ABSTRACT
The crude venom of the centipede Scolopendra subspinipes mutilans, injected with Escherichia coli K12D31 for 3-4 days showed broad-spectrum antimicrobial activity against Gram-positive. Gram-negative bacteria and fungi. It showed good antibacterial activity against E. coli K12D31 at different temperatures, pH, and ionic strengths. The crude venom was heated at 100 degrees C for 30 min, centrifuged at 10,000 rpm for 30 min at 4 degrees C and the supernatants were obtained, from which an antibacterial fraction having a molecular mass of 3000-5000 Da, was further separated by ultrafiltration. A homogeneous antibacterial peptide named scolopendrin I, having a molecular mass of 4,498 Da, was isolated using cation-exchange chromatography and two steps of reverse-phase high performance liquid chromatography (RP-HPLC). Scolopendrin I did not show any hemolytic and agglutination activities at the concentration below 30 microM.
Subject(s)
Animals , Anisoles/chemistry , Antimicrobial Cationic Peptides/biosynthesis , Arthropod Venoms/chemistry , Arthropods/chemistry , Indoles/chemistryABSTRACT
Objective Application of a new molecular marker to the identification of Dendrobium (Orchidaceae) species. Methods Complete sequences of the mitochondrial nad 1 intron 2 for nine species of Dendrobium Sw. were amplified and determined. Results Seventeen variable sites were found in the aligned 872 bp of nad 1 intron 2 sequences. Eight of the nine Dendrobium species except D. loddigesii could be identified by the nad 1 intron 2 sequences. Conclusion The mitochondrial nad 1 intron 2 sequences could be used as a new molecular marker for the identification of Dendrobium species.
ABSTRACT
Lipocalin-type prostaglandin D synthase (L-PGDS), an N-glycosylated dual functional monomeric protein, acts as a PGD2-producing enzyme and also as a lipophilic ligand-binding protein. L-PGDS is localized in the central nervous system, male genital organs of various mammals and in the human and monkey heart, and secreted into the cerebrospinal fluid, seminal plasma and blood plasma. The L-PGDS concentrations in these body fluids are useful for the diagnosis of several neurological disorders, dysfunction of sperm formation, cardiovascular and renal diseases.
Subject(s)
Animals , Humans , Amino Acid Sequence , Chromosome Mapping , Intramolecular Oxidoreductases , Chemistry , Genetics , Lipocalins , Molecular Sequence DataABSTRACT
AIM It is difficult to identify the Chinese crude drug snake gallbladder accurately by morphological and microscopical characteristics or chemical components only. In order to solve the problem, the technique based on DNA molecular marker was introduced into the authentication of snake gallbladder. METHODS DNA templates were extracted from the membrane or the bile of snake gallbladder, and also from the muscle of the original animal Elaphe schrenckii. About 400 bp DNA fragments of 12S rRNA gene were amplified from the templates and sequenced subsequently. RESULTS Enough amounts of DNA templates could be extracted from a bit of membrane or bile of snake gallbladder. The sequence of amplicons from the membrane, bile and muscle of the same individual were identical completely. CONCLUSION The technique of DNA molecular marker could be used for the authentication of snake gallbladder and bile. The results indicate that the technique could be used for the identification of crude drugs from other animal secretion. DNA sequence analysis also demonstrated that the origins of commercial snake gallbladder were complicated and more efficient quality control was necessary for supervising the crude drug in the market.
ABSTRACT
Object\ To develop a convenient and practical method for the identification of Carapax Trionycis Methods\ Based on the sequence variations of 12S rRNA gene between Pelodiscus sinensis and other softshell turtles, a pair of allele specific primers was designed to distinguish P. sinensis from other species of Trionychidae. DNA were extracted and anplified and Carapax Trionycis could be identified accurately by polymerase chain reaction (PCR) using the primers Results\ Ten samples of turtle shell from different sources were indentified by the allele specific PCR with the primers The result indicated that three samples were substitutes of Carapax Trionycis, consilient with the result from DNA sequence analysis The mitochondrial 12S rRNA gene fragment of P. maculatus and a faked imitation had also been sequenced Conclusion\ The primers could be used as key components in Carapax Trionycis identification kit
ABSTRACT
Objective:To prepare monoclonal antibodies against lipocalin type prostaglandin D synthase and to explore the distribution of lipocalin type prostaglandin D synthase on human sperm.Methods:BALB/c mice were immunized with lipocalin type prostaglandin D synthase,monoclonal antibodies were prepared by hybridoma technique,the biological characteristics of antibodies were analyzed by ELISA and Western blot,the distribution of lipocalin type prostaglandin D synthase on sperm were performed by immunohistochemistry.Results:A strain of hybridoma cell was obtain,and the IgG isotype of the antibody was identified as IgG1,with affinity constat(K) of 2?108 and titer over 103.Western blot demonstrated specific recognition of lipocalin type prostaglandin D synthase by the obtained monoclonal antibody.The result of immunohistochemistry displayed that lipocalin type prostaglandin D synthase was mainly distributed on the surface of human sperm.Conclusion:A hybridoma cell secreting anti-lipocalin type prostaglandin D synthase monoclonal antibodies stably was established.On the surface of human sperm,there is lipocalin type prostaglandin D synthase's distribution. [
ABSTRACT
Objective To explore the distribution and localization of LPGDS (Lipocalin-type prostaglandin D synthase) in rat testis and epididymidis and gain further insight into LPGDS potential function in male reproduction. Methods The expression of LPGDS in rat testis and epididymidis was assessed using RT-PCR and immunobloting. Using immunohistochemistry to explore the distribution of LPGDS in rat testis and epididymidis. Results None of the LPGDS studied was present in the testis of 2 month old SD rat. LPGDS was strikingly expressed in the caput epididymidis, while moderate to weak expressing was observed in the corpus and cauda epididymidis. The result of immunohistochemistry displayed that LPGDS was mainly localized within the cytoplasm and the cilia of the epithelial cells in rat epididymidis. Conclusion LPGDS was mainly distributed in epididymidis and localized with in the cytoplasm and the cilia of the epithelial cells. There was no LPGDS expressed in the testis of 2 month old SD rat, but it was strikingly expressed in the epididymidis, and the dose of expression was different, which suggested that LPGDS may play some kind of role during the process of the spermatozoa maturation.