Effect of yeast products supplementation during transition period on metabolic profile and milk production in dairy cows.

The aim of the current study was to assess the effect that supplementation with yeast culture plus enzymatically hydrolyzed yeast (YC-EHY) during the transition period and lactation had on the performance, somatic cell count (SCC), and metabolic profile of dairy cows. Thirty multiparous Holstein dairy cows were divided into two groups. The treatments were 0 supplementation (control) and supplementation with 28 g/cow/day of YC-EHY. The supplementation began 35 ± 5 days before the expected calving date. The cows were kept in their respective treatments for 50 days after the calving date. Body condition score (BCS), body weight, milk composition, SCC, and milk yield were assessed on a 2-weekly basis. Plasma samples were collected on days - 21st, - 7th, 0, 3rd, 7th, and weekly thereafter until 42 days postpartum and analyzed for albumin, ß-hydroxybutyrate (BHBA), urea, and non-esterified fatty acids (NEFA). There was an effect of treatment on milk yield in the supplemented animals in comparison to the control group (27.88 ± 0.98 vs 24.58 ± 0.99 kg/days, P = 0.03). There was no effect of treatment (P > 0.05) on variables like 3.5% fat-corrected milk (FCM) and energy-corrected milk (ECM), milk component (%), milk composition yield (kg/day), and SCC. There was an interaction between group × days on ECM (P = 0.04) and protein (P = 0.008). The supplementation had no effect on the analyzed metabolites and on body weight and BCS. The supplementation with YC-EHY during the transition period and lactation improved milk yield without altering the metabolic profile.

DNA barcoding reveals high levels of genetic diversity in the fishes of the Itapecuru Basin in Maranhão, Brazil.

DNA barcoding is a useful complementary tool for use in traditional taxonomic studies due to its ability to detect cryptic species, and may be particularly efficient in the identification of fish species. The fish fauna of the Itapecuru River represents an important fishery resource in the Brazilian State of Maranhão, although it is currently suffering increasing degradation as a result of anthropogenic impacts. Therefore, DNA barcoding was used in the present study to identify fish species and establish a database of the rich freshwater fish fauna of Maranhão. A total of 440 specimens were analyzed, corresponding to 64 species belonging to 59 genera, 31 families, and 10 orders. Overall, 92.19% of these species could be identified by DNA barcoding, and were characterized by low levels (average 0.80%) of intra-specific divergence. However, five species (Anableps anableps, Gymnotus carapo, Sciades couma, Pseudauchenipterus nodosus, and Leporinus piau) presented values of mean genetic divergence above 3%, indicating the existence of cryptic diversity in these fishes. The DNA barcoding approach permitted the analysis of a large number of specimens and facilitated the discrimination and identification of closely related fish species in the Itapecuru Basin.

Phylogeny of Marmosops and the occurrence of Marmosops pinheiroi (Pine, 1981) (Didelphimorphia, Didelphidae) in the Cerrado savanna of Maranhão, Brazil.

The present study reviews the phylogeny of the genus Marmosops and expands the geographic range of Marmosops pinheiroi to the Brazilian State of Maranhão. Five specimens of M. pinheiroi were collected from the Inhamum Municipal Environmental Protection Area in Caxias, Maranhão. Total DNA was extracted and the mitochondrial cytochrome b gene was sequenced in an ABI PRISM 3500. Additional sequences of Marmosops were obtained from GenBank for specimens from southeastern and northeast Brazil and the northern Amazon Forest. The results of the analysis indicated that M. incanus and M. paulensis form a well-supported clade, and that M. pinheiroi, M. pakaraimae, and M. parvidens are sister groups, with 71% support. The M. pinheiroi specimens from different regions grouped with high bootstrap values (99 and 100%). Interspecific genetic divergence varied from 8.3 to 21.5%, while intraspecific divergence ranged from 0.2 to 6.3% in M. pinheiroi, 0.3 to 2.9% in M. parvidens, and 0.4 to 6.9% in M. incanus. Given the taxonomic complexity of the genus, the results of the present study offer important insights into the taxonomic status of M. pinheiroi, as well as extend its known distribution 670 km south and east to the Brazilian State of Maranhão.

Fish diversity in the rivers that drain the Baixada Maranhense and the Mearim basin in northeastern Brazil.

