Tanjungides A and B: New Antitumoral Bromoindole Derived Compounds from Diazona cf formosa. Isolation and Total Synthesis

Tanjungides A (1) (Z isomer) and B (2) (E isomer), two novel dibrominated indole enamides, have been isolated from the tunicate Diazona cf formosa. Their structures were determined by spectroscopic methods including HRMS, and extensive 1D and 2D NMR. The stereochemistry of the cyclised cystine present in both compounds was determined by Marfey’s analysis after chemical degradation and hydrolysis. We also report the first total synthesis of these compounds using methyl 1H-indole-3-carboxylate as starting material and a linear sequence of 11 chemical steps. Tanjungides A and B exhibit significant cytotoxicity against human tumor cell lines.     

Extirpation, colonization, and habitat dynamics of a keystone species along an urban gradient

Using 5 years of patch occupancy data for 384 habitat fragments, we evaluated population and habitat dynamics of the black-tailed prairie dog in urban habitat remnants in the rapidly developing landscape of Denver, CO, USA. Specifically, we evaluated the landscape factors, including fragment area, age, and connectivity, that characterize the habitat fragments most likely to be colonized by prairie dogs, as well as those experiencing local extinctions. In addition, we determined which patch types were most often removed by land development. Sites in proximity to colonies were more likely to be colonized by prairie dogs. Local extinctions were most common on isolated colonies, and older and more isolated colonies were more likely to be extirpated by human activity. In general, smaller and older habitat patches were at the highest risk of being lost to land development. Our results provide observations of dynamic changes to the distribution of a potential keystone species in an urban area, which can be used to inform island biogeographic and metapopulation models for wildlife persistence in developing landscapes. Although populations are currently in decline, most local extinctions are the direct result of human activity, and we suggest that prairie dogs in this area can persist with appropriate management.     

Isolation of a cDNA from the virus responsible for enterically transmitted non-A, non-B hepatitis

Major epidemic outbreaks of viral hepatitis in underdeveloped countries result from a type of non-A, non-B hepatitis distinct from the parenterally transmitted form. The viral agent responsible for this form of epidemic, or enterically transmitted non-A, non-B hepatitis (ET-NANBH), has been serially transmitted in cynomolgus macaques (cynos) and has resulted in typical elevation in liver enzymes and the detection of characteristic virus-like particles (VLPs) in both feces and bile. Infectious bile was used for the construction of recombinant complementary DNA libraries. One clone, ET1.1, was exogenous to uninfected human and cyno genomic liver DNA, as well as to genomic DNA from infected cyno liver. ET1.1 did however, hybridize to an approximately 7.6-kilobase RNA species present only in infected cyno liver. The translated nucleic acid sequence of a portion of ET1.1 had a consensus amino acid motif consistent with an RNA-directed RNA polymerase; this enzyme is present in all positive strand RNA viruses. Furthermore, ET1.1 specifically identified similar sequences in complementary DNA prepared from infected human fecal samples collected from five geographically distinct ET-NANBH outbreaks. Therefore, ET1.1 represents a portion of the genome of the principal viral agent, to be named hepatitis E virus, which is responsible for epidemic outbreaks of ET-NANBH.     

Terrestrial leaf meals or freshwater aquatic fern as potential feed ingredients for farmed abalone Haliotis asinina (Linnaeus 1758)

Three terrestrial leaf meals, Carica papaya, Leucaena leucocephala, Moringa oliefera and a freshwater aquatic fern, Azolla pinnata were evaluated as potential ingredients for farmed abalone diet. All diets were formulated to contain 27% crude protein, 13% of which was contributed by the various leaf meals. Fresh seaweed Gracilariopsis bailinae served as the control feed. Juvenile Haliotis asinina (mean body weight=13.4±1.6 g, mean shell length= 38.8±1.4 mm) were fed the diets at 2–3% of the body weight day^–1. Seaweed was given at 30% of body weight day^–1. After 120 days of feeding, abalone fed M. oliefera, A. pinnata‐based diets, and fresh G. bailinae had significantly higher (P<0.01) specific growth rates (SGR%) than abalone fed the L. leucocephala‐based diet. Abalone fed the M. oliefera‐based diet had a better growth rate in terms of shell length (P<0.05) compared with those fed the L. leucocephala‐based diet but not with those in other treatments. Furthermore, protein productive value (PPV) of H. asinina was significantly higher when fed the M. oliefera‐based diet compared with all other treatments (P<0.002). Survival was generally high (80–100%) with no significant differences among treatments. Abalone fed the M. oliefera‐based diet showed significantly higher carcass protein (70% dry weight) and lipid (5%) than the other treatments. Moringa oliefera leaf meal and freshwater aquatic fern (A. pinnata) are promising alternative feed ingredients for practical diet for farmed abalone as these are locally available year‐round in the Philippines.     

