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1.
Chinese Journal of Medical Genetics ; (6): 1296-1300, 2023.
Article in Chinese | WPRIM | ID: wpr-1009293

ABSTRACT

OBJECTIVE@#To delineate the origin and content of a mosaicism small supernumerary marker chromosome (sSMC) in a fetus with combined chromosomal karyotyping, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH).@*METHODS@#The fetus of a 31-year-old pregnant woman who had presented at the Maternal and Child Health Care Hospital of Longhua District of Shenzhen City in 2022 was selected as the study subject. Non-invasive prenatal testing suggested that the fetus has harbored a 8.75 Mb duplication in 4q12q13.1. With informed consent, amniotic fluid and peripheral blood samples were taken from the couple for chromosomal karyotyping analysis. The origin and content of a sSMC was identified by CMA, and its proportion in amniotic fluid was determined with a FISH assay.@*RESULTS@#The karyotypes of the pregnant woman, her husband and the fetus were respectively determined as 46,XX, 46,XY,inv(9)(p12q12), and 47,XY,inv(9)(p12q12)pat,+mar[75]/ 46,XY,inv(9)(p12q12)pat[25]. CMA test of the amniotic fluid sample was arr[hg19]4p11q13.1(48978053_63145931)×3, which revealed no mosaicism. However, FISH analysis showed that 59% of interphase cells from the cultured amniotic fluid sample had contained three signals for the centromere of chromosome 4, whilst 65% of interphase cells from the re-sampled amniotic fluid had three such signals, which confirmed the existence of trisomy 8 mosaicism.@*CONCLUSION@#Chromosomal structural abnormality combined with mosaicism can be delineated with combined chromosomal karyotyping and molecular techniques such as FISH and CMA, which has enabled more accurate counseling for the family.


Subject(s)
Humans , Child , Female , Pregnancy , Adult , In Situ Hybridization, Fluorescence , Mosaicism , Genetic Techniques , Amniotic Fluid , Chromosomes, Human, Pair 4
2.
Chinese Journal of Biotechnology ; (12): 4708-4717, 2023.
Article in Chinese | WPRIM | ID: wpr-1008052

ABSTRACT

Plasmids are the most commonly used gene carriers in the field of gene synthesis and sequencing. However, the main problems faced by traditional plasmid DNA extraction technology are low extraction throughput and high production cost, so they cannot meet the growing demand. In this study, a double-magnetic-bead method (DMBM) for plasmid extraction was developed based on the principle of plasmid extraction. The effects of the input of magnetic beads, the size of plasmid DNA fragments, and the volume of bacterial on plasmid DNA extraction were explored. In addition, the quality, throughput, and cost of plasmid DNA extraction were also compared between this technique and the commercial plasmid DNA extraction kits. The results showed that the DMBM can meet the needs of extracting plasmid DNA with different cell densities and fragment lengths. Moreover, the sensitivity and quality of plasmid extraction by the DMBM method were both superior to those of the centrifugal adsorption column method. In addition, this technique could be applied on a 96-channel automated nucleic acid extractor, resulting in higher purity of the extracted plasmid DNA, 80% reduction in extraction time, and 57.1% reduction in cost. It also reduces manual operations, achieving high-throughput and low-cost plasmid DNA extraction, thus may facilitate gene synthesis and sequencing.


Subject(s)
Plasmids/genetics , DNA/genetics , Nucleic Acids , Genetic Techniques , Magnetic Phenomena
3.
Chinese Journal of Biotechnology ; (12): 4376-4396, 2023.
Article in Chinese | WPRIM | ID: wpr-1008031

ABSTRACT

Nowadays, engineered Komagataella phaffii plays an important role in the biosynthesis of small molecule metabolites and protein products, showing great potential and value in industrial productions. With the development and application of new editing tools such as CRISPR/Cas9, it has become possible to engineer K. phaffii into a cell factory with high polygenic efficiency. Here, the genetic manipulation techniques and objectives for engineering K. phaffii are first summarized. Secondly, the applications of engineered K. phaffii as a cell factory are introduced. Meanwhile, the advantages as well as disadvantages of using engineered K. phaffii as a cell factory are discussed and future engineering directions are prospected. This review aims to provide a reference for further engineering K. phaffii cell factory, which is supposed to facilitate its application in bioindustry.


