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Biomedical subjects

M Gulisano

Publications and source records attributed to M Gulisano.

At least 55 records · Page 3Linked to original sources

Emx1 and Emx2 show different patterns of expression during proliferation and differentiation of the developing cerebral cortex in the mouse.

Insights into the complex structure of the forebrain and its regulation have recently come from the analysis of the expression of genes that are likely to be involved in regionalization of this structure. We cloned four new homeo box genes, Emx1, Emx2, Otx1 and Otx2, and showed that in day 10 mouse embryos their expression domains are continuous regions of the developing brain contained within each other in the sequence Emx1 < Emx2 < Otx1 < Otx2. Recently Otx1 has been found to be specifically expressed during neurogenesis of layer 5 and 6 in the developing cerebral cortex. In order to better understand the role of Emx1 and Emx2 in the maturation of the cortex we analysed by in situ hybridization their expression patterns in the developing mouse cerebral cortex, from embryonic day 12.5 to adulthood. We found that Emx2 is expressed exclusively in proliferating cells of the ventricular zone whereas Emx1 is expressed in both proliferating and differentiated neurons, throughout the cortical layers and during all the developmental stages examined. Therefore, Emx2 gene products might control some biological parameters of the proliferation of cortical neuroblasts or of the subsequent cell migration of postmitotic neurons, leaving the cortical germinal zone. Conversely, Emx1 expression, which is confined exclusively to the dorsal telencephalon, characterizes most cortical neurons during proliferation, differentiation, migration and postnatal development and maturation.

Animals↗

Vascular endothelial growth factor messenger ribonucleic acid expression in human ovarian and endometrial cancer.

Vascular endothelial growth factor (VEGF) is a previously discovered angiogenic factor that seems to influence the neoangiogenesis of neoplastic and non-neoplastic tissues. Substantial experimental evidence links tumor growth and metastasis with blood vessel formation. Tumor angiogenesis can be induced by factors released by the tumor cells themselves. A variety of transformed cell lines expresses the VEGF transcript and secretes an EGF-like protein, suggesting that this angiogenic factor may be one of the mediators of tumor angiogenesis. By Northern blot analysis and in situ hybridization, we investigated the expression of VEGF transcript in human ovarian and endometrial neoplasms. Messenger RNA encoding VEGF was detected in all tissues studied and was more densely expressed in endometrial carcinoma. VEGF expression was also identified in cells obtained from ovarian and endometrial ascitic fluid. VEGF mRNA, detected by in situ hybridization, was identified in the epithelial cells of endometrial adenocarcinoma. This distribution was localized primarily in the apices of the papillae. The prominence of VEGF mRNA levels in human ovarian and endometrial tumors demonstrates that VEGF may be involved in promoting tumor angiogenesis and stroma generation, acting as an endothelial cell mitogen.

Adenocarcinoma↗

Emx and Otx gene expression in the developing mouse brain.

The homeobox genes Emx1, Emx2, Otx1 and Otx2 are all expressed in the rostral brain of embryos at E10. Their expression domains are continuous regions of the developing brain contained within each other, such that the expression domain of Otx2 contains that of the other three genes, the expression domain of Otx1 contains that of Emx1 and Emx2, and the expression domain of Emx2 contains that of Emx1. The Emx1 expression domain includes the dorsal telencephalon and it has a posterior boundary slightly anterior to that between the presumptive diencephalon and telencephalon, whereas the Otx2 expression domain covers almost the entire forebrain and midbrain. Starting from E10.75, Otx2 expression disappears progressively from the presumptive cerebral cortex, whereas Emx1, Emx2 and Otx1 are expressed in this structure until late gestation. In particular, Emx2 appears to be expressed exclusively in the germinal ventricular zone of the developing cerebral cortex.

Animals↗

c-otx2 is expressed in two different phases of gastrulation and is sensitive to retinoic acid treatment in chick embryo.

