Cloning of bradykinin precursor cDNAs from skin of Bombina maxima reveals novel bombinakinin M antagonists and a bradykinin potential peptide

Bombinakinin M (DLPKINRKGP-bradykinin) is a bradykinin-related peptide purified from skin secretions of the frog Bombina maxima. As previously reported, its biosynthesis is characterized by a tandem repeats with various copy numbers of the peptide and sometimes co-expressed with other structure-function distinguishable peptides. At present study, two novel cDNAs encoding bombinakinin M and its variants were cloned from a cDNA library from the skin of the frog. The encoded two precursor proteins are common in that each contains three repeats of a novel 16-amino acid peptide unit and one copy of kinestatin at their N- and C-terminal parts, respectively. They differ in that the first precursor contains two copies of bombinakinin M and the second one contains one copy of a novel bombinakinin M variant. Bombinakinin M was found to elicit concentration-dependent contractile effects on guinea pig ileum, with an EC50 value of 4 nM that is four times higher than that of bradykinin (1 nM). Interestingly, the synthetic peptide (DYTIRTRLH-amide), as deduced from the 16-amino acid peptide repeats in the newly cloned cDNAs, possessed weak inhibitory activity on the contractile effects of bombinakinin M, but not on that of bradykinin. Furthermore, the newly identified bombinakinin M variant (DLSKMSFLHG-Ile(1)-bradykinin), did not show contractile activity on guinea pig ileum, but showed potentiation effect on the myotropic activity of bradykinin. In a molar raito of 1:58, it augmented the activity of bradykinin up to two-fold. (C) 2004 Elsevier B.V. All rights reserved.

Humoral and protective response of Indian major carps to immersion vaccination with Aeromonas hydrophila

Fry of the Indian major carps, Catta catla (Ham.), Labeo rohita (Ham.) and Cirrhinus mrigala (Ham.) were immunized at 4 and 8 weeks post hatching (wph) by direct immersion in a suspension (10 super(8) cells ml super(-1))of heat inactivated Aeromonas hydrophila. Following the same procedure, booster dose was administered 20 days after the first immersion. Antibodies as well as protective response produced in both the groups after the first and the booster immersion were different and significant (P<0.05). No significant difference was found between the species in the two age groups. The specimens immunized 8 wph showed higher antibody titres and protection than the 4 wph group. C. catla had higher relative percent survival followed by L. rohita and C. mrigala.

Game Vertebrate Densities in Hunted and Nonhunted Forest Sites in Manu National Park, Peru

Manu National Park of southern Peru is one of the most renowned protected areas in the world, yet large-bodied vertebrate surveys conducted to date have been restricted to Cocha Cashu Biological Station, a research station covering <0.06 percent of the 1.7Mha park. Manu Park is occupied by >460 settled Matsigenka Amerindians, 300-400 isolated Matsigenka, and several, little-known groups of isolated hunter-gatherers, yet the impact of these native Amazonians on game vertebrate populations within the park remains poorly understood. On the basis of 1495 km of standardized line-transect censuses, we present density and biomass estimates for 23 mammal, bird, and reptile species for seven lowland and upland forest sites in Manu Park, including Cocha Cashu. We compare these estimates between hunted and nonhunted sites within Manu Park, and with other Neotropical forest sites. Manu Park safeguards some of the most species-rich and highest biomass assemblages of arboreal and terrestrial mammals ever recorded in Neotropical forests, most likely because of its direct Andean influence and high levels of soil fertility. Relative to Barro Colorado Island, seed predators and arboreal folivores in Manu are rare, and generalist frugivores specializing on mature fruit pulp are abundant. The impact of such a qualitative shift in the vertebrate community on the dynamics of plant regeneration, and therefore, on our understanding of tropical plant ecology, must be profound. Despite a number of external threats, Manu Park continues to serve as a baseline against which other Neotropical forests can be gauged.