The Baixada Maranhense Environmental Protection Area of the Brazilian state of Maranhão encompasses a biologically rich region that includes the basins of four principal rivers, the Mearim, Pericumã, Pindaré, and Turiaçu, which form a complex of wetlands. The present study provides a comprehensive inventory of the fish fauna of the rivers that drain the Baixada Maranhense based on the identification of voucher specimens that were deposited in a scientific collection. Expeditions were conducted between 2014 and 2020. The inventory identified a total of 100 fish species representing 79 genera, 34 families, and 11 orders, which are found in both freshwater and estuarine environments. Six (Potamotrygon orbignyi, Leporinus piau, Moenkhausia loweae, Serrasalmus spilopleura, Pachypops fourcroi and Peckoltia greedoi) of the 100 taxa identified here are considered to be new records for one or more of the basins surveyed during this study, based on the existing data, while four - Colossoma macropomum, Megaleporinus macrocephalus, Cichla sp. and Cichla kelberi - are not native. The most speciose orders were the Siluriformes and Characiformes, while the most diverse families were the Loricariidae, Characidae, Auchenipteridae, Cichlidae, and Serrasalmidae. These data are consistent with the findings of most of the previous studies of freshwater ecosystems in the Neotropical region, in particular those of the rivers of Maranhão. The results of the present study represent an important advance in the understanding of the diversity of the fish fauna of the Mearim, Pericumã, Pindaré, and Turiaçu rivers, which are still poorly studied, and have likely had their diversity underestimated up to now.

Bacteria associated with infectious keratoconjunctivitis outbreak in confined lambs.

Introduction. Infectious keratoconjunctivitis is a significant ocular disease found in confined sheep. Little information about the aetiological agents and their antimicrobial susceptibility is available.Gap Statement. There is limited information on the aetiological agents involved in keratoconjunctivitis outbreaks in sheep.Aim. The present research aimed to determine the bacterial aetiological factors involved in an outbreak of infectious keratoconjunctivitis in confined lambs.Methodology. Ocular swabs were collected from 23 randomly selected lambs, which were classified into three groups according to the severity of the lesion: group I (N=6; no ocular involvement), group II (N=8; less severe injuries) and group III (N=9; more severe injuries). Isolation of aerobic bacteria and antimicrobial susceptibility tests were carried out. Molecular detection of Mollicutes was performed, and positive samples were tested to confirm the presence of the following species: Mycoplasma conjunctivae, Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri.Results. Moraxella sp. and Mollicutes were detected in all groups, but we inferred that Moraxella sp. are only significant in the early stages of the disease. M. conjunctivae was detected in all tested groups, while M. agalactiae was detected in samples of group III only. One strain of Moraxella sp. was resistant to erythromycin and showed intermedite resistance to tetracycline.Conclusion. The presence of these species confirms their importance in the aetiology of this disease, and the low resistance profile observed in the studied farm suggested an increased cure success rate.

DNA Barcoding unveils cryptic lineages of Hoplias malabaricus from Northeastern Brazil.

The trahira or wolf fish - Hoplias malabaricus- is a valid species, although recent cytogenetic and molecular studies have indicated the existence of a species complex. In this context, the present study analyzed the mitochondrial COI marker to determine the levels of genetic diversity of specimens from the Brazilian state of Maranhão, and verify the occurrence of distinct lineages within the study area. Samples were collected from the basins of the Turiaçu, Pindaré, Mearim, Itapecuru, and Parnaíba rivers. A 630-bp fragment was obtained from 211 specimens, with 484 conserved and 108 variable sites, and 60 haplotypes (Hd = 0,947; π = 0,033). The phylogenetic analyses indicated the existence of three distinct lineages of H. malabaricus from Maranhão. Genetic distances of 1.5-8.2% were found between all the populations analyzed, while the variation between haplogroups ranged from 2.1% to 7.7%. The AMOVA indicated that most of the molecular variation was found among groups, with high FST values. The high levels of genetic variability found in the present study are supported by the available cytogenetic data. These findings reinforce the need for the development of effective programs of conservation and management independently for each river basin, in order to preserve the genetic variability found in this taxon.

New records of the occurrence of Megaleporinus macrocephalus () (Characiformes, Anostomidae) from the basins of the Itapecuru and Mearim rivers in Maranhão, Northeastern Brazil.

