9p21 Locus Polymorphisms: Risk and Severity Factors of Coronary Artery Disease in Venezuelan Patients

Abstract Background: The 9p21 region is the most relevant locus associated with coronary heart disease in different populations. However, there are no studies that prove that this region is a risk factor in the Venezuelan population. Objectives: To analyze whether or not the 9p21 rs1333049 polymorphism is a risk factor for acute myocardial infarction (AMI) in Venezuelan patients, as well as to investigate its correlation with cardiovascular risk factors (CVRF), age of occurrence, type and severity of infarction, and the correlation of the rs10757274 polymorphism with severity of coronary artery disease. Methods: This was an association study, including 487 unrelated Venezuelan individuals, grouped in 354 patients with AMI and 133 controls. The rs1333049 and rs10757274 polymorphisms were determined using the polymerase chain reaction (PCR) technique with sequence-specific primers. The analysis of association was determined using the SNPStats tool. The continuous variable description and the correlations were performed using the SPSS statistical software. Significance was established at p<0.05. Results: A positive correlation was observed between the rs1333049 polymorphism and the presence of hypertension ( r: 0.145, p: 0.006), and between hypertension and heart infarction ( r: 0.318, p: <0.0001). A positive correlation was found between the rs10757274 polymorphism and the number of coronary vessels that presented obstructive lesions in patients aged ≤ 55 years ( r: 0.276, p: 0.0078). Conclusion: The rs1333049 polymorphism at the 9p21 locus is correlated with hypertension in Venezuelan patients, while the rs10757274 polymorphism is associated with the progression of coronary atherosclerosis, suggested by the correlation with the number of coronary vessels that presented significant obstructive lesions.

Tipificación del cassette cromosómico estafilocócico de Staphylococcus aureus resistentes al meticilino en el estado de Aragua, Venezuela / Typification of the staphylococcal chromosome cassette of methicillin-resistant Staphylococcus aureus in the state of Aragua, Venezuela

RESUMEN Objetivos: Tipificar el casette SCCmec en cepas de Staphylococcus aureus resistentes a meticilino (SARM) en aislados clínicos de centros de salud del Estado Aragua-Venezuela y comparar la presencia de los genotipos SCCmec entre los centros de salud del estado y según el tipo de infección. Materiales y métodos: Durante enero y agosto de 2015 se estudiaron 81 cepas SARM de cuatro centros de salud del estado de Aragua en Venezuela. La resistencia al meticilino se midió con el método de Kirby-Bauer con discos de oxacilina (1 µgr) y cefoxitina (30 µgr). El gen mecA y el SCCmec se analizaron por la técnica de reacción en cadena de polimerasa múltiple. Resultados: 55 aislados (67,9%) amplificaron el gen mecA, y 24 cepas (43,6%) amplificaron el SCCmec. El SCCmec I fue el más frecuente, seguido de SCCmecIV y SCCmec III, representaron el 62,5%, 25% y 12,5%, respectivamente. El SCCmec I fue predominante en el centro de salud A (80%), mientras que el SCCmec IV se encontró en el centro de salud B (60%) y C (100%). En el centro de salud D, 50% resultó ser SCCmec I y 50% SCCmec IVd. Se encontró relación entre el SCCmec y el centro de salud con significancia estadística. En infecciones de piel y tejidos blandos y en las respiratorias predominó el SCCmec I con 63,2% y 50% respectivamente. Conclusiones: La frecuencia de SCCmec I y IV permitirá establecer nuevas medidas en el uso y control de la resistencia a los antibióticos.

ABSTRACT Objective: Typify the SCCmec cassette in methicillin-resistant strains of Staphylococcus aureus in clinical isolates from health centers in the State of Aragua-Venezuela and compare the presence of SCCmec genotypes among the state health centers and according to the type of infection. Materials and methods: 81 MRSA strains from four health centers of the Aragua-Venezuela State were studied. Methicillin resistance was performed with the Kirby-Bauer method with oxacillin (1 µg) and cefoxitin (30 µg) disks. The mecA gene and SCCmec were analyzed by the multiple PCR technique. Results: Only 55 isolates (67.9%) amplified the mecA gene, and 24 strains (43.6%) amplified SCCmec. SCCmec type I was the most frequency, followed by SCCmec IV and SCCmec III, representing 62.5%, 25% and 12.5%, respectively. SCCmec I was predominant in health center A (80%), while in B and C 60% and 100% respectively were SCCmec IV. At health center D, 50% turned out to be SCCmec I and 50% SCCmec IVd. A relationship was found between the SCCmec and the health center with statistical significance. SCCmec I predominated in skin and soft tissue and respiratory infections with 63.2% and 50%, respectively. There was no association between genotype and type of infection with a p value greater than 0.05. Conclusions: The prevalence of SCCmec I and IV will allow establishing new measures in the use of antibiotics and epidemiological control.