We cloned the chick homologue of the mouse Otx2 gene, c-otx2, and analyzed its expression pattern during gastrulation. During mouse embryogenesis, Otx2 expression is first detected in the entire epiblast and after the formation of the primitive streak becomes confined to the most anterior region of the embryo corresponding to presumptive fore- and mid-brain. Similarly, two distinct phases of c-otx2 expression were observed in the chick. c-otx2 transcripts were first detected in the unincubated egg and up to stage XIII, in all epiblast, and forming hypoblast and mesoblast cells. During primitive streak progression, c-otx2 expression becomes progressively restricted to anterior regions and is mainly associated with Hensen's node. When the extension of the streak is maximal, transcripts are only found in Hensen's node. A second phase of c-otx2 expression starts during streak regression. c-otx2 transcripts are lost from the node and present in higher abundance in anterior neuroectoderm and mesendoderm, with the exception of forming notochord and floor plate. The first phase of expression bears strong similarity with that of c-gsc, a gene shown to be a marker for cells that have organizer activity in the chick. Therefore, we compared the expression of the two genes by double staining on the same embryo. This analysis demonstrated that c-otx2 is transcribed first and its expression in the hypoblast precedes that of c-gsc. On the other hand, c-gsc is an earlier marker of primitive streak cells. The expression domains of the two genes transiently overlap in Hensen's node and anterior mesendoderm, whereas only c-otx2 is expressed in neuroectodermal areas. The second phase of c-otx2 expression is sensitive to an early treatment with retinoic acid. This treatment abolishes c-otx2 expression in mesendoderm and restricts it to most anterior regions in the forming neural plate. In conclusion, our results suggest that c-otx2 expression is first associated with cells with an anterior mesendoderm fate and subsequently extends to anterior neuroectoderm.

Amino Acid Sequence↗

Scanning electron microscopic evaluation of the alterations induced by polluted air in the rabbit bronchial epithelium.

With the aim of evaluating the influence of polluted air on the respiratory epithelium, ten New Zealand white rabbits, from a group of fifteen kept in the country, were transported to a site located in a metropolitan area. After 40 days, all the rabbits were killed, and the bronchial mucosa studied by scanning electron microscopy (SEM). In the animals exposed to polluted air, the ciliated cells, less numerous than in normal cases, show an evident decrease in the number and size of the cilia, exposing apical microvilli. It is therefore possible to hypothesize that a part of the non-ciliated cells is made up of cells that have lost their cilia. The number of non-ciliated elements and the amount of mucous secretion appear to have noticeably increased. The considerable response of the respiratory epithelium to inhaled agents appears to be confirmed, as is the irritant effect of polluted city air.

Air Pollutants↗

Tumor markers in breast cancer follow-up: a potentially useful parameter still awaiting definitive assessment. Forza Operativa Nazionale sul Carcinoma Mammario (FONCaM).

BACKGROUND: Although tumor markers are frequently used in the follow-up of patients with breast cancer, two points are still being debated: 1) their cost/effectiveness has been neither demonstrated nor disproved; 2) the reliability of the currently used dichotomous division into a positive/negative cut-off should be definitely validated. Dynamic criteria of interpretation based on serial serum samples would probably be more effective for early detection of relapse. PATIENTS AND METHODS: The aim of the present study was to compare the dichotomous cut-off based decision criteria to a dynamic serial sample based assessment of tumor markers. Since 1989, 794 patients have been followed in 11 institutions. CEA and CA15.3 were measured once a month for three months before every clinical examination. The present paper concerns the evaluation variability in 405 patients without evidence of disease in the first three institutions joining the study. RESULTS: In patients without evidence of disease, the coefficient of variation of all samples for every patient showed a median value of 19 for CEA and 21 for CA15.3. Variability was negatively associated with the antigen level and was most likely due to the analytical component. This was also confirmed by the significant difference in variability among the three institutions evaluated. The median value of the critical difference was 53% for CEA and 57% for CA15.3. CONCLUSIONS: 1) Individually tailored dynamic decision criteria are applicable in about 50% of the cases. 2) The problem of improving the precision of tumor marker assays in the low dose range must be urgently addressed to the manufacturers of tumor markers by the scientific community in order to apply individually tailored decision criteria for patients in whom the serum level of biological markers is low.

Biomarkers, Tumor↗

Plasmid content in different serovars of Salmonella isolated in Sicily: an ecology study.

