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Biomedical subjects

M J Soares

Publications and source records attributed to M J Soares.

At least 55 records · Page 3Linked to original sources
PubMed1999

Transcriptional activation of the decidual/trophoblast prolactin-related protein gene.

The decidual/trophoblast PRL-related protein (d/tPRP) is dually expressed by decidual and trophoblast cells during pregnancy. We have characterized the proximal d/tPRP promoter responsible for directing d/tPRP expression in decidual and trophoblast cells. We have demonstrated that the proximal 93 bp of d/tPRP 5'-flanking DNA are sufficient to direct luciferase gene expression in primary decidual and Rcho-1 trophoblast cells, but not in fibroblast, undifferentiated uterine stromal cells or trophoblast cells of a labyrinthine lineage. The 93-bp d/tPRP promoter was also sufficient to direct differentiation-dependent expression in trophoblast giant cells. Mutational analysis demonstrated the differential importance of activating protein-1 and Ets regulatory elements (located within the proximal 93 bp of d/tPRP 5'-flanking DNA) for activation of the d/tPRP promoter in decidual vs. trophoblast cells. Disruption of the activating protein-1 regulatory element inhibited d/tPRP promoter activity by more than 95% in decidual cells, and approximately 80% trophoblast cells. Disruption of the Ets regulatory element reduced d/tPRP promoter activity by approximately 50% in decidual cells, while inactivating the d/tPRP promoter in trophoblast cells. Protein interactions with the trophoblast Ets regulatory element were shown to be cell type specific and to change during trophoblast giant cell formation. In conclusion, a 93-bp region of the d/tPRP promoter is shown to contain regulatory elements sufficient for gene activation in decidual and trophoblast cells.

Animals↗
PubMed1999

Biological effects of lithium chloride on Herpetomonas muscarum muscarum and Blastocrithidia culicis (kinetoplastida: trypanosomatidae).

Lithium is widely used in medicine as an antidepressive drug and for myelosuppression attenuation during chemotherapy. In spite of abundant literature, questions on the biological action of lithium ions are far from being answered. We have here examined the effects of lithium (10-200 mM) on culture forms of the trypanosomatid protozoa Herpetornonas muscarum muscarum and Blastocrithidia culicis. Incubation of these parasites with LiCl inhibited cell growth in a concentration-dependent manner, but growth could be restored when the drug was removed from the medium. Furthermore, Li+ induced cell differentiation in H. m. muscarum. Light microscopy examination of cell viability, using erythrosin B staining, showed that all treated parasites remained viable with all drug concentrations used. Ultrastructural analysis by transmission electron microscopy showed that the cells presented no signs of degeneration. However, in H. m. muscarum the nuclei lost their peripheral heterochromatin and appeared filled with a homogeneous matrix, whereas in B. culicis an increased amount of lipid droplets was present in the cytoplasm. Our data show that LiCl treatment arrested the cell division process, stimulated cell differentiation, and affected the metabolism of these parasites.

Animals↗
PubMed1998

Differential regulation of lipopolysaccharide (LPS) activation pathways in mouse macrophages by LPS-binding proteins.

LPS binding to its receptor(s) on macrophages induces the synthesis of inflammatory mediators involved in septic shock. While the signaling mechanism(s) remains to be fully defined, the human LPS-binding protein (LBP) is known to regulate responses to LPS by facilitating its binding to CD14 on human monocytes. The structurally related bactericidal permeability increasing protein (BPI) differs from LBP by inhibiting LPS-induced human monocyte activation. We have demonstrated that, unlike the human monocyte response to LPS, both LBP and BPI inhibited LPS-stimulated TNF-alpha production in mouse peritoneal macrophages. In contrast, LPS-dependent nitric oxide release was not affected by LBP. LPS induces the phosphorylation of a number of proteins in a dose and time-dependent manner, however, the pattern of LPS-induced phosphorylation was not reduced by either LBP or BPI under conditions that result in selective TNF-alpha inhibition. Further, activation of the transcription factor NF-kappaB in response to LPS was also not modified by either LBP or BPI. Finally, no differences were detected in TNF-alpha or inducible nitric oxide synthase mRNA accumulations induced by LPS in the presence or absence of either protein, whereas a slight decreased mRNA stability was observed in the group with LPS treatment. These results would suggest that many of the early signaling events contribute to LPS-induced macrophage signaling at a point preceding the divergence of pathways that differentially regulate TNF-alpha and NO production.