Advances in three-dimensional genomics / 生物工程学报

Three-dimensional (3D) genomics is an emerging discipline that studies the 3D spatial structure and function of genomes, focusing on the 3D spatial conformation of genome sequences in the nucleus and its biological effects on biological processes such as DNA replication, DNA recombination and gene expression regulation. The invention of chromosome conformation capture (3C) technology speeds up the research on 3D genomics and its related fields. Furthermore, the development of 3C-based technologies, such as the genome-wide chromosome conformation capture (Hi-C) and chromatin interaction analysis using paired-end tag sequencing (ChIA-PET), help scientists get insight into the 3D genomes of various species. Aims of 3D genomics are to reveal the spatial genome organization, chromosomal interaction patterns, mechanisms underlying the transcriptional regulation and formation of biological traits of microorganism, plant, animal. Additionally, the identification of key genes and signaling pathways associated with biological processes and disease via chromosome 3C technology boosts the rapid development of agricultural science, life science and medical science. This paper reviews the research progress of 3D genomics, mainly in the concept of 3D genomics, the development of chromosome 3C technologies and their applications in agricultural science, life science and medical science, specifically in the field of tumor.

Three-dimensional chromosome conformation capture and its derived technologies / 生物工程学报

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.

Los cromosomas de las tortugas tropicales: Kinosternon leucostomum, Trachemys scripta y Staurotypus triporcatus (Testudines: Kinosternidae/Emydidae) / Tropical turtles chromosomes: Kinosternon leucostomum, Trachemys scripta and Staurotypus triporcatus (Testudines: Kinosternidae/Emydidae)

Mexico is a biodiverse country in several taxa as reptiles, that include several species of freshwater and marine turtles. Eventhough most of this group species are under protection, Tabasco State has nine native freshwater turtles, like Kinosternon leucostomum, Trachemys scripta and Staurotypus triporcatus that are very important in traditional dishes. This has resulted in a critical level of their populations, together with little biological knowledge for their conservation. Therefore, this study was dedicated to turtle cytogenetics. The study was conducted using the conventional methods for cytogenetics. The results showed the modal diploid and haploid number for K. leucostomum of 2n=56 (2n=56+3 microchromosomes “B”) and 1n=28 chromosomes in mitosis and meiosis, respectively. In T. scripta 2n=50 chromosomes (2n=50+2 microchromosomes “B”) and 1n=25 chromosomes were also characterized. Whereas in S. triporcatus we only report the 2=54 chromosomes (2n=54+2 microchromosomes “B”). The karyological formula for K. leucostomum was integrated by 12 metacentric-submetacentric chromosomes “msm”/”A”+22 subtelocentrictelocentric chromosomes “stt”/”B”+22 telocentric chromosomes “T”/”C” with fundamental number (FN) of 90 chromosome arms. While T. scripta karyotype was integrated by 32 “msm/”A”+10 “stt”/”B”+8”T/”C” chromosomes, with FN of 92 arms. S. triporcatus karyotype formula was built up by 20 chromosomes “msm”/”A”+34 chromosomes “T”/”C” with FN of 74. The variation in chromosome classification, the fundamental number and the presence of supernumerary microchromosomes “B” in the studied species, were evidence of a particular chromosome cytotypes in Tabasco. We considered that the presence of microchromosomes “B” probably has different origins, and they may be very important as a pattern for the formation or separation of new species. This study also showed the absence of heterologous chromosomes between the females and males karyotypes from the studied species. Rev. Biol. Trop. 62 (2): 671-688. Epub 2014 June 01.