The "piaussu", Megaleporinus macrocephalus is an anostomatid fish species native to the basin of the Paraguay River, in the Pantanal biome of western Brazil. However, this species has now been recorded in a number of other drainages, including those of the upper Paraná, Uruguay, Jacuí, Doce, Mucuri, and Paraíba do Sulrivers. This study presents two new records of the occurrence of M. macrocephalus, in the basins of the Itapecuru and Mearim rivers in the state of Maranhão, in the Brazilian Northeast. The piaussu is a large-bodied fish of commercial interest that is widely raised on fish farms, and its occurrence in the Itapecuru and Mearim rivers is likely the result of individuals escaping from fish tanks when they overflow during the rainy season. Morphological analyses and sequences of the Cytochrome Oxidase Subunit I (COI) gene confirmed the taxonomic identification of the specimens as M. macrocephalus. The COI sequences were 99.66% similar to those of M. macrocephalus deposited in the BOLDSystems database. These records extend the known distribution of M. macrocephalus to the basins of the Itapecuru and Mearim rivers in the Brazilian Northeast, highlighting a new case of introduction of exotic fish species into Brazilian river basins.

A case of esophageal carcinoma due to esophageal squamous papillomatosis.

INTRODUCTION: Squamous Papilloma is a rare benign tumor of the esophagus. Esophageal squamous papilloma is thought to arise from a chronic inflammatory reaction due to mechanical or chemical irritant. ESP is usually considered to have a benign clinical course; however, some reports highlight the potential development of a malignancy. The development of extensive esophageal squamous papillomas also known as squamous papillomatosis of the esophagus is even less frequent. PRESENTATION OF THE CASE: We present the case of an esophageal papillomatosis that developed into an invasive esophageal squamous cell carcinoma that was only diagnosed in the surgical specimen after minimally invasive Ivor-Lewis esophagectomy and in this case report, we discuss its etiology, diagnosis and treatment. CONCLUSION: Extensive papillomatosis with continuous symptoms, especially persistent dysphagia, should always prompt a thorough analysis into a possible underlying malignancy.

First record of Cynomops planirostris (Peters, 1865) (Chiroptera, Molossidae) from Maranhão state, Brazil, based on morphological and molecular data.

Based on morphological and molecular data, we present the first record of Cynomops planirostris for the State of Maranhão. The specimen was collected in the Inhamum Municipal Environmental Protection Area in Maranhão, Brazil and characterized morphologically as an adult male with scrotal testicles, dorsal pelage reddish chestnut, with ventral pelage slightly lighter in color, forearm length 34.70-34.80mm dental formula i:1/1, c:1/1, pm:1/2, m:3/3 = 26. The sequence of the Cytochrome Oxidase I subunit (COI) molecular marker confirmed the morphological diagnosis of the specimen as C. planirostris with significant similarities. The combined analysis of both morphological and molecular confirmed the occurrence of C. planirostris in the Brazilian state of Maranhão, in the Cerrado biome and records extends the known geographic distribution of the species by 411.30 km.

Genetic variability of populations of the white-eared opossum, Didelphis albiventris Lund 1840 (Didelphimorphia; Didelphidae) in Brazil.

Didelphis albiventris are found throughout Northeast and Central Brazil to central-southern Uruguay and it was subject of few studies in a population level. Given this, the present study investigated the genetic variability of the species using the mitochondrial molecular marker cytochrome oxidase c subunit I. We analyzed samples from the different biomes within three Brazilian regions: Northeast (Caatinga , Cerrado, and Atlantic Forest), Southeast (Cerrado , Atlantic Forest, Cerrado/Atlantic Forest, and Cerrado/Caatinga ecotones) and South (Pampa and Atlantic Forest). Software BAPs retrieved five distinct demes: dm 1, dm 2, and dm 5 that occurs in South, Northeast and Southeast regions respectively and the dm 3 and dm 4 are wide distributed in Northeast and Southeast. Population analysis performed with AMOVA, haplotype network and Mantel test estimated the veracity of the demes. The FST shows structuring for the five demes, with dm 1 (South region) isolated from the others, however the other analysis showed the Northeast/Southeast demes (dm 2-5) united, diagnosing gene flow between them, mainly at the transitional zones, in areas as far away as areas with similar latitude interval (Southeast vs South) that was not detected gene flow. In the haplotype network, the mutational steps was conclusive in split dm1 from dm 2-5 with 15 mutational steps and the Mantel test was moderated, which is explained by genetic similarity despite the great geographic distances (Northeast/Southeast). Thus, our analysis recognized two different lineages (South and Northeast/Southeast) and indicate that the biomes were not decisive in their isolation. The sharing of demes at the transitional zones and in areas with high latitudinal intervals reflects a recent ancestral polymorphism for D. albiventris. The plasticity in the occupation of the space by this species contributes in its wide dispersion capability, that is, geographical distribution. Our results revealed important implications for the management of D. albiventris in these transitional zones areas where demes were shared.

