Can differences between continental and insular habitats influence the parasites communities associated with the endemic frog Haddadus binotatus?

Habitats characterized by geographic isolation such as islands have been studied using different organisms as models for understanding the dynamic and insular patterns of biodiversity. Determinants of parasite richness in insular host populations have been conducted mainly with mammals and birds, showing that parasite richness decreases in insular areas. In the present study, we predicted that the type of environment (insular or continental) can influence the richness, diversity and abundance of parasites associated with the endemic frog Haddadus binotatus (Spix, 1824). We sampled frogs in two insular and two mainland fragments to survey their helminth parasites. The total richness was composed of 15 taxa of Nematoda and two of Acanthocephala, and the community composition of the two islands had more similarities between them than the two mainland localities. The insular effect was positive for richness and abundance of helminths, and no significant effect was observed on helminth diversity - even the mean diversity presented high numbers for the islands. We presumed that insular hosts could have lost some parasites in the colonization process when these continental islands were separated from the mainland, approximately 11,000 years ago. However, the high richness and abundance on islands can be explained by an epidemiological argument, which considers high population density due to insularity and other features of the host as factors that increase parasite transmission success among individuals.

Genome-wide association mapping and examination of possible maternal effect for the pace trait of horses.

Previously, a single nucleotide polymorphism (SNP) related to gait type was identified at position 22 999 655 of chromosome 23 in the coding region of DMRT3 (DMRT3:Ser301Ter) by showing that a cytosine (C) to adenine (A) mutation of this SNP induced pace in the Icelandic horse. We investigated the effect of DMRT3:Ser301Ter on the gait of Hokkaido Native Horses, a Japanese native breed, and examined genetic factors other than DMRT3 by exploring genome-wide SNPs related to gait determination. All animals exhibiting pace were AA for DMRT3:Ser301Ter, confirming the association of DMRT3:Ser301Ter with gait determination; however, 14.3% of the animals exhibiting trot also had AA for DMRT3:Ser301Ter, suggesting the presence of another factor(s) cooperatively working with DMRT3:Ser301Ter for gait determination. SNPs on chromosomes 13 and 23 were detected by genome-wide association analysis (false discovery rate <0.05), although SNPs on chromosome 23 were all located in the vicinity of DMRT3:Ser301Ter, confirming the association with DMRT3. A genome-wide association study targeting only animals with AA for DMRT3:Ser301Ter to examine genetic factors cooperatively working with DMRT3:Ser301Ter for gait determination suggested associations of 23 SNPs on six chromosomes. In a series of analyses of the effect of a maternal factor (dam's gait) on gait determination, the effect was suggested in comparison of the frequencies of exhibiting pace in gait checks in only two animal groups having dams with different DMRT3:Ser301Ter genotypes (P < 0.05), suggesting that the gait of the dam does not have a major effect on whether progeny homozygous for the DMRT3:Ser301Ter mutation will preferentially pace or trot.

Biological index of environmental lead pollution: accumulation of lead in liver and kidney in mice.

Lead (Pb) is known to be highly poisonous, and the acute poisoning of Cd causes the abdominal pains, vomiting, and shock. The digestive and nervous symptom is observed in the chronic lead poisoning. It was also known that the defect in hemoglobin synthesis by Pb produce anemia. The release of Pb into the environment presents a source of exposure for wild animals. In this study, we examined the utility of a new Pb-monitoring index in mice administered Pb. A solution containing 0.02, 0.2, 2, or 4 ppm lead chloride (PbCl2) was administered intraperitoneally to mice, and the Pb contents of the kidney and liver were determined at designated time points. The mean Pb content of both organs increased depending on the administered Pb dosage. Although the results of control was near the detection limits, the administration of 4 ppm in 4 weeks resulted in Pb levels of 260 mg ppm/wet weight and 110 ppm wet weight in the kidney and liver, respectively. However, there were no significant relationships among administered dose, duration of Pb treatment, and liver or kidney Pb content. Then, values in all mice administered control or 0.02 mg Pb were located inside the ellipse, representing the confidence area of the new index, and values in all mice administered more than 2 mg Pb were located outside the ellipse. These results confirm that animals exposed to high concentrations of Pb would be detected by this new index.

Gill ectoparasite assemblages of two non-native Cichla populations (Perciformes, Cichlidae) in Brazilian reservoirs.