Subject(s)
Saccharomycetales/genetics , Genetic Techniques
4.
Chinese Journal of Biotechnology ; (12): 605-619, 2022.
Article in Chinese | WPRIM | ID: wpr-927731

ABSTRACT

Acarbose is widely used as α-glucosidase inhibitor in the treatment of type Ⅱ diabetes. Actinoplanes sp. is used for industrial production of acarbose. As a secondary metabolite, the biosynthesis of acarbose is quite complex. In addition to acarbose, a few acarbose structural analogs are also accumulated in the culture broth of Actinoplanes sp., which are hard to remove. Due to lack of systemic understanding of the biosynthesis and regulation mechanisms of acarbose and its structural analogs, it is difficult to eliminate or reduce the biosynthesis of the structural analogs. Recently, the advances in omics technologies and molecular biology have facilitated the investigations of biosynthesis and regulatory mechanisms of acarbose and its structural analogs in Actinoplanes sp.. The genes involved in the biosynthesis of acarbose and its structural analogs and their regulatory mechanism have been extensively explored by using bioinformatics analysis, genetic manipulation and enzymatic characterization, which is summarized in this review.


Subject(s)
Humans , Acarbose/metabolism , Diabetes Mellitus, Type 2/drug therapy , Genetic Techniques
5.
Chinese Journal of Biotechnology ; (12): 2040-2050, 2020.
Article in Chinese | WPRIM | ID: wpr-878464

ABSTRACT

Linear chromatin is compacted into eukaryotic nucleus through a complex and multi-layered architecture. Consequently, chromatin conformation in a local or long-distance manner is strongly correlated with gene expression. Chromosome conformation capture (3C) technology, together with its variants like 4C/5C/Hi-C, has been well developed to study chromatin looping and whole genome structure. In this review, we introduce new technologies including chromosome capture combined with immunoprecipitation, nuclei acid-based hybridization, single cell and genome sequencing, as well as their application.


Subject(s)
Cell Nucleus , Chromatin/genetics , Chromosomes/genetics , Genetic Techniques , Genome/genetics
6.
Rev. Investig. Salud. Univ. Boyacá (En línea) ; 7(2): 138-172, 2020. tab, ilust
Article in Spanish | LILACS, COLNAL | ID: biblio-1292512

ABSTRACT

Introducción: el objetivo de la secuenciación es determinar la composición de los nucleótidos presentes en el ADN o el ARN. Desde la finalización del proyecto genoma humano, surgieron diversas tecnologías de secuenciación rápida como Roche 454, SOLiD, Illumina, Ion Torrent, PacBio y Oxford Nanopore, más precisas y costoeficientes, que permiten desarrollar proyectos a gran escala y estudiar genes y genomas, la composición de microbiomas, enfermedades metabólicas y enfermedades genéticas que afectan a la población. Objetivo: describir los fundamentos de los métodos de secuenciación de ADN y sus aplicaciones en las ciencias biomédicas. Métodos: revisión descriptiva de las principales estrategias de secuenciación de ADN de primera, segunda y tercera generación y su aplicación en el entorno biomédico, a partir de la búsqueda de artículos en bases de datos electró-nicas especializadas en investigación científica. Se encontraron 118 documentos, de los cuales se excluyeron 6, por no cumplir con los criterios de inclusión, y se seleccionaron 112, por cumplir con todos los requisitos. Conclusiones:el surgimiento de los métodos de secuenciación de siguiente generación arroja una gran canti-dad de datos, incluidos genomas secuenciados completamente de varias especies, con un rendimiento extenso, tiempos reducidos y costoeficiencia, que lleva a la completa transformación de las ciencias de la vida y logra un progreso sin precedentes en el análisis de genomas, la evaluación de la ecología microbiana y el diagnóstico de enfermedades.


Introduction: The purpose of sequencing is to determine the composition of the nucleotides present in DNA or RNA. Since the completion of the human genome project, several sequencing technologies such as Roche 454, SOLiD, Illumina, Ion Torrent, PacBio and Oxford Nanopore have emerged as tools for rapid sequencing, with greater precision and cost-efficiency, allowing the development of lar-ge-scale projects and the study of genes and genomes, along with the composition of microbiomes and the study of metabolic and genetic diseases that affect the population. Objective: To describe the foundations of the methods of DNA sequencing and their applications in the biomedical sciences. Methods: Descriptive review of the main strategies of first, second and third generation DNA sequencing and their application in the biomedical environment. This review was carried out by searching articles in electronic databases specialized in scientific research. A total of 118 papers were found, of which 6 were excluded as they did not meet the inclusion criteria and 112 were selected as meeting all the requirements. Conclusions: The emergence of next-generation sequencing methods yielding a wealth of data, including fully sequenced genomes of various species, with extensive throughput, reduced time and cost-effec-tiveness that has led to the complete transformation of the life sciences, achieving unprecedented progress in genome analysis, assessment of microbial ecology and disease diagnosis


