A possible role of Drosophila CTCF in mitotic bookmarking and maintaining chromatin domains during the cell cycle

BACKGROUND: The CCCTC-binding factor (CTCF) is a highly conserved insulator protein that plays various roles in many cellular processes. CTCF is one of the main architecture proteins in higher eukaryotes, and in combination with other architecture proteins and regulators, also shapes the three-dimensional organization of a genome. Experiments show CTCF partially remains associated with chromatin during mitosis. However, the role of CTCF in the maintenance and propagation of genome architectures throughout the cell cycle remains elusive. RESULTS: We performed a comprehensive bioinformatics analysis on public datasets of Drosophila CTCF (dCTCF). We characterized dCTCF-binding sites according to their occupancy status during the cell cycle, and identified three classes: interphase-mitosis-common (IM), interphase-only (IO) and mitosis-only (MO) sites. Integrated function analysis showed dCTCF-binding sites of different classes might be involved in different biological processes, and IM sites were more conserved and more intensely bound. dCTCF-binding sites of the same class preferentially localized closer to each other, and were highly enriched at chromatin syntenic and topologically associating domains boundaries. CONCLUSIONS: Our results revealed different functions of dCTCF during the cell cycle and suggested that dCTCF might contribute to the establishment of the three-dimensional architecture of the Drosophila genome by maintaining local chromatin compartments throughout the whole cell cycle.

Evaluation of liver regeneration with use of diet supplemented with l-arginine

PURPOSE: To compare controlled liver regeneration in rats submitted to 60% hepatic resection having L-arginine supplemented diet, based on weight changes of the regenerated liver, laboratory parameters of liver function and pathological findings. METHODS: Thirty-six rats were divided into two groups, control and L- arginine. The first received standard chow and saline solution by gavage. The second had supplementation with L- arginine. Animals were killed on postoperative period at 24h, 72h and seven days. For analysis of liver regeneration was used Kwon formula for weight, laboratory tests and mitosis. RESULTS: Weight, showed no benefit with L- arginine supplementation; however, intergroup comparison in the first 24h observed positive effect on supplementation (p=0.008). Alkaline phosphatase was increased in arginine group (p<0.04). The number of mitoses showed no difference between the two groups; however, in the first 24 hours, the supplemented group had higher number of mitoses within the groups (p=0.03). CONCLUSION: Supplementation with L-arginine did not show benefits in liver regeneration; however, supplemented group in the first 24 hours showed benefits over 72 hours and seven days of the evaluation by weight gain and number of mitosis. .

La proliferación de los miocitos ventriculares del corazón de mamífero adulto: un fenómeno esporádico pero factible / Proliferation of adult mammalian ventricular cardiomyocytes: A sporadic but feasible phenomenon

La proliferación de los miocitos que forman parte de los ventrículos cardíacos del mamífero adulto ha sido descartada por algunos investigadores con el argumento de que estas células están diferenciadas en forma terminal; sin embargo, este dogma ha sido puesto en duda a partir de los hallazgos de otros investigadores quienes han observado que estos miocitos pueden presentar los procesos necesarios para la proliferación, es decir síntesis de ADN, mitosis y citocinesis, cuando el miocardio se daña en forma experimental con estrategias de tipo farmacológico o quirúrgico, o debido a condiciones patológicas relacionadas con el sistema cardiovascular. Esta revisión integra algunos de los trabajos disponibles en la literatura que han evaluado la síntesis del ADN, mitosis y citocinesis en estas células, en el miocardio dañado, para saber si su proliferación puede ser considerada como un fenómeno factible. La revisión concluye con una reflexión sobre las perspectivas del conocimiento generado en esta área de estudio.

Proliferation of adult mammalian ventricular cardiomyocytes has been ruled out by some researchers, who have argued that these cells are terminally differentiated; however, this dogma has been rejected because other researchers have reported that these cells can present the processes necessary to proliferate, that is, DNA synthesis, mitosis and cytokinesis when the heart is damaged experimentally through pharmacological and surgical strategies or due to pathological conditions concerning the cardiovascular system. This review integrates some of the available works in the literature evaluating the DNA synthesis, mitosis and cytokinesis in these myocytes, when the myocardium is damaged, with the purpose of knowing if their proliferation can be considered as a feasible phenomenon. The review is concluded with a reflection about the perspectives of the knowledge generated in this area.