Acute-Phase Proteins↗
PubMed1998

Identification of two new members of the mouse prolactin gene family.

The prolactin (PRL) family consists of a collection of genes expressed in the uterus, placenta, and anterior pituitary. These cytokines/hormones participate in the control of maternal-fetal adaptations to pregnancy. In this report, we establish the presence of two new members of the mouse PRL family. Novel expressed sequence tags (ESTs) with significant homology to PRL were isolated from embryonic, ectoplacental cone, and placental cDNA libraries. The cDNAs were sequenced and compared to other members of the PRL family. The two new cDNAs were assigned to the PRL family based on sequence homology and were referred to as PRL-like protein-F (PLP-F) and PRL-like protein-G (PLP-G). PLP-F cDNA encodes for a predicted 267 amino acid protein containing a 30 amino acid signal peptide and three putative N-linked glycosylation sites. PLP-G cDNA encodes for a predicted 266 amino acid protein containing a 30 amino acid signal peptide and six putative N-linked glycosylation sites. Sequence alignments of these proteins with other members of the PRL family suggest some unique features. Both sequences contain an extra amino acid segment located between exons two and three of the prototypical PRL gene and a nine amino acid carboxy terminal extension. PLP-F contained an additional 15 amino acid region situated between exons four and five of the prototypical PRL gene. Both PLP-F and PLP-G mRNAs were expressed in the placenta but not in other tissues (uterus, brain, thymus, heart, lung, diaphragm, liver, kidney, and ovary). In summary, the two newly identified members share approximately 50% amino acid sequence identity, are specifically expressed in the placenta, and represent a new subfamily within the PRL family.

Amino Acid Sequence↗
PubMed1998

Gestational and smoking effects on peptidase activity in the placenta.

Angiotensin converting enzyme (ACE) and leucine aminopeptidase (LAP) regulate fetally and maternally generated peptides in the placenta. In this study, ACE-like activity was found to be decreased and LAP-like activity increased with increasing days of gestation in rat placental tissues forming the fetal:maternal interface. Membrane-associated ACE-like and LAP-like activities in the placenta of smokers were also found to be significantly higher than their respective activities in placenta of nonsmokers. Our collective findings suggest that gestational and environmentally-induced changes in placental peptidase activities may account for variable peptide hormone and/or therapeutic peptide metabolism in the placenta.

Animals↗
PubMed1998

No evidence for an ethnic influence on basal metabolism: an examination of data from India and Australia.

A lower BMR of Indians, when compared with Westerners matched for age, sex and either surface area or body weight, has often been reported in the literature and has been interpreted to reflect an ethnic influence on BMR. To determine the contribution of body composition to these observed differences in BMR, we analysed the data on ninety-six Indians and eighty-one Caucasian Australians of both sexes, aged 18-30 years, studied in Bangalore, India and Melbourne, Australia. Absolute BMR and BMR adjusted for body weight were significantly lower in Indians when compared with Australians of the corresponding sex. However, BMR adjusted for fat-free mass (FFM) in men, and BMR adjusted for FFM and fat mass (FM) in women, were not significantly different between the two groups. Stepwise regression of FFM, FM, sex (0 = women; 1 = men) and ethnicity (0 = Indian; 1 = Australian) on BMR, resulted in the following relationship for the combined data on all subjects: BMR = 88.7 x FFM (kg) + 1713 (n 177; r 0.92; r2 0.85; SEE 425 kJ). The Indian equations of Hayter & Henry (1994), based on body weight, resulted in a significant bias (measured-predicted BMR) of 318 (SE 54) kJ/d in Indian men and -409 (SE 70) kJ/d in Indian women. The equation of Cunningham (1991), based on FFM, accurately predicted the BMR of Indian men, Indian women and Australian men. The small but significant bias of 185 (SE 61) kJ/d in Australian women, may be explained by the significant contribution of FM to BMR in this group. The present study does not provide any evidence for an ethnic influence on basal metabolism. The results strongly support the use of FFM, rather than body weight, for the prediction of BMR in population groups of varying body size and composition. This would allow an accurate estimation of BMR and hence energy requirements in population groups worldwide.

Adolescent↗
PubMed1998

Homologues for prolactin-like proteins A and B are present in the mouse.