Introdução: o objetivo do sequenciamento é determinar a composição dos nucleotídeos presentes no DNA ou RNA. Desde a conclusão do projeto do genoma humano, surgiram diversas tecnologias de sequenciamento rápida como Roche 454, SOLiD, Illumina, Ion Torrent, PacBio e Oxford Nanopore, mais precisas e econômicas, que permitem o desenvolvimento de projetos de grande escala e estudo de genes e genomas, composição de microbiomas, doenças metabólicas e genéticas que afetam a popula-ção. Objetivo: descrever os fundamentos dos métodos de sequenciamento de DNA e suas aplicações nas ciências biomédicas. Métodos: revisão descritiva das principais estratégias de sequenciamento de DNA de primeira, segunda e terceira geração e sua aplicação no ambiente biomédico, a partir da busca de artigos em bases de dados eletrônicas especializadas em pesquisa científica. Foram encontrados 118 documen-tos, dos quais 6 foram excluídos por não atenderem aos critérios de inclusão e 112 fo-ram seleciona-dos por atenderem a todos os requerimentos. Conclusões: o surgimento de métodos de sequenciamento de próxima geração rende uma riqueza de dados, incluindo genomas totalmente se-quenciados de várias espécies, com produção extensa, tempos reduzidos e eficiência de custo, levando à transformação completa das ciências da vida e alcançando um progresso sem precedentes no genoma análise, avaliação de ecologia microbiana e diagnóstico de doenças.


Subject(s)
High-Throughput Nucleotide Sequencing , DNA , Genome, Human , Genetic Techniques , Sequence Analysis
7.
Rev. peru. med. exp. salud publica ; 36(3): 423-432, jul.-sep. 2019. tab, graf
Article in Spanish | LILACS | ID: biblio-1058763

ABSTRACT

RESUMEN Objetivos . Comparar diferentes métodos de extracción de ADN a partir de quistes y trofozoítos de Giardia spp. mediante la técnica de reacción en cadena de la polimerasa (PCR) convencional. Materiales y métodos. Se aislaron quistes de Giardia spp. a partir de 65 muestras coprológicas procedentes de hospitales de referencia nacional, obteniéndose una carga promedio de 5x104 parásitos. Asimismo, se cultivaron trofozoítos de Giardia intestinalis (ATCC® 30957™) obteniéndose una carga parasitaria de 5x106. Se compararon once métodos de extracción para quistes y seis para trofozoítos. La concentración y pureza del ADN extraído se determinó por espectrofotometría y el rendimiento de la extracción se evaluó mediante la amplificación de los genes beta giardina (bg) y glutamato deshidrogenasa (gdh) por PCR semi-anidada. Resultados. Se observó que el método I mostró la mayor concentración de ADN a partir de quistes (12,24 ng/µL), pureza (1,4) y mejor rendimiento (100% amplificación bg, 60% gdh) en comparación con los otros métodos evaluados. En el caso de los trofozoítos el método que no tuvo pretratamientos presentó la mayor concentración de ADN, pureza y rendimiento (26,56 ng/µL; 1,85; 100% amplificación bg y gdh). Conclusiones. Los pretratamientos mecánicos, de choque térmico y enzimáticos son necesarios para la ruptura de la pared quística de Giardia spp., siendo el marcador molecular bg de mayor rendimiento para detección de ADN de quistes. Los trofozoítos no requieren pretratamientos para lograr resultados satisfactorios. Se cuenta con una metodología reproducible para la extracción de ADN de Giardia spp. a partir de cualquier estadio evolutivo.


ABSTRACT Objectives. To compare different methods of DNA extraction from cysts and trophozoites of Giardia spp. using the conventional polymerase chain reaction (PCR) technique. Materials and Methods. Cysts of Giardia spp. were isolated from 65 coprological samples from national reference hospitals, obtaining an average load of 5x104 parasites. In addition, Giardia intestinalis trophozoites (ATCC® 30957™) were cultured obtaining a 5x106 parasitic load. Eleven extraction methods for cysts and six for trophozoites were compared. The concentration and purity of the extracted DNA were determined by spectrophotometry and the extraction yield was assessed by amplification of the ß-giardin (bg) and glutamate dehydrogenase (gdh) genes with a semi nested PCR assay. Results. It was observed that method 1 showed the highest concentration of DNA from cysts (12.24 ng/µL), purity (1.4) and best performance (bg: 100% amplification; gdh: 60% amplification) compared to the other methods evaluated. In the case of trophozoites, the method without pre treatment showed the highest level of DNA concentration, purity, and yield (26.56 ng/µL; 1.85; 100% amplification of bg and gdh, respectively). Conclusions . Mechanical, thermal shock, and enzymatic pre-treatments are necessary for the rupture of the cystic wall of Giardia spp. making it the highest-yielding bg molecular marker for detecting cyst DNA. Trophozoites do not require pre-treatment to achieve satisfactory results. A reproducible methodology for the extraction of DNA from Giardia spp. from any evolutionary stage is available.