This study determined plasmid relative abundance and distribution among a representative collection of Sicilian strains of Salmonella, with the purpose of gaining more insight into bacterial and plasmid ecology. 161 Salmonella strains of different serovars, isolated in Sicily from human and environmental sources between 1981 and 1985 were characterized by plasmid profile typing. Two main sets of Salmonella serovars were identified, one composed by plasmid-free serovars, rarely associated with clinical salmonellosis but typical of environmental niches; the other comprising the serovars usually encountered as major agents of human and animal salmonellosis. The biological relevance of the baseline information so provided on the genetic profiles of different serovars in Sicily is discussed.

Animals↗

Cloning and characterization of two members of the vertebrate Dlx gene family.

A number of vertebrate genes of the Dlx gene family have been cloned in mouse, frog, and zebrafish. These genes contain a homeobox related to that of Distalless, a gene expressed in the developing head and limbs of Drosophila embryos. We cloned and studied the expression of two members of this family, which we named Dlx5 and Dlx6, in human and mouse. The two human genes, DLX5 and DLX6, are closely linked in an inverted convergent configuration in a region of chromosome 7, at 7q22. Similarly, the two human genes DLX1 and DLX2 are closely linked in a convergent configuration at 2q32, near the HOXD (previously HOX4) locus. In situ hybridization experiments in mouse embryos revealed expression of Dlx5 and Dlx6 mRNA in restricted regions of ventral diencephalon and basal telencephalon, with a distribution very similar to that reported for Dlx1 and Dlx2 mRNA. A surprising feature of Dlx5 and Dlx6 is that they are also expressed in all skeletal structures of midgestation embryos after the first cartilage formation. The expression pattern of these genes, together with their chromosome localization, may provide useful cues for the study of congenital disorders in which there is a combination of craniofacial and limb defects.

Amino Acid Sequence↗

Cloning and characterization of a new human Xq13 gene, encoding a putative helicase.

We describe the cloning and characterization of a new human Xq13 gene (XH2), extending over a 220 kb genomic stretch between MNK and DXS56. The gene, which undergoes X-inactivation, contains a 4 kb open reading frame and encodes a putative NTP-binding nuclear protein homologous to several members of the helicase II superfamily. The murine homologue maps to the syntenic genetic interval, between Pgk1 and Xist. In situ hybridization studies in mouse reveal precocious, widespread expression of the murine homologue of XH2 at early stages of embryogenesis, and more restricted expression during late developmental stages and at birth. XH2 is a new member of an expanding family of proven and putative helicases, sharing six conserved, collinear domains. In particular, the XH2 protein shows homology with yeast RAD54. Type II helicases have been implicated in nucleotide excision repair and the initiation of transcription. This new gene, represents a potential candidate for several genetic disorders mapped to human Xq13.

Amino Acid Sequence↗

Histochemical study of human nasopharyngeal epithelium by horseradish peroxidase conjugated lectins.

This study proposes a simple method for assessing whether non-respiratory epithelia of the nasopharynx (i.e. cuboidal, squamous) represent a modified differentiation from the normal pseudo-stratified lining, or whether they possess common characteristics suggesting they are just in a different morphological stage. Peroxidase-labelled lectins were used, since they detect sugar residues of membrane glycoconjugates which play an important role in cell differentiation. Biopsies were taken from healthy controls and from patients affected by upper airway inflammation of varying degree and duration. While pseudostratified and cuboidal epithelia possess similar histochemical characteristics, squamous epithelium has a different behaviour; in particular it is the only type to show intense reactivity to Lotus tetragonolobus lectin (LTA). This would seem to indicate that squamous epithelium is in a different state of differentiation, possibly suggesting dysplasia. Lectins may be valuable both in morphological studies and in the clinical evaluation of airway epithelia.

Adolescent↗

Emx and Otx homeobox genes in the developing mouse brain.

