Biomedical subjects

J Tolivia

Publications and source records attributed to J Tolivia.

At least 19 recordsLinked to original sources

PubMed1995

Ultrastructural study of a special type of ependymal cell at paraventricular level of the golden hamster third ventricle.

The present paper describes the presence of a special cell located in the ependymal wall at the level of the paraventricular nucleus. At this level, ultrastructural observation of these ependymal cells, unlike most other mammalian species, shows the presence of nucleolus-like bodies in their cytoplasm and occasionally basal processes. These processes appear perpendicular to the ependymal surface and end in contact with the basal membrane of hypothalamic capillaries. Mitochondria, endoplasmic reticulum and numerous filaments are present in the basal processes. Nucleolus-like bodies or nematosomes consist of round or ovoid unbound masses of granular appearing material of variable density located in the apical cytoplasm of the cells. Some of their ultrastructural characteristics are similar to other ependymal specialized cells which are classically termed tanycytes. These findings point out the possibility that those special cells may also be implicated in a ventricle-blood vessel communication.

Aging↗

PubMed1994

Apolipoprotein D gene induction by retinoic acid is concomitant with growth arrest and cell differentiation in human breast cancer cells.

We have examined the regulation by retinoic acid of the gene encoding apolipoprotein D (apoD), a human plasma protein belonging to the superfamily of the lipocalins that is produced by a specific subtype of highly differentiated breast carcinomas. Northern blot analysis revealed that all-trans-retinoic acid (RA) strongly induced the accumulation of apoD mRNA in T-47D and ZR-75-1 estrogen receptor-positive human breast cancer cells in a time- and dose-dependent manner, while no inductive effect was observed in estrogen receptor-negative cell lines, including MDA-MB-231 and MDA-MB-435. The effect of RA on apoD expression by T-47D cells was at least 12-fold more potent than the effect of the steroids dihydrotestosterone and dexamethasone, which had been previously described as hormonal up-regulators of apoD expression in these cells. A time course study demonstrated that the induction of apoD mRNA reached a level of 15-fold over the untreated control cells after 48 h of incubation in the presence of a 10(-7) M concentration of RA. A dose-response analysis showed that as little as 10(-13) M RA produced an accumulation of 5-fold over the control, while incubation of the cells in the presence of 10(-5) M RA induced a maximal accumulation of 24-fold over the control untreated cells. The induction of apoD mRNA was independent of the synthesis of proteins de novo, as demonstrated by the fact that the induction was also detected in the presence of cycloheximide. The incubation of the cells in the presence of RA did not affect significantly the stability of apoD mRNA. By contrast, treatment of the T-47D cells with RA produced an increase of approximately 8-fold in the rate of transcription of the apoD gene. Furthermore, treatment of the T-47D cells with RA induced the synthesis and secretion to the culture medium of apoD. This increased expression of apoD was accompanied by an inhibition of cell proliferation and a progression through a more differentiated phenotype, suggesting that the mechanisms controlling RA-induced growth arrest, cell differentiation, and apoD synthesis may be directly coordinated in human breast cancer cells.

Apolipoproteins↗

PubMed1994

Molecular cloning and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas.

A cDNA coding for a new human matrix metalloproteinase (MMP) has been cloned from a cDNA library derived from a breast tumor. The isolated cDNA contains an open reading frame coding for a polypeptide of 471 amino acids. The predicted protein sequence displays extensive similarity to the previously known MMPs and presents all the structural features characteristic of the members of this protein family, including the well conserved PRCGXPD motif, involved in the latency of the enzyme and the zinc-binding domain (HEXGHXXXXXHS). In addition, this novel human MMP contains in its amino acid sequence several residues specific to the collagenase subfamily (Tyr-214, Asp-235, and Gly-237) and lacks the 9-residue insertion present in the stromelysins. According to these structural characteristics, the MMP described herein has been tentatively called collagenase-3, since it represents the third member of this subfamily, composed at present of fibroblast and neutrophil collagenases. The collagenase-3 cDNA was expressed in a vaccinia virus system, and the recombinant protein was able to degrade fibrillar collagens, providing support to the hypothesis that the isolated cDNA codes for an authentic collagenase. Northern blot analysis of RNA from normal and pathological tissues demonstrated the existence in breast tumors of three different mRNA species, which seem to be the result of the utilization of different polyadenylation sites present in the 3'-noncoding region of the gene. By contrast, no collagenase-3 mRNA was detected either by Northern blot or RNA polymerase chain reaction analysis with RNA from other human tissues, including normal breast, mammary fibroadenomas, liver, placenta, ovary, uterus, prostate, and parotid gland. On the basis of the increased expression of collagenase-3 in breast carcinomas and the absence of detectable expression in normal tissues, a possible role for this metalloproteinase in the tumoral process is proposed.