Subject(s)
Humans , DNA/isolation & purification , Polymerase Chain Reaction , Trophozoites/genetics , Giardia/genetics , Parasitology/methods , Polymerase Chain Reaction/methods , Genetic Techniques
8.
Rev. Asoc. Méd. Argent ; 132(1): 7-19, Mar. 2019. ilus, tab, graf
Article in Spanish | LILACS | ID: biblio-1009926

ABSTRACT

Se presenta el caso clínico de un lactante fallecido a los siete meses de edad con cuadro intersticial persistente. Objetivos: describir detalladamente el camino diagnóstico; alertar sobre posibles confusiones en recién nacidos con diagnósticos más frecuentes; detallar los hallazgos clínicos, radiológicos y de anatomía patológica (consultas en el exterior). Metodología: sumatoria de estudios complejos para descartar causas más frecuentes de patología intersticial en el lactante; consultas radiológicas, de anatomía patológica y genética en el exterior del país. Resultado: con diagnóstico de PAP (proteinosis alveolar pulmonar) se encontró una duplicación de material genético a nivel de cromosoma X, correspondiente al gen CSF2RA (colony stimulating factor 2-subunidad a). Este gen codifica al receptor CSF2 cuya citoquina controla la producción, diferenciación y función de granulocitos/macrófagos. (AU)


A clinical case of a deceased seven month old infant presenting persistent interstitial lung compromise is presented. Objectives. Detailed description of the diagnostic pathway used; to alert about possible confusion with other ­ more frequent ­ pathologies in the new borninfant age; to present clinical, radiological, genetic and pathology findings (consultations abroad). Methodology. A complete description of complex studies to rule out other more frequent pathologies is presented together with radiological, pathological and genetic results from consultations abroad. Results. A diagnosis of PAP (pulmonary alveolar proteinosis) was confirmed with duplication of genetic material at CSF2RA gene (colony stimulating factor 2-subunit a). This gene codifies the CSF2 receptor whose cytokine controls production, differentiation and function of granulocytes/macrophages. (AU)


Subject(s)
Humans , Male , Infant, Newborn , Infant , Lung Diseases, Interstitial/diagnosis , Lung Diseases/diagnosis , Lung Diseases/genetics , Lung Diseases/pathology , Lung Diseases/diagnostic imaging , Sex Chromosome Aberrations , Pulmonary Surfactants , Tomography, X-Ray Computed , Follow-Up Studies , Genetic Techniques , Lung Diseases, Interstitial/genetics , Diagnosis, Differential , Lung/pathology , Mutation/genetics
9.
Journal of Zhejiang University. Science. B ; (12): 728-739, 2019.
Article in English | WPRIM | ID: wpr-1010480

ABSTRACT

As one of the most important aquatic fish, Micropterus salmoides suffers lethal and epidemic disease caused by rhabdovirus at the juvenile stage. In this study, a new strain of M. salmoides rhabdovirus (MSRV) was isolated from Yuhang, Zhejiang Province, China, and named MSRV-YH01. The virus infected the grass carp ovary (GCO) cell line and displayed virion particles with atypical bullet shape, 300-500 nm in length and 100-200 nm in diameter under transmission electron microscopy. The complete genome sequence of this isolate was determined to include 11 526 nucleotides and to encode five classical structural proteins. The construction of the phylogenetic tree indicated that this new isolate is clustered into the Vesiculovirus genus and most closely related to the Siniperca chuatsi rhabdovirus. To explore the potential for a vaccine against MSRV, a glycoprotein (1-458 amino acid residues) of MSRV-YH01 was successfully amplified and cloned into the plasmid pFastBac1. The high-purity recombinant bacmid-glycoprotein was obtained from DH10Bac through screening and identification. Based on polymerase chain reaction (PCR), western blot, and immunofluorescence assay, recombinant virus, including the MSRV-YH01 glycoprotein gene, was produced by transfection of SF9 cells using the pFastBac1-gE2, and then repeatedly amplified to express the glycoprotein protein. We anticipate that this recombinant bacmid system could be used to challenge the silkworm and develop a corresponding oral vaccine for fish.


