Effects of chronic morphine on biliary tract responses to cholecystokinin-octapeptide in male guinea pigs.

Opioid peptides share the spasmogenic action of acutely administered morphine on the sphincter of Oddi. In this study, gallbladder function was assessed following chronic opioid administration. Implantation of morphine pellets (400 mg) in male guinea pigs depressed cholecystokinin-octapeptide(CCK)-induced emptying of gallbladder bile (monitored via a duodenal cannula). Gallbladder muscle strips, isolated from the morphine treated animals, showed depressed contractile responses to CCK. This antagonism was non-specific and indirectly mediated, as ACh contractions were also depressed, whereas CCK-induced contractions of gallbladder strips from untreated animals were unaffected by direct exposure to morphine (3 x 10(-6)M). The depression of CCK stimulation of bile flow by chronic morphine administration in male guinea pigs suggests that chronic exposure to opioids can impede gallbladder emptying.     

Alternative transcriptional initiation and splicing of mouse Lamc2 message.

To investigate the mechanism of two forms of messages (5.1 and 2.4 kb) in mouse Lamc2, a gene encoding for the gamma2 chain of epithelial cell-specific laminin 5, we analyzed approximately 40 kb of genomic DNA containing the sequences from the first intron to the 23rd exon to the 3' untranscribed region. With the 5' rapid amplification of cDNA end (RACE) and primer extension technique using RNA from mouse kidney and thymic epithelial cells, we found that the smaller message starts at the 2895th base of the cDNA within the 19th exon. Upstream sequences of this alternative start site showed the promoter activity in the reporter assay with the secreted form of alkaline phosphatase (SEAP). We also identified a novel alternatively spliced exon (exon 19B) that includes two stop codons. These results revealed two possible additional open reading frame that are different from the previously described alternative human gamma2 peptide. Therefore, the mechanism for generating smaller message and the event of alternative splicing are quite distinct between mouse and human Lamc2, although genomic organization is highly conserved. The significance of the presence of alternative message is discussed.     

Prostaglandin synthesis inhibited by formamidine pesticides.

The formamidine pesticides, chlordimeform and amitraz, were shown to have both antipyretic and anti-inflammatory activity when given intraperitoneally to rats at 5 to 80 mg per kg. They reduced yeast-induced fever in rats with potencies intermediate between those of indomethacin and aspirin, and antagonized the carageenin-induced swelling of the hind paw. In both these actions, chlordimeform was more potent than amitraz. Both formamidines also inhibited the synthesis of prostaglandin E₂ from arachidonic acid by bovine seminal vesicle microsomes. At an arachidonic acid concentration of 0.4 μM, the I₅₀ values for chlordimeform and amitraz were 34 and 880 μM respectively, compared to 0.4 μM and 790 μM for indomethacin and aspirin. These aspirin-like actions may provide a clue to some of the physiological effects of the formamidines, which represent a new and unsual group of prostaglandin synthetase inhibitors.     

High resolution crystal structure of PedB: a structural basis for the classification of pediocin-like immunity proteins

Background  

Pediocin-like bacteriocins, ribosomally-synthesized antimicrobial peptides, are generally coexpressed with cognate immunity proteins in order to protect the bacteriocin-producer from its own bacteriocin. As a step for understanding the mode of action of immunity proteins, we determined the crystal structure of PedB, a pediocin-like immunity protein conferring immunity to pediocin PP-1.     

Detection of hemi/homozygotes through heteroduplex formation in high-resolution melting analysis

Heteroduplex formation, required for the complete detection of hemi/homozygotes using high-resolution melting analysis, can be induced either by pre-PCR mixing of genomic DNAs or by post-PCR mixing of PCR products from unknown and reference samples. This study investigates the effects of both methods using two single nucleotide polymorphisms in X-linked DMD gene. The results show that both methods resulted in the same effect when mixing samples with the same gene copy number. Mixing samples with different gene copy numbers has not been previously explored and we show that post-PCR mixing is insensitive to gene copy number differences as compared to pre-PCR mixing.     

Hypoxic and nitrosative stress conditions modulate expression of myoglobin genes in a carcinogenic hepatobiliary trematode,Clonorchis sinensis

Despite recent evidence suggesting that adult trematodes require oxygen for the generation of bioenergy and eggshells, information on the molecular mechanism by which the parasites acquire oxygen remains largely elusive. In this study, the structural and expressional features of globin genes identified in Clonorchis sinensis, a carcinogenic trematode parasite that invades the hypoxic biliary tracts of mammalian hosts, were investigated to gain insight into the molecules that enable oxygen metabolism. The number of globin paralogs substantially differed among parasitic platyhelminths, ranging from one to five genes, and the C. sinensis genome encoded at least five globin genes. The expression of these Clonorchis genes, named CsMb (CsMb1—CsMb3), CsNgb, and CsGbX, according to their preferential similarity patterns toward respective globin subfamilies, exponentially increased in the worms coinciding with their sexual maturation, after being downregulated in early juveniles compared to those in metacercariae. The CsMb1 protein was detected throughout the parenchymal region of adult worms as well as in excretory-secretory products, whereas the other proteins were localized exclusively in the sexual organs and intrauterine eggs. Stimuli generated by exogenous oxygen, nitric oxide (NO), and nitrite as well as co-incubation with human cholangiocytes variously affected globin gene expression in live C. sinensis adults. Together with the specific histological distributions, these hypoxia-induced patterns may suggest that oxygen molecules transported by CsMb1 from host environments are provided to cells in the parenchyma and intrauterine eggs/sex organs of the worms for energy metabolism and/or, more importantly, eggshell formation by CsMb1 and CsMb3, respectively. Other globin homologs are likely to perform non-respiratory functions. Based on the responsive expression profile against nitrosative stress, an oxygenated form of secreted CsMb1 is suggested to play a pivotal role in parasite survival by scavenging NO generated by host immune cells via its NO dioxygenase activity.     

Benthic Diatoms as Indicators of Stream Sediment Concentrationin Hong Kong

Diatoms are photosynthetic unicellular, eukaryotic, microorganisms (algae) that are distinguished by their silicified (SiO₂‐nH₂O) cell walls. They have often been employed to assess salinity, pH, and nutrient conditions. Our data suggest that, they may also be used to assess suspended solid levels in streams. The ratio of motile to attached benthic diatoms from five different stream sites in Hong Kong was correlated with the level of suspended solids for each of these sites. Sites with high concentrations of suspended solids had a high percentage of motile diatoms on their rocks while sites with low concentrations of suspended solids had a high percentage of attached (non‐motile) diatoms on their rocks. When water carried by a stream has a high concentration of suspended solids, benthic diatoms in the stream are often covered in a layer of silt. Those diatoms that are motile are able to get on top of this silt layer while attached diatoms cannot. If the silt layer is not washed away, the attached diatoms perish and the percentage of motile diatoms increases. Thus, streams with high concentrations of suspended solids display a higher proportion of motile diatoms than do clear water streams with low concentrations of suspended solids. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)