The gills of 41 Cichla piquiti and 39 C. kelberi from Itaipu and Lajes reservoirs, respectively, Brazil, were examined to describe the ectoparasite assemblages of these two non-native peacock-bass populations. All ectoparasite species of the two studied hosts (C. piquiti and C. kelberi) were dominant, but Ascocotyle sp. (metacercariae) was the prevalent (58.53%) and most abundant helminth species in C. piquiti hosts, while Sciadicleithrum ergensi was the dominant species in C. kelberi hosts. Gill ectoparasites of C. piquiti and C. kelberi showed a typical pattern of overdispersion or aggregation, which is commonly reported for many other freshwater fishes. Ectoparasite prevalence and abundance did not vary between host sexes of the two Cichla populations. The prevalence and abundance of Ascocotyle sp. were positively correlated with C. piquiti standard length (SL), but only the abundance of S. ergensi showed a positive correlation with C. kelberi SL. Although environmental differences between reservoirs might also have influenced the results, we anticipated that the presence of a close congener in Itaipu reservoir and the lack of other Cichla species in Lajes reservoir were the key factors to explain the contrasts between C. piquiti and C. kelberi gill ectoparasites. Overall, our results suggest that the trend of parasite species loss through the invasion process may have contributed to the establishment of non-native C. piquiti and C. kelberi populations in Brazilian reservoirs.

Diversity of parasites of fish from the Upper Paraná River floodplain, Brazil.

The aim of the present study was to provide a current survey of the species of parasites found in the Upper Paraná River floodplain, as well as to investigate which strategies and mechanisms used by parasites, are favored and which environmental factors influence the parasite community in the studied environments. During a seven-year period from February 2000 to September 2007, 3,768 fish belonging to 72 species were collected and analyzed for the purpose of studying the parasite fauna. A total of 337 species of parasites were reported, including 12 new descriptions: one myxosporid, Henneguya paranaensis Eiras, Pavanelli and Takemoto, 2004; eight monogeneans, Kritskyia annakohnae Boeger, Tanaka and Pavanelli, 2001; Kritskyia boegeri Takemoto, Lizama and Pavanelli, 2002; Kritskyia eirasi Guidelli, Takemoto and Pavanelli, 2003; Demidospermus labrosi França, Isaac, Pavanelli and Takemoto, 2003; Demidospermus mandi França, Isaac, Pavanelli and Takemoto, 2003; Pseudovancleaveus paranaensis França, Isaac, Pavanelli and Takemoto, 2003; Tereancistrum curimba Lizama, Takemoto and Pavanelli, 2004 and Tereancistrum toksonum Lizama, Takemoto and Pavanelli, 2004; two digeneans, Sanguinicola platyrhynchi Guidelli, Isaac and Pavanelli, 2002 and Dadayius pacupeva Lacerda, Takemoto and Pavanelli, 2003 and one cestode, Nomimoscolex pertierrae Chambrier, Takemoto and Pavanelli, 2005. In addition, several other species were reported for the first time in new hosts or in the floodplain. Monogeneans presented the highest number of species, followed by digeneans. The infection site with the highest species richness was the intestine, with 127 species.

Diversity of parasites of fish from the Upper Paraná River floodplain, Brazil / Diversidade dos parasitos de peixes da planície de inundação do Alto Rio Paraná, Brasil

The aim of the present study was to provide a current survey of the species of parasites found in the Upper Paraná River floodplain, as well as to investigate which strategies and mechanisms used by parasites, are favored and which environmental factors influence the parasite community in the studied environments. During a seven-year period from February 2000 to September 2007, 3,768 fish belonging to 72 species were collected and analyzed for the purpose of studying the parasite fauna. A total of 337 species of parasites were reported, including 12 new descriptions: one myxosporid, Henneguya paranaensis Eiras, Pavanelli and Takemoto, 2004; eight monogeneans, Kritskyia annakohnae Boeger, Tanaka and Pavanelli, 2001; Kritskyia boegeri Takemoto, Lizama and Pavanelli, 2002; Kritskyia eirasi Guidelli, Takemoto and Pavanelli, 2003; Demidospermus labrosi França, Isaac, Pavanelli and Takemoto, 2003; Demidospermus mandi França, Isaac, Pavanelli and Takemoto, 2003; Pseudovancleaveus paranaensis França, Isaac, Pavanelli and Takemoto, 2003; Tereancistrum curimba Lizama, Takemoto and Pavanelli, 2004 and Tereancistrum toksonum Lizama, Takemoto and Pavanelli, 2004; two digeneans, Sanguinicola platyrhynchi Guidelli, Isaac and Pavanelli, 2002 and Dadayius pacupeva Lacerda, Takemoto and Pavanelli, 2003 and one cestode, Nomimoscolex pertierrae Chambrier, Takemoto and Pavanelli, 2005. In addition, several other species were reported for the first time in new hosts or in the floodplain. Monogeneans presented the highest number of species, followed by digeneans. The infection site with the highest species richness was the intestine, with 127 species.