[Prognostic factors associated with postoperative complications in liver transplant]. / Factores pronósticos de complicaciones postoperatorias en el transplante hepático.

OBJECTIVES: the postoperative evolution of patients submitted to orthotopic liver transplant (OLT) is frequently associated with the appearance of different types of complications such as renal failure, graft rejection, infections, and neurological disorders. These complications are the most significant causes of early morbidity and mortality in patients undergoing OLT. The purpose of the present study was the identification of factors related to the different postoperative complications after OLT. EXPERIMENTAL DESIGN: a prospective study was carried out. PATIENTS: seventy-eight variables were analyzed in 32 consecutive patients undergoing OLT. The factors independently associated with the appearance of postoperative complications were identified using a stepwise logistic regression analysis. RESULTS: the multivariate analysis showed that malondialdehyde and creatinine pretransplant serum levels were associated with the development of renal dysfunction. The pretransplant levels of haemoglobin and the units of platelets administered during surgery were prognostic factors of infections. Acute graft rejection was predicted by ?-glutamyl transpeptidase and total bilirubin serum levels. The pretransplant sodium and glutaredoxin levels in serum were associated with neurological complications. CONCLUSIONS: we propose these markers for the identification of high-risk patients allowing an early surveillance and/or treatment to improve morbidity and survival in patients submitted to OLT.

New records of the occurrence of Megaleporinus macrocephalus (Garavello & Britski, 1988) (Characiformes, Anostomidae) from the basins of the Itapecuru and Mearim rivers in Maranhão, Northeastern Brazil / Novos registros da ocorrência de Megaleporinus macrocephalus (Garavello & Britski, 1988) (Characiformes, Anostomidae) nas bacias dos rios Itapecuru e Mearim no Maranhão, Nordeste, Brasil

The "piaussu", Megaleporinus macrocephalus is an anostomatid fish species native to the basin of the Paraguay River, in the Pantanal biome of western Brazil. However, this species has now been recorded in a number of other drainages, including those of the upper Paraná, Uruguay, Jacuí, Doce, Mucuri, and Paraíba do Sulrivers. This study presents two new records of the occurrence of M. macrocephalus, in the basins of the Itapecuru and Mearim rivers in the state of Maranhão, in the Brazilian Northeast. The piaussu is a large-bodied fish of commercial interest that is widely raised on fish farms, and its occurrence in the Itapecuru and Mearim rivers is likely the result of individuals escaping from fish tanks when they overflow during the rainy season. Morphological analyses and sequences of the Cytochrome Oxidase Subunit I (COI) gene confirmed the taxonomic identification of the specimens as M. macrocephalus. The COI sequences were 99.66% similar to those of M. macrocephalus deposited in the BOLDSystems database. These records extend the known distribution of M. macrocephalus to the basins of the Itapecuru and Mearim rivers in the Brazilian Northeast, highlighting a new case of introduction of exotic fish species into Brazilian river basins.

Megaleporinus macrocephalus é uma espécie de peixe anostomatídeo nativa da bacia do rio Paraguai, no bioma Pantanal do oeste do Brasil. No entanto, essa espécie já foi registrada em várias outras drenagens, incluindo as dos rios Alto Paraná, Uruguai, Jacuí, Doce, Mucuri e Paraíba do Sul. Este estudo apresenta dois novos registros da ocorrência de M. macrocephalus, nas bacias dos rios Itapecuru e Mearim, no estado do Maranhão, no nordeste brasileiro. O piaussu é um peixe de grande porte, de interesse comercial, amplamente criado em pisciculturas, e sua ocorrência nos rios Itapecuru e Mearim é provavelmente o resultado de indivíduos que escapam dos tanques quando transbordam durante a estação chuvosa. Análises morfológicas e sequências do gene da subunidade I do citocromo oxidase (COI) confirmaram a identificação taxonômica dos espécimes como M. macrocephalus. As sequências de COI foram 99,66% semelhantes às de M. macrocephalus depositadas no banco de dados BOLDSystems. Esses registros estendem a conhecida distribuição de M. macrocephalus às bacias dos rios Itapecuru e Mearim, no nordeste brasileiro, destacando um novo caso de introdução de espécies exóticas de peixes nas bacias hidrográficas brasileiras.