We have analyzed the expression of four mouse homeobox genes related to two Drosophila genes expressed in the developing head of the fly. Two of these genes, Emx1 and Emx2, are related to empty spiracles, and two genes, termed Otx1 and Otx2, are related to orthodenticle. These genes are all expressed in the developing rostral brain of E10 mouse embryos and their expression domains can be compared. Otx2 is expressed in all dorsal and most ventral regions of telencephalon, diencephalon, and mesencephalon. The Otx1 expression domain is similar to that of Otx2, but smaller and contained within it. The Emx2 expression domain is comprised of dorsal telencephalon and small diencephalic regions, both dorsally and ventrally. Finally, Emx1 expression is exclusively confined to the dorsal telencephalon. At the time when regional specification of major brain regions takes place, the expression domains of the four genes appear to be continuous regions contained within each other in the sequence Emx1 < Emx2 < Otx1 < Otx2. The first appearance of transcripts of the four genes is also sequential: Otx2 is expressed first (E5.5), followed by Otx1 and Emx2 (E8-8.5), and finally by Emx1 (E9.5). It is tempting to speculate about a possible role of the four genes in establishing and/or signalling the limits of the various embryonic brain regions in a discrete progressive process with its center in the dorsal telencephalon.

Animals↗

Variability of tumor markers in the follow-up of patients radically resected for breast cancer.

The biological and analytical components of variability of tumor markers should be distinguished from the variations due to tumor progression. The aim of the present study was to evaluate tumor marker variability in the follow-up of patients resected for breast cancer. So far, we have carried out 2,085 CEA and 2,550 CA 15-3 determinations in 435 patients. The total variability of both CEA and CA 15-3 was widely scattered among different subjects (CEA coefficient of variation 0-105%; CA 15-3 coefficient of variation 0-89.2%). The biological variability of CA 15-3, which was calculated in a limited number of cases, was scattered between 0 and 23% and was higher than the intra-assay variability. From these findings we conclude that when evaluating serial marker assays the intra-individual variability should be assayed initially to obtain a reference value of individual variability in relapse-free conditions.

Analysis of Variance↗

Conserved homeobox genes in the developing brain.

We analysed the expression of four mouse homeobox genes related to two Drosophila genes expressed in the developing head of the fly: Emx1 and Emx2, related to empty spiracles and Otx1 and Otx2, related to orthodenticle. The four genes are all expressed in the developing rostral brain of E10 mouse embryos in a specific manner. Otx2 is expressed in every dorsal and most ventral regions of telencephalon, diencephalon and mesencephalon. The Otx1 expression domain is similar to that of Otx2, but contained within it. The Emx2 expression domain is comprised of dorsal telencephalon and small diencephalic regions, both dorsally and ventrally. Finally, Emx1 expression is exclusively confined to the dorsal telencephalon. Thus, at this stage the expression domains of the four genes appear to be continuous regions contained within each other in the sequence Emx1 < Emx2 < Otx1 < Otx2. The first appearance of transcripts of these genes is also sequential: Otx2 is expressed first (E5.5), followed by Otx1 and Emx2 (E8-8.5) and finally by Emx1 (E9.5). These findings suggest a role of the four genes in establishing cell fates within the limits of the various embryonic brain regions in a discrete progressive process with its center in the dorsal telencephalon. The structural organization and the regulation of these genes appear remarkably conserved in evolution.

Animals↗

Blood and/or lymphatic connection of laryngo-pharynx and thyroid: an anatomic and clinical problem.

Three patients, undergoing surgery for pharyngeal carcinoma, were studied after their informed consence in the aim of ascertaining: 1) whether the routinary hemithyroidectomy in case of surgical treatment of the cancer of pyriform sinus is justified by a real probability of neoplastic invasion of the gland, and 2) which pathway the neoplasia follows in the metastatic process. In each patient a small amount of methylene blue solution was injected in the pyriform sinus submucosa, just before starting the surgical operation, currently including hemithyroidectomy. The stain spreads to the superior portion of the thyroid lobe, and seems to follow the periarterial lymphatic vessels. Even if further researches are necessary, the obtained data seem to justify routinary thyroidectomy in course of surgical treatment of the hypopharyngeal cancer.

Humans↗

Homeobox genes in the developing central nervous system.

A number of genes containing sequences coding for DNA-binding domains homologues to homeobox sequences in Drosophila have been isolated in vertebrate and their mechanism of action have been studied. In Particular Hox family genes share with Drosophila homeotic genes a genomic organization in gene clusters and an expression pattern that is similar in a number of important respects. In situ hybridization experiment have shown that there is a strict correspondence, or colinearity, between the order of the Hox genes (3' to 5') within the chromosomal cluster and that of their expression domains (anterior to posterior) in the embryo. Recent data obtained in embryonal carcinoma (EC) cells induced to differentiate by retinoic acid (RA) cast some light on the molecular mechanisms underlying the colinear expression of the Hox genes.

