Optimization of Laccase Production, and Characterization of Lignin Degradation Products by Fusarium oxysporum JUMAD-053

The objective of this work was to optimize the culture conditions for laccase production in the presence of Kraft lignin by Fusarium oxysporum JUMAD-053, and to evaluate the biodegradation products of lignin. The fungal isolate that presented highest laccase activity had its production optimized by a statistical factorial design 33 in 15 experimental runs. F. oxysporum presented the highest constitutive laccase titer (5.37 U/mL). Statistical factorial design demonstrated a maximum laccase titer of 9.8 U/mL when assayed against ABTS under the conditions optimized: 1.125% (w/v) yeast extract, 0.5% (w/v) Kraft lignin and 10 days of cultivation. The maximum laccase titer when assayed on DMP was 8.4 U/mL, following the conditions optimized: 1.125% yeast extract, 0.25% Kraft lignin and 7 days of cultivation. The analysis of cultures led to identification of metabolites; two being aromatic: 2,6-dimethoxy benzoic acid and sesamin; also, fumonisin and long-chain fatty acids. As a result of the study, the maximum laccase activities of 9.8 and 8.4 U/mL measured from ABTS and DMP substrates, respectively. The search shows new sources of fungal laccase for obtaining new metabolites of biodegradation from Kraft lignin in culture medium. DOI: http://dx.doi.org/10.17807/orbital.v13i5.162

Optimization of Laccase Production, and Characterization of Lignin Degradation Products by Fusarium oxysporum JUMAD-053

The objective of this work was to optimize the culture conditions for laccase production in the presence of Kraft lignin by Fusarium oxysporum JUMAD-053, and to evaluate the biodegradation products of lignin. The fungal isolate that presented highest laccase activity had its production optimized by a statistical factorial design 33 in 15 experimental runs. F. oxysporum presented the highest constitutive laccase titer (5.37 U/mL). Statistical factorial design demonstrated a maximum laccase titer of 9.8 U/mL when assayed against ABTS under the conditions optimized: 1.125% (w/v) yeast extract, 0.5% (w/v) Kraft lignin and 10 days of cultivation. The maximum laccase titer when assayed on DMP was 8.4 U/mL, following the conditions optimized: 1.125% yeast extract, 0.25% Kraft lignin and 7 days of cultivation. The analysis of cultures led to identification of metabolites; two being aromatic: 2,6-dimethoxy benzoic acid and sesamin; also, fumonisin and long-chain fatty acids. As a result of the study, the maximum laccase activities of 9.8 and 8.4 U/mL measured from ABTS and DMP substrates, respectively. The search shows new sources of fungal laccase for obtaining new metabolites of biodegradation from Kraft lignin in culture medium. DOI: http://dx.doi.org/10.17807/orbital.v13i5.162

Overexpression of KLC2 due to a homozygous deletion in the non-coding region causes SPOAN syndrome

SPOAN syndrome is a neurodegenerative disorder mainly characterized by spastic paraplegia, optic atrophy and neuropathy (SPOAN). Affected patients are wheelchair bound after 15 years old, with progressive joint contractures and spine deformities. SPOAN patients also have sub normal vision secondary to apparently non-progressive congenital optic atrophy. A potential causative gene was mapped at 11q13 ten years ago. Here we performed next-generation sequencing in SPOAN-derived samples. While whole-exome sequencing failed to identify the causative mutation, whole-genome sequencing allowed to detect a homozygous 216-bp deletion (chr11.hg19:g.66,024,557_66,024,773del) located at the non-coding upstream region of the KLC2 gene. Expression assays performed with patient’s fibroblasts and motor neurons derived from SPOAN patients showed KLC2 overexpression. Luciferase assay in constructs with 216-bp deletion confirmed the overexpression of gene reporter, varying from 48 to 74%, as compared with wild-type. Knockdown and overexpression of klc2 in Danio rerio revealed mild to severe curly-tail phenotype, which is suggestive of a neuromuscular disorder. Overexpression of a gene caused by a small deletion in the non-coding region is a novel mechanism, which to the best of our knowledge, was never reported before in a recessive condition. Although the molecular mechanism of KLC2 up-regulation still remains to be uncovered, such example adds to the importance of non-coding regions in human pathologyFil: Melo, Uira S.. Universidade de Sao Paulo; BrasilFil: Macedo Souza, Lucia I.. Universidade de Sao Paulo; BrasilFil: Figueiredo, Thalita. Federal University of Paraiba; Brasil. Paraiba State University; BrasilFil: Muotri, Alysson R. University of California at San Diego; Estados UnidosFil: Gleeson, Joseph G.. The Rockefeller University; Estados UnidosFil: Coux, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Armas, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Calcaterra, Nora Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Kitajima, João P.. Mendelics Genomic Analysis; BrasilFil: Amorim, Simone. Universidade de Sao Paulo; BrasilFil: Olávio, Thiago R.. Universidade de Sao Paulo; BrasilFil: Griesi Oliveira, Karina. Universidade de Sao Paulo; BrasilFil: Coatti, Giuliana C.. Universidade de Sao Paulo; BrasilFil: Rocha, Clarissa R.R. Universidade de Sao Paulo; BrasilFil: Martins Pinheiro, Marinalva. Universidade de Sao Paulo; BrasilFil: Menck, Carlos F.M.. Universidade de Sao Paulo; BrasilFil: Zaki, Maha S.. National Research Center. EL Cairo; EgiptoFil: Kok, Fernando. Universidade de Sao Paulo; BrasilFil: Zatz, Mayana. Universidade de Sao Paulo; BrasilFil: Santos, Silvana. Federal University of Paraiba; Brasil. Paraiba State University; Brasi

