Biomedical subjects

S Ramos

Publications and source records attributed to S Ramos.

At least 55 records · Page 3Linked to original sources

PubMed1998

The mouse tumor necrosis factor receptor 2 gene: genomic structure and characterization of the two transcripts.

The mouse TNFR2 gene has been cloned, sequenced, and characterized as a gene spanning >44 kb of the genome. By alignment of five genomic clones we have established that TNFR2 consists of 10 exons and 9 introns with exons ranging in size from 35 bp to 2.6 kb and introns ranging from 322 bp to >16 kb. All splice acceptor and donor sites conform to the canonical AG/GT rule. The translation initiation and termination sites are located in exon 1 and 10, respectively. Although TNFR2 lacks a canonical TATA box, the gene is transcribed from a unique start site located 70 bp upstream of the ATG initiation codon that conforms to the consensus Inr motif. Several cis-elements for transcription factors were identified in the 5' flanking region, including NF-1, Sp-1, AP2, gamma-IRE, and NF-kappaBeta motifs. Functional analysis indicates that the region -705/-412 contains a negative cis-acting element and that the minimal promoter contains motifs that confer LPS inducibility. Two mouse TNFR2 mRNAs of 3.2 and 4.1 kb are detected by Northern blot analysis, but until now their origin has not been explained. No evidence of alternative splicing of the coding exons was found. However, hybridization studies and amplification of cDNA ends suggest the use of a noncanonical polyadenylation signal in the untranslated region of exon 10. A comparative analysis of the 3' untranslated regions of the human and mouse TNFR2 genes shows highly divergent 3' ends. The possibility of an ancestral mouse TNFR2 mRNA similar to the short transcript is discussed.

Amino Acid Sequence↗

PubMed1998

Hamartoma of the mitral valve with blood cysts: a rare tumor detected by echocardiography.

A 16-year-old boy was submitted to a cardiac examination after an episode of faintness. A transthoracic echocardiogram was performed, which revealed a very mobile multicystic tumor attached to the mitral valve. A small solid mass adherent to the cysts was better defined by transesophageal echocardiography. The patient was submitted to cardiac surgery consisting of tumor resection and a mitral prosthesis implant. The surgery was successful. The tumor consisted of three bright red tense cysts with hematic content, each 1 to 2 cm in diameter. The cysts were coalescent and adherent to a solid mass attached to the posterior papillary muscle head. Histopathologic examination revealed a hamartoma with a cystic part composed of the proliferation of myofibroblast cells in a stroma with vessels, collagen, and elastin fibers. Valvular hamartoma with blood cysts is a very rare cardiac tumor both for its histopathology and its localization.

Adolescent↗

PubMed1998

Percutaneous techniques for the treatment of cerebrovascular disease.

Stroke is the third leading cause of death in the United States. Carotid artery stenting is being investigated as a therapeutic strategy for the management of extracranial bifurcation stenosis and has the potential to prevent stroke in thousands of patients. Carotid endarterectomy, although effective, does have limitations, and percutaneous techniques may offer an alternative method of treatment, especially for those who are at highest risk. Although the technique is still evolving, this article describes the protocol and technique of stent-supported carotid angioplasty and care for patients undergoing this procedure at Washington Adventist Hospital in Takoma Park, MD.

Aged↗

PubMed1998

Mechanism of hypothyroidism action on insulin-like growth factor-I and -II from neonatal to adult rats: insulin mediates thyroid hormone effects in the neonatal period.

The effects of thyroid hormone deficiency on serum levels and liver messenger RNA (mRNA) expression of insulin-like growth factors (IGFs) were studied in neonatal (until 20 days of life), weaned (22-37 days), and adult (72-87 days) rats, short periods (5, 10, and 15 days) after thyroidectomy (T). Serum levels and liver mRNA expression of IGF-I, plasma and pituitary GH, plasma insulin, and glycemia were measured in all populations; and serum levels and liver mRNA expression of IGF-II were measured only in the neonatal populations. Surprisingly, plasma insulin and GH and serum and liver mRNA expression of IGF-I were found elevated in T neonatal rats, and they decreased in weaned and adult rats and in neonatal rats rendered hypothyroid by mercapto-1-methylimidazole (MMI) treatment (MMI-hypothyroid). T and MMI-treatment of neonatal rats disturbed the normal pattern of progressive decrease of IGF-II with age. A positive correlation between insulin and IGF-I and a poor correlation between GH and IGF-I were found in both hypothyroid neonates (T and MMI-hypothyroid). On the contrary, a positive correlation between GH and IGF-I and a poor correlation between insulin and IGF-I were found for control and T adult rats. Because plasma insulin and GH changed in the same direction in all groups, insulin secretion in T neonatal was suppressed by streptozotocin, and insulin was given to T adult rats. The combined results of these experiments support the idea that the effects of thyroid hormones on IGF-I secretion are age-dependent, and they are mediated mainly by insulin during the neonatal period and by GH during adulthood.

