Mitochondrial transmembrane potential and reactive oxygen species generation regulate the enhanced effect of CCCP on TRAIL-induced SNU-638 cell apoptosis

TRAIL is a member of the tumor necrosis factor family and engages apoptosis via recruitment and rapid activation of caspase-8. This study investigated the effect of carbonyl cyanide m-chlorophenylhydrazone (CCCP), a classic uncoupler of oxidative phosphorylation, on TRAIL-induced apoptosis in SNU-638 cells derived from human gastric cancer cells. It was found that treatment with CCCP followed by incubation with TRAIL markedly enhanced apoptosis by 2 fold compared with treatment with TRAIL alone. This effect was accompanied by reduction in mitochondrial transmembrane potential and generation of reactive oxygen species. This sensitization was inhibited by N-acetyl-l-cysteine, which restored the mitochondrial transmembrane potential and reduced reactive oxygen species generation. Treatment with N-acetyl-L-cysteine also inhibited expression of apoptotic proteins such as Bax and Smac and abrogated caspase-8 activation. Moreover, treatment with N-acetyl-L-cysteine prior to induction with TRAIL increased expression of the anti-apoptotic Bcl-2 protein. These data indicate that CCCP enhanced TRAIL-induced apoptosis by dissipation of mitochondrial transmembrane potential and reactive oxygen species, suggesting that treatment with CCCP combined with that with TRAIL can be an efficient method to induce death of tumor cells, particularly cells that are resistant to TRAIL-induced apoptosis.     

Echinochrome A Treatment Alleviates Atopic Dermatitis-like Skin Lesions in NC/Nga Mice via IL-4 and IL-13 Suppression

Atopic dermatitis (AD) is a chronic inflammatory skin disease in which skin barrier dysfunction leads to dryness, pruritus, and erythematous lesions. AD is triggered by immune imbalance and oxidative stress. Echinochrome A (Ech A), a natural pigment isolated from sea urchins, exerts antioxidant and beneficial effects in various inflammatory disease models. In the present study, we tested whether Ech A treatment alleviated AD-like skin lesions. We examined the anti-inflammatory effect of Ech A on 2,4-dinitrochlorobenzene (DNCB)-induced AD-like lesions in an NC/Nga mouse model. AD-like skin symptoms were induced by treatment with 1% DNCB for 1 week and 0.4% DNCB for 5 weeks in NC/Nga mice. The results showed that Ech A alleviated AD clinical symptoms, such as edema, erythema, and dryness. Treatment with Ech A induced the recovery of epidermis skin lesions as observed histologically. Tewameter^® and Corneometer^® measurements indicated that Ech A treatment reduced transepidermal water loss and improved stratum corneum hydration, respectively. Ech A treatment also inhibited inflammatory-response-induced mast cell infiltration in AD-like skin lesions and suppressed the expression of proinflammatory cytokines, such as interferon-γ, interleukin-4, and interleukin-13. Collectively, these results suggest that Ech A may be beneficial for treating AD owing to its anti-inflammatory effects.     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

Epidemiological and microbiological study of an outbreak of infectious keratoconjunctivitis in sheep

After several thousand sheep had been imported from Australia and New Zealand to Croatia during 1995, many native sheep that had been in contact with the imported animals acquired a severe ocular disease closely resembling infectious keratoconjunctivitis. In affected flocks glucose-fermenting mycoplasma were isolated from 48 per cent of conjunctival swabs and Branhamella ovis from 58 per cent. Twelve of 42 culturally and biochemically identical isolates were identified as Mycoplasma conjunctivae by polymerase chain reaction. From the conjunctivae of two animals M conjunctivae and M arginini were isolated in mixed culture. For many reasons most farmers removed the imported animals from their flocks and only sporadic cases of the disease were recognised in 1996. At the end of 1997, six flocks which were clinically free of the disease but had been affected during 1995, and five flocks with no history of the severe ocular disease were examined clinically and microbiologically, and were found to be free of M conjunctivae infection. At the time, B ovis was cultured almost exclusively from sheep originating from flocks which had been affected during 1995 and/or 1996. It was usually isolated in pure culture or as the predominant bacterial species, and was often accompanied by mild conjunctivitis. There were no microbiologically confirmed new cases of infectious keratoconjunctivitis during 1998 and 1999.     

Global metabolomic analysis of blood from mice infected with Brucella abortus

To better understanding Brucella abortus infection, serum metabolites of B. abortus-infected and -uninfected mice were analyzed and twenty-one metabolites were tentatively identified at 3 and 14 days post-infection (d.p.i.). Level of most lysophosphatidylcholines (LPCs) was found to increase in infected mice at 3 d.p.i., while it was decreased at 14 d.p.i. as compared to uninfected mice. In contrast, acylcarnitines were initially reduced at 3 d.p.i then elevated after two-weeks of infection, while hydroxysanthine was increased at 14 d.p.i. in infected mice. Our findings suggest that the significant changes in LPCs and other identified metabolites may serve as potential biomarkers in acute phase of B. abortus infection.     