Amino Acid Sequence↗

Molecular characterization of Yersinia strains isolated from human and environmental sources.

Thirty-seven strains of Yersinia isolated from human and environmental sources were investigated for virulence and molecular characteristics. Clinical isolates of Y. enterocolitica serotypes 0:3 and 0:9, harbouring the 45 MDa virulence plasmid were Ca++ dependent and responded positively to the CV-binding assay. The environmental Y. kristensenii strain, although harbouring a 45 MDa plasmid proved to be negative for both assays. All the environmental strains showed pyrazinamidase activity, while isolates from human sources proved negative. Two different plasmid profiles were disclosed: one Y. enterocolitica strain (serotype 0:7, 8) showed one plasmid of 67 MDa and one plasmid of 1.5 MDa. Eleven human isolates of Y. enterocolitica and one environmental strain of Y. krinstensenii showed only one plasmid of 45 MDa. Restriction endonuclease analysis of the 45 MDa plasmids supported the hypothesis that the environmental Y. krinstensenii plasmid is unrelated to the family of homologous virulence plasmids of Y. enterocolitica.

Amidohydrolases↗

Characterization of the glycoconjugate sugar residues in developing chick esophageal epithelium.

The development of the esophagus in the chick embryo is characterized by remarkable morphological changes especially at the level of the epithelium. Using horseradish peroxidase-conjugated lectins (DBA, PNA, SBA, WGA, ConA, LTA, UEAI) we have studied, at the level of the esophagus of chick embryos from the 8th to the 21st day of incubation and of 1- and 2-day-old chicken, the evolution of the saccharidic moieties of glycoconjugates, which precedes and/or is concomitant with the epithelial morphological transformations. We have found differences in content and cellular distribution of oligosaccharides during the histogenetic processes which characterize the lining and glandular epithelium. Before the appearance of cilia and mucus secretion at the bathyprismatic epithelial cells, the sugar residues D-galactose-(beta 1-->3)-N-acetyl-D-galactosamine, beta-N-acetyl-D-galactosamine and alpha-L-fucose were detected only at the luminal cell surface. These oligosaccharides were probably involved in giving rise to the polarization of the esophageal epithelial cells. The esophageal gland mucus was first characterized by the presence of alpha-L-fucose and afterwards also by the presence of D-galactose-(beta 1-->3)-N-acetyl-D-galactosamine, D-glucosamine and sialic acid.

Animals↗

Nested expression domains of four homeobox genes in developing rostral brain.

Insight into the genetic control of the identity of specific regions along the body axis of vertebrates has resulted primarily from the study of vertebrate homologues of regulatory genes operating in the Drosophila trunk, but little is known about the development of most anterior regions of the body either in flies or vertebrates. Three Drosophila genes have been identified that are important in controlling the development of the head, two of which, empty spiracles and orthodenticle, have been cloned and shown to contain a homeobox. We previously cloned and characterized Emx1 and Emx2, two mouse genes related to empty spiracles that are expressed in restricted regions of the developing forebrain, including the presumptive cerebral cortex and olfactory bulbs. Here we report the identification of Otx1 and Otx2, which are related to orthodenticle. We have compared the expression domains of the four genes in the developing rostral brain of mouse embryos at a developmental stage, day 10 post coitum, when they are all expressed. Otx2 is expressed in every dorsal and most ventral regions of telencephalon, diencephalon and mesencephalon. The Otx1 expression domain is similar to that of Otx2, but contained within it. The Emx2 expression domain is comprised of dorsal telencephalon and small diencephalic regions, both dorsally and ventrally. Finally, Emx1 expression is exclusively confined to the dorsal telencephalon. Thus at the time when regional specification of major brain regions takes place, the expression domains of the four genes seem to be continuous regions contained within each other in the sequence Emx1 less than Emx2 less than Otx1 less than Otx2.

Amino Acid Sequence↗