The prolactin (PRL) family consists of a collection of proteins expressed in the uterus, placenta, and anterior pituitary. These cytokines/hormones are hypothesized to control maternal-fetal adaptations to pregnancy. Establishment of mouse models for members of the PRL family expands the experimental repertoire available for investigations on their biological activities. In this report, we establish the presence of mouse homologues for two rat members, PRL-like protein-A (PLP-A) and PLP-B. We present data on their cDNAs and describe aspects of their expression in uteroplacental tissues. A mouse genomic DNA fragment was found to hybridize with a rat PLP-A cDNA. Perusal of the National Center for Biotechnology Information dbEST database resulted in the identification of several putative mouse PLP-A cDNAs and a single putative mouse PLP-B cDNA. The cDNAs were obtained from the IMAGE consortium and Research Genetics and sequenced, and their corresponding mRNAs and proteins were characterized. Overall, mouse PLP-A and PLP-B showed considerable similarities with rat PLP-A and PLP-B in both structure and expression. PLP-A was expressed in both trophoblast giant cells and spongiotrophoblast cells, whereas PLP-B was expressed in decidual and spongiotrophoblast cells. However, some notable exceptions were evident. Mouse PLP-A contained a single putative N-linked glycosylation site and consisted of a single 29-kDa protein species, whereas rat PLP-A contained two putative N-linked glycosylation sites and consisted of two protein species, of 29 and 33 kDa. Subtle differences in the expression patterns in the mouse and rat are also apparent. In summary, we have established the presence of PLP-A and PLP-B in the mouse. The findings expand our knowledge of these two cytokines/hormones and provide additional strategies for studying their function.

Amino Acid Sequence↗
PubMed1998

Time-dependent physiological regulation of rodent and ovine placental glucose transporter (GLUT-1) protein.

To examine the in vivo and in vitro time-dependent effects of glucose on placental glucose transporter (GLUT-1) protein levels, we employed Western blot analysis using placenta from the short-term streptozotocin-induced diabetic pregnancy (STZ-D), uterine artery ligation-intrauterine growth restriction (IUGR) rat models, pregnant sheep exposed to chronic maternal glucose and insulin infusions, and the HRP.1 rat trophoblastic cell line exposed to differing concentrations of glucose. In the rat, 6 days of STZ-D with maternal and fetal hyperglycemia caused no substantive change, whereas 72 h of IUGR with fetal hypoglycemia and ischemic hypoxia resulted in a 50% decline in placental GLUT-1 levels (P < 0.05). In late-gestation ewes, maternal and fetal hyperglycemia caused an initial threefold increase at 48 h (P < 0.05), with a persistent decline between 10 to 21 days, whereas maternal and fetal hypoglycemia led to a 30-50% decline in placental GLUT-1 levels (P < 0.05). Studies in vitro demonstrated no effect of 0 mM, whereas 100 mM glucose caused a 60% decline (P < 0.05; 48 h) in HRP.1 GLUT-1 levels compared with 5 mM of glucose. The added effect of hypoxia on 0 and 100 mM glucose concentrations appeared to increase GLUT-1 concentrations compared with normoxic cells (P < 0.05; 100 mM at 18 h). We conclude that abnormal glucose concentrations alter rodent and ovine placental GLUT-1 levels in a time- and concentration-dependent manner; hypoxia may upregulate this effect. The changes in placental GLUT-1 concentrations may contribute toward the process of altered maternoplacentofetal transport of glucose, thereby regulating placental and fetal growth.

Animals↗
PubMed1998

Is there evidence for an age-related reduction in metabolic rate?

To determine whether the age-related reduction in basal metabolic rate (BMR) is explained by a quantitative and/or qualitative change in the components of lean tissue, we conducted a cross-sectional study in groups of young (n = 38, 18-35 yr) and older (n = 24, 50-77 yr) healthy individuals. BMR was measured by indirect calorimetry. Body composition was obtained by using dual-energy X-ray absorptiometry (DEXA), which permitted four compartments to be quantified [bone mineral mass, fat mass (FM), appendicular lean tissue mass (ALTM), and nonappendicular lean tissue mass (NALTM)]. Absolute BMR and ALTM were lower, whereas FM was significantly higher in the older, compared with young, subjects. BMR, adjusted for differences in FM, ALTM, and NALTM, was significantly lower in the older subjects by 644 kJ/day. In separate regression analyses of BMR on body compartments, older subjects had significantly lower regression coefficients for ALTM and NALTM, compared with young subjects. Hence, the age-related decline in BMR is partly explained by a reduction in the quantity, as well as the metabolic activity, of DEXA-derived lean tissue components.

