PubMed Health⌕ Search

Biomedical subjects

M Gulisano

Publications and source records attributed to M Gulisano.

At least 37 records · Page 2Linked to original sources
PubMed2000

Effect of a vitamin D3 analogue on keratinocyte growth factor-induced cell proliferation in benign prostate hyperplasia.

Prostate enlargement and function is under the dual control of androgens and intraprostatic growth factors. They regulate, in concert, prostate cell proliferation and apoptosis. An increased signaling of both growth factors and androgens are supposed to underlie benign prostate hyperplasia (BPH), one of the more common disorders of the aging male. Since, in clinical practice, androgen ablation resulted in a rather limited decrease in prostate volume, therapeutic strategies targeting intraprostatic growth factors are emerging. The activated form of vitamin D, vitamin D3, and some of its analogues have been described as potent regulators of cell growth and differentiation. In this study, we report the effects of one of these vitamin D3 analogues, 1,25-dihydroxy-16ene-23yne D3, or analogue (V), on the fate of isolated epithelial cells derived from patients with BPH. We essentially found that analogue (V), as well as vitamin D3, inhibited BPH cell proliferation and counteracted the mitogenic activity of a potent growth factor for BPH cells, such as keratinocyte growth factor (KGF). Moreover, analogue (V) induced bcl-2 protein expression, intracellular calcium mobilization, and apoptosis in both unstimulated and KGF-stimulated BPH cells. Since a short-term (5-min) incubation with analogue (V) reduced the KGF-induced tyrosine phosphorylation of a 120-kDA protein, corresponding to the KGF receptor, a rapid and direct cross-talk between these two molecules is suggested. Such a rapid effect of analogue (V), together with the transient induction of intracellular calcium waves, seems to indicate the partial involvement of a membrane, nongenomic receptor for vitamin D3. In conclusion, we demonstrated the antiproliferative and proapoptotic effect of analogue (V) in BPH cells and speculated on its possible use in the therapy of BPH.

Blotting, Western↗
PubMed1999

Effect of 0.2 T static magnetic field on human neurons: remodeling and inhibition of signal transduction without genome instability.

We describe the effect of the static magnetic field generated by a 0.2 T magnetic resonance tomograph on a normal human neuronal cell culture (FNC-B4). After 15 min exposure cells showed dramatic changes of morphology: they formed vortexes of cells and exposed branched neurites featuring synaptic buttons. At the same time, thymidine incorporation and inositol lipid signaling were significantly reduced. Control (sham exposed) or non-neuronal cells (mouse leukemia, and human breast carcinoma cells) did not show any alteration following exposure. Endothelin-1 release from FNC-B4 cells was also dramatically reduced after 5 min exposure. However, PCR analysis of 12 DNA microsatellites selected as gauges of genome instability, did not reveal any alteration following exposure, thus ruling out a direct effect of the magnetic field on DNA stability. These data can be interpreted as a specific effect of the static magnetic field on human neuronal cells and are consistent with the induction of remodeling and differentiation; they demonstrate that fields below 0.5 T have significant biological effects on human neurons.

Cell Line↗
PubMed1999

Otx genes and the genetic control of brain morphogenesis.

Understanding the genetic mechanisms that control brain patterning in vertebrates represents a major challenge for developmental neurobiology. The cloning of genes likely to be involved in the organization of the brain and an analysis of their roles have revealed insights into the molecular pathways leading to neural induction, tissue specification, and regionalization of the brain. Among these genes, both Otx1 and Otx2, two murine homologs of the Drosophila orthodenticle (otd) gene, contribute to several steps in brain morphogenesis. Recent findings have demonstrated that Otx2 plays a major role in gastrulation and in the early specification of the anterior neural plate while Otx1 is mainly involved in corticogenesis, and Otx1 and Otx2 genes cooperate in such a way that a minimal level of OTX proteins are required for proper regionalization and subsequent patterning of the developing brain. Finally, experiments have shown functional equivalence between Drosophila otd and vertebrate Otx genes, suggesting a surprising conservation of function required in brain development throughout evolution.

Animals↗
PubMed1999

Lack of association between body weight, bone mineral density and vitamin D receptor gene polymorphism in normal and osteoporotic women.