Subject(s)
Animals , Female , Baculoviridae/metabolism , Bass/metabolism , Carps/virology , Cell Line , Genetic Techniques , Genome, Viral , Glycoproteins/biosynthesis , Insecta , Ovary/virology , Phylogeny , Plasmids/metabolism , Recombinant Proteins/biosynthesis , Rhabdoviridae/metabolism
10.
Journal of Zhejiang University. Science. B ; (12): 713-727, 2019.
Article in English | WPRIM | ID: wpr-1010479

ABSTRACT

Production of reactive oxygen species (ROS) is a conserved immune response primarily mediated by NADPH oxidases (NOXs), also known in plants as respiratory burst oxidase homologs (RBOHs). Most microbe-associated molecular patterns (MAMPs) trigger a very fast and transient ROS burst in plants. However, recently, we found that lipopolysaccharides (LPS), a typical bacterial MAMP, triggered a biphasic ROS burst. In this study, we isolated mutants defective in LPS-triggered biphasic ROS burst (delt) in Arabidopsis, and cloned the DELT1 gene that was shown to encode RBOHD. In the delt1-2 allele, the antepenultimate residue, glutamic acid (E919), at the C-terminus of RBOHD was mutated to lysine (K). E919 is a highly conserved residue in NADPH oxidases, and a mutation of the corresponding residue E568 in human NOX2 has been reported to be one of the causes of chronic granulomatous disease. Consistently, we found that residue E919 was indispensable for RBOHD function in the MAMP-induced ROS burst and stomatal closure. It has been suggested that the mutation of this residue in other NADPH oxidases impairs the protein's stability and complex assembly. However, we found that the E919K mutation did not affect RBOHD protein abundance or the ability of protein association, suggesting that the residue E919 in RBOHD might have a regulatory mechanism different from that of other NOXs. Taken together, our results confirm that the antepenultimate residue E is critical for NADPH oxidases and provide a new insight into the regulatory mechanisms of RBOHD.


Subject(s)
Humans , Agrobacterium tumefaciens/metabolism , Alleles , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Gene Expression Regulation, Plant , Genetic Techniques , Lipopolysaccharides/metabolism , Luminescence , Mutation , NADPH Oxidase 2/chemistry , NADPH Oxidases/genetics , Plant Stomata/metabolism , Protein Domains , Reactive Oxygen Species/metabolism , Nicotiana/metabolism
11.
Yonsei Medical Journal ; : 1-9, 2019.
Article in English | WPRIM | ID: wpr-719693

ABSTRACT

For the past three decades, a large number of genetic studies have been performed to examine genetic variants associated with asthma and its subtypes in hopes of gaining better understanding of the mechanisms underlying disease pathology and to identify genetic biomarkers predictive of disease outcomes. Various methods have been used to achieve these objectives, including linkage analysis, candidate gene polymorphism analysis, and genome-wide association studies (GWAS); however, the degree to which genetic variants contribute to asthma pathogenesis has proven to be much less significant than originally expected. Subsequent application of GWAS to well-defined phenotypes, such as occupational asthma and non-steroidal anti-inflammatory drugexacerbated respiratory diseases, has overcome some of these limitations, although with only partial success. Recently, a combinatorial analysis of single nucleotide polymorphisms (SNPs) identified by GWAS has been used to develop sets of genetic markers able to more accurately stratify asthma subtypes. In this review, we discuss the implications of the identified SNPs in diagnosis of asthma and its subtypes and the progress being made in combinatorial analysis of genetic variants.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal , Aspirin , Asthma , Asthma, Occupational , Biomarkers , Diagnosis , Genetic Association Studies , Genetic Markers , Genetic Techniques , Genome-Wide Association Study , Hope , Pathology , Phenotype , Polymorphism, Single Nucleotide
12.
Journal of Zhejiang University. Science. B ; (12): 620-629, 2018.
Article in English | WPRIM | ID: wpr-1010399