Aging↗

PubMed1998

[The influence of therapy (sympathomimetic amines versus vasodilators) in the indexes of ejection fraction derived from an analysis of Doppler aortic flow].

PURPOSE: To assess the influence of inotropic IV therapy--dobutamine and/or dopamine--versus vasodilator therapy--nitroprusside, captopril or hydralazine--in aortic flow ejection phase indexes obtained by pulsed Doppler echocardiography. METHODS: In 17 patients admitted to the ICU (11 males, 62 +/- 14 years, 9 with ischemic cardiomyopathy and 8 with dilated cardiomyopathy, all in sinus rhythm), with congestive heart failure, and submitted to tailored therapy, 53 serial pulsed Doppler and hemodynamic evaluations were made as the therapy changed the hemodynamic and clinical status. Considering serial consecutive evaluations, a hemodynamic improvement was obtained only with inotropics in 13 (group A), and with vasodilators in only 15 (group B). The following ejection phase indexes were calculated from Doppler registers: average acceleration (AvAc) and ejection force (EFor), calculated according to the formula: Efor = 1.06 x Aortic Orifice Area x AvAc x Acceleration Velocity Time Integral. RESULTS: Arterial blood pressure increased in gr A (76.2 +/- 14.1 to 81.4 +/- 14.8 mm Hg, p < 0.05) and decreased in gr B (85.1 +/- 12.6 to 76.2 +/- 9.7 mm Hg (p < 0.05). In both groups there was a significant (p < 0.05) increase in cardiac output (CO)-from 3.9 +/- 1.1 to 4.9 +/- 1.4 L/min in group A, and from 3.9 +/- 1.2 to 4.4 +/- 1.2 L/min in group B. CO increased 18.7% in group A and 13.8% in group B (NS). There was a decrease in pulmonary capillary wedge pressure (PCWP) from 19.5 +/- 6.0 to 15.1 +/- 5.8 mm Hg in group A (p < 0.05), and from (16.9 +/- 5.7 to 12.1 +/- 4.6 mm Hg in group B (p < 0.05). PCWP decreased 19.7% in group A and 27.8% in group B (NS). Systemic vascular resistance (SVR) changed from 18.2 +/- 7.0 to 16.2 +/- 7.1 Wood U in group A (p < 0.05), and from 22.3 +/- 9.3 to 17.7 +/- 5.7 Wood U in group B (p < 0.05). In group A, AvAc increased-from 1347 +/- 611 cm.s-2 (p < 0.05), as did Efor-from 15.4 +/- 10.7 to 20.2 +/- 11.0 g.cm,s-2 (p < 0.05), whereas in group B there was no significant change in either AvAc-from 1337 +/- 284 to 1277 +/- 256 cm.s-2, or Efor-from 22.7 +/- 17.0 to 23.8 +/- 15.0 g.cm.s-2. CONCLUSIONS: Vasodilator therapy, although inducing hemodynamic changes similar to inotropics, does not alter the ejection phase indexes. Therefore, AvAc and Efor, in spite of being calculated from the aortic flow, are independent of the changes in PCWP and SVR and seem to reflect changes in inotropism in the clinical setting.

Captopril↗

PubMed1997

[Community-acquired pneumonia (CAP) with hospital treatment. The value of the clinical picture and complementary exams in predicting its etiology].

We studied 162 patients with community-acquired pneumonia admitted for hospital treatment, in order to determine the utility of clinical and ancillary examinations for predicting etiology and guiding the most appropriate empirical treatment. Acute first appearance of symptoms, purulent expectoration, chest sounds indicating lung condensation, pleuritic chest pain and leukocytosis over 12,500/ml were statistically significant in differentiating typical pneumonias from those with atypical behavior patterns. The last two features were the most relevant according to multivariate analysis. We conclude that careful taking of case histories and basic blood testing continue to be relevant and must not be considered anachronistic for the differential diagnosis of community-acquired pneumonias.

Adolescent↗

PubMed1997

Effects of melatonin on mammary gland lesions in transgenic mice overexpressing N-ras proto-oncogene.

