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.

Simultaneous silencing of VEGF and KSP by siRNA cocktail inhibits proliferation and induces apoptosis of hepatocellular carcinoma Hep3B cells

BACKGROUND: Vascular endothelial growth factor (VEGF) is involved in the growth of new blood vessels that feed tumors and kinesin spindle protein (KSP) plays a critical role in mitosis involving in cell proliferation. Simultaneous silencing of VEGF and KSP, an attractive and viable approach in cancer, leads on restricting cancer progression. The purpose of this study is to examine the therapeutic potential of dual gene targeted siRNA cocktail on human hepatocellular carcinoma Hep3B cells. RESULTS: The predesigned siRNAs could inhibit VEGF and KSP at mRNA level. siRNA cocktail showed a further downregulation on KSP mRNA and protein levels compared to KSP-siRNA or VEGF-siRNA, but not on VEGF expression. It also exhibited greater suppression on cell proliferation as well as cell migration or invasion capabilities and induction of apoptosis in Hep3B cells than single siRNA simultaneously. This could be explained by the significant downregulation of Cyclin D1, Bcl-2 and Survivin. However, no sigificant difference in the mRNA and protein levels of ANG2, involving inhibition of angiogenesis was found in HUVECs cultured with supernatant of Hep3B cells treated with siRNA cocktail, compared to that of VEGF-siRNA. CONCLUSION: Silencing of VEGF and KSP plays a key role in inhibiting cell proliferation, migration, invasion and inducing apoptosis of Hep3B cells. Simultaneous silencing of VEGF and KSP using siRNA cocktail yields promising results for eradicating hepatocellular carcinoma cells, a new direction for liver cancer treatment.

Caracterización cariotípica en mitosis y meiosis del robalo blanco Centropomus undecimalis (Pisces: Centropomidae)

Karyotypic characterization in mitosis and meiosis of the common snook Centropomus undecimalis (Pisces: Centropomidae). The common snook Centropomus undecimalis inhabits marine, brackish and freshwater habitats in the Western Central Atlantic Ocean, including the Gulf of Mexico. Common snook is an economically important fish in many localities, nevertheless the number of studies on its biology and genetics are still few. The present study attempts to establish the cytogenetic profiles of the specimens collected in Paraiso Municipality Tabasco, Mexico. Tissue of five females and eight male organisms were processed by conventional cytological techniques to obtain chromosome slides of high quality in order to assemble the karyotype. The results from the kidney tissue analysis showed that 85.1% of 288 mitosis had a 2n=48 chromosomes, and 52.8% of 104 meiosis exhibited the haploid number 1n=24. The diploid karyotype showed 48 monoarmed chromosomes of the telocentric (T) type. There was no chromosome heteromorphism between females and males. The diploid karyotype was very similar to that observed in the majority of marine fishes. Rev. Biol. Trop. 59 (2): 683-692. Epub 2011 June 01.

El robalo blanco Centropomus undecimalis, vive en hábitats marinos, salobres y dulceacuícolas en el océano Atlántico occidental, incluyendo el golfo de México. La especie, es económicamente importante en varias localidades, no obstante los estudios sobre su biología y genética son hasta el momento pocos. El presente estudio tiene como propósito, la caracterización citogenética de especímenes recolectados en el municipio de Paraíso, Tabasco, México. Cinco hembras y ocho machos fueron procesados por técnicas citológicas convencionales para la obtención de preparaciones cromosómicas de buena calidad para elaborar el cariotipo. Los resultados del análisis del tejido del riñón, mostraron que 85.1% de 288 mitosis tienen 2n=48 cromosomas y 52.8% de 104 meiosis exhiben el número haploide de 1n=24. El cariotipo diploide mostro 48 cromosomas monorrámeos de tipo telocéntrico (T). No se observó heteromorfismo cromosómico entre hembras y machos. El cariotipo diploide fue similar a los observados en la mayoría de peces marinos.