The jejunoileal bypass provokes morphological changes in the large intestine? An experimental study in rats / A derivação jejuno-ileal provoca alterações morfológicas no intestino grosso? Estudo experimental em ratos

PURPOSE: To analyse histopathological alterations characterized by the mitotic index in the mucosa of the large intestine in Wistar rats submitted to jejunoileal bypass operation after continued administration of sodium nitrite and vitamin C to different groups. METHODS: Eighty male Wistar rats were employed and separated into 12 groups. In the control group (20 rats): five animals ingested only water; five animals received vitamin C; five animals received sodium nitrite and five received sodium nitrite + vitamin C. In the sham group (20 rats), the animals were anesthetized and underwent midline laparotomy and only intestinal manipulation was performed: five animals ingested only water; five animals received vitamin C; five animals received sodium nitrite and five received sodium nitrite + vitamin C. In the operated group 40 rats underwent a jejunoileal bypass surgery: ten animals ingested only water; ten animals received vitamin C; ten animals received sodium nitrite and ten received sodium nitrite + vitamin C. The mean weight of the animals was measured weekly. The large intestine was subdivided into cecum (S1), ascending colon (S2), transverse colon (S3), descending colon (S4) and rectum (S5) for histopathological analysis and mitotic counts. The statistical analysis was used to compare the mitotic indices. The level of significance was 5%. RESULTS: The mean of all the segments indicates that the sodium nitrite+vitamin C group obtained the lowest mitotic index compared to the other treatments in the control group. The segments S1 and S2 showed a statistical difference with the vitamin C treatment: a higher mitotic index and better preservation of the mucosa in the operated group. In the sham group the main statistical difference occurred only in the sodium nitrite+vitamin C group between the means of the segments. CONCLUSIONS: The comparison of all the colonic segments of the various groups revealed a lower mitotic index in the animals treated with sodium nitrite+vitamin C. In addition, it was found that vitamin C did not present a statistically significant inhibiting effect on the preservation of the mucosa and the mitotic index.

OBJETIVO: Analisar as alterações histopatológicas caracterizada pelo índice mitótico na mucosa do intestino grosso em ratos Wistar submetidos a operação de bypass jejunoileal após a administração continuada de nitrito de sódio e vitamina C para diferentes grupos. MÉTODOS: Oitenta ratos Wistar foram utilizados e separados em 12 grupos. No grupo controle (20 ratos): cinco animais ingeriram apenas água; cinco animais receberam vitamina C, cinco animais receberam nitrito de sódio e cinco receberam nitrito de sódio + vitamina C. No grupo sham (20 ratos), os animais foram anestesiados e submetidos a laparotomia mediana e só a manipulação intestinal foi realizada: cinco animais ingeriram apenas água; cinco animais receberam vitamina C, cinco animais receberam nitrito de sódio e cinco receberam nitrito de sódio + vitamina C. No grupo operado 40 ratos foram submetidos a uma cirurgia de bypass jejunoileal: dez animais ingeridos apenas água; dez animais receberam vitamina C, dez animais receberam nitrito de sódio e dez nitrito de sódio + vitamina C. O peso médio dos animais foi medido semanalmente. O intestino grosso foi subdividido em ceco (S1), cólon ascendente (S2), cólon transverso (S3), cólon descendente (S4) e reto (S5) para análise histopatológica e contagem das mitoses. A análise estatística foi utilizado para comparar os índices mitóticos. O nível de significância foi de 5%. RESULTADOS: A média de todos os segmentos indica que o grupo que ingeriu nitrito de sódio + vitamina C obteve o menor índice mitótico em relação aos demais tratamentos no grupo controle. Os segmentos S1 e S2 mostraram uma diferença estatística com a vitamina C de tratamento: um maior índice mitótico e melhor preservação da mucosa no grupo operado. No grupo sham a principal diferença estatística ocorreu apenas no grupo que ingeriu nitrito de sódio + vitamina C entre as médias dos segmentos. CONCLUSÕES: A comparação de todos os segmentos do colon dos vários grupos revelaram um menor índice de mitose nos animais tratados com nitrito de sódio + vitamina C. Além disso, a vitamina C não apresentou efeito inibidor, estatísticamente significativo, na preservação da mucosa e do índice de mitoses.