Monoculture of three native air breathing fishes in homestead ditches and plastic barrels

Six on-farm trials were conducted from 1 August to 23 November 2004 in two different environments such as homestead ditches (10 to 17 square meters) and plastic barrels (240 liters) to develop techniques for nonoculture [sic] of climbing perch, Anabas testudineus, stinging catfish, Heteropneustes fossilis and walking catfish, Clarias batrachus for poor and landless people who have no access to pond. Stocking density for ditch was 10 fry/square meter while that for barrel was 20 fry/cubic meter. The fishes were fed with 3-test diets viz. low-cost formulated feed (rice bran 20%, wheat meal 10%, mustered [sic] oil cake 35%, poultry offal 35%), live foods (chopped snails and clams), and a commercial feed (Saudi-Bangla feed, starter 3: first month and grower-1: subsequent two months) and designated as T1, T2 and T3, respectively. Feeding rate was the same in all the treatments viz. 10% of body weight (first two months), 8% (third month) and 6% (fourth month). T1 and T2 had three replications while T3 had two replications. Water temperature was recorded weekly while fish growth was monitored monthly. After 4 months' rearing, H. fossilis and C. batrachus in ditches and barrels attained higher average weight in T2 followed by T3 and T1 while A. testudineus in barrels also attained higher average weight in T2. The variation in net weight gain by A. testudineus in ditch fed test diets T2 and T3 was not significantly different (P>0.05) though the net gain in both T2 and T3 was significantly (P<0.05) higher than that of T1. The yield of climbing perch as obtained from T1, T2 and T3 was 988, 1136 and 1185 kg/ha, respectively while that stinging catfish was 395, 242 and 444 kg/ha and walking catfish was 1605, 2,099 and 1,654 kg/ha respectively. All the three species showed significantly lower growth rate in barrels than in ditches.

Influence of sex, spawning, starvation and water temperature on fatty acid composition in Tilapia mossambica

The influence of sex, spawning, starvation and water temperature on the fatty acid composition of Tilapia mossambica has been studied. Tilapia egg lipid was found to have unusually high percentage of C sub(22:6) fatty acids (9.09%) compared to body and intestinal lipids. The C sub(16:1) acid was much less in the egg lipids (3.5%) whereas it was 11% in the body lipids. There was no significant difference in the fatty acid composition of body and intestinal lipids of male and female tilapia. Starvation caused the presence of high content of lower fatty acids (C sub(6), C sub(8), C sub(30), C sub(12) and C sub(33)) in the body lipids. Water temperature also influenced the fatty acid composition of Tilapia; the difference was more significant in body lipids than in intestinal lipids.

Hydrobiological studies of an irrigation tank with reference to the culture of large murrel, Channa marulius (Hamilton) and striped murrel Channa striatus (Bloch)

The importance of selection of species for culture according to the ecological niches and fish food organisms is highlighted with respect to the Fox Sagar, an irrigation take. The tank was infested with submerged vegetation as well as minnows and weed fishes, which rendered the tank unsuitable for the culture of Indian major carps. The tank was stocked with 8000 fingerlings of Channa marulius and C. striatus during 1981 by the local fisherman co-operative society. Only partial harvest was possible during 1982 because of high water level. The final harvest was in April-May, 1983. The yield obtained was 3640 kg during the culture period of about 20 months.

Determination of acute mercury toxicity to developing stages of Cyprinus carpio and Cirrhinus mrigala

Toxicity of inorganic mercury to different life history stages of fresh water fishes, Cyprinus carpio and Cirrhinus mrigala were demonstrated by static bioassays. 48 and 94% of egg hatching occurred in controls at 72 and 24h of experimentation in C. carpio and C. mrigala respectively. While fish eggs in water containing mercuric chloride showed delayed development as compared to the control. LC50, LC100 and safe concentrations of hatchling, fry and fingerling were calculated. Hatchling and fry were observed to be more susceptible as compared to fingerlings of C. carpio and C. mrigala.

Growth and survival of Penaeus monodon Fabricius zoeae on different diatom feeds

A study was undertaken to determine the most suitable food for Penaeus monodon zoea out of mixed diatoms, Skeletonema costatum, Chaetoceros calcitrans and C. gracilis. Results indicate that mixed diatoms and C. gracilis are the better food. Addition of zooplankton, e.g.rotifers, is also recommended as early as Z3 since the larvae start to be omnivorous at this stage.