In an ethnically homogeneous population of women living in Tuscany, Italy, the relationships between age, body weight, bone mineral density and the vitamin D receptor (VDR) gene polymorphism were studied, with the objective of recognizing patients at risk for osteoporosis. In 275 women bone mineral density was measured by Dual Energy X-rays Absorptiometry (DEXA). In 50 of them the individual genetic pattern for VDR was evaluated by DNA extraction followed by PCR amplification of the VDR gene, and digestion with the restriction enzyme BsmI. Age and bone mineral density were inversely related (R2 = 0.298). Body weight was associated with bone mineral density (R2 = 0.059), but not with age. In osteoporotic women, mean (+/- SD) body weight was 59.9 +/- 6.5 Kg, lower than that recorded in non osteoporotic women (64.2 +/- 9.4 Kg), even though not significantly different (p = 0.18). No association was found between VDR gene polymorphism, bone density or body weight. The performance of anthropometric and genetic components appear to be poor, and, at least for the time being, bone mineral density measurement by means of MOC-DEXA represents the optimal method to detect women at risk for postmenopausal osteoporosis.

Adult↗
PubMed1999

Sex steroids and odorants modulate gonadotropin-releasing hormone secretion in primary cultures of human olfactory cells.

Olfactory neurons and GnRH neurons share a common origin during development. In the nasal epithelia, GnRH neurons persist throughout fetal life and adulthood. The fate and function of these neurons in vivo have remained unknown. In a previous in vitro study, we isolated, cloned, and propagated primary long term cell cultures from the olfactory neuroepithelium of 8- to 12-week-old human fetuses. These cells expressed both neural proteins as well as olfactory genes and were responsive to odorant stimuli. We now report that these human olfactory cells also express the GnRH gene and protein. Combined HPLC and RIA studies have indicated that these cells release authentic GnRH in spent media. The release of GnRH was time dependent and was positively affected by sex steroids and odorants. Immunohistochemical data demonstrated the presence of sex steroid receptors in these cells. The presence of the alpha- and beta-subtypes of the estrogen receptor was also demonstrated by RT-PCR and Western blot analysis. When the cells were stimulated with increasing concentrations of 17beta-estradiol in the presence of a fixed concentration of progesterone (10(-7) mol/L), the combination of the two steroids induced a 3- to 4-fold increase in GnRH secretion. This stimulatory effect was completely blunted by tamoxifen. Neither 17beta-estradiol nor progesterone was effective when tested separately. Treatment with increasing concentrations of the odorant, l-carvone, induced a time- and dose-dependent dramatic increase in GnRH protein release (1000-fold increase) and gene expression. Repeated application of the stimulus resulted in a progressive lower responsiveness of the cells. To our knowledge, this is the first time that primary cell cultures from human fetal olfactory neuroepithelium have been shown to express and release GnRH. Our results also demonstrate that these cultures, which are sensitive to sex steroids and odorants, can be useful models in the study of the complex array of regulatory factors that finely tune GnRH secretion in humans.

Acyclic Monoterpenes↗
PubMed1999

Influence of static magnetic field on the antiproliferative effects of vitamin D on human breast cancer cells.

We describe the effect of a 0.2 tesla (T) static magnetic field generated by a magnetic resonance tomograph and of vitamin D treatment on a human breast cancer cell line (MCF-7). Cell damage and proliferation were monitored by measuring the incorporation of [3H]thymidine in duplicating DNA and by the clonogenic assay. [3H]Thymidine incorporation in MCF-7 was stimulated by vitamin D at low doses (10(-12)-10(-10) M), whereas it was inhibited at higher concentrations (10(-9)-10(-6) M). Magnetic field treatment (0.2 T) decreased [3H]thymidine incorporation in human breast cancer cells, eliminating the proproliferative effect of low doses of vitamin D, and enhanced the vitamin D antiproliferative effect, further reducing [3H]thymidine incorporation, from -12.5% (P < 0.05) to -66.7% (P < 0.001), over the range of 10(-9) to 10(-6) M. In the clonogenic assay, ability of MCF-7 to form colonies was inhibited by vitamin D 10(-9) M and above, whereas 3-h exposure to 0.2 T magnetic field had no effect on the number of cell colonies formed. In conclusion, vitamin D treatment yields a permanent antiproliferative effect, while magnetic field exposure only temporarily slows down cellular growth. These findings suggest that therapy with vitamin D may prove beneficial for chemoprevention or treatment of breast cancer. Static magnetic field, alone or in combination, does not appear to represent an effective candidate for breast cancer therapy, at least at the intensity used in the present study.