The oncostatic effects of melatonin on the mammary gland have been studied in transgenic mice carrying the N-ras proto-oncogene under the control of the MMTV-LTR. Female (4-week-old) virgin mice with positive transgenic pedigrees were injected with melatonin (200 micrograms/mouse/ day, five times a week) or vehicle late in the evening. After 5 months of treatment, animals were sacrificed and the mammary glands were dissected for whole mounts, histology, and immunohistochemical analysis with a mouse monoclonal antibody specific for N-ras protein. Mammary glands of control transgenic mice showed different densities of hyperplastic alveolar nodules (HANs) consisting primarily of dysplastic epithelial cells with nuclear atypia and prominent nucleoli. The epithelial cells of HANs showed a high expression of N-ras while no immunostaining was detected in the unaffected mammary parenchyma. Only one (10%) of the control transgenic mice presented an infiltrating ductal carcinoma with the neoplastic cells overexpressing N-ras protein. The mammary glands of melatonin treated mice had a lower density of HANs, absence of epithelial dysplastic cells, and weak immunostaining of N-ras protein in comparison to the vehicle-treated group. None of the melatonin treated animals developed mammary carcinomas during the observation period. The lymph nodes of the inguinal mammary glands of all the vehicle-treated transgenic mice presented hyperplasia and two animals even had lymphomas, whereas in melatonin-treated animals there was less hyperplasia (two cases were atrophic) and a lack of lymphomas. We conclude that in the mammary glands of MMTV-LTR/N-ras transgenic female virgin mice, melatonin a) reduces the incidence of HANs and the expression of N-ras protein in focal hyperplastic lesions, b) completely prevents the development of epithelial cell atypia and mammary adenocarcinomas, and c) also reduces the hyperplasia of the mammary lymphoid tissue and prevents the development of lymphomas.

Animals↗

PubMed1997

Rotavirus and reovirus interaction with mouse peritoneal resident phagocytic cells.

Rotaviruses and reoviruses are involved in human and animal diseases. It is known that both viruses penetrate the gastrointestinal tract but their interaction with phagocytic cells is unknown. To study this interaction, peritoneal resident phagocytic cells were used and rotavirus and reovirus replication in peritoneal phagocytic cells was observed. However, rotavirus replication in these cells led to the production of defective particles since MA-104 cells inoculated with rotavirus phagocytic cell lysate did not show any evidence of virus replication. On the basis of these results, we suggest that, although reovirus dissemination may be helped by these phagocytic cells, these cells may control rotavirus infection and probably contribute to the prevention of its dissemination.

Animals↗

PubMed1996

Molecular cloning of a mouse homologue for the Drosophila splicing regulator Tra2.

We report the identification of a mouse cDNA, SIG41, encoding a protein of 288 amino acids that is 45% identical (58% similar) to the Drosophila splicing regulator Tra2. SIG41 cDNA contains four polyadenylation signals whose alternative use gives rise to four types of transcripts (2.1, 2.0, 1.5, and 1.4 kb) in mouse cells. Northern analysis and RT-PCR assays showed that SIG41 mRNA is present in virtually all the cell lines and tissues studied, with remarkable levels of expression in uterus and brain tissues. Differential stability of the SIG41 mRNAs was detected in mouse macrophage cells.

Amino Acid Sequence↗

PubMed1996

Identification of an additional member of the cytochrome c oxidase subunit VIIa family of proteins.

We report the cloning, nucleotide sequence, evolutionary analysis, and intracellular localization of SIG81, a silica-induced cDNA from mouse macrophages. The cDNA encodes a 111-amino acid protein with extensive sequence identity with members of the mammalian cytochrome c oxidase subunit VIIa (COX7a) family. A human SIG81 sequence >80% identical with the mouse cDNA was deducted from homologous sequences in the human expressed tags data base. The deduced aminoterminal region shows features common to mitochondrial targeting sequences. A phylogenetic analysis of the carboxyl-terminal domain homologous to COX7a identifies SIG81 as a divergent member of the family with an ancient origin. Southern blot analysis showed that the mouse genome contains two to three copies of the SIG81 gene. Northern blot analysis revealed that the SIG81 transcript is approximately 1 kb and expressed in every tissue tested, with higher levels of expression observed in kidney and liver. Antibodies raised against a glutathione S-transferase SIG81 fusion protein detected a 13.5-kDa protein that co-fractionates with mitochondrial localized enzymatic activity. Taken together, our data suggest that SIG81 is a novel member of the COX7a family that is constitutively expressed in mouse cells.

Amino Acid Sequence↗

PubMed1996

Molecular mechanisms of TNFalpha cytotoxicity: activation of NF-kappaB and nuclear translocation.