Amino Acid Sequence↗

PubMed1994

Hamster supraoptic nucleus: cytoarchitectural, morphometric, and three-dimensional reconstruction.

BACKGROUND: The present paper describes the cytoarchitectonic, morphometric, and three-dimensional characteristics of the golden hamster supraoptic nucleus (SON) in order to provide an anatomical basis for subsequent morphofunctional studies that use this species as an experimental animal. The dimensions (volume and length) and the number of cells of each part of the supraoptic nucleus were obtained, as well as morphometric parameters of their neurons (cross-sectional area and maximum and minimum diameters). A three-dimensional reconstruction of hamster SON has been made in order to know the spatial morphology of this nucleus and to reveal the structural differences between both parts. METHODS: Ten male adult golden hamsters (Mesocricetus auratus) were used. Animals were anaesthetized and transcardially perfused with 4% paraformaldehyde in 0.1 M phosphate buffer at pH 7.2. The hypothalamic area from seven animals was dissected out, dehydrated, and embedded in paraffin. Serial sections of 10 microns were cut in a coronal plane. Sections were stained with thionin, dehydrated, cleared in eucalyptol, and mounted with Eukitt. To prove the neurosecretory nature to the SON, every fourth section was immunostained against neurophysin by using the peroxidase-antiperoxidase method. To study the neuronal morphometric parameters, all magnocellular neurons of the SON were drawn in sections separated 80 microns with the aid of a camera lucida under 500x magnification. Serial 50 microns thick frozen sections of the hypothalamus from three animals were drawn with camera lucida to determine the volume of the two parts of the SON and to make the three-dimensional reconstruction. RESULTS: The SON extends rostrocaudally 1.98 +/- 0.03 mm from the preoptic area to the tuberal hypothalamic area. Two classical parts can be clearly delimited: principal (SONp) and retrochiasmatic (SONr). The neuronal population of the two parts of the SON appears constituted only by magnocellular neurons. The volume of the SONp is 0.039 +/- 0.03 mm3 and contains about 762 +/- 93 magnocellular cells, with a density of 19,151.8 cells/mm3. The volume of the SONr is 0.126 +/- 0.03 mm3 and contains about 1,296 +/- 132 neurons with a density of 10,536.6 cells/mm3. The three-dimensional reconstruction reveals that the SONp appears located in a more cephalic, lateral, and dorsal position than the SONr, and a clear discontinuity between the two parts is observed. CONCLUSIONS: The present study shows that the classically termed SON, in the hamster, clearly consists of two spatially separated neural populations. The SONr is longer than SONp and has the larger volume and higher number of neurons; however, the neurons of the SONr are smaller in cell area than those of the SONp.

Animals↗

PubMed1994

Differential staining of nerve cells and fibres for sections of paraffin-embedded material in mammalian central nervous system.

A differential staining method is described of myelinated fibres and nerve cell bodies applicable to sections of mammalian, including human, central nervous system specimens embedded in paraffin wax. Experimental and human necropsy material fixed in acetic paraformaldehyde in phosphate buffer was used. Sections of 15-20 microns in thickness were obtained, attached to slides, deparaffinized and hydrated. After hydration, sections are oxidized (30 s) in 2% potassium permanganate, bleached (1 min) in 5% oxalic acid and rinsed in distilled water. Staining is for 2-5 h in the following solution: 0.06% thionin, 1% formaldehyde, 10% acetic acid in distilled water. Sections are subsequently washed in distilled water, dehydrated through 96% and absolute ethanol, cleared in eucalyptol and mounted in Eukitt. Using the method described in the present paper, a differential coloration of myelin and neurons is obtained. Myelinated fibres appear red, whereas nerve cell bodies and glial nuclei are stained blue. This procedure provides a high contrast between myelin and cells suitable for observation and photography of sections. Simultaneous and differential coloration of both myelin and cells is easily and directly obtained with constant and homogeneous results.

Animals↗

PubMed1994

Polychromatic staining of epoxy semithin sections: a new and simple method.