Western Blot Analysis of Proacrosin/Acrosin in Frozen Dog Sperm During In Vitro Capacitation

Acrosin is an acrosomal protease synthesized as an inactive precursor, proacrosin, which is processed via autoproteolysis into active forms alpha- and beta-acrosin. In this paper, a comparative study on the immunoreactivity of acrosin during in vitro capacitation of frozen and fresh (control) canine sperm using Western blot analysis is reported. Semen samples were obtained by digital stimulation and ejaculates processed as fresh and frozen samples and then capacitated for 0, 30, 60 and 90 min. At each time period, samples were analyzed with monoclonal antibody C5F10 by Western blot. The antibody specifically recognized, in fresh and frozen/thawed spermatozoa, a 40-, 32- and 27-kDa bands corresponding to proacrosin, alpha- and beta-acrosin, respectively, during capacitation. Western immunoblots showed that the beta-acrosin reactivity in fresh sperm was directly proportional to the time of capacitation, whereas a decreased reactivity of active form of acrosin was observed with frozen-thawed sperm (p   < 0.05). These results suggest that proacrosin is activated to beta-acrosin earlier in frozen/thawed dog spermatozoa than in fresh dog spermatozoa.     

Giant cell tumour of bone in a cat with extraskeletal metastases: pathological and immunohistochemical study

A case of giant cell tumour of bone (GCTb) in the lung and in a subcutaneous mass located in the right flank, with a probable primary origin in the mid-diaphysis of the right tibia, was described in a 8-year-old female cat. Numerous multinucleated giant cells were homogeneously distributed among a population of ovoid or spindle-shaped mononuclear cells. All of them were positive for vimentin suggesting a mesenchymal origin. Spindle-shaped tumour cells resemble fibroblastic cells, showing collagen fibres in their vicinity. Ovoid mononuclear cells are similar to macrophages, with a cytoplasm rich in electron-dense lysosomes. Multinucleated giant cells appear morphologically similar to osteoclasts. These findings are supported for the positive reaction to tartrate-resistant acid phosphatase (TRAP) and lysozyme, encountered only in ovoid and multinucleated giant cells. No immunoreactivity against human oestrogen receptors was observed in the nuclei of any neoplastic cells.     

Enrofloxacin assay validation and pharmacokinetics following a single oral dose in chickens

The pharmacokinetics of enrofloxacin (ENRO), a fluoroquinolone antimicrobial agent, was studied in male broiler chickens (Cobb) after single oral administration of 10 mg of ENRO/kg b.w. A high-performance liquid chromatography-photodiode array detector (DAD) (HPLC-DAD) method was developed and validated and used for quantitation of ENRO and its major metabolite ciprofloxacin in plasma. The HPLC analyses were carried out using a cationic-octadecyl mixed column and 0.05 mol/L phosphate buffer (pH 2.5)/acetonitrile as mobile phase. The sample preparation of plasma consisted of the precipitation of proteins followed by solid phase extraction on cationic-octadecyl mixed cartridges. The method was validated considering linear range, linearity, selectivity, sensitivity, limit of detection (LOD), limit of quantitation (LOQ), intra- and inter-day precisions and accuracy. The LOD and LOQ for both fluoroquinolones were 60 and 200 ng/mL for plasma. The plasma concentration vs. time graph was characteristic of a two-compartment open model. The maximal plasma concentration of 1.5 +/- 0.2 mg/mL was achieved at 9 +/- 2 h. The elimination half-life and the mean residence time of ENRO were 1.5 +/- 0.2 and 15.64 h, respectively. The area under the concentration-time curve was calculated as 35 +/- 4 mgxh/mL.