New records of the occurrence of Megaleporinus macrocephalus (Garavello & Britski, 1988 (Characiformes, Anostomidae) from the basins of the Itapecuru and Mearim rivers in Maranhão, Northeastern Brazil / Novos registros da ocorrência de Megaleporinus macrocephalus (Garavello & Britski, 1988) (Characiformes, Anostomidae) nas bacias dos rios Itapecuru e Mearim no Maranhão, Nordeste, Brasil

The "piaussu", Megaleporinus macrocephalus is an anostomatid fish species native to the basin of the Paraguay River, in the Pantanal biome of western Brazil. However, this species has now been recorded in a number of other drainages, including those of the upper Paraná, Uruguay, Jacuí, Doce, Mucuri, and Paraíba do Sulrivers. This study presents two new records of the occurrence of M. macrocephalus, in the basins of the Itapecuru and Mearim rivers in the state of Maranhão, in the Brazilian Northeast. The piaussu is a large-bodied fish of commercial interest that is widely raised on fish farms, and its occurrence in the Itapecuru and Mearim rivers is likely the result of individuals escaping from fish tanks when they overflow during the rainy season. Morphological analyses and sequences of the Cytochrome Oxidase Subunit I (COI) gene confirmed the taxonomic identification of the specimens as M. macrocephalus. The COI sequences were 99.66% similar to those of M. macrocephalus deposited in the BOLDSystems database. These records extend the known distribution of M. macrocephalus to the basins of the Itapecuru and Mearim rivers in the Brazilian Northeast, highlighting a new case of introduction of exotic fish species into Brazilian river basins.

Megaleporinus macrocephalus é uma espécie de peixe anostomatídeo nativa da bacia do rio Paraguai, no bioma Pantanal do oeste do Brasil. No entanto, essa espécie já foi registrada em várias outras drenagens, incluindo as dos rios Alto Paraná, Uruguai, Jacuí, Doce, Mucuri e Paraíba do Sul. Este estudo apresenta dois novos registros da ocorrência de M. macrocephalus, nas bacias dos rios Itapecuru e Mearim, no estado do Maranhão, no nordeste brasileiro. O piaussu é um peixe de grande porte, de interesse comercial, amplamente criado em pisciculturas, e sua ocorrência nos rios Itapecuru e Mearim é provavelmente o resultado de indivíduos que escapam dos tanques quando transbordam durante a estação chuvosa. Análises morfológicas e sequências do gene da subunidade I do citocromo oxidase (COI) confirmaram a identificação taxonômica dos espécimes como M. macrocephalus. As sequências de COI foram 99,66% semelhantes às de M. macrocephalus depositadas no banco de dados BOLDSystems. Esses registros estendem a conhecida distribuição de M. macrocephalus às bacias dos rios Itapecuru e Mearim, no nordeste brasileiro, destacando um novo caso de introdução de espécies exóticas de peixes nas bacias hidrográficas brasileiras.

New records of the occurrence of Megaleporinus macrocephalus () (Characiformes, Anostomidae) from the basins of the Itapecuru and Mearim rivers in Maranhão, Northeastern Brazil

Abstract The piaussu, Megaleporinus macrocephalus is an anostomatid fish species native to the basin of the Paraguay River, in the Pantanal biome of western Brazil. However, this species has now been recorded in a number of other drainages, including those of the upper Paraná, Uruguay, Jacuí, Doce, Mucuri, and Paraíba do Sulrivers. This study presents two new records of the occurrence of M. macrocephalus, in the basins of the Itapecuru and Mearim rivers in the state of Maranhão, in the Brazilian Northeast. The piaussu is a large-bodied fish of commercial interest that is widely raised on fish farms, and its occurrence in the Itapecuru and Mearim rivers is likely the result of individuals escaping from fish tanks when they overflow during the rainy season. Morphological analyses and sequences of the Cytochrome Oxidase Subunit I (COI) gene confirmed the taxonomic identification of the specimens as M. macrocephalus. The COI sequences were 99.66% similar to those of M. macrocephalus deposited in the BOLDSystems database. These records extend the known distribution of M. macrocephalus to the basins of the Itapecuru and Mearim rivers in the Brazilian Northeast, highlighting a new case of introduction of exotic fish species into Brazilian river basins.