A simple, rapid method is described for the polychromatic coloration of semithin sections, which is applicable to material routinely processed for transmission electron microscopy. Material fixed with a glutaraldehyde-paraformaldehyde mixture and postfixed in osmium tetroxide with or without potassium ferrocyanide and embedded in different types of resin (Durkupan-ACM, Spurr resin, Taab resin) can be used. Constant and homogenous results are obtained with this technique, the staining procedure being achieved at room temperature in no more than 10 min. Sections of 0.5-1 microns in thickness are oxidised and bleached. After washing, sections are stained in two steps with carbol methylene blue/carbol gentian violet solution and pararosaniline solution. Using the method described in this paper, a polychromatic coloration of the different cells and tissues was obtained (epithelial cells in various shades of blue-violet, connective tissue and elastic laminae of blood vessels in pink or red, etc.). This procedure provides greater contrast between cytoplasm and nuclei, and among the different types of cells and tissues than is seen with toluidine blue, which is very useful for observation and photography of semithin sections. Polychromatic methods found in the literature are normally complex and require a lengthy staining time or cannot be applied on material routinely processed for transmission electron microscopy. Our method is simple, rapid and can be used on any type of material routinely processed for transmission electron microscopy and embedded in epoxy resins.

Animals↗

PubMed1994

Glucocorticoids and androgens up-regulate the Zn-alpha 2-glycoprotein messenger RNA in human breast cancer cells.

We have studied the hormonal regulation of the gene encoding Zn-alpha 2-glycoprotein (Zn-alpha 2-gp), a human protein with a high degree of amino acid sequence similarity to class I histocompatibility antigens that is produced by a specific subset of breast carcinomas. Northern blot analysis revealed that dexamethasone and 5 alpha-dihydrotestosterone strongly induced the accumulation of Zn-alpha 2-gp mRNA in T-47D human breast cancer cells. Furthermore, the effect of these two hormones was shown to be additive, since the combination of both hormones produced a stimulation of Zn-alpha 2-gp mRNA of at least 3-fold over that produced by either hormone alone. By contrast, the addition of 5 beta-dihydrotestosterone, 17 beta-estradiol, or progesterone failed to induce the expression of Zn-alpha 2-gp. The stimulatory effect of glucocorticoids and androgens on Zn-alpha 2-gp expression was produced in a time and dose dependent manner, without significantly affecting the cell proliferation rate. A time-course study demonstrated that the induction of Zn-alpha 2-gp mRNA by androgens and glucocorticoids reached a level of 4 or 3.2-fold over the untreated control after seven days of incubation in the presence of a 10(-7) M concentration of 5 alpha-dihydrotestosterone or dexamethasone, respectively. A dose-response study showed that as little as 10(-11) M of 5 alpha-dihydrotestosterone or dexamethasone produced an accumulation of Zn-alpha 2-gp mRNA of 2.4 or 2.1-fold over the control, respectively. On the basis of these results, we propose that Zn-alpha2-gp may be useful as a biochemical marker of breast carcinomas with a specific pattern of hormone responsiveness in whose development glucocorticoids and/or androgens may play a significant role.

Androgens↗

PubMed1994

Expression and prognostic significance of apolipoprotein D in breast cancer.

Apolipoprotein D (apo D) is a glycoprotein involved in the human plasma lipid transport system and present at large amounts in cyst fluid from women with gross cystic disease of the breast. Apo D expression in breast carcinomas was examined by immunoperoxidase staining of a series of 163 tumors. A total of 60 (36.8%) tumors were negative for apo D immunostaining, 28 (17.2%) carcinomas were weakly positive, 33 (20.2%) were moderately stained, whereas the remaining 42 (25.8%) tumors were strongly stained with the specific antibodies. No significant correlation was found between apo D content and tumor size, lymph node involvement, or biochemical parameters such as estrogen receptors, cathepsin D, or pS2 protein. However, the finding of a significant association between apo D and menopausal status of patients or differentiation grade of tumors, with apo D values being lower in tumors from premenopausal women or in poorly differentiated carcinomas, suggested a potential value of this glycoprotein as a prognostic factor in breast cancer. Preliminary analysis of relapse-free survival and overall survival in a subgroup of 152 women with a mean follow-up of 42 months confirmed that low apo D values were significantly associated to a shorter relapse-free survival and poorer survival. According to these data, we propose that apo D in combination with other well-established prognostic factors may contribute to more accurately identify subgroups of breast cancer patients with low or high risk for relapse and death.

Adult↗

PubMed1993

Rostrocaudal changes in neuronal cell size in human lateral vestibular nucleus.