México es un país biodiverso en varios grupos taxonómicos incluyendo a los reptiles, por ello en el país existen varias especies de tortugas dulceacuícolas y marinas. Las especies que integran dicho grupo se encuentran dentro del listado de especies sujetas a protección. El estado de Tabasco cuenta con nueve especies de tortugas de agua dulce, de las cuales Kinosternon leucostomum, Trachemys scripta y Staurotypus triporcatus son de las más importantes dentro de la tradición culinaria, hecho que las ha llevado a niveles críticos en sus poblaciones; aunado al poco conocimiento biológico que sobre dichas especies existe para conservarlas. Por lo anterior, el presente estudio de citogenética es el primero en tortugas de agua dulce en la región. El estudio se realizó, empleando el método convencional de citogenética. Los resultados muestran, el número modal diploide y haploide de K. leucostomum de 2n=56 (2n=56+3 microcromosomas “B”) y 1n=28 cromosomas 686 en mitosis y meiosis, respectivamente. En T. scripta de 2n=50 cromosomas (2n=50+2 microcromosomas “B”) y 1n=25 cromosomas. Mientras que en S. triporcatus solo se reporta el 2n=54 cromosomas (2n=54+2 microcromosomas “B”). La fórmula cromosómica en K. leucostomum, fue de 12 cromosomas metacéntricos submetacéntricos “msm”/“A”+22 cromosomas subtelocéntricos-telocéntricos “stt”/“B”+22 cromosomas telocéntricos “T”/“C”, con número fundamental (NF) de 90 brazos cromosómicos. En T. scripta fue de 32 cromosomas “msm”/“A”+10 cromosomas “stt”/“B”+8 cromosomas “T”/“C”, con NF de 92 y en S. triporcatus 20 cromosomas “msm”/“A”+34 cromosomas “T”/“C” con NF de 74. La variación en la clasificación cromosómica, el número fundamental y la presencia de microcromosomas “B” supernumerarios en las tres especies, son evidencia de citotipos cromosómicos particulares de las tortugas de Tabasco. Se argumenta que la presencia de los microcromosomas “B” tiene diferentes orígenes y de su importancia como pauta para la formación o separación de nuevas especies. En el estudio se descarta la presencia de cromosomas heterólogos entre las hembras y los machos de las especies estudiadas.

Pattern of chromosome involvement in childhood hyperdiploid pre-B-cell acute lymhoblastic leukemia cases from India.

BACKGROUND: Hyperdiploid pre‑B‑cell acute lymhoblastic leukemia (pre‑B‑ALL) is a common form of childhood leukemia with very good prognosis with present day chemotherapy. However, the chromosomal composition of the hyperdiploidy has not been extensively studied and possible mechanism for this pathology remains so far conjectural. OBJECTIVE: To analyze the pattern of chromosome involvement in a cohort of childhood hyperdiploid pre‑B‑ALL from India and from this pattern to develop an understanding on the causation of such pathology. Whether such patients also carry translocations and FLT3 mutations in addition to their hyperdiploid karyotype. MATERIALS AND METHODS: One hundred and twenty‑six childhood pre‑B‑ALL patients were studied. Bone marrow aspirate of these patients were evacuated for morphology, FAB classification, immunophenotyping and both conventional and molecular cytogenetics. RESULTS: Of 126 patients with pre‑B‑ALL (age 2-15 years), 90 patients with abnormal karyotype showed 50 with hyperdiploid karyotype (50/90 i.e. 55.5%). Chromosomes 9, 10, 14, 17, 18, 20 and 21 were more often involved in hyperdiploidy. Chromosome 21 duplication was present in 92% of the cases. Chromosomes 5, 15, 16, 17 and Y were less often involved (18-20%) in hyperdiploidy. About 44% of patients with hyperdiploidy had additional karyotypic abnormality of which TEL‑AML1 was predominant (24%). Chromosome loss was rare and accounted for 20% of the cases only. We did not find any FLT3 mutation in our patients. CONCLUSION: In this study, the pattern of chromosome involvement in hyperdiploid karyotype of ALL patients is same as other studies except some chromosomes like 1, 6, 11, 12, 19 and 22 have some more frequent involvement than other studies. This study also showed the occurrence of TEL/AML1 fusion is more (19.8%) than other reports from India.

Alterações genéticas em casais com antecedentes de aborto recorrente no primeiro trimestre da gestação / Genetic alterations in couples with a history of recurrent miscarriage in the first trimester of pregnancy

O abortamento é considerado um problema multifatorial, cujas principais causas envolvidas na sua etiologia são os fatores ambientais (como exposição a substâncias tóxicas), genéticos, anatômicos, endócrinos, imunológicos, trombofílicos e doenças infecciosas (como toxoplasmose, rubéola). No entanto, os fatores genéticos são atribuídos principalmente aos abortamentos de primeiro trimestre da gestação. As alterações cromossômicas, o polimorfismo C677T, no gene da metilenotetrahidrofolato redutase (MTHFR677C>T); o polimorfismo G1691A, no gene do Fator V de Leiden (FVL1691G>A), e o polimorfismo G20210A, no gene da protrombina (PRT20210G>A), têm sido associados a problemas obstétricos, incluindo aborto recorrente. O objetivo deste trabalho foi investigar associação entre as mutações relacionadas à trombofilia, presença de alterações cromossômican e a ocorrência de aborto espontâneo recorrente e avaliar possíveis interações entre as referidas mutações e as alterações cromossômicas. A casuística foi composta por 151 mulheres com história de aborto recorrente, 94 parceiros e 100 controles (mulheres sem histórico de aborto). A investigação das mutações foi realizada pela técnica de Reação em Cadeia da Polimerase- Polimorfismo de Tamanho de Fragmento de Restrição. As alterações cromossômicas foram investigadas pela cariotipagem com banda–G. A frequência das alterações cromossômicas foi de 7,3% nas mulheres com abortamento recorrente e 1% nos controles (p=0,022), e de 2,1% nos parceiros. No entanto, a frequência dos alelos MTHR677C>T (23% versus 22,5%), FVL1691G>A (1,5% versus 1% ) e PRT20210G>A (1,45% versus 0%) foi similar entre casos e controles, respectivamente. No grupo investigado, foi observada associação entre aborto recorrente e alterações cromossômicas, mas não foi encontrada associação com os polimorfismos gênicos investigados.