Cytogenetical and ultrastructural effects of copper on root meristem cells of Allium sativum L

Different copper concentrations, as well as different exposure times, were applied to investigate both cytogenetical and ultrastructural alterations in garlic (Allium sativum L.) meristem cells. Results showed that the mitotic index decreased progressively when either copper concentration or exposure time increased. C-mitosis, anaphase bridges, chromosome stickiness and broken nuclei were observed in the copper treated root tip cells. Some particulates containing the argyrophilic NOR-associated proteins were distributed in the nucleus of the root-tip cells and the amount of this particulate material progressively increased with increasing exposure time. Finally, the nucleolar material was extruded from the nucleus into the cytoplasm. Also, increased dictyosome vesicles in number, formation of cytoplasmic vesicles containing electron dense granules, altered mitochondrial shape, disruption of nuclear membranes, condensation of chromatin material, disintegration of organelles were observed. The mechanisms of detoxification and tolerance of copper are briefly discussed.

p53 and MDM2 protein expression in actinic cheilitis

Actinic cheilitis is a potentially malignant lip lesion caused by excessive and prolonged exposure to ultraviolet radiation, which can lead to histomorphological alterations indicative of abnormal cell differentiation. In this pathology, varying degrees of epithelial dysplasia may be found. There are few published studies regarding the p53 and MDM2 proteins in actinic cheilitis. Fifty-eight cases diagnosed with actinic cheilitis were histologically evaluated using Banóczy and Csiba (1976) parameters, and were subjected to immunohistochemical analysis using the streptavidin-biotin method in order to assess p53 and MDM2 protein expression. All studied cases expressed p53 proteins in basal and suprabasal layers. In the basal layer, the nuclei testing positive for p53 were stained intensely, while in the suprabasal layer, cells with slightly stained nuclei were predominant. All cases also tested positive for the MDM2 protein, but with varying degrees of nuclear expression and a predominance of slightly stained cells. A statistically significant correlation between the percentage of p53 and MDM2-positive cells was established, regardless of the degree of epithelial dysplasia. The expression of p53 and MDM2 proteins in actinic cheilitis can be an important indicator in lip carcinogenesis, regardless of the degree of epithelial dysplasia.

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.

Induction of mitotic crossing-over in diploid strains of Aspergillus nidulansusing low-dose X-rays

As a contribution towards detecting the genetic effects of low doses of genotoxic physical agents, this paper deals with the consequences of low-dose X-rays in the Aspergillus nidulans genome. The irradiation doses studied were those commonly used in dental clinics (1-5 cGy). Even very low doses promoted increased mitotic crossing-over frequencies in diploid strains heterozygous for several genetic markers including the ones involved in DNA repair and recombination mechanisms. Genetic markers of several heterozygous strains were individually analyzed disclosing that some markers were especially sensitive to the treatments. These markers should be chosen as bio-indicators in the homozygotization index assay to better detect the recombinogenic/carcinogenic genomic effects of low-dose X-rays.

Inestabilidad cariológica durante la formación de células madres del polen en Aloe vera (Aloaceae)

Con el objetivo de esclarecer la posible existencia de anomalías citogenéticas que aminoren la fertilidad del polen de Aloe vera, se analizó la etapa de proliferación celular que lleva a la formación de células madres del polen (CMPs). Se recolectaron botones florales (BF) en 25 plantas de una población ubicada a 10°34’15’’ N, 64°12’08’’ W, los cuales fueron fijados en Carnoy I por 24 h y almacenados en etanol (70 % v/v). Las observaciones se realizaron en preparaciones temporales obtenidas por la tinción del contenido de las anteras suspendidas en orceína acética (1.5 % p/v) por 5 minutos. De las 9 411 células analizadas, 17 % mostraron 1-8 puentes entre cromátidas hermanas, 13 % 1-7 micronúcleos de 0.9-4.8 µm, 8.1 % estaban unidas por puentes y 0.1 % no contenían cromatina. El resto de las células (61.8 %) presentó configuraciones aparentemente normales y sin variaciones morfométricas. La proliferación irregular de una fracción de CMPs (39.2 %) sugiere que las condiciones ambientales de la zona árida donde se realizaron los muestreos inducen inestabilidad cromosómica y cambios fisiológicos que afectan el normal desarrollo de la mitosis premeiótica, generando pérdida o adición de fragmentos, asociados a deficiencias y duplicaciones génicas.