Proliferative and morphologic characterization of buccal mucosal fibroblasts in areca nut chewers: A cell culture study.

Objective: To isolate, culture and characterize fibroblasts from the buccal mucosa of areca nut chewers with and without oral submucous fibrosis (OSF). Materials and Methods: Primary fibroblast cultures were established by the collagenase disaggregation technique and the phenotypic and growth characteristics were studied. Results: Cells cultured from OSF showed a statistically significant increase in both the post-mitotic fibroblast subpopulation and the population doubling time when compared with controls. Conclusion: There was a significant increase in the pro-fibrotic, post-mitotic subpopulation of fibroblasts in areca nut chewers with OSF.

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.

Changes in the corpora allata and epidermal proliferation along the fourth instar of the Chagas disease vector Triatoma infestans

Triatoma infestans, a blood-feeding insect, synchronises physiological mechanisms leading to moult with food intake. Since the corpora allata are important in moult and metamorphosis regulation, we have studied morphological changes in 4th instar nymphs (gland size, cell density, percent of animals showing mitoses and cell size). Changes were correlated with the effect of precocene II, epidermal proliferation, and with the extent of the [quot ]head critical period[quot ]. Based on morphological grounds, three stages can be defined in the gland along the 4th instar: Stage 1 (days 0-2 after feeding) showed small corpora allata, composed by a small number of cells, and in which mitoses were absent; Stage 2 (days 3-9) showed growing corpora allata, in which cell number was increasing and proliferation was apparent; and Stage 3 (days 10-13) showed no mitotic activity, and a sharply diminishing size of the gland, as a consequence of the diminishing size of their cells. The ability of precocene II to induce abnormal moulting disappeared during stage 2 correlating with the termination of the head critical period and suggesting that corpora allata are essential during days 3 to 5 to determine normal growth. Epidermal cell number was increasing as a consequence of more frequent mitotic activity, beginning after the finalization of the head critical period and after a first increment in the size of the gland.

Characterization of mitosis-specific phosphorylation of tumor-associated microtubule-associated protein

Tumor-associated microtubule-associated protein (TMAP), also known as cytoskeleton associated protein 2 (CKAP2), has been recently shown to be involved in the assembly and maintenance of mitotic spindle and also plays an essential role in maintaining the fidelity of chromosome segregation during mitosis. We have previously reported that TMAP is phosphorylated at multiple residues specifically during mitosis, and characterized the mechanism and functional importance of phosphorylation at one of the mitosis-specific phosphorylation residues (i.e., Thr-622). However, the phosphorylation events at the remaining mitotic phosphorylation sites of TMAP have not been fully characterized in detail. Here, we report on generation and characterization of phosphorylated Thr-578- and phosphorylated Thr-596-specific antibodies. Using the antibodies, we show that phosphorylation of TMAP at Thr-578 and Thr-596 indeed occurs specifically during mitosis. Immunofluorescent staining using the antibodies shows that these residues become phosphorylated starting at prophase and then become rapidly dephosphorylated soon after initiation of anaphase. Subtle differences in the kinetics of phosphorylation between Thr-578 and Thr-596 imply that they may be under different mechanisms of phosphorylation during mitosis. Unlike the phosphorylation-deficient mutant form for Thr-622, the mutant in which both Thr-578 and Thr-596 had been mutated to alanines did not induce significant delay in progression of mitosis. These results show that the majority of mitosis-specific phosphorylation of TMAP is limited to pre-anaphase stages and suggest that the multiple phosphorylation may not act in concert but serve diverse functions.