Breast Neoplasms↗
PubMed1998

Presence and location of endothelin and endothelin-binding sites in human nasal mucosa under normal and metaplastic conditions.

The presence and site of production of endothelin-1 (ET-1) was investigated in biopsies obtained from the nasal mucosa of 10 healthy human subjects and 10 patients affected by chronic rhinitis. The presence and localization of receptors for ET-1 was also investigated. Bioptic fragments were examined by scanning electron microscopy. ET-1 was present in the vessels and in the respiratory epithelium of normal subjects, whereas in patients affected by epithelial metaplasia induced by chronic rhinitis, it was absent in the metaplastic epithelium and present in the endothelium and vascular wall. Receptors for ET (A- and B-receptor subtypes) were localized in the vessels of the nasal mucosa, both in normal and in pathological subjects. In particular, A-receptors were identified in the vascular wall, whereas B-receptors were mainly distributed in the endothelium. We suggest that ET-1 is involved in the homeostasis of nasal blood flow (shunting the blood toward the deep cavernous plexus and inducing mucosal swelling) by an autocrine and/or paracrine mechanism. Normal epithelium seems to be important in this mechanism, since it is able to produce ET. However, when pathologic conditions induce squamous or cuboidal metaplasia, the epithelium is no longer able to play this role.

Adult↗
PubMed1998

Mutations in GDI1 are responsible for X-linked non-specific mental retardation.

Rab GDP-dissociation inhibitors (GDI) are evolutionarily conserved proteins that play an essential role in the recycling of Rab GTPases required for vesicular transport through the secretory pathway. We have found mutations in the GDI1 gene (which encodes uGDI) in two families affected with X-linked non-specific mental retardation. One of the mutations caused a non-conservative substitution (L92P) which reduced binding and recycling of RAB3A, the second was a null mutation. Our results show that both functional and developmental alterations in the neuron may account for the severe impairment of learning abilities as a consequence of mutations in GDI1, emphasizing its critical role in development of human intellectual and learning abilities.

Brain↗
PubMed1997

Expression and activation of SH2/PTB-containing ShcA adaptor protein reflects the pattern of neurogenesis in the mammalian brain.

The adult mammalian brain comprises many functionally distinct neuronal types, which are generated during development as a result of a coordinated signaling cascade that drives neuroblasts from proliferation into differentiation. We investigated whether and how ShcA adaptor proteins, which are known to function as initiators of the Ras signaling cascade in various nonneuronal systems where they have been considered to be expressed ubiquitously, are involved in the proliferative and differentiative phases of the developing brain. We found that in the forebrain expression and activation of ShcA proteins were strictly regulated during embryonic development, both temporally and spatially. The mRNAs encoded by the ShcA gene were expressed exclusively within an area to which active proliferation of immature neuroblasts was confined, the ventricular zone. In postnatal and adult brain, ShcA mRNAs and proteins were present only faintly. In the adult olfactory epithelium, in which neuronal cell renewal occurs throughout life, ShcA remained strongly expressed. These phenomena were peculiar to ShcA, since Grb2 adaptor protein remained expressed at constant level throughout development. The embryonically expressed ShcA proteins were functionally active, since p52(ShcA) became phosphorylated on tyrosine and associated with Grb2 following intraventricular injection of epidermal growth factor in the embryonic brain. Our data indicate that, through an orderly pattern of expression, ShcA gene products may play a role in the control of the switch between proliferation and differentiation of brain neuroblasts.

Adaptor Proteins, Signal Transducing↗
PubMed1997

Branchial HOX gene expression and human craniofacial development.

Members of the Antennapedia class of homeobox genes, known as Hox genes, are believed to be pivotal in vertebrate craniofacial development. Here we show that eight members of paralogous groups 1, 2, 3, and 4 are expressed in the human embryonic hindbrain and branchial arches at 4 weeks of development. The combinatorial patterns of expression of genes representing the first three paralogous groups parallel the patterns described for their homologues in various animal models, demonstrating a high degree of conservation of the branchial Hox code. Arch expression of group 4 genes is defined for the first time in any vertebrate. Furthermore, as development proceeds, individual paralogues of a single paralogous group (group 3), which initially share a common expression domain, are differentially down-regulated in a tissue-, organ-, or site-specific fashion.