A cytoarchitectonic and morphometric study of the human lateral vestibular nucleus (LVN) is presented. In sagittal sections, the LVN appears as a triangular cell group rostrally located near the motor trigeminal nucleus and caudally near the vestibular root. The estimated volume is 13.49 mm3 with a neuronal population of 25,046 cells and 1855 neurons/mm3 in density. The average neuronal cross-sectional area changes from a minimum caudally (380.02 +/- 7.23 microns 2) to a maximum rostrally (825.16 +/- 25.10 microns 2). Four types of neurons can be observed: small (< 200 microns 2), medium (200-500 microns 2), large (500-100 microns 2) and giant or Deiter's cells (> 1000 microns 2). The small and medium cells constitute 62%, large cells 26% and the giant cells only 12% of the neuronal population.

Adult↗

PubMed1993

An electron microscopic study of the harderian gland of the Syrian hamster with particular reference to the processes of formation and discharge of the secretory vacuoles.

The lipid-secreting cells of the Harderian gland of the Syrian hamster were studied using light, transmission, and scanning electron microscopy. Three morphologically different secretory cell types are identified in the gland: type I and II cells of the male gland and, distinct from either, the female gland cell. In all secretory cell types, lipid droplets in the cytoplasm were surrounded by unit membranes. Ultrastructural evidence of the involvement of the Golgi apparatus in the formation of the secretory vacuoles was obtained. The process of secretion involved the fusion of the boundary unit membrane of the vacuole with the plasma membrane and the release of the vacuolar content alone into the lumen. No evidence of holocrine processes was observed in this study. In addition to lipids, vacuoles contained materials whose solubility properties clearly differed from those of lipids. There appear to be variations in the ultrastructural characteristics of the vacuole content of the different types of secretory cell.

Animals↗

PubMed1993

Expression of pepsinogen C in human breast tumours and correlation with clinicopathologic parameters.

We have examined by immunohistochemistry the ability of breast carcinomas to produce pepsinogen C, an aspartyl proteinase usually involved in the digestion of proteins in the stomach. A total of 113 out of 245 breast tumours (46%) were positive for pepsinogen C immunostaining. There was a significant association between pepsinogen C and oestrogen receptors with proteinase levels higher (HSCORE) in oestrogen receptor positive tumours than in oestrogen receptor negative. There was also a significant association between pepsinogen C and histological grade, pepsinogen C levels being higher in well and moderately differentiated breast carcinomas than in poorly differentiated tumours. On the basis of these results, we suggest that pepsinogen C may be useful as a marker of good prognosis in breast cancer.

Adult↗

PubMed1993

Morphometric analysis of the vestibular complex in the rat.

The characteristics of the four major vestibular nuclei and accessory cell groups in the rat have been studied in serially cut horizontal sections. Camera lucida drawings of the vestibular nuclei and their neurons were made in these sections and subjected to a computerized image analysis. The dimensions (volume and length) and the number of cells of each vestibular nucleus were obtained, as well as morphometric parameters of their neurons (cross-sectional area, maximum and minimum diameter, and shape). These parameters were statistically analyzed by comparing the cell population from different nuclei and different parts of each nucleus. Of the major nuclei, the medial, which is the largest, has the greatest number of cells, its neurons being the smallest of all the nuclei, with the size of cells decreasing in a rostrocaudal direction. In contrast, the lateral nucleus contains the fewest cells but also the largest ones. Neurons of the superior and descending nuclei are of an intermediate size and number, with a rostrocaudal decrease in the size of the descending nucleus cells. In addition, minor nuclei are identified and described in their relationship to the other structures in the brain stem. The relevant aspects of the anatomical information in regard to functional roles are reviewed.

Animals↗

PubMed1992

Postnatal development of the harderian gland in the Syrian golden hamster (Mesocricetus auratus): a light and electron microscopic study.

The main objective of the present investigation was to study the morphological and chronological aspects of the development of the Harderian gland in the Syrian golden hamster. Tissues were obtained from male and female hamsters at days 1,3,5,7,10,12,15,17,20,27,37,46, and 90 after birth and processed for light and transmission electron microscopy. The present observations indicate that a well-defined temporal sequence in microscopic and ultrastructural modification is recognizable in the development of the hamster Harderian gland. Four stages of development were proposed. Between days 1-5 (first stage), the gland shows characteristics of an immature structure. The glandular cells contain many free ribosomes, few and small organelles, and large irregular-shape nuclei. Between days 7-17 (second stage), there is a marked increase of organelles involved in synthesis and secretion. The gland begins the secretion of lipids and porphyrins, but no morphological differences between male and female glands are observed. Between days 20-36 (third stage), the morphological differences between the two sexes appear and progressively develop. In 45-day-old hamsters, the Harderian gland possesses the structural characteristics of adult glands, and further developmental changes are essentially quantitative in nature (fourth stage). At all stages of development, the population of secretory cells has a uniform appearance. The morphological results are discussed as well as the possible relationship of this temporal sequence with hormonal changes.