Resumo Megaleporinus macrocephalus é uma espécie de peixe anostomatídeo nativa da bacia do rio Paraguai, no bioma Pantanal do oeste do Brasil. No entanto, essa espécie já foi registrada em várias outras drenagens, incluindo as dos rios Alto Paraná, Uruguai, Jacuí, Doce, Mucuri e Paraíba do Sul. Este estudo apresenta dois novos registros da ocorrência de M. macrocephalus, nas bacias dos rios Itapecuru e Mearim, no estado do Maranhão, no nordeste brasileiro. O piaussu é um peixe de grande porte, de interesse comercial, amplamente criado em pisciculturas, e sua ocorrência nos rios Itapecuru e Mearim é provavelmente o resultado de indivíduos que escapam dos tanques quando transbordam durante a estação chuvosa. Análises morfológicas e sequências do gene da subunidade I do citocromo oxidase (COI) confirmaram a identificação taxonômica dos espécimes como M. macrocephalus. As sequências de COI foram 99,66% semelhantes às de M. macrocephalus depositadas no banco de dados BOLDSystems. Esses registros estendem a conhecida distribuição de M. macrocephalus às bacias dos rios Itapecuru e Mearim, no nordeste brasileiro, destacando um novo caso de introdução de espécies exóticas de peixes nas bacias hidrográficas brasileiras.

Adjuvant chemotherapy for prevention of recurrence of invasive hepatocellular carcinoma after orthotopic liver transplantation.

UNLABELLED: Orthotopic liver transplantation (OLT) is the best treatment for nonresectable hepatocellular carcinoma (HCC), but tumor recurrence reduces long-term and medium-term survival. The effectiveness of adjuvant chemotherapy to prevent tumor recurrence has not been fully established. METHODS: Three hundred eighty-seven consecutive patients, including 43 with HCC superimposed on liver cirrhosis, underwent OLT. Twelve patients with one or more prognostic criteria for HCC recurrence were entered into a prospective prophylaxis protocol with monthly cycles of cisplatin (60 mg/m(2)) and adriamycin (30 mg/m(2)), beginning the fourth week post-OLT for a maximum of seven sessions. RESULTS: The 5-year survival of the non-HCC patients was 65.7% and that of the HCC patients was 60.46% (P = NS). Chemotherapy was reasonably well tolerated, but the 9 patients with hepatitis C- or B-associated cirrhosis showed viral and histological recurrence of the primary disease. A high proportion of patients (7 of 12) developed tumor recurrence during the first year after OLT. Six of these patients died, all but one due to HCC relapse. Five patients remain healthy and tumor free at 58 to 130 months. Post-OLT adjuvant chemotherapy does not avoid tumor recurrence and its fatal consequences but may contribute to prolonged tumor-free survival among a significant proportion of patients with high-risk HCC. However, the uncertain implications on viral recurrence and the lack of control groups do not allow post-OLT chemotherapy to be recommended outside controlled clinical trials, which are clearly warranted.

Treatment of refractory cholestatic pruritus with molecular adsorbent recirculating system (MARS).

UNLABELLED: Pruritus is a common complication of cholestatic liver diseases or liver graft dysfunction. Current medical therapies lack efficacy. The molecular adsorbent recirculating system (MARS) represents an interesting therapeutic option. Our objective was to report our experience in the management of four patients with intractable pruritus with MARS. PATIENTS AND METHODS: The MARS treatment cycle included three consecutive treatments, each of 8 hours duration. The four patients with intractable pruritus who were treated had primary biliary cirrhosis/autoimmune hepatitis overlap syndrome (n = 1), ductopenic allograft rejection (n = 2), or posttransplant cholestatic HCV recurrence (n = 1). Intensity of pruritus was documented 24 hours before as well as 24 hours, 7 and 30 days after MARS therapy, and at the end of follow-up. We measured complete blood cell counts, glucose, BUN, creatinine, sodium, potassium, AST, ALT, GGT, alkaline phosphatase, bilirubin, prothrombin activity, and activated partial thromboplastin time. RESULTS: MARS therapy was well tolerated. Patient 1 experienced temporal relief of pruritus, but needed another MARS cycle because of relapse. Patient 2 experienced partial and temporary relief of pruritus, was listed for retransplantation, and received a liver graft 2 months later. Patient 3 showed a dramatic reduction in the degree of pruritus with MARS. Pruritus in patient 4 decreased promptly with MARS therapy and conversion of immunosuppression to tacrolimus, thereby avoiding retransplantation. CONCLUSION: MARS therapy is a promising, safe therapeutic option to treat refractory pruritus caused by cholestatic liver disorders.