Abortion is considered a multifactorial problem, the most important causes involved in its etiology are, environmental factors ( as exposure to toxic chemicals), genetic, anatomic, endocrine, immunological, thrombophilic and infectious diseases (such as toxoplasmosis, rubella). However, genetic factors are mainly attributed to abortions of the first trimester of pregnancy. Chromosomal abnormalities, MTHFR 677C>T, factor V Leiden 1691G>A and prothrombin 20210G>A mutations have been associated with obstetric problems, including recurrent miscarriage. The objective of this research was to investigate associations between mutations in three genes commonly associated to thrombophilic events, chromosomal abnormalities and the occurrence of recurrent miscarriage. As well evaluate possible interactions between these mutations and chromosomal abnormalities. The sample was comprised of 151 women with history of recurrent miscarriages, 94 partners and 100 control (women with no history of abortion). The investigation of the mutations was performed by Polymerase Chain Reaction (PCR)/ Restriction Fragment Length Polymorphism (RFLP). Chromosomal aberrations were investigated by karyotyping with G-banda. The frequency of chromosomal abnormalities was 7.3% in women with recurrent miscarriage and 1% in controls (p = 0.022), and 2.1% in the partners. However, the frequency of allele MTHR677C> T (23% versus 22.5%), FVL1691G> A (1.5% vs. 1%) and PRT20210G> A (1.45% vs. 0%) was similar for cases and controls, respectively. In the investigated group was found association between recurrent miscarriage and chromosomal abnormalities, but no association was found with the genetic polymorphisms investigated.

Progress with schistosome transgenesis

Genome sequences for Schistosoma japonicum and Schistosoma mansoni are now available. The schistosome genome encodes ~13,000 protein encoding genes for which the function of only a minority is understood. There is a valuable role for transgenesis in functional genomic investigations of these new schistosome gene sequences. In gain-of-function approaches, transgenesis can lead to integration of transgenes into the schistosome genome which can facilitate insertional mutagenesis screens. By contrast, transgene driven, vector-based RNA interference (RNAi) offers powerful loss-of-function manipulations. Our laboratory has focused on development of tools to facilitate schistosome transgenesis. We have investigated the utility of retroviruses and transposons to transduce schistosomes. Vesicular stomatitis virus glycoprotein (VSVG) pseudotyped murine leukemia virus (MLV) can transduce developmental stages of S. mansoni including eggs. We have also observed that the piggyBac transposon is transpositionally active in schistosomes. Approaches with both VSVG-MLV and piggyBac have resulted in somatic transgenesis and have lead to integration of active reporter transgenes into schistosome chromosomes. These findings provided the first reports of integration of reporter transgenes into schistosome chromosomes. Experience with these systems is reviewed herewith, along with findings with transgene mediated RNAi and germ line transgenesis, in addition to pioneering and earlier reports of gene manipulation for schistosomes.

Octaploidy in idiopathic thrombocytopenic purpura.

We report a case of an elderly 68-year-old male who presented in our hospital with chief complaints of petechial rashes and ecchymosis over extremities and bleeding from the oral cavity since 3–4 days prior to hospitalization. He saw a physician before coming to our hospital and received one dose of IV methylprednisolone and oral wysolone. He had come to our hospital for further management. Bone marrow karyotyping was done and chromosomal analysis revealed two cell lines. Eighty percent of the cells analyzed revealed apparently normal male karyotype. However, 20% cells analyzed revealed a total of 184 chromosomes, suggesting octaploidy.