The murine fibrosarcoma cell line WEHI 164 is well known for its susceptibility to tumor necrosis factor (TNFalpha). We have studied the activation of the transcription factor NF-kappaB when WEHI 164 cells are challenged with TNFalpha. NF-kappaB is retained in the cytoplasm of unchallenged cells by its inhibitor IkappaB-alpha. Upon cellular stimulation, IkappaB-alpha is functionally inactivated and NF-kappaB translocated to the nucleus. The extent of the cytotoxic effect and that of nuclear translocation of NF-kappaB show the same TNFalpha dependence. TNFalpha induces a rapid and transient activation of NF-kappaB in WEHI 164 cells which is followed by a second, long lasting phase in which the amount of NF-kappaB complex in the nucleus remains at about 50% of maximum. Upon TNFalpha treatment, IkappaB-alpha is rapidly degraded. However, newly synthesized IkappaB-alpha can be demonstrated later in the cell cytosol. A persistent nuclear localization of NF-kappaB is an obligatory step for the cytotoxic effect to take place. Thus, WEHI 164 cells treated with TNFalpha for up to 6 h can be rescued as long as NF-kappa relocalizes to the cytoplasm in its inactive form. On the other hand, TNFalpha treatments as short as 15 min cause the cytotoxic effect provided that NF-kappaB remains in the nucleus. The activation of NF-kappaB is controlled by both phosphorylation and proteolysis. The activation of NF-kappaB can be blocked by the cysteine protease inhibitor calpain inhibitor I and the serine protease inhibitor TPCK. Signal-induced phosphorylation of IkappaB-alpha does not lead to the dissociation of the inhibitor from NF-kappaB. Phosphorylation appears to regulate the inhibitory activity of IkappaB-alpha both positively and negatively. since inhibitors of protein kinases have opposite effects. Thus, treatment of cells with staurosporin induced a partial activation of NF-kappaB and was synergistic with TNFalpha induced activation. Calphostin C, on the other hand, can block the activation of NF-kappaB by TNFalpha, also blocking its proteolytic degradation.

Animals↗

PubMed1996

Measurement of cytotoxicity by propidium iodide staining of target cell DNA. Application to the quantification of murine TNF-alpha.

A rapid and sensitive method is described for the determination of murine tumor necrosis factor (TNF-alpha), which can be performed in microtiter plates using a fluorescence plate scanner. The method is based on the binding of propidium iodide (PI), a membrane-impermeant dye, to nucleic acids of WEHI 164 cells, whose plasma membrane permeable due to TNF-alpha-induced cell damage. The analytical range for the proposed method is 0.3-200 pg/ml of TNF-alpha after 5 h of incubation. The optimal number of target cells was found to be 4-5X10(4)/well. The variability obtained for the PI assay was 7.6%; lower than that obtained with a commonly employed method in which MTT is used to determine cell viability (11.3%). Thus, the PI assay appears to be a reliable and reproducible method for the determination of biologically active TNF-alpha. The assay can be performed in a few hours and has the advantage over the current MTT and 51Cr-released assays that kinetic studies of TNF-alpha toxicity are possible since it permits multiple, sequenced measurements of cell viability during the incubation of the sample. The method can also be used for the determination of human TNF-alpha.

3T3 Cells↗

PubMed1996

MYC amplification in squamous cell carcinomas of the head and neck.

OBJECTIVES: To establish the frequency of MYC amplification in squamous cell carcinoma (SCC) of the head and neck to evaluate its correlation with clinicopathologic variables that are used in clinical practice. DESIGN: Cohort analytic study. SETTING: University Hospital. PATIENTS: Fifty-nine consecutive patients with SCC of the head and neck. INTERVENTION: Oncologic surgery. MAIN OUTCOME MEASURES: The MYC copy number in tumor samples was estimated with the polymerase chain reaction. The presence or absence of amplification was correlated with the anatomic site, T stage, nodal involvement, pathologic grade, recurrence, distant metastases, and survival. RESULTS: Six SCC specimens (11%) showed MYC amplification. A highly statistical correlation between MYC amplification and T stage was noted (P < .005). Amplification was also significantly correlated with a hypopharyngeal primary site (P < .05). No correlation among amplification status, nodal involvement, pathologic grade, relapse, metastases, and survival was observed. CONCLUSIONS: Amplification of MYC is associated with advanced primary tumors, and it appears to be a late event in the tumorigenesis of SCCS of the head and neck. However, there is no correlation between MYC amplification and prognosis.

Adult↗

PubMed1996

Differential regulation of the murine ribosomal protein L26 gene in macrophage activation.