DNA Barcoding unveils cryptic lineages of Hoplias malabaricus from Northeastern Brazil / DNA Barcoding revela linhagens crípticas de Hoplias malabaricus no nordeste do Brasil

Abstract The trahira or wolf fish - Hoplias malabaricus- is a valid species, although recent cytogenetic and molecular studies have indicated the existence of a species complex. In this context, the present study analyzed the mitochondrial COI marker to determine the levels of genetic diversity of specimens from the Brazilian state of Maranhão, and verify the occurrence of distinct lineages within the study area. Samples were collected from the basins of the Turiaçu, Pindaré, Mearim, Itapecuru, and Parnaíba rivers. A 630-bp fragment was obtained from 211 specimens, with 484 conserved and 108 variable sites, and 60 haplotypes (Hd = 0,947; π = 0,033). The phylogenetic analyses indicated the existence of three distinct lineages of H. malabaricus from Maranhão. Genetic distances of 1.5-8.2% were found between all the populations analyzed, while the variation between haplogroups ranged from 2.1% to 7.7%. The AMOVA indicated that most of the molecular variation was found among groups, with high FST values. The high levels of genetic variability found in the present study are supported by the available cytogenetic data. These findings reinforce the need for the development of effective programs of conservation and management independently for each river basin, in order to preserve the genetic variability found in this taxon.

Resumo A traíra - Hoplias malabaricus- é uma espécie válida, embora recentes estudos citogenéticos e moleculares tenham indicado a existência de um complexo de espécies. Neste contexto, o presente estudo analisou o marcador mitocondrial COI para determinar os níveis de diversidade genética dos espécimes do estado do Maranhão e verificar a ocorrência de linhagens distintas dentro da área de estudo. As amostras foram coletadas nas bacias dos rios Turiaçu, Pindaré, Mearim, Itapecuru e Parnaíba. As análises filogenéticas indicaram a existência de três linhagens distintas nas populações do Maranhão. Obteve-se um fragmento de 630 pb de 211 espécimes, com 484 sítios conservados, 108 variáveis e 60 haplótipos (Hd = 0,947; π = 0,033). As análises filogenéticas indicaram a ocorrência de três linhagens distintas de H. malabaricus do Maranhão. Distâncias genéticas de 1.5 a 8.2% foram encontradas entre todas as populações analisadas, enquanto a variação entre os haplogrupos variou de 2.1% a 7.7%. A AMOVA indicou que a maior variação molecular foi entre os grupos, com altos valores de FST. Os altos níveis de variabilidade genética encontrados no presente estudo são suportados pelos dados citogenéticos disponíveis. Essas descobertas reforçam a necessidade de desenvolver programas de conservação e manejo independentemente para cada bacia hidrográfica, a fim de preservar a variabilidade genética encontrada neste táxon.

Study on the efficacy and safety of adefovir dipivoxil treatment in post-liver transplant patients with hepatitis B virus infection and lamivudine-resistant hepatitis B virus.

Hepatitis B virus (HBV) recurrence and de novo HBV infection are frequent events in liver transplantation recipients. Treatment with lamivudine is initially efficient in both infections but the incidence of lamivudine-resistant HBV emergence increases over time. Adefovir appears to be promising in post-liver transplantation patients with recurrent HBV infection and lamivudine-resistant HBV. This study analyzed adefovir treatment in 42 post-liver transplantation patients who developed recurrent HBV or de novo HBV infection with lamivudine-resistant HBV (54.8% HCV-coinfected). Patients received 10 mg of oral adefovir once daily for a mean period of time of 21.5 months (range from 12 to 31 months). In 62.9% of patients, ALT levels decreased significantly. Serum HBV-DNA was undetectable in 64% of the cases. Twenty percent of patients lost HBeAg marker and 13.3% of them developed anti-HBe. In 9.5% of recipients, HBsAg became negative. There was no significant change in serum creatinine levels. In only one patient was worsening of the renal function detected, making dose adjustment necessary. No other side effects were reported. Our results confirm the efficacy and safety of adefovir treatment in post-liver transplantation patients with lamivudine-resistant HBV, neither were adefovir-resistant mutations identified in patients after 21 months of therapy, nor were there adverse events, especially renal toxicity.

