Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp

The growth responses of nine human intestinal bacteria to liquid culture of Cordyceps militaris Link. Pt. (Ascomycotina: Clavicipitaceae) collected from a pupa of Bombyx mori L. (Lepidoptera: Bombycidae) were examined using spectrophotometric and impregnated paper disk methods and compared to those of tetracycline and chloramphenicol, as well as those of Coptis japonica root-derived berberine chloride. The biologically active constituent of the cultures was characterized as cordycepin (3'-deoxyadenosine) by spectroscopic analysis. This compound revealed potent growth-inhibiting activity toward Clostridium paraputrificum and Clostridium perfringens at 10 microgram/disk without adverse effects on the growth of Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Lactobacillus casei, whereas tetracycline and chloramphenicol inhibited the growth of these lactic acid-producing bacteria, clostridia and Escherichia coli. However, C. militaris-derived materials revealed no growth stimulation on the bifidobacteria and lactobacilli. These results may be an indication of at least one of the pharmacological actions of C. militaris. As a naturally occurring antibacterial agent, cordycepin could be useful as a new preventive agent against various diseases caused by clostridia.     

Effect of central corneal thickness on intraocular pressure with the rebound tonometer and the applanation tonometer in normal dogs

Objective To evaluate the effect of central corneal thickness (CCT) on the measurement of intraocular pressure (IOP) with the rebound (TonoVet^®) and applanation (TonoPen XL^®) tonometers in beagle dogs. Animal studied Both eyes of 60 clinically normal dogs were used. Procedures The IOP was measured by the TonoVet^®, followed by the TonoPen XL^® in half of the dogs, while the other half was measured in the reverse order. All CCT measurements were performed 10 min after the use of the second tonometer. Results The mean IOP value measured by the TonoVet^® (16.9 ± 3.7 mmHg) was significantly higher than the TonoPen XL^® (11.6 ± 2.7 mmHg; P < 0.001). The IOP values obtained by both tonometers were correlated in the regression analysis (γ² = 0.4393, P < 0.001). Bland–Altman analysis showed that the lower and upper limits of agreement between the two devices were ?0.1 and +10.8 mmHg, respectively. The mean CCT was 549.7 ± 51.0 μm. There was a correlation between the IOP values obtained by the two tonometers and CCT readings in the regression analysis (TonoVet^® : P = 0.002, TonoPen XL^® : P = 0.035). The regression equation demonstrated that for every 100 μm increase in CCT, there was an elevation of 1 and 2 mmHg in IOP measured by the TonoPen XL^® and TonoVet^®, respectively. Conclusions The IOP obtained by the TonoVet^® and TonoPen XL^® would be affected by variations in the CCT. Therefore, the CCT should be considered when interpreting IOP values measured by tonometers in dogs.     

Expression and regulation of Enpp2 in rat uterus during the estrous cycle

Ectonucleotide pyrophosphatase/phosphodiestrase 2 (Enpp2) isolated from the supernatant of human melanoma cells is a lysophospholipase D that transforms lysophosphatidylcholine into lysophospatidic acid. Although multiple analyses have investigated the function of Enpp2 in the hypothalamus, its role in the uterus during the estrous cycle is not well understood. In the present study, rat uterine Enpp2 was analyzed by RT-PCR, Western blotting, and immunohistochemistry. Quantitative PCR analysis demonstrated that uterine Enpp2 mRNA was decreased during estrus compared to proestrus and diestrus. To determine whether uterine Enpp2 expression is affected by sex steroid hormones, immature rats were treated with 17β-estradiol (E2), progesterone, or both on postnatal days 14 to 16. Interestingly, the expression of Enpp2 mRNA and protein were down-regulated by E2 in the uterus during estrus but not during proestrus or diestrus, suggesting that Enpp2 may play a role in uterine function during estrus. Enpp2 is primarily localized in the stromal cells of the endometrium during proestrus and estrus. During diestrus, Enpp2 was highly expressed in the epithelial cells of the endometrium. Taken together, these results suggest that uterine Enpp2 may be regulated by E2 and plays a role in reproductive functions during female rat development.     

Astaxanthin Confers a Significant Attenuation of Hippocampal Neuronal Loss Induced by Severe Ischemia-Reperfusion Injury in Gerbils by Reducing Oxidative Stress

Astaxanthin is a powerful biological antioxidant and is naturally generated in a great variety of living organisms. Some studies have demonstrated the neuroprotective effects of ATX against ischemic brain injury in experimental animals. However, it is still unknown whether astaxanthin displays neuroprotective effects against severe ischemic brain injury induced by longer (severe) transient ischemia in the forebrain. The purpose of this study was to evaluate the neuroprotective effects of astaxanthin and its antioxidant activity in the hippocampus of gerbils subjected to 15-min transient forebrain ischemia, which led to the massive loss (death) of pyramidal cells located in hippocampal cornu Ammonis 1-3 (CA1-3) subfields. Astaxanthin (100 mg/kg) was administered once daily for three days before the induction of transient ischemia. Treatment with astaxanthin significantly attenuated the ischemia-induced loss of pyramidal cells in CA1-3. In addition, treatment with astaxanthin significantly reduced ischemia-induced oxidative DNA damage and lipid peroxidation in CA1-3 pyramidal cells. Moreover, the expression of the antioxidant enzymes superoxide dismutase (SOD1 and SOD2) in CA1-3 pyramidal cells were gradually and significantly reduced after ischemia. However, in astaxanthin-treated gerbils, the expression of SOD1 and SOD2 was significantly high compared to in-vehicle-treated gerbils before and after ischemia induction. Collectively, these findings indicate that pretreatment with astaxanthin could attenuate severe ischemic brain injury induced by 15-min transient forebrain ischemia, which may be closely associated with the decrease in oxidative stress due to astaxanthin pretreatment.     