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

KARYOTYPE AND BANDING-PATTERNS OF FEA TREE RAT (CHIROMYSCUS-CHIROPUS)

Fea's tree rat (Chiromyscus chiropus) is a very rare species which there are only a few specimens in the world. The chromosomes of two male specimens, collected from Xishuanbanna, Yunnan, are analysed by several banding technique (G-, C-bands, as well as Ag-staining). The diploid chromosome number is 22, and autosomes comprise 5 pairs of metacentrics, 2 pairs of subacrocentrics, and 3 pairs of acrocentrics. The X chromosome is a acrocentric, and Y is a micro-chromosome, almost a point, which could be a marker chromosome of the species and the genus. The centromeric C-bands are very faint, and C-bands of Nos. 1, 2, 9 and Y chromosome are negative. Only one pair Ag-NORs was found on No. 10 in the silver-stained karyotype. The relationship between morphologic and chromosomal features was discussed, and C-banded karyotype evolutionary trend has also been discussed. Moreover, the conventional karyotype of Niviventer confucianus was described.

Survey of biosystematic and sexual reproductive physiology in Holothuroidea on Hormozgan Province

Sea cucumbers belong to phylum Echinodermata, order Holothuroidea are an abundant and diverse group of Invertebrates, with over 1400 species occuring from the intertidal to the deepest oceanic trenches. Sea cucumbers are important components of the food chain in temperate and coral reef ecosystems and they play an important role as deposite feeders and suspension feeders. Rapid decline in populations may have serious consequences for the survival of other species that are part of the same complex food web,as the eggs, larve and juveniles constitute an important food source for the other marine species including crustaceans, fish and mollusks. In addition sea cucumbers are often called the earthworms of the sea, because they are responsible for the extensive shifting and mixing of the substrate, and recycling of detrital matter. Sea cucumbers consume and grind sediment and organic material into finer particles , turning over the top layers of sediment in lagoons , reefs and other habitats and allowing the penetration of oxygen. While the taxonomy of the holothurian families is generally well known , the distinction of similar species is difficult. There are relatively few holothurian taxonomist.Most sea cucumber species can be identified by Holothurin taxonomists by using the calcareous skeletal ossicles found in the body wall. In this study , at first a sea cucumber from Kish island in Persian gulf has recognized. Individuals collected from west and east extend far away into north and south of coral reefs by diving. I have checked them morphologically and anatomically.Then with key to the orders of the Holothuroidea, They belong to the Aspidochirotida with key to the families of Aspidochirotida, they were in Stichopodidae families and with key to the genus of Stchopodidae, they were Stichopus. Then ossicles were extracted at National Museum of Natural History, by Dr David Pawson. The ossicles were measured on a transect across a slide prepared from the mid-dorsal region of each specimen.The one we have in the shallow waters of Kish island, is Stichopus hermanni, a massive holothurian, body broad, considerably flattened ventraly ,the dorsal side slightly arched and the lateral sides almost vertical; body wall fairy thick and soft ; mouth subterminal; anus central; tentacles usually 20 in number of length and leaf shaped. Numerous ossicles consisting of table with large discs having usually 7 to 15 peripheral holes, but often irregular or incomplete and spire of moderate height ending in a group of spinelets, rosettes of variable development, and c-shaped rods. Color (exept papillae)partly remained after preservation in alcohol which is found at the depth of 4 to 8 meters, on coral reef. Furthermore, the sexual reproductive cycle was described using standard methods. Gonads were removed and transferred to Bouin's fixative for four weeks and then processed according to standard embedding technique. To prevent the loss of tubule contents during embedding, the tubule sections, were cut well beyond the segment selected for sectioning. For each individual, six sections, each section with 5µm diameter by microtome were cut from tubules. These sections were first placed on gelatin coated slides (the gelatin was heated to 42°c) and then transferred to the oven at 37°c for one hour. This technique usually prevents the fragil tubules from breaking and the loss of gametes. The slides were stained with Eosin and Hematoxylin, and good resolution of the various cell types achieved.A second series of slides was stained with the Periodic Acid Schiff(PAS) to identify polysaccharides(glycogen). Monthly sampling was occurred.The sexual reproductive cycle was defined through the combined use of these criteria: Monthly percentages of the gonad stages for each sex, the monthly gonad index (GI) , given as the ratio of the wet gonad weight (G) to the dray weight (DW)and the monthly percentage of individuals that undetermined sex. The gonad consists of two tufts of tubules on which saccules develop. Gonadal development was classified into five stages: post spawning, recovery, growth, advanced growth, and mature stage that were adapted from the earlier studies of holothurians. Histological preparations showed that the sex of larger individuals could be identified by the presence of oogonia and young oocytes in females, and spermatogonic stages in males.The mean diameter of the tubules and gonadal mass follow annual cycles, increasing from late winter through spring, and dropping abruptly after spawning in the summer. Gametogenesis is generally a prolongate process and begins in March. By summer the ovarian tubules contain oocytes with diameter of 120-240 pm and the testicular tubules contain an abundance of spermatozoa (diameter 5-6 gm ).Following spawning the predominant activity within the spent tubules is phagocytosis of the residual gamets.The active phase of gametogenesis (March to July), coincides with an increasing photoperiod regim, and an accelerated gametogenesis occurs in July when temperature is high. Throughout the year, the gonad of Stichopus hermanni is larger in males than in females, and this is due to the number of tubules in the testis rather than to tubules length or diameter.