Evidence for immunodominance between closely related epitopes in the selection of T cell repertoire: hierarchy of T cell epitopes in a repeating sequence.

In a protein antigen the number of epitopes that are presented by the MHC molecules to the T cells is generally limited. This phenomenon of immunodominance determines the T cell response to a given antigen. To understand the molecular basis of epitope selection we have analysed the hierarchy of T cell epitopes in a repeating synthetic polypeptide antigen Poly 18, Poly EYK(EYA)5 in the mice of H-2d haplotype. Because of its repeating nature, all the potential epitopes of Poly EYK(EYA)5 generated as a result of antigen processing will have extensive sequence overlap, therefore, providing an excellent system to investigate the molecular basis of T cell peptide epitope selection in vivo. We have synthesized a series of 12 and 15 amino acid peptides to mimic these epitopes. In H-2d mice Poly EYK(EYA)5 elicits T cell clones that optimally react with 15 amino acid peptides EYK(EYA)4 and/or (EYA)5. Similar results were obtained when related 12 amino acid peptides EYK(EYA)3 and/or (EYA)4 are used. (EYA)5 reactive T cell clones appear to be very heterogenous and much larger in number than EYK(EYA)4 reactive clones. (EYA)5 reactive clones could be elicited by at least three short Poly-18 derived epitopes (EYA)4, EYK(EYA)3 and (EYA)3EYK while EYK(EYA)4 reactive clones elicited only by the EYK(EYA)3 epitope. However, we observed the dominance of (EYA)5 reactive clones even when EYK(EYA)3 was used as an immunogen and this could be related to the degeneracy of their antigen specificity. Our earlier antigen competition studies suggest that (EYA)5 does not compete with EYK(EYA)4 epitope in binding to I-Ad. Therefore, there is no intramolecular competition between these epitopes to activate T cells. The epitope (EYA)3EYK appears to be subdominant since it can elicit Poly EYK(EYA)5 specific T cells upon immunization but does not appear to be part of Poly EYK(EYA)5 repertoire. Peptides such as (EYA)2EYKEYA or EYAEYK(EYA)2 with lysine substitution in the middle of the sequence were non immunogeneic. Similar results were obtained when the larger 15 amino acid peptides were used as antigen. Another level of epitope immunodominance is seen when substituted peptides of the two immunodominant epitopes are used. Some of these epitopes have potential to be part of the Poly 18 repertoire but they are greatly under represented when intact Poly 18 is used as antigen. The unusual hierarchy observed for immunodominance in these overlapping epitopes of EYK(EYA)5 sequence suggest a bias in the selection of T cell repertoire based upon the crossreactivity between potential epitopes generated as a result of antigen processing.

Fine-specificity analysis of antibodies directed to the C-terminal peptides of cytochrome c recognized by T-lymphocytes.

Recognition of cytochrome c by T-lymphocytes seems to involve the amino acid residues in the C-terminal region of the molecule. Lys-99 has particularly been identified as one of the critical residues in the recognition process. We have now raised antibodies against the C-terminal region of the cytochrome c molecule to map the residues that may be recognized by B-lymphocytes. These antibodies were generated in high-responder B10.A mice against either the 81-104 CNBr fragment of pigeon cytochrome c or against the synthetic spliced fragment (86-90)-(94-103) of the tobacco hornworm moth cytochrome c. A good antibody response was obtained for both fragments as measured by solid-phase radioimmunoassay. A series of peptides related to these fragments were synthesized for competitive inhibition to assess the antigenic sites on these molecules. In spite of substantial homology between the moth (86-90)-(94-103) and pigeon (81-104) fragments, the antibody populations raised against each fragment differed in their recognition patterns. Residues 99 (Lys), 103 (Ala) and 104 (Lys) were found to be crucial for binding of the anti-pigeon antibody to the pigeon 81-104 fragment. The fine specificity mapping of the antigenic sites on the moth (86-90)-(94-103) fragment indicated that along with some of the residues in the N-terminus (86-90), residue 99 (Lys) was involved in recognition of the moth (86-90)-(94-103) fragment by its antibody. This residue (Lys-99) also acts as a T-cell receptor contact site for both pigeon and moth cytochrome c. We therefore conclude that common patterns of recognition must exist between T and B-cells that recognize the C-terminal region of cytochrome c. Since Lys-99 is also present in mouse cytochrome c, the antigenic site must involve both self and non-self residues.