Acaricidal activities of paeonol and benzoic acid from Paeonia suffruticosa root bark and monoterpenoids against Tyrophagus putrescentiae (Acari: Acaridae)

The acaricidal activities of paeonol (2'-hydroxy-4'-methoxyacetophenone) and benzoic acid identified in the root bark of tree peony, Paeonia suffruticosa Andrews, against copra mite, Tyrophagus putrescentiae (Schrank), adults were examined using direct contact and vapour phase toxicity bioassays and compared with those of cinnamyl acetate, cinnamyl alcohol and 37 monoterpenoids as well as the acaricides benzyl benzoate, dibutyl phthalate and N,N-diethyl-m-toluamide (DEET). Based on LD(50) values in fabric piece contact toxicity bioassays, the acaricidal activities of benzoic acid (4.80 microg cm(-2)) and paeonol (5.29 microg cm(-2)) were comparable to that of benzyl benzoate (4.46 microg cm(-2)) but more pronounced than those of DEET (30.03 microg cm(-2)) and dibutyl phthalate (25.23 microg cm(-2)). In vapour phase toxicity bioassays, paeonol and benzoic acid were much more effective in closed containers than in open ones, indicating that the effects of these compounds were largely due to action in the vapour phase. As judged by 24 h LD(50) values, (1S)-(-)-verbenone (7.42 mg per disc) was the most toxic fumigant, followed by (1S)-(-)-camphor, (S)-(+)-carvone, (R)-(-)-linalool and (+/-)-camphor (10.45-18.18 mg). Potent fumigant toxicity was also observed with paeonol, (2S,5R)-(-)-menthone, (+/-)-citronellal, benzoic acid, (1S,4R)-(-)-alpha-thujone and (R)-(+)-pulegone (25.10-34.63 mg). Neither benzyl benzoate, DEET nor dibutyl phthalate caused fumigant toxicity. Paeonia root bark-derived materials, particularly paeonol and benzoic acid, as well as the monoterpenoids described, merit further study as potential acaricides or as leads for the control of T. putrescentiae.     

Activation of Akt/protein kinase B mediates the protective effects of mechanical stretching against myocardial ischemia-reperfusion injury

Akt/protein kinase B is a well-known cell survival factor and activated by many stimuli including mechanical stretching. Therefore, we evaluated the cardioprotective effect of a brief mechanical stretching of rat hearts and determined whether activation of Akt through phosphatidylinositol 3-kinase (PI3K) is involved in stretch-induced cardioprotection (SIC). Stretch preconditioning reduced infarct size and improved post-ischemic cardiac function compared to the control group. Phosphorylation of Akt and its downstream substrate, GSK-3β, was increased by mechanical stretching and completely blocked by wortmannin, a PI3K inhibitor. Treatment with lithium or SB216763 (GSK-3β inhibitors) before ischemia induction mimicked the protective effects of SIC on rat heart. Gadolinium (Gd³⁺), a blocker of stretch-activated ion channels (SACs), inhibited the stretch-induced phosphorylation of Akt and GSK-3β. Furthermore, SIC was abrogated by wortmannin and Gd³⁺. In vivo stretching induced by an aorto-caval shunt increased Akt phosphorylation and reduced myocardial infarction; these effects were diminished by wortmannin and Gd³⁺ pretreatment. Our results showed that mechanical stretching can provide cardioprotection against ischemia-reperfusion injury. Additionally, the activation of Akt, which might be regulated by SACs and the PI3K pathway, plays an important role in SIC.     

Biological evaluation of neopeptins isolated from a Streptomyces strain

BACKGROUND: Microbial secondary metabolites are a rich source of antifungal agents and have merit as alternatives to synthetic fungicides. To develop disease control agents against powdery mildew, the lipopeptide antibiotic neopeptins were identified from the culture broth of a Streptomyces sp., and in vivo control efficacy of the compounds was evaluated on cucumber plants under glasshouse conditions. RESULTS: The Streptomyces sp. KNF2047 antagonistic against powdery mildew development in cucumber plants was isolated from a soil sample. Antifungal compounds were purified from the culture broth and identified as neopeptin A and B. In vitro microtitre assays revealed the inhibitory activities of the compounds in the range 128-512 microg mL(-1) against the mycelial growth of Alternaria mali, Botrytis cinerea, Cladosporium cucumerinum, Colletotrichum lagenarium, Didimella bryoniae and Magnaporthe grisea. Although neither compound showed remarkable in vitro antifungal activity against other plant pathogenic fungi, a mixture of neopeptins (484 mg of neopeptin A and 290 mg of neopeptin B per gram of partially purified powder) showed potent protective and curative activity against cucumber powdery mildew in vivo. The disease control activity of the neopeptins at a concentration of 2.4 mg L(-1) was 92.1%, which was similar to that of the commercial fungicide fenarimol (89.3% at 63 mg L(-1)) and that of the commercial biocontrol agent Actinovate (67.4% at 2 x 10(7) cfu L(-1)). CONCLUSION: Neopeptin mixtures isolated from Streptomyces sp. KNF2047 showed potent disease control activity against powdery mildew development on cucumber plants. .