Branchial Region↗
PubMed1997

The morphology of the prostatic capsule with particular regard to the posterosuperior region: an anatomical and clinical problem.

The composition of the prostatic capsule is important from a clinical point of view, since it is considered to be a barrier against the spread of prostatic tumours. Clinical follow-up demonstrates that apparently intracapsular tumours, particularly in the posterosuperior region of the gland, are frequently understaged at clinical diagnosis. The morphology of the prostatic capsule was studied in 6 cadavers. In each case the prostate was obtained in one block together with the basal portion of the bladder, the seminal vesicles, the anterior wall of the rectum and the periprostatic connective tissue. Part of the material was plastinated, while the remainder was studied using histological and immunohistochemical methods. The prostate was found to be surrounded by connective tissue abundant with smooth muscle cells, and continuous with the stromal septa which subdivide the glandular tissue. A rich network of blood vessels was identifiable. In some regions, particularly in the posterosuperior region, a real capsule was not identifiable. The connective tissue seemed to constitute a continuum between the prostate gland and neighbouring organs, in particular, a connective tissue barrier between the prostate and the seminal vesicles was completely absent. Therefore, due to the absence of a capsular barrier and to the presence of a rich vascular network a prostatic tumour which begins in the posterior region of the gland should be considered as potentially extracapsular.

Adult↗
PubMed1997

Damage to the nasopharyngeal mucosa induced by current levels of urban air pollution: a field study in lambs.

This study concerns the effects of urban air pollution on the nasopharyngeal epithelium, with the aim of evaluating the possible harmful activity of levels of atmospheric pollution which are not currently considered to be dangerous. Over a 3 month period, 10 lambs kept in a zone characterized by numerous vehicles were sacrificed at regular intervals, and their nasopharyngeal mucosa was examined by scanning electron microscopy and image analysis. Two lambs kept in a rural area were used as controls. The local levels of some airborne contaminants (NO(x), NO2, NO, SO2, CO and particulate matter with aerodynamic diameter < or =10 microm (PM10)) were monitored throughout the experiment. The urban air had an irritating effect, inducing hypersecretion of mucus and morphological damage to the ciliated epithelium. These alterations increased with the duration of exposure to urban air and with increasing pollution levels, although the levels remained below current legal levels. We conclude that the harmful effects of airborne contaminants are probably underestimated. Moreover, physicochemical evaluation of pollution parameters should be complemented by morphological study of upper respiratory epithelium in exposed animals, since this mucosa is a sensitive target for irritating agents.

Air Pollutants↗
PubMed1997

Kinking of the human internal carotid artery: a statistical study in 100 healthy subjects by echocolor Doppler.

BACKGROUND: Kinking of the internal carotid artery is a rather frequent anomaly. It is still controversial whether it has a clinical relevance, causing cerebral vascular disturbances, or if it is a mere morphologic anomaly. A study concerning its frequency in asymptomatic subjects and an evaluation of its possible correlations were, therefore, considered interesting. METHODS: The internal carotid artery was studied in 100 apparently healthy subjects (50 men and 50 women; age range: 18 to 98 years) with no history of atherosclerotic disease of the supra-aortic trunks, or diabetes mellitus, or hypertension. Each patient underwent color Doppler ultrasonography. RESULTS: In 38 subjects out of 100, and in 53 out of 200 examined arteries (100 right and 100 left arteries), a kinking was found (26.5%). In 23 kinkings (43% of all the kinkings), the maximal systolic velocity (MSV) was greater than 1 m/sec at the level of the kinking. The kinkings were more frequent in women; such prevalence was statistically significant. The frequency of kinkings was uncorrelated with the side of the artery, the age, the body weight, the height and the length of the neck. Moreover, the presence of the kinkings was not significantly correlated with the lumen of the internal carotid artery. CONCLUSIONS: Hypotheses regarding the genesis of the kinkings were discussed, and the embryogenic hypothesis was emphasized. The importance of the MSV in the diagnosis of kinkings was stressed.

Adolescent↗
PubMed1996

cDNA sequence, map, and expression of the murine homolog of GTBP, a DNA mismatch repair gene.