Chromosomal analyses in Megalonema platanum (Siluriformes: Pimelodidae), an endangered species from South American rivers

Este estudo apresenta dados cromossômicos de Megalonema platanum do rio Tibagi, Paraná, Brasil e do rio Paraná, Argentina. O número diploide foi igual 54 com composição cariotípica de 24m+16sm+2st+12a em ambas populações. Os sítios AgNORs foram detectados na posição terminal de um par submetacêntrico das duas populações analisadas, coincidindo com constrição secundária no braço curto do par 15. CMA3 e FISH com sonda de DNAr 18S exibiram sinais fluorescentes que correspondem aos sítios AgNORs e à constrição secundária. A presença de um pequeno cromossomo supranumerário acrocêntrico foi observado em M. platanum do rio Tibagi, com heterocromatina centromérica. Outros blocos heterocromáticos foram evidenciados na posição terminal de alguns cromossomos e um par cromossômico submetacêntrico grande, provavelmente o primeiro par, mostrou heterocromatina intersticial. Na população do rio Paraná foram observados ainda blocos heterocromáticos em ambas regiões terminais em alguns cromossomos. Este trabalho mostra pela primeira vez dados citogenéticos de M. platanum, que é uma espécie muito rara na bacia do rio Paraná e pode estar ameaçada de extinção.

This study presents chromosomal data of Megalonema platanum from rio Tibagi, Paraná, Brazil and from rio Paraná, Argentina. The diploid number was equal 54 with karyotype composition of 24m+16sm+2st+12a in both populations. The AgNOR siteswere detected in the terminal position of a submetacentric pair of the two analyzed populations, coinciding with secondary constrictions on the short arm of pair 15. CMA3 and FISH with 18S rDNA probe displayed fluorescent signals that correspond to the AgNOR sites and secondary constriction. The presence of a small acrocentric supernumerary chromosome can be observed in M. platanum from rio Tibagi, with centromeric heterochromatin. Others heterochromatic blocks were evidenced in the terminal position of some chromosome and one metacentric large chromosome pair, probably the first pair, showed an interstitial heterochromatin. In the population of the rio Paraná were still observed heterochromatic blocks in both ends in some chromosomes. This work brings for the first time cytogenetic date of M. platanum, which is a very rare species in the rio Paraná basin and may be endangered.

Los cromosomas meióticos de la rana arborícola Smilisca baudinii (Anura: Hylidae)

Meiotic chromosomes of the tree frog Smilisca baudinii (Anura: Hylidae). The Mexican tree frog Smilisca baudinii, is a very common frog in Central America. In spite their importance to keep the ecological equilibrium of the rainforest, its biology and genetics are poorly known. In order to contribute with its biological knowledge, we described the typical meiotic karyotype based in standard cytogenetic protocols to specimens collected in Tabasco, Mexico. The study was centered in the analysis of 131 chromosome spreads at meiotic stage from two adults of the species (one female and one male). The metaphase analysis allowed the establishment of the modal haploid number of 1n=12 bivalent chromosomes. The chromosomic formulae from the haploid bivalent karyotype was integrated by 12 biarmed chromosomes characterized by twelve pairs of metacentric-submetacentric (msm) chromosomes. The meiotic counting gives the idea that diploid chromosome number is integrated by a complement of 2n=24 biarmed chromosomes. The presence of sex chromosomes from female and male meiotic spreads was not observed. Current results suggest that S. baudinii chromosome structure is well shared among Hylidae family and "B" chromosomes are particular structures that have very important evolutionary consequences in species diversification. Rev. Biol. Trop. 59 (1): 355-362. Epub 2011 March 01.

La rana arborícola mexicana Smilisca baudinii, es una especie de rana común en Centroamérica. Sin embargo, la biología y genética de la especie, es pobremente conocida a pesar de su importancia para mantener en equilibrio ecológico las selvas tropicales. Con el propósito de contribuir con el conocimiento biológico de esta especie, establecimos el cariotipo típico en meiosis en especímenes recolectados en Tabasco, México, mediante procedimientos citogenéticos estándares. El estudio, se fundamentó en el análisis de 131 dispersiones cromosómicas en estadio meiótico de dos adultos de la especie (una hembra y un macho). El análisis de las metafases, permitió establecer el número modal haploide de 1n=12 cromosomas bivalentes. La fórmula cromosómica del cariotipo haploide, se integró por 12 cromosomas birrámeos caracterizado por 12 pares de cromosomas bivalentes metacéntricos-submetacéntricos (msm). Los conteos en meiosis, hacen suponer como número diploide de cromosomas a un complemento integrado por 2n=24 cromosomas birrámeos. No fue posible observar presencia de cromosomas sexuales, entre las dispersiones meióticas del espécimen hembra y macho. Los resultados sugieren que la estructura cromosómica de S. baudinii, es compartida ampliamente entre las especies de la familia Hylidae y los cromosomas "B" son estructuras importantes en la diversificación de las especies.

Biologia molecular: aspectos básicos da genética: parte I / Molecular biology: genetic basic aspects: part I

A presente revisão pretende analisar os mais recentes métodos existentes na Biologia molecular-genética médica, com direcionamento quanto à detecção e ao acompanhamento das diferentes neoplasias ginecológicas. Inicialmente, neste estudo serão discutidos os aspectos básicos e as características celulares importantes, as diversas fases celulares e os métodos de análise existentes para avaliação dos cromossomos e dos genes. São revistos os mais importantes fatores oncogênicos responsáveis pela carcinogênese, bem como a ação dos oncogenes e dos genes supressores dos tumores.