Absorptiometry, Photon↗
PubMed1998

Placental lactogen-I (PL-I) target tissues identified with an alkaline phosphatase-PL-I fusion protein.

The rat placenta expresses a family of genes related to prolactin (PRL). Target tissues and physiological roles for many members of the PRL family have yet to be determined. In this investigation we evaluated the use of an alkaline phosphatase (AP) tag for monitoring the behavior of a prototypical member of the PRL family, placental lactogen-I (PL-I). A probe was generated consisting of a fusion protein of human placental AP and rat PL-I (AP-PL-I). The AP-PL-I construct was stably expressed in 293 human fetal kidney cells, as was the unmodified AP vector that served as a control. AP activity was monitored with a colorimetric assay in conditioned medium from transfected cells. Immunoreactivity and PRL-like biological activities of the AP-PL-I fusion protein were demonstrated by immunoblotting and the Nb2 lymphoma cell proliferation assay, respectively. AP-PL-I specifically bound to tissue sections known to express the PRL receptor, including the ovary, liver, and choroid plexus. Binding of AP-PL-I to tissues was specific and could be competed with ovine PRL. The results indicate that AP is an effective tag for monitoring the behavior of PL-I and suggest that this labeling system may also be useful for monitoring the actions of other members of the PRL family.

Alkaline Phosphatase↗
PubMed1998

A new member of the mouse prolactin (PRL)-like protein-C subfamily, PRL-like protein-C alpha: structure and expression.

In this study, we establish the presence of a unique member of the PRL-like protein-C (PLP-C) subfamily in the mouse, PLP-C alpha, characterize its complementary DNA and gene, and map its chromosomal location and pattern of expression during pregnancy. Mouse PLP-C alpha encodes for a 239 amino acid protein and possesses from 69-71% identity with rat PLP-C, PLP-Cv, PLP-D, and PLP-H. Another feature characteristic of PLP-C subfamily members that is also present in mouse PLP-C alpha is a 6-exon/5-intron gene structure including an aromatic domain encoded by exon 3. Southern analysis with mouse and rat PLP-C subfamily probes suggested the existence of a single mouse PLP-C alpha gene. Mouse PLP-C alpha maps to chromosome 13 along with other members of the mouse PRL family. Expression of mouse PLP-C alpha increases dramatically as gestation advances and is restricted to spongiotrophoblast and trophoblast giant cells of the junctional zone. In summary, we have established the presence of a new PLP-C subfamily member in the mouse and demonstrated its similarity in structure and expression to rat PLP-C subfamily members. This level of conservation between species expands the biological significance of the PLP-C subfamily and provides additional opportunities for genetically evaluating its function.

Amino Acid Sequence↗
PubMed1998

[Left atrial myxoma and isquemic stroke in a child].

The authors present a case of left atrial myxoma with stroke in an 8-year-old female child. The tumor was removed and two months after the surgery a left atrial septum mass was observed suggesting recurrence. The child was asymptomatic. Literature review emphasizes the rarity and clinically aggressive behavior of this tumor at this age group and demonstrates rare recurrence rates after surgery.

Brain Ischemia↗
PubMed1998

Mapping of bovine pericardium: physical and histopathologic tests.

BACKGROUND AND AIMS OF THE STUDY: This study was performed to identify the physical and histopathologic characteristics of different sections of glutaraldehyde-tanned bovine pericardium. METHODS: Ten pericardial sacs were obtained from animals aged from 18 to 36 months. Physical tests included shrinkage and mechanical resistance (rupture, elongation, tenacity index). Collagen and elastic fibers were evaluated in Gomori's trichrome-stained sections, hematoxylin and eosin, by PAS and Verhoeff's method. Studied areas were proximal to the great arteries, and the right atrial, right ventricular, left ventricular and left atrial regions. RESULTS AND CONCLUSIONS: Results showed that bovine pericardium does not have enough regional differences to identify any single region for bioprosthesis manufacture. However, histopathology showed better preservation of collagen and elastic fibers in the right ventricular region, implying that this area is more adequate as bioprosthetic material.

Animals↗
PubMed1997

Permeability and metabolic properties of a trophoblast cell line (HRP-1) derived from normal rat placenta.