O objetivo do presente trabalho foi realizar um novo levantamento das espécies de parasitos encontradas na planície de inundação do Alto Rio Paraná, bem como investigar quais estratégias e mecanismos utilizados pelos parasitos são mais favorecidos e quais fatores ambientais estão influenciando a comunidade parasitária nos ambientes estudados. Durante o período de fevereiro de 2000 a setembro de 2007, 3.768 espécimes de peixes pertencentes a 72 espécies foram coletados e analisados, visando o estudo da fauna parasitária. Até o momento foram registradas, 337 espécies de parasitos, sendo 12 novas espécies: uma de mixosporídeo, Henneguya paranaensis Eiras, Pavanelli e Takemoto, 2004; oito de monogenéticos, Kritskyia annakohnae Boeger, Tanaka e Pavanelli, 2001; Kritskyia boegeri Takemoto, Lizama e Pavanelli, 2002; Kritskyia eirasi Guidelli, Takemoto e Pavanelli, 2003; Demidospermus labrosi França, Isaac, Pavanelli e Takemoto, 2003; Demidospermus mandi França, Isaac, Pavanelli e Takemoto, 2003; Pseudovancleaveus paranaensis França, Isaac, Pavanelli e Takemoto, 2003; Tereancistrum curimba Lizama, Takemoto e Pavanelli, 2004 e Tereancistrum toksonum Lizama, Takemoto e Pavanelli, 2004; duas de digenéticos, Sanguinicola platyrhynchi Guidelli, Isaac e Pavanelli, 2002 e Dadayius pacupeva Lacerda, Takemoto e Pavanelli, 2003 e uma de cestóide, Nomimoscolex pertierrae Chambrier, Takemoto e Pavanelli, 2005. Além dessas novas espécies, várias outras foram registradas pela primeira vez em novos hospedeiros ou na planície. O grupo dos monogenéticos foi o que apresentou maior número de espécies encontradas, seguido pelos digenéticos. O intestino foi o sítio de infecção que apresentou a maior riqueza, com 127 espécies.

Swift and definite serotyping for isolated Listeria monocytogenes strains.

The serotype is most important for molecular epidemiological analysis of Listeria monocytogenes (L.m.) contaminating marketed meats. An improvement on the traditional method was thus attempted in the present study because of the requirement of swift and definite serotyping. In the determination of O-antigen, definite judgement was allowed by an immediate cooling at 80 degrees C after autoclaving the bacteria. In the determination of H-antigen, use of a culture plate without Craigie's tube yielded the active bacteria only by single culture. The stable and clear agglutination in many samples was also obtained with a microplate using less antiserum. The availability was confirmed with 123 strains and the serovar 1/2b was dominant in the Japanese strains.

[Basic study on the sensitivity test for inhibiting fibroblast proliferation (II)].

The results of in vitro tests against solid tumors have been generally influenced by the mixing in of fibroblasts, and so have SDI tests. We demonstrated that the influence of fibroblasts can be excluded by performing the SDI test on an agar layer, thus significantly inhibiting the growth of fibroblasts in the liquid top layer. In the agar SDI test, SDS is used to dissolve formazan; however, this lowers the sensitivity to some extent. We added additional DMSO to SDS and to raise the sensitivity.

The chemistry of indoles. CVII. A novel synthesis of 3,4,5,6-tetrahydro-7-hydroxy-1H-azepino[5,4,3-cd]indoles and a new finding on Pictet-Spengler reaction.

Serotonins were found to produce 3,4,5,6-tetrahydro-7-hydroxy-1H-azepino[5,4,3-cd]indoles by simple heating with amines under an oxygen atmosphere. Serotonins also reacted with various aldehydes to provide 3,4,5,6-tetrahydro-7-hydroxy-1H-azepino[5,4,3-cd]indoles rather than beta-carbolines under basic conditions. In these novel reactions, the presence of the 5-hydroxy group on the indole nucleus was suggested to be essential. Possible mechanisms are discussed.

[Fundamental study on the sensitivity test inhibiting fibroblast proliferation].

Although many kinds of chemosensitivity test are available for the selection of suitable anticancer agents, the results of in vitro tests against solid tumors have been generally influenced by mixed in fibroblasts, as have SDI tests. We demonstrated that the influence of fibroblasts can be excluded by performing the SDI test on an agar layer, thus significantly inhibiting the growth of fibroblasts in the liquid top layer. The sensitivity of the agar SDI test was determined on the 3-4th day after incubation because cell proliferation tended to be delayed, and there were also somewhat higher cell counts of about 40,000 per well. The inhibition indices with and without agar in vitro were the same, showing no significant differences. With nude mice, the transplanted tumor index value of the agar SDI test is higher than that of SDI, and approaches in value the SDI of a pure tumor cell, which means that the sensitivity of a solid tumor might be more accurately shown by an agar SDI test than by an SDI test.

The chemistry of indoles. CIII. Simple syntheses of serotonin, N-methylserotonin, bufotenine, 5-methoxy-N-methyltryptamine, bufobutanoic acid, N-(indol-3-yl)methyl-5-methoxy-N-methyltryptamine, and lespedamine based on 1-hydroxyindole chemistry.