This review intends to analyze the latest existing methods in Molecular Biology-Medical genetics, focusing on the detection and tracking of different gynecological neoplasias. This study will initially discusses basic aspects and important cellular characteristics, as well as the different cellular phases and existing analytical methods for the study of chromosomes and genes. It also reviews the most important oncogenic factors which are responsible for carcinogenesis, as well as the action of oncogenes and tumor-supressing factors.

Síndrome de Coffin-Siris: relato de caso / Coffin-Siris syndrome: case report

Objetivo: Apresentar o caso clínico de ummenino de 4 anos com Síndrome de Coffin-Siris.Relato do caso: O paciente apresentava retardo decrescimento pós-natal, face grosseira, sobrancelhascheias, nariz alargado, filtro longo, boca larga, criptorquidiae hipoplasia da unha do quinto dedo. Ocariótipo em sangue periférico foi normal, 46 XY.Conclusão: As características clínicas do pacientesão consistentes com o diagnóstico da Síndromede Coffin-Siris, uma doença genética em que hámenos de 100 casos descritos.

Objective: To report the clinical case of 4 yearsold boy with Coffin-Siris syndrome. Case report:The patient presented post natal growth retardation,coarse facies, brushy eyebrows, broad nose, long philtrum,wide mouth, cryptorchidism and hypoplastic 5thfingernails. The blood cariotype was normal, 46XY.Conclusions: The clinical characteristics are consistentwith the diagnosis of Coffin-Siris syndrome, agenetic disorder of which there are less than 100 casesreported.

Mutación G47R en el gen SBSN causa síndrome de Bartter con sordera en dos hermanas venezolanas / Bartter syndrome with deafness in two venezuelan sisters

El Síndrome de Bartter (SB) es un grupo heterogéneo de tubulopatías autosómicas recesivas, perdedoras de sal e hipokalémicas. Se han identificado cinco tipos de SB causados por diferentes defectos genéticos, uno de ellos está asociado con sordera neurosensorial (SBSN). Recientemente se han descrito mutaciones en el gen SBND, mapeado en el cromosoma 1p31, asociadas con BSNS. El gen Barttin, codifica para una subunidad B esencial, subunidad de los canales ClC-ka y ClC-kb. Ambas subunidades están co- expresadas en la membrana basolateral de los túbulos renales, en las ramas delgada y gruesa del asa de Henle, y en la vascularización del oído interno. En el presente trabajo se describen los casos clínicos de dos hermanas venezolanas hijas de padres consanguíneos (primo-hermanos) de Jadacaquiva en la Península de Paraguaná, estado Falcón. La secuencia de análisis del gen SBSN mostró que las niñas afectadas eran homocigotas para una transición C-T en axón 1. Esta alteración resulta en una mutación ausente, G47R, la cual suprime el efecto estimulante sobre el barttin de la subunidad del canal ClC-KB. Estas niñas con la mutación G47R presentaron polihidramnios, partoprematuro y pérdida de sal. Sin embargo, la tasa de filtración glomerular de las pacientes es normal. Las manifestaciones clínicas son más moderadas en pacientes con mutación G47R, en relación a otros pacientes publicados con SBSN. Éste es el primer reporte de casos con SBSN en Venezuela.

Bartter syndrome (BS) is a heterogeneous group of autosomal recessive hypokalemic salt-losing tubulopathies. Five types of BS caused by different genetic defects have been identified, and one of them is associated with sensorineural deafness (BSND). Mutations in the recently described BSND gene, mapped in chromosome 1p31, have been reported to be associated with BSNS. This gene encodes barttin, an essential B-subunit ClC-ka and ClC-kb channels. Both subunits are co-expressed in basolateral membranes of renal tubules in the thin and thick ascending limb of Henle’s loop and in the stria vascularis of the inner ear. We studied two venezuelan sisters, daughters of consanguineous parents from a small town called Jadacaquiva, in the peninsula of Paraguaná, Venezuela. Sequence analysis of the BSND gene showed that the affected members were homozygous for C to T transition in axon 1. This alteration results in a missense mutation, G47R that has been previously shown to abolish the stimulatory effect on the subunit barttin of the ClC-Kb channel. The patients with the G47R mutation presented polyhidramnios, premature birth and salt loss. Nevertheless, glomerular filtration rate is normal. Clinical manifestations are moderate in patients with G47R mutation with regard to other patients reported with BSND. This is the first report of BSND in Venezuela.