Aging↗

PubMed1992

An ultrastructural study of myoepithelium maturation during postnatal development of the hamster Harderian gland.

This study reports the ultrastructural cell modifications in the myoepithelium of the Harderian gland during the postnatal development of the Syrian hamster. Tissues were obtained from male and female hamsters at days 1, 3, 5, 7, 10, 12, 15, 17, 20, 27, 37, 46 and 90 after birth, and processed for transmission electron microscopy. Electron microscopy was coupled with point counting methods to quantitate changes in several subcellular organelles during the course of myoepithelial cell maturation. The myoepithelial cells in this gland remained immature at birth. The earliest age of development when organized bundles of microfilaments were observed was 7 days. By the 12th day, the myoepithelial cells had developed most of their specific characteristics and resembled the mature form. Myoepithelial cells mature synchronously with each other and with the secretory cells. No undifferentiated myoepithelial and myoepithelial cells, the secretory endpieces of the adult hamster Harderian gland contain a third cell type which resembles the myoepithelial cell in shape and has an extremely electron-lucent cytoplasm lacking microfilament bundles.

Animals↗

PubMed1991

A new rapid silver impregnation for neuronal bodies on methacrylate sections.

A simple and rapid method for the impregnation of neuronal bodies applicable to methacrylate embedded sections is described in the present paper. Sections of 10-12 microns in thickness were attached to slides, placed in mordant for 1 min, rinsed in distilled water and impregnated in ammoniacal silver solution for 1 min. They were then rinsed in absolute ethanol for 30 s and developed in 50% formalin. Sections were toned in 0.25% gold chloride, reduced in 10% oxalic acid and fixed in 5% sodium thiosulfate. After washing, the sections were dehydrated through 90% and absolute ethanol, cleared in eucalyptol, and mounted in the usual way. When this method is used most of the neuronal somata and proximal dendritic trees are impregnated. Frequently some glial cell are also weakly impregnated but their density does not obscure the neurons.

Animals↗

PubMed1990

Ultrastructural study of lamellar and nucleolus-like bodies in the harderian gland during postnatal development of the hamster (Mesocricetus auratus).

Membranous structures identified as lamellar bodies (LBs) and dense intracytoplasmic bodies referred to as nucleolus-like bodies (NLBs) have been found in the hamster Harderian gland during neonatal stages. Both structures appear between 8 and 12 postnatal days, coinciding with the beginning of secretory activity. LBs in males and NLBs in both sexes gradually decrease in number with further differentiation of the glandular cells. The morphological features of these cytoplasmic structures are described, and their origin as well as their possible functional significance are discussed.

Animals↗

PubMed1990

Circadian changes in synaptic ribbons and spherules in pinealocytes of the Syrian hamster (Mesocricetus auratus).

In the present study, "synaptic" ribbons were studied morphologically and quantitatively in hamster pineal gland. The number of ribbons and spherules of hamster pinealocytes was counted over a 24-h period. The 24-h variations in the quantity of "synaptic" ribbons were found to parallel fluctuations in pineal melatonin concentrations. No significant circadian changes were observed for "synaptic" spherules, indicating different roles for these two structures.

Animals↗

PubMed1989

Differential thionin block staining of nerve cells and fibers for paraffin-embedded material in mammalian central nervous system.

A differential staining method of myelinated fibers and nerve cell bodies applicable to whole blocks of mammalian central nervous tissue is described. Experimental material fixed by perfusion or necropsy material fixed in block can be used. Blocks of 2 mm in thickness were obtained with a vibratome, immersed in 50% ethanol for 7 h and stained for 1 week in the following solution: 0.3% thionine, 5% formaldehyde and 5% acetic acid. After the staining period the blocks were washed in distilled water, dehydrated through graded alcohols, cleared in butyl acetate and infiltrated in paraffin. Sections of 10 microns in thickness were obtained, attached to slides, dewaxed in xylene and coverslipped with mounting media in the usual manner.

Animals↗