ABSTRACT

Targeting Induced Local Lesions IN Genomes (TILLING) is a reverse genetics strategy for the high-throughput screening of induced mutations. γ radiation, which often induces both insertion/deletion (Indel) and point mutations, has been widely used in mutation induction and crop breeding. The present study aimed to develop a simple, high-throughput TILLING system for screening γ ray-induced mutations using high-resolution melting (HRM) analysis. Pooled rice (Oryza sativa) samples mixed at a 1:7 ratio of Indel mutant to wild-type DNA could be distinguished from the wild-type controls by HRM analysis. Thus, an HRM-TILLING system that analyzes pooled samples of four M2 plants is recommended for screening γ ray-induced mutants in rice. For demonstration, a γ ray-mutagenized M2 rice population (n=4560) was screened for mutations in two genes, OsLCT1 and SPDT, using this HRM-TILLING system. Mutations including one single nucleotide substitution (G→A) and one single nucleotide insertion (A) were identified in OsLCT1, and one trinucleotide (TTC) deletion was identified in SPDT. These mutants can be used in rice breeding and genetic studies, and the findings are of importance for the application of γ ray mutagenesis to the breeding of rice and other seed crops.


Subject(s)
Crops, Agricultural/radiation effects , Gamma Rays , Genetic Techniques , Genome, Plant , Homozygote , INDEL Mutation , Mutagenesis , Oryza/radiation effects , Plant Breeding , Polymerase Chain Reaction , Seeds , Sequence Analysis, DNA , Sequence Deletion
13.
Dementia and Neurocognitive Disorders ; : 131-136, 2018.
Article in English | WPRIM | ID: wpr-718822

ABSTRACT

Alzheimer's disease (AD) related genes have been elucidated by advanced genetic techniques. Familial autosomal dominant AD genes founded by linkage analyses are APP, PSEN1, PSEN2, ABCA7, and SORL1. Genome-wide association studies have found risk genes such as ABCA7, BIN1, CASS4, CD33, CD2AP, CELF1, CLU, CR1, DSG2, EPHA1, FERMT2, HLA-DRB5-HLA-DRB1, INPP5D, MEF2C, MS4A6A/MS4A4E, NME8, PICALM, PTK2B, SLC24A4, SORL1, and ZCWPW1. ABCA7, SORL1, TREM2, and APOE are proved to have high odds ratio (>2) in risk of AD using next generation sequencing studies. Thanks to the promising genetic techniques such as CRISPR-CAS9 and single-cell RNA sequencing opened a new era in genetics. CRISPR-CAS9 can directly link genetic knowledge to future treatment. Single-cell RNA sequencing are providing useful information on cell biology and pathogenesis of diverse diseases.


Subject(s)
Alzheimer Disease , Apolipoproteins E , CRISPR-Cas Systems , Genetic Techniques , Genetics , Genome-Wide Association Study , Odds Ratio , Sequence Analysis, RNA , Single-Cell Analysis
14.
Rev. Inst. Adolfo Lutz (Online) ; (77): 1-7, 2018. graf
Article in Portuguese | LILACS, SES-SP, SESSP-ACVSES, SESSP-IALPROD, SES-SP, SESSP-IALACERVO | ID: biblio-1119212

ABSTRACT

O conceito de Saúde Única surgiu para ressaltar a união indissociável entre a saúde animal, humana e ambiental. Nesse contexto, a leishmaniose visceral americana (LVA) é considerada uma importante doença de saúde pública no Brasil, devido a sua crescente expansão geográfica e aumento na incidência de casos humanos. A LVA é uma doença parasitária, zoonótica, causada pela Leishmania (Leishmania) infantum (syn. chagasi) e transmitida por flebotomíneos do gênero Lutzomyia. Os cães são considerados os principais reservatórios do parasito nas áreas urbanas. O diagnóstico da LVA é baseado em aspectos epidemiológicos, clínicos e laboratoriais. A demonstração da presença do parasito através de exames diretos em tecidos biológicos do hospedeiro é o diagnóstico de escolha, principalmente, em municípios em que a transmissão de LVA ainda não tenha sido confirmada. Diversas metodologias podem ser aplicadas com essa finalidade. O objetivo desse trabalho é apresentar as técnicas citológicas, anatomo-patológicas e moleculares em amostras fixadas em formalina e incluídas em parafina para o diagnóstico da leishmaniose visceral em humanos e cães. Esses dados são complementares à apresentação realizada no I Simpósio Internacional de Leishmaniose Visceral, realizado nos dia 23 e 24 de Abril de 2018, e organizado pelo Instituto Adolfo Lutz em São Paulo-SP, Brasil. (AU)