Cariotipo del pejelagarto tropical Atractosteus tropicus (Lepisosteiformes: Lepisosteidae) y variación cromosómica en sus larvas y adultos

Karyotype of the tropical gar Atractosteus tropicus Lepisosteiformes: Lepisosteidae) and chromosomal variation in their larval and adults. The karyotype of the tropical gar Atractosteus tropicus is described from conventional Giemsa-staining of 295 mitotic chromosome slides from 120 larvae and 15 adults (five females and ten males) from Tabasco, southern of Mexico. The diploid number 2n = 56 chromosomes was calculated (73 spreads from 206 larval and 208 adult metaphases). Variation on chromosome number was from 46 (4.4%) to 64 (3.9%) chromosome elements on larval samples, whereas 58 (11.7 %) chromosomes were the second most abundant after the diploid number of 2n = 56 (35%). Such variation was related with the presence of mobile microchromosomes. The karyotype was determined from six clear chromosome spreads photographed from three females and three males. The averaged karyotype was integrated by eight pairs of metacentric (m) chromosomes, four submetacentric (sm) pairs, eight telocentric (t) pairs and eight pairs of telocentric micro-chromosomes (*t). The fundamental number was FN = 80 chromosome arms. We saw no sexual differences on chromosome structure. Rev. Biol. Trop. 57 (3): 529-539. Epub 2009 September 30.

El cariotipo del pejelagarto Atractosteus tropicus se describe por medio de tinción Giemsa de 295 preparaciones cromosómicas en mitosis a partir de 120 larvas y 15 adultos (5 hembras y diez machos) de la población que habita en Tabasco, sureste de México. El número diploide 2n=56 cromosomas se dio en 281 dispersiones cromosómicas de un total de 445 muestras, de ellas 73 dispersiones provinieron de 206 metafases larvales, y 208 dispersiones de 239 metafases de adultos. Se observó variación en el número de cromosomas desde 46 hasta 64 elementos cromosómicos (larvas). Las metafases con 58 cromosomas fueron el segundo valor más abundante, después del número diploide 2n=56 (35%). En los adultos esa variación no fue relevante. Esta variación cromosómica se relaciona con microcromosomas móviles. El cariotipo se determinó de seis dispersiones cromosómicas bien definidas de tres hembras y tres machos; y el cariotipo promedio se integró de ocho pares de cromosomas metacéntricos (m), cuatro pares de cromosomas submetacéntricos (sm), ocho pares de telocéntricos (t), y ocho pares de microcromosomas telocéntricos (*t) El número fundamental observado en el cariotipo típico promedio fue NF = 80 brazos cromosómicos. No se detectaron diferencias sexuales.

Chromosomal organization of simple sequence repeats in the Pacific oyster (Crassostrea gigas): (GGAT)(4), (GT)(7) and (TA)(10) chromosome patterns.

Chromosome identification is essential in oyster genomic research. Fluorescence in situ hybridization (FISH) offers new opportunities for the identification of oyster chromosomes. It has been used to locate satellite DNAs, telomeres or ribosomal DNA sequences. However, regarding chromosome identification, no study has been conducted with simple sequence repeats (SSRs). FISH was used to probe the physical organization of three particular SSRs, (GGAT)(4), (GT)(7) and (TA)(10) onto metaphase chromosomes of the Pacific oyster, Crassostrea gigas. Hybridization signals were observed in all the SSR probes, but the distribution and intensity of signals varied according to the oligonucleotide repeat. The intercalary, centromeric and telomeric bands were observed along the chromosomes, and for each particular repeat every chromosome pair presented a similar pattern, allowing karyotypic analysis with all the SSRs tested. Our study is the first in mollusks to show the application of SSR in situ hybridization for chromosome identification and karyotyping. This technique can be a useful tool for oyster comparative studies and to understand genome organization in different oyster taxa.

Taxonomic reappraisal of Antithamnion sparsum Tokida (Ceramiaceae, Rhodophyta).

The taxonomic criterion of Antithamnion sparsum was reappraised in comparison with A. densum and A. defectum based on crossing experiments, morphological observation, chromosome study and Random Amplified Polymorphic DNA (RAPD) analysis. These species had a very similar morphology but were sexually isolated. The chromosome number was n = ca. 24 for A. densum, n = ca. 21 for A. defectum, and n = ca. 44 for A. sparsum. All isolates of A. sparsum and A. densum showed polysiphonia-type life history Asexual reproduction was induced by favorable environmental conditions. In A. sparsum, 1-2% of male plants developed mitotic tetrasporangia together with spermatangia. In A. densum, 5-10% of tetraspores developed into asexual tetrasporophytes. Phylogenetic relationships between these species were examined using RAPD analysis, and A. glanduliferum was used as an outgroup. A total of 167 polymorphic RAPD markers amplified from 15 different primers were analyzed. Results suggested that these species were closely related, with A. defectum placed in the middle of A. sparsum and A. densum. Chromosome study and RAPD analysis implied that A. sparsum first separated from A. defectum through polyploidization and later A. densum evolved. These species may present another example of the narrow species concept in the genus Antithamnion.