Application of regioselective nucleophilic substitution reactions of 1-hydroxytryptamines to novel and simple syntheses of serotonin (1a), N-methylserotonin (1b), bufotenine (1c), 5-methoxy-N-methyltryptamine (2a), bufobutanoic acid (3a), N-(indol-3-yl)methyl-5-methoxy-N-methyltryptamine (4), and lespedamine (5) are described. Effective syntheses of 5-benzyloxytryptamine and 1-methoxy-2-oxindoles are also reported.

[Hepatic arterial injection therapy (HAI) for metastatic liver tumor from gastric cancer].

The results and problems of hepatic artery infusion therapy (HAI) for gastric carcinoma with synchronous liver metastasis were evaluated. The response rate of HAI with CDDP and 5-FU for metastatic liver tumor was 55% (1 CR + 5 PR/11). The median survival time for responders was 16.5 months, which was statistically longer than that of non-responders at only 5.5 months. Histologically, most responder cases were with AFP producing tumors and NSE positive tumors without distant lymph node involvement. Non-responder cases developed marked distant lymph node involvement besides the liver metastasis. Most of responder patients died of lymph node recurrence or distant metastasis other than liver tumor. It may be concluded that additional therapy to HAI is needed to improve the prognosis of gastric cancer patients with multiple liver metastases.

Specific immune response to a synthetic peptide derived from outer surface protein C of Borrelia burgdorferi predicts protective borreliacidal antibodies.

In a previous study, we described the development of a new specific serodiagnostic test for Lyme disease involving enzyme-linked immunosorbent assay and a synthetic peptide, OspC-I. The OspC-I peptide is derived from part of the outer surface protein C (OspC) amino acid sequence of Borrelia burgdorferi and is located in the region conserved among B. burgdorferi sensu stricto or sensu lato isolates. In this study, we demonstrate that sera containing antibodies against OspC-I from patients with early Lyme disease had borreliacidal activity against isolates of three genospecies of Lyme disease spirochete, B. burgdoreferi B31, B. garinii HPI and B. afzelii HT61. However, the borreliacidal activity against B. burgdorferi, which has not been isolated in Japan, was weaker than that against the other species. Vaccination of mice with OspC-I induced the production of anti-OspC-I antibodies in serum with borreliacidal activity. The immune mouse serum had significantly higher levels of borreliacidal activity against HP1 and HT61, than against B31. Neutralization of borreliacidal activity with anti-IgM antibodies showed that the borreliacidal activity of anti-OspC-I antibodies in serum was due to IgM. Furthermore. mice vaccinated with OspC-I were protected against challenge with HPI and HT61. but not fully protected against infection with B31. These results suggest that OspC-I is not only a specific antigen for use in serodiagnostic tests for Lyme disease, but is also a potential candidate for a Lyme disease vaccine in Japan.

[Development of synthetic methods for 4-substituted indoles and their applications for the syntheses of natural products].

In our continuing work on synthesizing 4-substituted indole alkaloids as simple as possible by creating suitable reactions, we developed synthetic methods for 4-substituted indoles having a nitrogen or oxygen functional group at the 4-position starting from indole-3-carbaldehyde and one step conversion method of indole-3-carbaldehydes into indole-3-acetonitriles. Utilizing them, we could establish a practical synthetic route to 1,3,4,5-tetrahydropyrrolo[4,3,2-de]-quinoline in three steps from indole-3-carbaldehyde. On the basis of these results, short step total syntheses of marine indole alkaloids, such as isobatzelline C, batzelline C, makaluvamine A, and damirones A and B, were achieved. Furthermore, a novel preparative method of psilocin was established in only five steps from indole-3-carbaldehyde. The syntheses of psilocin analogs having a formyl group or bromine in the benzene part were also achieved.

Mucin ball-producing extrahepatic bile duct carcinoma.

The characteristic features of surgically curable mucin-producing extrahepatic bile duct carcinoma (MPEBC) have not previously been elucidated. Three (6.5%) of 46 patients who underwent surgery in our department for bile duct carcinoma between 1986 and 1997 had MPEBC. Clinicopathological features, diagnostic procedures and operative methods for patients with MPEBC were investigated. Tumors in the bile duct were identified by cholangioscopy combined with cholangiography after removal of mucin balls. Tumors were located close to the hepatic confluence in these patients. Two patients underwent hepatic lobectomy together with caudate lobectomy while the other underwent resection of the hepatic confluence. Absence of residual tumors was confirmed histologically in these patients. All three patients remain alive without evidence of recurrence, 22-54 months after surgery. MPEBC is a curable disease. Accurate localization in the biliary tree is essential and can only be obtained after, i) removal of mucin balls, and ii) extensive diagnostic work-up including cholangiography, cholangioscopy and intraoperative pathological examination.