Alteraciones cromosómicas en linfocitos de sangre periférica / Chromosomic alterations in peripherical blood lymphocytes

Background: The chromosomic alterations prevalence is about 1 percent at birth; therefore, cytogenetics is an important tool for the Neonatology and Pediatrics Units at all health centers. Objective: Evaluate cytogenetics studies performed at the Pathology Unit in the Hospital of Antofagasta between 1997 and 2006. Method: Evaluation of 534 chromosome analysis results in peripherial blood lymphocytes obtained from patients belonging to the Neonatology and Pediatrics Units from Hospital Regional of Antofagasta. The sample included newborn, infants, pre-scholars and scholars with Down Syndrome, congenital malformations, dysmorphic syndromes and other genetic alterations. An optical microscope and international nomenclature (ISCN) were used; chromosome analysis included 35 or more mitosis per case. Results: 22,50 percent (n = 120) of exams showed chromosome alterations: 76,67 percent of these cases were numerical alterations and 23,33 percent corresponded to structural alterations. Conclusions: In the studied population, the cytogenetics study is an important tool together with clinical aspects, especially regarding genetic counsil.

La prevalencia estimativa de las alteraciones cromosómicas en el nacimiento es de alrededor de 1 por ciento, por esta razón es que la citogenética se ha transformado en una herramienta importante para el equipo de Neonatología y Pediatría de cualquier servicio hospitalario. Objetivo: Evaluar los estudios citogénicos realizados en el servicio de Anatomía Patológica de Hospital de Antofagasta entre los años 1997 y 2006. Paciente y Métodos: Se evaluaron 534 análisis cromosómicos en linfocitos de sangre periférica obtenidos en el Hospital de Antofagasta, de los Servicios de Pediatría y Neonatología. La muestra incluyó a recién nacidos lactantes, preescolares y escolares con síndrome de Down, malformaciones congénitas, síndrome dismórfico, ambigüedad genital, genopatías en estudio y otras. Se usó un microscopio óptico y el análisis cromosómico incluyó un mínimo de 35 mitosis por caso, se utilizó la nomenclatura internacional ISCN. Resultados: Un 22,50 por ciento (n = 120) de los exámenes mostraron alteraciones cromosómicas, de estos casos un 76,67 por ciento fueron alteraciones numéricas y 23,33 por ciento correspondieron a alteraciones estructurales. Conclusión: El estudio citogénico ha resultado de gran importancia como complemento de la clínica, y especialmente en relación al consejo genético, en la población estudiada.

Asexual Recombination in a uvsH Mutant of Aspergillus nidulans

Mutations in the gene uvsH of Aspergillus nidulans result in increased spontaneous chromosome instability and increased intragenic and intergenic mitotic recombination in homozygous diploids. The aim of the present work was to obtain a uvs mutant of A. nidulans and to use it for the isolation of asexual recombinants (parameiotic segregants). The mutant uvsH, named B511, showed normal frequency of meiotic recombination in sexual crosses and high frequency of parameiotic segregants in the parasexual crossings with master strains (B511//A757 and B511//A288). Asexual haploid recombinants (parameiotic segregants), diploid and aneuploid segregants were recovered directly from the uvs//uvs+ heterokaryons (B511//A757 and B511// A288). Parameiotic segregants originated through mitotic crossing-over and independent assortment of chromosomes.

Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae)

Microsporogenesis and pollen development were analyzed in a tetraploid (2n = 4x = 36) accession of the forage grass Brachiaria jubata (BRA 007820) from the Embrapa Beef Cattle Brachiaria collection that showed partial male sterility. Microsporocytes and pollen grains were prepared by squashing and staining with 0.5 percent propionic carmine. The meiotic process was typical of polyploids, with precocious chromosome migration to the poles and laggards in both meiosis I and II, resulting in tetrads with micronuclei in some microspores. After callose dissolution, microspores were released into the anther locule and appeared to be normal. Although each microspore initiated its differentiation into a pollen grain, in 11.1 percent of them nucleus polarization was not observed, i.e., pollen mitosis I was symmetric and the typical hemispherical cell plate was not detected. After a central cytokinesis, two equal-sized cells showing equal chromatin condensation and the same nuclear shape and size were formed. Generative cells and vegetative cells could not be distinguished. These cells did not undergo the second pollen mitosis and after completion of pollen wall synthesis each gave rise to a sterile and uninucleate pollen grain. The frequency of abnormal pollen mitosis varied among flowers and also among inflorescences. All plants were equally affected. The absence of fertile sperm cells in a considerable amount of pollen grains in this accession of B. jubata may compromise its use in breeding and could explain, at least in part, why seed production is low when compared with the amount of flowers per raceme.

Benznidazole-induced genotoxicity in diploid cells of Aspergillus nidulans

Genotoxic effects of benznidazole were studied by the induction of homozygosis of genes previously present in heterozygous. UT448//A757 diploid strain was used in the benznidazole's recombinagenesis test. Although toxic effects on growth of colonies were not observed, 75 and 100 æM benznidazole induced an increasing of mitotic recombination events in diploid strain. Results were related to the induction of chromosomal breaks by the antiparasitic drug.

Clastogenic effects of different Ureaplasma urealyticum serovars on human chromosomes

The possibility that Ureaplasma urealyticum might play an important role in human infertility was first raised more than 20 years ago, but this association remains speculative. Considering the hypothesis that the pathogenicity of Ureaplasma urealyticum may depend on its serotypes, the clastogenic effcts of different strains of Ureaplasma urealyticum, at concentrations of 10(3) CCU (color changing units)/ml, 10(4) CCU/ml and 10(5) CCU/ml, were evaluated in vitro in short-term cultures of human lyphocytes. Total or partial mitotic inhibition was produced by Ureaplasma urealyticum serotypes 2,3 and 10 independent of the concentration (10(3) CCU/ml, 10(4) CCU/ml or 10 (5) CCU/ml) of the microorganisms employed. In contrast, the clastogenic effects observed with serotypes 1,7 and 12 varied according to the concentration employed in the test. Mitotic alterations were observed in Ureaplasma urealyticum serotypes 5,6,7,8,9,11 and 12. Chromatid gaps (53.0 percent) and chromatid breaks (13.9 percent) were the most frequent types of alterations observed. The results of this in vitro assay demonstrated that the clastogenic effects varied with the Ureaplasma urealyticum serotypes evaluated.

Ensayo de la proliferación celular en cultivo de linfocitos de cerdo / Assay kinetics cell proliferations lymphocytes from pigs

Se reporta el cultivo de linfocitos de sangre periférica completa de cerdos sanos e infectados, para evaluar la cinética de proliferación celular; 0.5 ml de sangre fue cultivada en 6.0 ml de RPMI-11640 suplementado, en presencia de fitohemaglutinina y 5-BrdU, a 37§C. Al cabo de 48 horas de incubación se cosecharon y fueron teñidos de acuerdo a la técnica de fluorescencia más Giemsa, obteniéndose así células en diferentes etapas de diferenciación, identificándolas en primera, segunda y tercera división. La actividad proliferativa se midió bajo dos parámetros: índice mitótico e índice de replicación. Los resultados son reproducibles y se pueden diferenciar los efectos citotóxicos de los citostáticos sobre los linfocitos del huésped