The HRP-1 cell line is derived from normal rat placenta and appears morphologically similar to and retains characteristic expression of cellular markers of labyrinthine trophoblast cells. In this study, monolayers of HRP-1 cells grown on permeable supports were evaluated as a potential in vitro system to study trophoblast transport and metabolism. The cell line was shown to express and retain functional activity of the predominant placental cytochrome P450 isozyme, CYP1A1. Additionally, the HRP-1 cells retain functional activity of angiotensin I converting enzyme and carboxypeptidase N-like enzyme, peptidases characteristic of the trophoblast. The permeation of several hydrophilic, inert markers across the HRP-1 monolayers was observed to be dependent on effective molecular size and to be passive in nature. Functional asymmetry of the HRP-1 cells was illustrated by the predominant permeation of linoleic acid in the apical-to-basolateral direction across the monolayers. Transferrin passage across HRP-1 monolayers was concentration-dependent, was bidirectional, and could be inhibited by unlabeled transferrin, features typical of the trophoblast transport system for transferrin. Collectively, these properties suggest that the HRP-1 cell line may provide a useful tool for evaluating some of the permeability and metabolic properties of the trophoblast.

Animals↗
PubMed1997

Developmental regulation of a pregnancy-specific oligosaccharide structure, NeuAcalpha2,6GalNAcbeta1,4GlcNAc, on select members of the rat placental prolactin family.

Successful pregnancy is dependent upon an array of signaling proteins secreted by the trophoblast cells of the placenta. Among these is a group of proteins related to pituitary prolactin, known as the prolactin/growth hormone family. These proteins are expressed at specific times during gestation and synthesized in distinct trophoblast cell types in the rat placenta. We report here that select members of this family, prolactin-like protein (PLP-A), PLP-B, PLP-C, decidual/trophoblast PRP, and placental lactogen I variant, only which are expressed in the spongiotrophoblast, late in rat placental development bear Asn-linked oligosaccharides terminating with NeuAcalpha2,6GalNAcbeta1,4GlcNAcbeta-R. This reflects the concurrent expression of these prolactin/growth hormone family members with the peptide-specific beta1,4GalNAc-transferase and an alpha2,6-sialyltransferase, which can add sialic acid to terminal beta1,4-linked GalNAc. We have determined that at least one of the prolactin-like proteins, PLP-A, is recognized by the protein-specific GalNAc-transferase. The presence of NeuAcalpha2, 6GalNAcbeta1,4GlcNAcbeta-R on only a limited number of glycoproteins synthesized by the spongiotrophoblasts between mid gestation and birth reflects the need for both the GalNAc-transferase and the peptide recognition determinant for efficient addition of GalNAc. Thus, expression of the GalNAc-transferase and specific members of the prolactin/growth hormone family is developmentally regulated in the rat placenta, suggesting a physiological role for the terminal NeuAcalpha2,6GalNAcbeta1,4GlcNAcbeta-R sequence on Asn-linked oligosaccharides of these proteins.

Animals↗
PubMed1997

Urea kinetics varies in Jamaican women and men in relation to adiposity, lean body mass and protein intake.

OBJECTIVE: We have measured urea kinetics in normal adult men and women of different body composition to determine whether adiposity is associated with differences in the rate of urea production or endogenous urea hydrolysis. DESIGN: Urea kinetics were determined from the excretion of [15N15N]urea in urine over a period of 48 h following a single oral dose of [15N15N]urea, in nine lean and nine obese women and in seven light and seven heavy males while they were consuming their habitual diets. Urinary 5-L-oxoproline was measured as an index of glycine metabolic status. SETTING: The studies were carried out in the research ward of the Tropical Metabolism Research Unit, University of the West Indies. RESULTS: Successful studies were completed in eight obese and five lean women and in six heavy and five light men. When compared with lean women, in obese women the rate of urea production and hydrolysis was significantly greater and this difference could not be accounted for by the greater fat-free mass alone, and was in part associated directly with the increase in fat mass. The rate of urea production and hydrolysis was greater in heavy men than in light men, a difference which was attributed to an increase in dietary protein. In obese women and heavy men there was a significantly higher rate of excretion of 5-L-oxoproline in urine when compared with lean women and lean men respectively. CONCLUSION: This paper highlights the difficulty in identifying an appropriate reference with which to express results in people of different body composition. In obese women urea production and the hydrolysis of urea are increased, in part related to the increased fat-free mass, but also related to the increased fat mass itself. In obese women and men on high protein diets the greater rate of hydrolysis urea may be a reflection of an increased demand for the synthesis of non-essential amino acids, especially glycine.

Adult↗