DNA mismatch repair genes (MMR) have been actively studied in humans for their role in cancer susceptibility. Recently, GTBP, a new member of the MMR gene family, has been cloned and found to encode a 160-kDa protein that forms a heterodimer with hMSH2. Our group has isolated and characterized the mouse homolog of GTBP. The transcript, termed Gtmbp (G/T mismatch-binding protein), encodes a protein highly similar to its human counterpart. The gene is markedly conserved across species and maps to the distal portion of mouse chromosome 17, in a region homologous to human 2p. One Gtmbp-related sequence maps to proximal mouse chromosome 4. Studies of the expression of Gtmbp suggest that its regulated transcription is tightly linked to the level of DNA replication, consistent with the protein's DNA proofreading function.

Animals↗
PubMed1996

A family of transmembrane proteins with homology to the MET-hepatocyte growth factor receptor.

In hunting for unknown genes on the human X chromosome, we identified a cDNA in Xq28 encoding a transmembrane protein (SEX) of 1871 amino acids. SEX shares significant homology with the extracellular domain of the receptors encoded by the oncogenes MET, RON, and SEA [hepatocyte growth factor (HGF) receptor family]. Further screenings of cDNA libraries identified three additional sequences closely related to SEX: these were named SEP, OCT, and NOV and were located on human chromosomes 3p, 1, and 3q, respectively. The proteins encoded by these genes contain large cytoplasmic domains characterized by a distinctive highly conserved sequence (SEX domain). Northern blot analysis revealed different expression of the SEX family of genes in fetal tissues, with SEX, OCT, and NOV predominantly expressed in brain, and SEP expressed at highest levels in kidney. In situ hybridization analysis revealed that SEX has a distinctive pattern of expression in the developing nervous system of the mouse, where it is found in postmitotic neurons from the first stages of neuronal differentiation (9.5 day postcoitus). The SEX protein (220 kDa) is glycosylated and exposed at the cell surface. Unlike the receptors of the HGF family, p220SEX, a MET-SEX chimera or a constitutively dimerized TPR-SEX does not show tyrosine kinase activity. These data define a gene family (SEX family) involved in the development of neural and epithelial tissues, which encodes putative receptors with unexpected enzymatic or binding properties.

Amino Acid Sequence↗
PubMed1996

In vitro effects of some differentiation inductors in metaplastic epithelium of the human nasal cavity.

The alterations in the mucociliary unit in the course of chronic inflammation of the upper respiratory tract correspond to morphologic anomalies of the respiratory epithelium and induce cuboidal and squamous metaplasia. While the squamous pattern is most probably irreversible, it is still not clear whether it is possible to restore ciliogenesis in cuboidal metaplasia. In the present study, the action of different inductors of differentiation was evaluated in vitro in isolated cells and explants from human nasal metaplastic epithelium. Polar/apolar compounds induced secretory activity, whereas retinoic acid was able to induce ciliogenesis in some cases. Therefore, the cuboidal metaplastic condition appears to be reversible, and two distinct pathways of differentiation, secretory and ciliogenetic, are identifiable.

Acetamides↗
PubMed1996

EMX1 homeoprotein is expressed in cell nuclei of the developing cerebral cortex and in the axons of the olfactory sensory neurons.

We analyzed the distribution of EMX1 during mouse development. EMX1 is a homeoprotein encoded by Emx1, a regulatory homeobox gene expressed in the developing forebrain. Its distribution essentially overlaps the expression domains of Emx1 transcripts. The EMX1 protein is present in the developing dorsa telencephalon, that is in the cerebral cortex, olfactory bulb and hippocampus. In the cerebral cortex EMX1 is present in nuclei of proliferating, differentiating and most mature neurons belonging to all cortical layers. In the olfactory bulb it is present in all proliferating cells during development, whereas postnatally it is faintly expressed in some mitral cells. Non-cerebral localizations include a transient expression in branchial pouches, in the apical ectodermal ridge of the developing limbs and in the developing kidney. Of particular interest is the presence of EMX1 in the olfactory nerve from its first appearance during embryogenesis to birth. The protein is present in axons of olfactory sensory neurons along their entire length, including their terminals in spherical regions of neuropil in the olfactory bulb called glomeruli.

Animals↗