The One Health concept emerged to highlight the inseparable link between animal, human and environmental health. In this context, American Visceral Leishmaniasis (AVL) is considered an important public health disease in Brazil, due to its increasing geographic expansion and in the incidence of human cases. AVL is a parasitic and zoonotic disease caused by Leishmania (Leishmania) infantum (syn. chagasi) and transmitted by sandflies of the genus Lutzomyia. Dogs are considered the main reservoirs of the parasite in urban areas. The diagnosis of AVL is based on epidemiological, clinical and laboratory aspects. The demonstration of the presence of the parasite through direct examinations in biological tissues of the host is the diagnosis of choice, mainly in municipalities where the transmission of AVL has not yet been confirmed. Several methodologies can be applied for this purpose. The objective of this work is to present the cytological, anatomopathological and molecular techniques in formalin fixed and paraffin embedded samples for the diagnosis of visceral leishmaniasis in humans and dogs. These data are complementary to the present study at the First International Symposium on Visceral Leishmaniasis, held on April 23 and 24, 2018, and organized by Adolfo Lutz Institute in São Paulo, Brazil. (AU)


Subject(s)
Animals , Dogs , Immunohistochemistry , Genetic Techniques , Public Health Surveillance , Leishmaniasis, Visceral/diagnosis
15.
Psychiatry Investigation ; : 300-305, 2018.
Article in English | WPRIM | ID: wpr-713459

ABSTRACT

OBJECTIVE: Autism spectrum disorders (ASD) have a complex pathophysiology including genetic, inflammatory and neurodevelopmental components. We aim to investigate the relationship between ASD and gene polymorphisms of stromal cell-derived factor-1 (SDF-1) and its receptor CXC chemokine receptor-4 (CXCR4), which may affect inflammatory and neurodevelopmental processes. METHODS: 101 children diagnosed with ASD aged 2–18 and their biological parents were included in the study. All participants were assessed using an information form and the Children were assessed using Childhood Autism Rating Scale (CARS). SDF-1 G801→A and CXCR4 C13→T polymorphisms were detected by genetic techniques. The results were evaluated using the transmission disequilibrium test (TDT) and haplotype relative risk (HRR). RESULTS: Following TDT evaluation for CXCR4, the assumption of equality was not rejected (χ²=1.385, p=0.239). HRR for the C allele was 1.037 [HRR (95%CI)=0.937 (0.450–2.387), χ²=0.007, p=0.933] and HRR for the T allele was 0.965 [HRR (95%CI)=0.965 (0.419– 2.221), χ²=1.219, p=0.270], but the findings were statistically insignificant. Based on TDT evaluation for SDF1, the assumption of equality cannot be rejected (χ²=0, p=0.999). HRR for the A allele was 0.701 [HRR (95%CI)=0.701 (0.372–1.319), χ²=1.219, p=0.270] and HRR for the G allele was 1.427 [HRR (95%CI)=1.427 (0.758–2.686), χ²=1.219, p=0.270], but the findings were statistically insignificant. CONCLUSION: The genetic screening of blood samples from mother, father and child trios could not show a significant association between SDF1/CXCR4 genes and ASD on the basis of TDT and HRR tests. More extensive genetic studies are now needed to investigate the relationship between SDF1/CXCR4 gene polymorphisms and ASD.


Subject(s)
Child , Humans , Alleles , Autism Spectrum Disorder , Autistic Disorder , Fathers , Genetic Techniques , Genetic Testing , Haplotypes , Mothers , Parents
16.
Korean Journal of Pediatrics ; : 55-63, 2017.
Article in English | WPRIM | ID: wpr-197571

ABSTRACT

Advances in podocytology and genetic techniques have expanded our understanding of the pathogenesis of hereditary steroid-resistant nephrotic syndrome (SRNS). In the past 20 years, over 45 genetic mutations have been identified in patients with hereditary SRNS. Genetic mutations on structural and functional molecules in podocytes can lead to serious injury in the podocytes themselves and in adjacent structures, causing sclerotic lesions such as focal segmental glomerulosclerosis or diffuse mesangial sclerosis. This paper provides an update on the current knowledge of podocyte genes involved in the development of hereditary nephrotic syndrome and, thereby, reviews genotype-phenotype correlations to propose an approach for appropriate mutational screening based on clinical aspects.


