Enantioselective 2-hydroperoxylation of long-chain fatty acids in marine algae

SUMMARY: When long-chain saturated and unsaturated fatty acids were incubated with crude enzyme of marine green alga Ulva pertusa , the corresponding ( R )-2-hydroperoxy acids were found to have high enantiomeric excess (> 99%). In a similar administration experiment, the ( R )-2-hydroperoxy acid was obtained from the incubation of palmitic acid with crude enzymes of a variety of marine algae. Thus, we found that not only green algae but also brown and red algae are capable of enantioselective 2-hydroperoxylation of palmitic acid.     

A cardiac rhabdomyoma in a Guinea pig

A guinea pig (9-week-old) that had been placed in a control group for a pharmacological test was found to have a single nodule on the surface of the right ventricular wall. In a transverse section of the heart after fixation, a whitish mass was found that extended from the subendocardium to the subepicardium of the right ventricular wall. Histopathological examination revealed a spongy network consisting of vacuolated spaces in the myocardium of the right ventricle extending to the myocardium and subepicardium of the right atrium. The vacuolated space was PAS-positive. Immunohistochemical examinations revealed that the lesions contained striated fibers that were positive for anti-desmin and anti-myoglobin. Electron micrographs revealed the lesions resulting in affected striated muscle fibers and accumulations of many glycogen granules. Based on the findings, the lesions were diagnosed as a cardiac rhabdomyoma. This is the first report of application of immunohistochemical examinations to diagnosis of cardiac rhabdomyoma in the guinea pig.     

The first assessment of cyanobacterial and diazotrophic diversities in the Japan Sea

The diversity of cyanobacteria and diazotrophs in the Japan Sea was investigated by analyzing sequences of cyanobacterial 16S rRNA genes and nitrogen fixation genes (nifH) from seawater sampled at depths ranging from the surface to 100?m at two stations. Of the 107 cyanobacterial 16S rRNA gene sequences obtained, 97 and three sequences were assigned to Synechococcus sub-cluster 5.1 and Prochlorococcus HL (II), respectively. Unlike other oceanic regions, at our two sampling stations the composition ratio of the sequences assignable to Synechococcus sub-cluster 5.3 was relatively high (8?%). No sequences of diazotrophic cyanobacteria were found in the cyanobacterial 16S rRNA genes. In the nifH clone library (36 sequences), ten sequences were identified as a UCYN-A group of diazotrophic cyanobacteria; the other 26 sequences (72?%) were assigned to proteobacteria. These results suggest that heterotrophic bacteria, including UCYN-A, dominate the diazotrophic community in the Japan Sea. Our study reveals the dominance of Synechococcus in cyanobacterial community and (photo)heterotrophic diazotrophs in the diazotrophic community in the Japan Sea, suggesting its unique characteristics.     

Purification and characterization of sulfotransferase specific to O-22 of 11-hydroxy saxitoxin from the toxic dinoflagellate Gymnodinium catenatum (dinophyceae)

ABSTRACT: A novel sulfotransferase (O-ST), which transferred the sulfate group of 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to O-22 of 11-α,β-hydroxy saxitoxin (STX) and produced GTX2 + 3, was purified to homogeneity from the cytosolic fraction of clonal-axenic vegetative cells of the toxic dinoflagellate Gymnodinium catenatum GC21V. After four purification steps, including affinity chromatography and anion exchange chromatography, the enzyme was purified 500-fold and the yield was 4%. On affinity chromatography with a PAP-agarose column, O-ST was observed in the bound fraction, and N-ST specific to N-21 of STX and GTX2 + 3 was found in the unbound fraction. The molecular mass of the purified enzyme was determined by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) to be 65 kDa. Gel filtration chromatography showed a native molecular mass of 67 kDa, indicating that O-ST is a monomeric enzyme. The enzyme was optimally active at pH 6.0 and 35°C. O-ST did not require metal cations for its activity. O-ST required PAPS as the sole source of sulfate. O-ST transferred a sulfate group from PAPS to only O-22 of 11-α,β-hydroxy STX and not to N-21 of these toxins. These observations suggested that two ST, N-ST and O-ST, participate in the sulfation of PSP toxins.     

Follicular development after ovum pick-up and fertilizability of retrieved oocytes in postpartum dairy cattle

This study aimed to evaluate gonadotropin secretion and the developmental competence of follicular oocytes in dairy cattle during the early postpartum (PP) period. The number of follicles developed after transvaginal ultrasound-guided ovum pick-up (OPU) and fertilizability of retrieved oocytes were compared between cows in which the first dominant follicle (DF) ovulated (ovulated group, n=4) and did not ovulate (non-ovulated group, n=3), and between early PP (early PP group, n=2) and after the resumption of the estrous cycle (cyclic group, n=2). Follicular ablation was performed 2-4 days after the detection of DF in the second follicular wave PP. OPU was repeated 3-5 times at 3 or 4-day intervals from 3-4 days after the follicular ablation. At OPU, the follicles were enumerated and all those > or = 5 mm in diameter were aspirated. Recovered oocytes were subjected to in vitro maturation and fertilization. Both criteria were similar between ovulated and non-ovulated groups, and between early PP and cyclic groups. These results suggest that FSH/LH secretions required for follicle recruitment and subsequent follicular growth during the early PP period are similar to those after resumption of the estrous cycle. They also indicate that follicular oocytes during the early PP period have developmental competence.     

Sperm motility, plasma membrane integrity, and binding capacity to homologous zona pellucida of cryopreserved epididymal spermatozoa in the domestic cat

We examined motility, plasma membrane integrity, and binding capacity to homologous zona pellucidae (ZP) of frozen/thawed epididymal cat sperm as a model species for endangered felines. Epididymal spermatozoa from 20 domestic cats were frozen with freezing egg-yolk extender containing 3.0% glycerol in 0.25-ml straws. Post-thaw motility and plasma membrane integrity of the frozen/thawed spermatozoa were 31.8 +/- 2.4% and 32.2 +/- 4.2%, respectively. The frozen/thawed spermatozoa were co-cultured with frozen/thawed immature homologous oocytes with intact ZP for 3 h to examine their ability to bind to the ZP. Sixteen of the 20 frozen/thawed sperm samples demonstrated the ability to bind to ZP. These results indicated that the freezing system for epididymal sperm used in the present study gives appropriate information for banking the genetic resources of wild felid species.     

Ventrolateral temporal lobectomy in normal dogs as a counterpart to human anterior temporal lobectomy: a preliminary study on the surgical procedure and complications

Anterior temporal lobectomy (ATL) is a surgical procedure for drug-resistant mesial temporal lobe epilepsy that is commonly performed in human medicine. The purpose of this study was to determine whether ATL-like surgery, i.e., removal of the amygdala and hippocampal head, is possible in dogs, and to investigate its safety and postoperative complications. Eight healthy beagles underwent ATL-like surgery and were observed for 3 months postoperatively. Samples from the surgically resected tissues and postmortem brain were evaluated pathologically. The surgical survival rate was 62.5%. The major postoperative complications were visual impairment, temporal muscle atrophy on the operative side, and a postoperative acute symptomatic seizure. Due to the anatomical differences between dogs and humans, the surgically resected area to approach the medial temporal structures in dogs was the ventrolateral part of the temporal lobe. Therefore, the ATL-like surgery described in this study was named “ventrolateral temporal lobectomy” (VTL). This study is the first report of temporal lobectomy including amygdalohippocampectomy in veterinary medicine and demonstrates its feasibility. Although it requires some degree of skill, VTL could be a treatment option for canine drug-resistant epilepsy and lesions in the mesial temporal lobe.