expression of certain genes (egl, pehC, phcA, phcB, pilT, and hrpB). The significantly lower levels of phcA and phcB expression in φRSM3-infected cells suggested that functional PhcA was insufficient to activate many virulence genes. Tomato plants injected with φRSM3-infected cells of different R. solanacearum strains did not show wilting symptoms. The virulence and virulence factors were restored when φRSM3-encoded orf15, the gene for a putative repressor-like protein, was disrupted. Expression levels of phcA as well as other virulence-related genes in φRSM3-ΔORF15-infected cells were comparable with those in wild-type cells, suggesting that orf15 of φRSM3 may repress phcA and, consequently, result in loss of virulence.