Subject(s)
Humans , Genetic Association Studies , Genetic Techniques , Genetics , Glomerulosclerosis, Focal Segmental , Mass Screening , Nephrotic Syndrome , Podocytes , Sclerosis , Wills
17.
The Korean Journal of Internal Medicine ; : 984-995, 2017.
Article in English | WPRIM | ID: wpr-67287

ABSTRACT

Although debate on the concept of fibromyalgia (FM) has been vigorous ever since the classification criteria were first published, FM is now better understood and has become recognized as a disorder. Recently, FM has come to be considered a major health problem, affecting 1% to 5% of the general population. As familial aggregations have been observed among some FM patients, genetic research on FM is logical. In fact, genome-wide association studies and linkage analysis, and studies on candidate genes, have uncovered associations between certain genetic factors and FM. Genetic susceptibility is now considered to influence the etiology of FM. At the same time, novel genetic techniques, such as microRNA analysis, have been used in attempts to improve our understanding of the genetic predisposition to FM. In this article, we review recent advances in, and continuing challenges to, the identification of genes contributing to the development of, and symptom severity in, FM.


Subject(s)
Humans , Classification , Fibromyalgia , Genetic Predisposition to Disease , Genetic Research , Genetic Techniques , Genetics , Genome-Wide Association Study , Logic , MicroRNAs
18.
Annals of Pediatric Endocrinology & Metabolism ; : 226-230, 2017.
Article in English | WPRIM | ID: wpr-169583

ABSTRACT

Newly developed genetic techniques can reveal mosaicism in individuals diagnosed with monosomy X. Noninvasive prenatal diagnosis using maternal blood can detect most fetuses with X chromosome abnormalities. Low-dose and ultralow-dose estrogen replacement therapy can achieve a more physiological endocrine milieu. However, many complicated and controversial issues with such treatment remain. Therefore, lifetime observation, long-term studies of health problems, and optimal therapeutic plans are needed for women with Turner syndrome. In this review, we discuss several diagnostic trials using recently developed genetic techniques and studies of physiological hormone replacement treatment over the last 5 years.


Subject(s)
Female , Humans , Diagnosis , Estrogen Replacement Therapy , Fetus , Genetic Techniques , Hormone Replacement Therapy , Mosaicism , Prenatal Diagnosis , Turner Syndrome , X Chromosome
19.
Journal of Experimental Hematology ; (6): 1284-1288, 2016.
Article in Chinese | WPRIM | ID: wpr-246774

ABSTRACT

CRISPR/Cas genome editing technology is a newly developed powerful tool for genetic manipulation, which can be used to manipulate the genome at specific locations precisely, to restore the function of genetic defect cells, and to develop various disease models. In recentl years, with the advances of precise genome manipulation, CRISPR/Cas technology has been applied to many aspects of diseases research and becomes an unique tool to investigate gene function and discover new therapeutic targets for genetic diseases. Nowadays, CRISPR/Cas technology has been a hot research point in agriculture, graziery, biotechnology and medicine. This review focuses on the recent advances in CRISPR/Cas technology and its application in hematological diseases.


Subject(s)
Humans , CRISPR-Cas Systems , Gene Editing , Genetic Techniques , Genome , Hematologic Diseases , Phenotype
20.
International Journal of Stem Cells ; : 9-20, 2016.
Article in English | WPRIM | ID: wpr-196827

ABSTRACT

Forced expression of lineage-specific transcription factors in somatic cells can result in the generation of different cell types in a process named direct reprogramming, bypassing the pluripotent state. However, the introduction of transgenes limits the therapeutic applications of the produced cells. Numerous small-molecules have been introduced in the field of stem cell biology capable of governing self-renewal, reprogramming, transdifferentiation and regeneration. These chemical compounds are versatile tools for cell fate conversion toward desired outcomes. Cell fate conversion using small-molecules alone (chemical reprogramming) has superiority over arduous traditional genetic techniques in several aspects. For instance, rapid, transient, and reversible effects in activation and inhibition of functions of specific proteins are of the profits of small-molecules. They are cost-effective, have a long half-life, diversity on structure and function, and allow for temporal and flexible regulation of signaling pathways. Additionally, their effects could be adjusted by fine-tuning concentrations and combinations of different small-molecules. Therefore, chemicals are powerful tools in cell fate conversion and study of stem cell and chemical biology in vitro and in vivo. Moreover, transgene-free and chemical-only transdifferentiation approaches provide alternative strategies for the generation of various cell types, disease modeling, drug screening, and regenerative medicine. The current review gives an overview of the recent findings concerning transdifferentiation by only small-molecules without the use of transgenes.


Subject(s)
Biology , Drug Design , Genetic Techniques , Half-Life , Mass Screening , Regeneration , Regenerative Medicine , Stem Cells , Transcription Factors , Transgenes
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