Back-fat thickness as a primary index reflecting the yield and overall acceptance of pork meat

Predicting aspects of lean meat yield and eating quality of pork based on some particular carcass traits become increasingly important from an economic profitability point of view. The present study aimed to determine the relationship between the back-fat (BF) thickness with lean meat yield and meat quality traits of crossbred pigs. A total of 220 crossbred pigs [(Landrace × Yorkshire) ♀ × Duroc ♂] reared under identical conditions and harvested at 180 days old were slaughtered and screened for BF thickness. Four BF thickness groups: 12–15, 16–20, 21–25, and 26–30 mm were classified, and longissimus dorsi (LD) muscles from these groups were used to analyze meat quality characteristics. Results showed that increasing BF thickness decreased the lean meat yield and loin lean area. Increasing BF thickness up to 26–30 mm increased intramuscular fat content and pH of LD muscles. The LD muscles of the thicker BF (21–30 mm) groups received significantly higher flavor, juiciness, and overall acceptability scores than those of the thinner back-fat (12–20 mm) groups. Overall, when all the pre-harvest factors are kept the same, BF thickness could be considered as a primary index for predicting lean meat yield, and the minimal back-fat thickness of 21–30 mm is required to improve the overall acceptability of pork meat.     

Clove oil as an anaesthetic for common octopus (Octopus minor, Sasaki)

We evaluated the anaesthetic effect of clove oil [2‐methoxy‐4‐2‐(2‐propenyl)‐phenol] on the common octopus (Octopus minor), in terms of the time required to become anaesthetized (‘anaesthetic time’) and recovery time. We used a factorial experimental design and administered clove oil at different temperatures (15, 20 and 25°C) and concentrations (50, 100, 150, 200, 250 and 300 mg L⁻¹). We observed a significant (P<0.05) relationship between concentration and temperature, and each variable was effective (P<0.05). Anaesthetic time linearly decreased as the concentration and temperature increased. However, recovery time increased as the concentration increased and temperature decreased. There was no mortality. A concentration of 200 mg L⁻¹ clove oil showed rapid anaesthetic and recovery times in the common octopus, indicating its suitability for this species.     

Yersiniosis in zoo marmosets (Callitrix jacchuss) caused by Yersinia enterocolitica 4/O:3

The aim of this research was to describe two fatal cases of Yersinia enterocolitica bioserotype 4/O:3 infection in non-human primates and to characterise the isolates by PCR and PFGE. In July 2004, two marmosets (Callitrix jacchuss) born in captivity in Zagreb Zoo, died following a few days of intermittent diarrhoea in intervals of 2 weeks. The pathomorphological diagnosis of the female (born in 1997) and the male (born in 1995) marmoset, was disseminated miliary necrosis of the liver. Y. enterocolitica 4/O:3 was isolated from both livers showing that monkeys are susceptible to this bioserotype. The ail gene, which is an essential chromosomal virulence factor in pathogenic Y. enterocolitica isolates, was present in the marmoset isolates. Two different PFGE patterns were obtained from the isolates of the male liver with NotI enzyme. One genotype of the male marmoset isolate was indistinguishable from the genotype of the female marmoset isolate when NotI, ApaI and XhoI enzymes were used indicating a common infection source for the marmosets. The genotypes of the marmoset isolates differed only slightly from one human (of seven Croatian isolates) and from one pig isolate (representing a common genotype found among human and porcine isolates in Germany) suggesting that raw pork fed to the marmoset could have been the infection source.     

密闭空间雾滴沉积状态参数的显微图像解析

针对植保机械喷雾取样技术存在的不足,提出了利用显微图像结合数字图像处理技术对密闭空间中雾滴的沉积状态参数进行取样和分析的方法.利用该方法对棚室内雾滴覆盖率、沉积密度和均匀性等参数进行了试验和分析.试验结果表明,随距离增加,雾滴覆盖率先上升后下降,而雾滴沉积密度的变化趋势正好相反;栅室内距地面高度0.4～0.8m范围内,雾滴的覆盖率和沉积密度基本不随高度变化.用雾滴覆盖率、沉积密度和均匀性来表示整个喷雾场雾滴的沉积状态是有效、可行的.     

PCR-based sensitive and specific detection of Pectobacterium atrosepticum using primers based on Rhs family gene sequences

A sensitive and specific assay was developed to detect potato blackleg caused by Pectobacterium atrosepticum in potato. Primers PEAF and PEAR from the Rhs family gene homologous to RhsA (cell envelope biogenesis – outer membrane) were used to amplify a 904 bp DNA fragment. PCR was used to detect the pathogen in artificially inoculated potato seed tubers. The PCR product was only produced from 12 isolates of P. atrosepticum from various countries among 36 isolates of other species of Pectobacterium , Pseudomonas , Xanthomonas , as well as Escherichia coli and the soilborne fungus Fusarium oxysporum f.sp. dianthi .