Los cromosomas mitóticos y meióticos del pez tropical Petenia splendida (Cichlidae)

The mitotic and meiotic chromosomes of the tropical fish Petenia splendida (Cichlidae). The karyotype of bay snook, Petenia splendida, is described based on mitotic and meiotic stages of sixty larvae and twelve juveniles from Tabasco, Mexico. Standard cytological procedures with minor modifications were followed to obtain mitotic and meiotic chromosome spreads. One hundred chromosome slides were analyzed and 290 chromosome spreads were counted. High-quality spreads in mitosis and meiosis were used for karyotype analysis. Mitotic chromosome spreads showed 76.7 % of such cells with 2n=48 chromosomes, while meiotic spreads revealed 55.2 % with 24 chromosomes in haploid stage. Photographic documentation of eight highquality pictures showed that the karyotype consists of three pairs of bi-armed metacentric-submetacentric chromosomes (msm) and 21 pairs with uni-armed subtelocentric-acrocentric chromosomes (sta), with a fundamental number (FN) of 54 arms. Karyotype chromosomes were verified by analysis of haploid and diploid metaphases at meiotic stage I. Abundant chromosome spreads were observed more frequently on slides from larvae. No evidence of heteromorphism to discriminate sexual chromosomes was detected. There were "dot-like" chromatic bodies in both sexes and they were classified as "B" chromosomes. The karyotype of P. splendida is type "A", i.e. primitive in the Cichlid family, similar to other species of Cichlasoma. The occurrence of supernumerary chromosomes is still unknown: studies on the effects of pollution and hybridization might be important to understand that phenomenon. Rev. Biol. Trop. 56 (2): 895-907. Epub 2008 June 30.

Para describir los cromosomas del cariotipo en mitosis y meiosis de la mojarra tenguayaca P. splendida, se procesaron 60 larvas y doce jóvenes (seis hembras y seis machos) procedentes de Tabasco, México. Se emplearon los procedimientos citológicos clásicos para peces pequeños y grandes, con algunas modificaciones que permitieron obtener campos cromosómicos en meiosis y mitosis. Analizamos al microscopio 100 laminillas, contando 290 dispersiones cromosómicas. En mitosis, 76.7 % de los conteos dieron número modal diploide de 2N=48 cromosomas, mientras en meiosis el 55.2 % mostró 24 cromosomas en condición haploide. Se analizaron ocho de las mejores fotografías para establecer el cariotipo y se identificaron tres pares de cromosomas birrámeos metacéntricos-submetacéntricos (msm) y 21 pares de cromosomas monorrámeos subtelocéntricos-acrocéntricos (sta) con número fundamental (N.F) de 54 brazos. Se corroboró el cariotipo mediante el análisis de campos cromosómicos en estadio haploide y diploide de la meiosis I. Las dispersiones cromosómicas tuvieron un número mayor en larvas que en jóvenes. No hubo diferencias heteromórficas para distinguir cromosomas sexuales. Sin embargo, se observó la presencia de cuerpos cromáticos en forma de puntos, como una característica propia de los microcromosomas "B". Para esta familia, el cariotipo de P. splendida es primitivo o tipo "A"; y es estrechamente parecido al del género Cichlasoma. El origen de los cromosomas supernumerarios es un fenómeno aun desconocido en los cíclidos por lo que faltan estudios relacionados con el daño causado por la contaminación y la hibridación.

Chromosome variability in the Chagas disease vector Rhodnius pallescens (Hemiptera, Reduviidae, Rhodniini)

Rhodnius pallescens is the main vector of Trypanosoma cruzi in Panama and one of the most relevant secondary vectors in Colombia. Despite the importance of this species, there is limited knowledge about the genetic variability along its geographical distribution. In order to evaluate the degree of karyotype variability we analyzed the meiotic behavior and banding pattern of the chromosomes of 112 males of R. pallescens coming from different regions of Colombia and Panama. Using the C-banding technique we identified two chromosomal patterns or cytotypes characterized by differences in the amount, size and distribution of constitutive heterochromatic regions in the chromosome complement (2n = 20 autosomes plus XY in males). The individuals can be easily classified in each cytotype by the analysis of the chromosomes during first meiotic prophase. The frequencies of the cytotypes are variable according to the geographic origin of the populations. This chromosomal divergence together with morphological data supports the existence of three genetically different populations of R. pallescens and provides new information to understand the distribution dynamics of this species.