Matrilineal Components and Genetic Relationship of Silkies from China and Japan

Silkie is a famous black-bone chicken breed with beautiful silky feather. The unique medical property of this chicken was recorded in Chinese traditional medicine dictionary about 700 years ago. In this study, we analyzed the mtDNA D-loop sequence variation of 26 Bairong Silkies from Fujian Province, China, together with 100 reported Silkie mtDNAs from China and Japan, and studied their matrilineal components and genetic relationship. A total of 21 haplotypes were detected, which could be assigned to six haplogroups (A-E, G). Among them, haplogroups D and G were exclusively presented in Japanese Silkies and Chinese Silkies, respectively. Chinese Silkies had higher frequency of lineages belonging to haplogroups A, B, and E, and lower frequency of haplogroup C than Japanese Silkies. For the four Chinese Silkie populations, most of samples of Taihe, Chengdu, and Hubei Silkies were grouped in haplogroups A, B, and C, whereas most of Bairong Silkies were grouped in haplogroup E. Five haplotypes were shared by Japanese and Chinese Silkies. The genetic diversity of each Silkie population varied, but the overall diversity of Chinese Silkies was similar to that of Japanese Silkies. Taken together, our results confirmed the genetic connection between Chinese and Japanese Silkies, but also clearly showed that the matrilineal genetic structures of Chinese and Japanese Silkies had some differences.

蝙蝠科三种蝙蝠的G-带和C-带

对肺、心等进行组织培养,用空气干燥法制作染色体标本,对贵州3种蝙蝠即中华鼠耳蝠(Myotischinensis)、西南鼠耳蝠(M.altarium)和亚洲长翼蝠(Miniopterus fuliginosus)进行了G-带、C-带带型分析.结果表明,2种鼠耳蝠的G-带基本相同,亚洲长翼蝠的G-带与两种鼠耳蝠有一定同源性;C-带核型中,中华鼠耳蝠和亚洲长翼蝠只有着丝粒带,而西南鼠耳蝠有的染色体有插入C-带和端位C-带.根据带型异同分析讨论了鼠耳蝠和长翼蝠间的进化关系.

Parameter extraction and calorimetric validation for a silicon carbide jfet pspice model

This paper focuses on the PSpice model of SiC-JFET element inside a SiCED cascode device. The device model parameters are extracted from the I-V and C-V characterization curves. In order to validate the model, an inductive test rig circuit is designed and tested. The switching loss is estimated both using oscilloscope and calorimeter. These results are found to be in good agreement with the simulated results.

Comparison of manual and user guided methodologies for the classification and retrieval of construction site images

The amount of original imaging information produced yearly during the last decade has experienced a tremendous growth in all industries due to the technological breakthroughs in digital imaging and electronic storage capabilities. This trend is affecting the construction industry as well, where digital cameras and image databases are gradually replacing traditional photography. Owners demand complete site photograph logs and engineers store thousands of images for each project to use in a number of construction management tasks like monitoring an activity's progress and keeping evidence of the "as built" in case any disputes arise. So far, retrieval methodologies are done manually with the user being responsible for imaging classification according to specific rules that serve a limited number of construction management tasks. New methods that, with the guidance of the user, can automatically classify and retrieve construction site images are being developed and promise to remove the heavy burden of manually indexing images. In this paper, both the existing methods and a novel image retrieval method developed by the authors for the classification and retrieval of construction site images are described and compared. Specifically a number of examples are deployed in order to present their advantages and limitations. The results from this comparison demonstrates that the content based image retrieval method developed by the authors can reduce the overall time spent for the classification and retrieval of construction images while providing the user with the flexibility to retrieve images according different classification schemes.