Evaluación morfométrica de la relación embrio-uterina de las etapas pre y post implantacional en conejo (Oryctolagus cuniculus) / Morphometric evaluation of the embryo-uterine relationship of the pre and post implantational stages in rabbit (Oryctolagus cuniculus)

El conejo ha demostrado ser un excelente modelo de estudio de implantación. Como ovulador obligado, el tiempo de preñez se puede establecer en forma precisa. La observación morfológica de cortes de úteros de 7, 8, 9 y 10 días pos coito, permitió en este estudio reconstruir una secuencia de los eventos morfométricos que ocurren durante la implantación, en el conejo. Se utilizaron 20 conejas neozelandesas blancas adultas (Oryctolagus cuniculus), nulíparas, no gestantes. Para la cruza, se utilizaron 5 machos de probada fertilidad. Los conejos fueron obtenidos del Bioterio de la Facultad de Medicina de la Universidad de La Frontera, Temuco, Chile. Determinamos como día 0, el momento del coito, sacrificando las hembras los días 7, 8, 9 y 10 de cada cruza. Una vez sacrificados, se disecó macro y mesoscópicamente la región pélvica, seleccionándose vesículas uterinas para sus estudios histológico e inmunocitoquímico. Se realizaron estudios morfométricos y de cinética celular con las técnicas de Túnel y PCNA. Se efectuó estadística descriptiva en base a promedio y desviación estándar (p< 0.001), utilizando el programa estadístico Stata 9.0. Las mediciones morfométricas obtenidas fueron de diámetro y volumen de la vesícula uterina, altura de la pared uterina y lumen glandular de regiones mesometrial y antimesometrial, para los días 7 al 10 post coito. Los análisis inmunocitoquímicos permitieron determinar: índice apoptótico de los núcleos celulares y el índice celular mitótico. Para el día 8 post coito se observaron los cambios morfométricos más significativos a nivel del lumen vesicular, altura de pared uterina de la región mesometrial, y lumen glandular. El mismo día se observaron cambios importantes en los índices celular mitótico y apoptótico. El conejo podría ser una especie usada para predecir el normal desarrollo embrionario, tras la comprensión morfológica y morfométrica de la implantación. Presenta una forma poco invasiva de implantación...

The rabbit has demonstrated to be an outstanding implanting model. As an obligated ovulating animal, its pregnancy time can be established exactly. The morphologic observation of uterus cuts of 7, 8, 9 and 10 days post mating, allowed this research to rebutid an apparent sequence morphometric events, that take place during rabbit implantation. Twenty New Zealand, white, adult, female rabbits were used in this research. (Oryctolagus cunículus), nulipara. And 4, proven, fertile, males were used for mating. These males were obtained from the Bioterio of the Faculty of Medicine at the Universidad de La Frontera, Temuco, Chile. Mating day was defined as day cero, sacrificing the females days 7, 8, 9 and 10 in each mating. Once sacrificed, the pelvic region was macro and mesoscopically dissected, and the vesicles were selected for histological and immunocitochemical study. Histological studies were performed with H.E. technique; morphometric and cellular kinetics with Tunnel and PCNA techniques. Descriptive statistics was used based on an average and standard deviation (p< 0.001). The Stata 9.0 statistical program was used. The morphometric measurements obtained were: diameters and volume of the uterine vesicle, height of the uterine wall and glandular lumen of the mesometrial and antimesometrial regions to 7-10 days post coitus. The immunocitochemical analysis identified: apoptotic Index of the cellular nucleous and cellular mitotic Índex. By day 8 post coitus were observed significant morphometric changes at the vesicular lumen, height of the uterine wall of the mesometrial region and glandular lumen. For the same day important changes of the cellular mitotic Índex and apoptotic Index were observed. The rabbit could be a species used to predict normal embryo development, after the morphologic comprehension of the implantation, given its not so invasive form of implantation and its capacity of early placenta formation. Providing updated information...

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.

Transient phosphorylation of tumor associated microtubule associated protein (TMAP)/cytoskeleton associated protein 2 (CKAP2) at Thr-596 during early phases of mitosis

Tumor associated microtubule associated protein (TMAP), also known as cytoskeleton associated protein 2 (CKAP2) is a mitotic spindle-associated protein whose expression is cell cycle-regulated and also frequently deregulated in cancer cells. Two monoclonal antibodies (mAbs) against TMAP/CKAP2 were produced: B-1-13 and D-12-3. Interestingly, the reactivity of mAb D-12-3 to TMAP/CKAP2 was markedly decreased specifically in mitotic cell lysate. The epitope mapping study showed that mAb D-12-3 recognizes the amino acid sequence between 569 and 625 and that phosphorylation at T596 completely abolishes the reactivity of the antibody, suggesting that the differential reactivity originates from the phosphorylation status at T596. Immunofluorescence staining showed that mAb D-12-3 fails to detect TMAP/CKAP2 in mitotic cells between prophase and metaphase, but the staining becomes evident again in anaphase, suggesting that phosphorylation at T596 occurs transiently during early phases of mitosis. These results suggest that the cellular functions of TMAP/CKAP2 might be regulated by timely phosphorylation and dephosphorylation during the course of mitosis.