In mouse RAW 264.7 macrophages, the gene for ribosomal protein L26 is positively regulated by silica. In order to study L26 gene expression a near full-length cDNA for mouse L26 was isolated and characterized. Sequence analysis revealed that mouse L26 is a 145 amino acid protein highly homologous to other vertebrate L26 proteins. The treatment of RAW 264.7 cells with the inflammatory mediators LPS and IFN gamma induced the expression of L26 mRNA, but the patterns of expression obtained differed markedly from silica. On the contrary, TNF alpha acted as a down-regulator of L26 gene. Our results suggest that the synthesis of ribosomal components in response to macrophage activators is inducer-specific. Mouse genomic DNA analysis revealed the presence of multiple (10-12) sequences related to the L26 gene.

Amino Acid Sequence↗

PubMed1996

Amplification of ERBB oncogenes in squamous cell carcinomas of the head and neck.

The activation of ERBB oncogenes has been described in various human tumours, including squamous cell carcinomas of the head and neck (SCCHN), and, in some of them, it has been correlated with a poor prognosis. Tissue samples from 59 patients with SCCHN were studied. After DNA extraction, the ERBB1, ERBB2 and ERBB3 copy number in tumour samples was estimated with the polymerase chain reaction (PCR) method. The PCR products were analysed by agarose gel electrophoresis and quantified by image analysis techniques. 9 (15%) cases presented with ERBB1 amplification, which was correlated with lymph node involvement (P = 0.04), poorly differentiated tumours (P = 0.03) and a hypopharyngeal primary site (P = 0.035). No correlation among amplification status, recurrence, metastases and survival was observed, although this may be due to the small number of patients in the amplified group. None of the 59 cases presented amplification of ERBB2 and ERBB3 oncogenes.

Adult↗

PubMed1996

Basement membrane thickening in small intramyocardial vessels in eosinophilic myocarditis.

The case reported in this paper, besides illustrating several aspects of the eosinophilic myocarditis, documents a yet undescribed feature of the cardiac involvement in idiopathic hypereosinophilic syndrome characterized by marked basement membrane thickening of small intramyocardial vessels. We are unaware of previous reports showing the presence of an extensive thickening of the basement membrane of arteriolar and capillary intramyocardial vessels in eosinophilic myocarditis. Considering the findings in the present case, i.e., the focal nature of the lesions, the type of necrosis, and the diffuse microangiopathy, it can be hypothesized that the microvascular disease could play a role in the development of the myocardial changes.

Basement Membrane↗

PubMed1995

Isolation of nine gene sequences induced by silica in murine macrophages.

Macrophage activation by silica is the initial step in the development of silicosis. To identify genes that might be involved in silica-mediated activation, RAW 264.7 mouse macrophages were treated with silica for 48 h, and a subtracted cDNA library enriched for silica-induced genes (SIG) was constructed and differentially screened. Nine cDNA clones (designated SIG-12, -14, -20, -41, -61, -81, -91, -92, and -111) were partially sequenced and compared with sequences in GenBank/EMBL databases. SIG-12, -14, and -20 corresponded to the genes for ribosomal proteins L13a, L32, and L26, respectively. SIG-61 is the mouse homologue of p21 RhoC. SIG-91 is identical to the 67-kDa high-affinity laminin receptor. Four genes were not identified and are novel. All of the mRNAs corresponding to the nine cloned cDNAs were inducible by silica. Steady-state levels of mRNAs in RAW 264.7 cells treated with various macrophage activators and inducers of signal transduction pathways were determined. A complex pattern of induction and repression was found, indicating that upon phagocytosis of silica particles, many regulatory mechanisms of gene expression are simultaneously triggered.

Animals↗

PubMed1995

Histologic and genetic assessment of explanted allograft valves.

A possible way of analyzing the immune response triggered by the allograft and the cellular viability is to compare immunocompetent and immunosuppressed patients, such as those having valve replacement and heart transplantation, respectively. These groups differ in immunosuppression therapy, preparation methods, valve hemodynamics. In the present study, we investigated polymerase chain reaction-amplified DNA flanking hypervariable (CA)n regions obtained from valve leaflets taken from patients having valve replacement or heart transplantation and performed a histologic analyses of the cells. In addition, we assessed an autograft valve to compare the hemodynamic effects on the cellular composition of the valve leaflet. We conclude that leaflet cellularity of the heart transplantation and autograft patients is superior to that of the valve replacement patients. These differences were consistent with the occurrence of an immune response in the valve replacement group, which was prevented or abrogated by immunosuppressive therapy administered to the heart transplantation group. However, it cannot be excluded that preparation procedures have a long-term effect on the extracellular matrix, leading to deterioration of cell adhesion and homing conditions.

Adult↗