Dengue Virus Type 4 Phylogenetics in Brazil 2011: Looking beyond the Veil

Dengue Fever and Dengue Hemorrhagic Fever are diseases affecting approximately 100 million people/year and are a major concern in developing countries. In the present study, the phylogenetic relationship of six strains of the first autochthonous cases of DENV-4 infection occurred in Sao Paulo State, Parana State and Rio Grande do Sul State, Brazil, 2011 were studied. Nucleotide sequences of the envelope gene were determined and compared with sequences representative of the genotypes I, II, III and Sylvatic for DEN4 retrieved from GenBank. We employed a Bayesian phylogenetic approach to reconstruct the phylogenetic relationships of Brazilian DENV-4 and we estimated evolutionary rates and dates of divergence for DENV-4 found in Brazil in 2011. All samples sequenced in this study were located in Genotype II. The studied strains are monophyletic and our data suggest that they have been evolving separately for at least 4 to 6 years. Our data suggest that the virus might have been present in the region for some time, without being noticed by Health Surveillance Services due to a low level of circulation and a higher prevalence of DENV-1 and DENV- 2

Evaluation of the antioxidant and phototoxic potentials of Bauhinia microstachya var. massambabensis Vaz leaf extracts

Four different leaf extracts of B. microstachya var. massambabensis were studied to evaluate their antioxidant capacity by using three in vitro methods, with Ginkgo biloba and Trolox® as the standards. With the DPPH and ABTS·+ methods, the antioxidant activity of the extracts was in the following order, from maximum to minimum: AcEt > WAc > raw EtOH > EtOH CA > EGb, while with the ORAC method, it was as follows: EtOH CA > raw EtOH > AcEt > WAc > EGb. Phototoxic analysis was performed in yeast cultures of Saccharomyces cerevisiae. From the ethyl acetate extract, 2 flavonoids kaempferol-3-O-rhamnoside and astragalin-2”,6”-O-digallate were isolated and identified by HPLC and 1H- and 13C-NMR; to our knowledge, this is the first report of the occurrence of astragalin-2”,6”-O-digallate in the Bauhinia genus.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Genomics and epidemiology for gastric adenocarcinomas (GE4GAC): a Brazilian initiative to study gastric cancer

Abstract Gastric cancer (GC) is the fifth most common type of cancer worldwide with high incidences in Asia, Central, and South American countries. This patchy distribution means that GC studies are neglected by large research centers from developed countries. The need for further understanding of this complex disease, including the local importance of epidemiological factors and the rich ancestral admixture found in Brazil, stimulated the implementation of the GE4GAC project. GE4GAC aims to embrace epidemiological, clinical, molecular and microbiological data from Brazilian controls and patients with malignant and pre-malignant gastric disease. In this letter, we summarize the main goals of the project, including subject and sample accrual and current findings

Genome of the Avirulent Human-Infective Trypanosome—Trypanosoma rangeli

Background: Trypanosoma rangeli is a hemoflagellate protozoan parasite infecting humans and other wild and domestic mammals across Central and South America. It does not cause human disease, but it can be mistaken for the etiologic agent of Chagas disease, Trypanosoma cruzi. We have sequenced the T. rangeli genome to provide new tools for elucidating the distinct and intriguing biology of this species and the key pathways related to interaction with its arthropod and mammalian hosts.  Methodology/Principal Findings: The T. rangeli haploid genome is ,24 Mb in length, and is the smallest and least repetitive trypanosomatid genome sequenced thus far. This parasite genome has shorter subtelomeric sequences compared to those of T. cruzi and T. brucei; displays intraspecific karyotype variability and lacks minichromosomes. Of the predicted 7,613 protein coding sequences, functional annotations could be determined for 2,415, while 5,043 are hypothetical proteins, some with evidence of protein expression. 7,101 genes (93%) are shared with other trypanosomatids that infect humans. An ortholog of the dcl2 gene involved in the T. brucei RNAi pathway was found in T. rangeli, but the RNAi machinery is non-functional since the other genes in this pathway are pseudogenized. T. rangeli is highly susceptible to oxidative stress, a phenotype that may be explained by a smaller number of anti-oxidant defense enzymes and heatshock proteins.  Conclusions/Significance: Phylogenetic comparison of nuclear and mitochondrial genes indicates that T. rangeli and T. cruzi are equidistant from T. brucei. In addition to revealing new aspects of trypanosome co-evolution within the vertebrate and invertebrate hosts, comparative genomic analysis with pathogenic trypanosomatids provides valuable new information that can be further explored with the aim of developing better diagnostic tools and/or therapeutic targets