Comparison of staining of mitotic figures by haematoxylin and eosin-and crystal violet stains, in oral epithelial dysplasia and squamous cell carcinoma.

Mitosis of cells gives rise to tissue integrity. Defects during mitosis bring about abnormalities. Excessive proliferation of cells due to increased mitosis is one such outcome, which is the hallmark in precancer and cancer. The localization of proliferating cells or their precursors may not be obvious and easy. Establishing an easy way to distinguish these mitotic cells will help in grading and understanding their biological potential. Although immunohistochemistry is an advanced method in use, the cost and time factor makes it less feasible for many laboratories. Selective histochemical stains like toluidine blue, giemsa and crystal violet have been used in tissues including the developing brain, neural tissue and skin. AIM OF THE STUDY: 1) To compare the staining of mitotic cells in haematoxylin and eosin with that in crystal violet. 2) To compare the number of mitotic figures present in normal oral mucosa, epithelial dysplasia and oral squamous cell carcinoma in crystal violet-stained sections with that in H and E-stained sections. MATERIALS AND METHODS: Ten tissues of normal oral mucosa and 15 tissues each of oral epithelial dysplasia seen in tobacco-associated leukoplakia and squamous cell carcinoma were studied to evaluate the selectivity of 1% crystal violet for mitotic figures. The staining was compared with standard H and E staining. Statistical analysis was done using Mann-Whitney U test. RESULTS: A statistically significant increase in the mean mitotic count was observed in crystal violet-stained sections of epithelial dysplasia as compared to the H and E-stained sections (p=0.0327). A similar increase in the mitotic counts was noted in crystal violet-stained sections of oral squamous cell carcinoma as compared to the H and E-stained sections.(p=0.0443). No significant difference was found in the mitotic counts determined in dysplasia or carcinoma by either the crystal violet (p=0.4429) or the H and E-staining techniques (p=0.2717). CONCLUSION: One per cent crystal violet provides a definite advantage over the H and E-stained sections in selectively staining the mitotic figures.

Immunohistochemical expression of h-telomerase reverse transcriptase in canine and feline meningiomas

Telomere length maintenance is regarded as a fundamental step in tumorigenesis, as most human brain tumors, including meningiomas, stabilize the ends of their chromosomes using telomerase. This investigation represents an introduction to telomerase expression in canine and feline meningiomas. Twenty-five archived cases (14 dogs and 11 cats) were immunohistochemically tested for human-telomerase reverse transcriptase (h-TERT), scored, and quantified; furthermore, mitoses were counted on sections stained with a modified toluidine blue. The h-TERT antibody immunolabelled the nucleus and nucleolus of meningeal neoplastic cells, with an intensity ranging from mild to strong and a speckled distribution; a significantly higher expression in cats was noted, while no significant association between h-TERT immunolabelling and sex or histotype was evident in dogs or cats. The telomerase enzyme represents a fundamental parameter of potential malignant transformation, which may occur independently of the signal to proliferate, thereby supplying the cells with unlimited growth capabilities. Telomerase expression could be a prognostic indicator independent of the kinetic parameters, although this should be evaluated using a larger dataset with available clinical information.

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.

Sertoli cell proliferation during the post hatching period in domestic fowl

There has been no study aimed at directly determiningof the periods of Sertoli cell proliferation in birds evendomestic fowl. The aims of this study were to observe thecessation of post-hatching mitotic proliferation of Sertolicells in domestic fowl, and to determine the volumedensity of Sertoli and germ cells during this period. Atotal of 50 Leghorn chicks were used in this study. Thetestes sections of the animals were immunostained withBrdU to observe the proliferation of cells from one to 10weeks of age. The volume density of the Sertoli and germcells were determined using the standard point countingmethod. The volume density of the germ cell nuclei wasinitially less than that of the Sertoli cells but the volumedensity converged by week 6, and remained relativelyconstant until the commencement of meiosis. Clearlabeling of Sertoli and germ cells was observed from week1 to week 7. The only those cells still labeled after 8 weekswere germ cells, indicating that Sertoli cell proliferationhad ceased. Therefore, it is recommended that anyresearch into the testes of domestic fowl should considerthe cessation of Sertoli cell proliferation by approximately8 weeks.

Role of HGF/c-Met in Serum-Starved ARPE-19 Cells

PURPOSE: Hepatocyte growth factor (HGF) and its receptor (HGFR/c-Met) regulate motility, mitogenesis, and morphogenesis in a cell type-dependent fashion. We report the role of HGF and c-Met on stress-induced ARPE-19 human retinal pigment epithelial (RPE) cells in this study. METHODS: The cells were cultured either with or without serum. Southern and Western blot analyses were done to determine the expression patterns of HGF/c-Met in serum-starved ARPE-19 cells. The cell proliferation pattern in serum-starved condition was analyzed using MTS assay. Inhibition level of cell proliferation was analyzed using a neutralizing monoclonal antibody against c-Met (2 microgram/ml). RESULTS: Abnormal cell proliferation and scattering of ARPE-19 cells was observed under serum starvation. HGF/c-Met were expressed in serum-starved ARPE-19 cells. ARPE-19 cell proliferation was also enhanced with recombinant HGF treatment. Neutralization against c-Met inhibited the proliferation of serum-deprived ARPE-19 by 64.5% (n=9, S.D. 5.5%). Serum starvation appears to induce epithelial-mesenchymal transition of ARPE-19 cells, resulting in scatter, and the expression of alpha-smooth muscle actin (alpha-SMA), a marker for fibrosis. CONCLUSIONS: In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.

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.

Sec13 induces genomic instability in U2OS cells

Sec13p has been known as an endoplasmic reticulum-Golgi transport protein. Recently, it has also been shown to be required for the formation of septation in the fission yeast Schizosaccharomyces pombe. In the present study, we focused on the role of a human homolog of Saccharomyces cerevisiae SEC13, Sec13 protein during mitosis in U2OS cells. We found that the expression of Sec13 was constant throughout the cell cycle, and localized to the kinetochores at metaphase during mitosis. By using green fluorescent protein technology, we observed that Sec13 is required for evasion of mitotic arrest in response to spindle damage, leading to G1-like phase and apoptotic cell death. In addition, cells expressing exogenous Sec13 showed giant nuclei compared to endogenous ones in the absence of nocodazole. These results demonstrate that Sec13 is involved in the regulation of the metaphase/anaphase transition and may be functionally associated with mitotic machinery to maintain genomic stability during mitosis.

Estudo laboratorial da cicatrização de córneas humanas após debridamento epitelial / Laboratory study of the wound healing response to epithelial scrape injury in the human cornea

Objetivo: Verificar resposta após debridamento epitelial de córneas humanas. Métodos: Córneas normais foram submetidas a debridamento antes da cirurgia de enucleação. Realizou-se histologia, TUNEL, Ki67, SMA e microscopia eletrônica. Resultados: Seis córneas foram debridadas e preservadas entre ½ e 65 horas, apresentando apoptose nos ceratócitos do estroma anterior. Células estromais em proliferação foram observadas apenas no tempo de 65 horas. Miofibroblastos não foram encontrados. Uma córnea serviu de controle. /Purpose: To examine the early wound healing response to epithelial scrape in human corneas. Methods: Normal corneas had epithelial scrape prior to enucleation. Histology, TUNEL assay, Ki67, SMA and transmission electron microscopy were performed. Results: Epithelial scrape was performed in six corneas from ½ to 65 hours prior to preservation. Keratocyte apoptosis was detected in the anterior stroma in all scraped corneas...

Entry, dispersion and differentiation of microglia in the developing central nervous system

Microglial cells within the developing central nervous system (CNS) originate from mesodermic precursors of hematopoietic lineage that enter the nervous parenchyma from the meninges, ventricular space and/or blood stream. Once in the nervous parenchyma, microglial cells increase in number and disperse throughtout the CNS; these cells finally differentiate to become fully ramified microglial cells. In this article we review present knowledge on these phases of microglial development and the factors that probably influence them.