Search PubMed⌕ Search

Biomedical subjects

S Ramirez

Publications and source records attributed to S Ramirez.

At least 19 recordsLinked to original sources
PubMed2006

Diversity of human rotaviruses detected in Sicily, Italy, over a 5-year period (2001-2005).

Rotavirus infection was detected in 39.9% of 1030 children hospitalized with gastroenteritis in Palermo, Italy, in the period 2001-2005. Rotavirus strains belonging to G1, G4 and G9 types were continually detected, with G1 being the most common type in 2001, 2002 and 2004. A G4 epidemic occurred in 2003, while G9 was predominant in 2005. G2 strains displayed a low prevalence, except in 2003. G3 rotaviruses accounted for 2.7-17% of the gastroenteritis episodes in 2002-2005. The P-type of a subset of 166 strains confirmed the circulation of the usual G/P combinations, but single G1P[6], G9P[9] and G6P[9] strains were also found.

Antigens, Viral↗
PubMed2005

Motor neuron pathology in experimental autoimmune encephalomyelitis: studies in THY1-YFP transgenic mice.

Using adult male C57BL/6 mice that express a yellow fluorescent protein transgene in their motor neurons, we induced experimental autoimmune encephalomyelitis (EAE) by immunization with myelin oligodendrocyte glycoprotein peptide 35-55 (MOG peptide) in complete Freund's adjuvant (CFA). Control mice of the same transgenic strain received CFA without MOG peptide. Early in the course of their illness, the EAE mice showed lumbosacral spinal cord inflammation, demyelination and axonal fragmentation. By 14 weeks post-MOG peptide, these abnormalities were much less prominent, but the mice remained weak and, as in patients with progressive multiple sclerosis, spinal cord atrophy had developed. There was no significant loss of lumbar spinal cord motor neurons in the MOG peptide-EAE mice. However, early in the course of the illness, motor neuron dendrites were disrupted and motor neuron expression of hypophosphorylated neurofilament-H (hypoP-NF-H) immunoreactivity was diminished. By 14 weeks post-MOG peptide, hypoP-NF-H expression had returned to normal, but motor neuron dendritic abnormalities persisted and motor neuron perikaryal atrophy had appeared. We hypothesize that these motor neuron abnormalities contribute to weakness in this form of EAE and speculate that similar motor neuron abnormalities are present in patients with progressive multiple sclerosis.

Animals↗
PubMed2000

Functional interplay between nuclear factor-kappaB and c-Jun integrated by coactivator p300 determines the survival of nerve growth factor-dependent PC12 cells.

Nerve growth factor (NGF) activates the transcription factors nuclear factor kappaB (NF-kappaB) and activator protein-1 (AP-1) in sympathetic neurons. Whereas NGF-inducible NF-kappaB is required for the survival of neurons, c-Jun has the ability to promote neuronal death. In this report, we have examined the effect of NGF withdrawal on c-Jun and NF-kappaB transcription factors in PC12 cells differentiated to a neuronal phenotype. We show that the withdrawal of NGF from these cultures results in de novo synthesis of c-Jun, increase in AP-1 activity, and down-regulation of NF-kappaB activity. To investigate how the signal transduction pathways activating c-Jun and NF-kappaB are differentially regulated by NGF, we performed transcriptional analyses. Expression of ReIA (NF-kappaB) suppressed the c-Jun-dependent transcription of c-jun, and this effect was reversed by overexpression of the coactivator p300. RelA's effects on c-Jun transcription were mediated by competitive binding of the carboxy-terminal region of RelA to the CH1 domain of p300, which also binds to c-Jun; deletion of this region abrogated the ability of RelA to inhibit c-Jun activity. Furthermore, the inhibition of endogenous NF-kappaB in NGF-maintained neuronal PC12 cells led to the induction of c-Jun synthesis and a marked increase in cell death. Together, these studies demonstrate a functional interaction between NF-kappaB and c-Jun and suggest a novel mechanism of NF-kappaB-mediated neuroprotection.

Animals↗
PubMed2000

Androgen receptor gene CAG trinucleotide repeats in anovulatory infertility and polycystic ovaries.

Hyperandrogenism is currently thought to be central to the pathogenesis of polycystic ovarian syndrome (PCOS), a common endocrine disorder in premenopausal women characterized by irregular menstruation and anovulatory infertility. Although hyperandrogenism is characteristic, some women with PCOS have normal serum androgen levels. All androgens act through the X-linked androgen receptor (AR), the N-terminal domain of which contains a polyglutamine tract encoded by a highly polymorphic CAG trinucleotide repeat tract. Recently, variations in this CAG microsatellite tract, while remaining within the normal polymorphic range (11-38 CAGs), have been inversely correlated with receptor activity. Thus, short tracts are associated with high intrinsic AR activity and increased severity and earlier age of onset of the androgen-regulated tumor prostate cancer, whereas longer CAG tracts are associated with low AR activity and oligospermic infertility. To investigate the role of the CAG repeat tract in PCOS, we measured its length in 91 patients with ultrasound diagnosis of polycystic ovaries, irregular menstrual cycles, and anovulatory infertility and compared them to 112 control subjects of proven fertility with regular menses. Fluorescent-labeled DNA fragments containing the CAG repeat tract were amplified from leucocytic DNA, and their lengths were compared with internal size markers on an automated DNA Sequencer. There were no differences in the mean CAG length between patients and controls when both alleles were considered together or separately. Because there is a subset of PCOS patients whose serum androgens are normal, we compared differences in CAG length between patients whose serum testosterone (T) levels were below the normal laboratory mean, to those that were higher. There was a trend for a lower mean CAG biallelic length among anovulatory patients with T less than 1.73 nmol/L compared with those whose T was more than 1.73 nmol/L (22.47 +/- 0.36 vs. 23.25 +/- 0.29). This difference in CAG length between patients with low and high T levels (20.38 +/- 0.51 vs. 21.98 +/- 0.29) was highly significant (P = 0.004) when only the shorter allele of each individual was considered. Ethnic differences were also evident in our data; Indian subjects had a significantly shorter AR-CAG length compared with Chinese, being 22.08 +/- 0.50 and 23.16 +/- 0.17, respectively. Our data indicate an association between short CAG repeat length and the subset of anovulatory patients with low serum androgens, suggesting that the pathogenic mechanism of polycystic ovaries in these patients could be due to the increased intrinsic androgenic activity associated with short AR alleles.

Adult↗
PubMed1999

Phosphorylation by p44 MAP Kinase/ERK1 stimulates CBP histone acetyl transferase activity in vitro.

The transcriptional coactivator CBP displays an intrinsic histone acetyl transferase (HAT) activity which seems to participate in transcriptional activation through the destabilization of nucleosome structure. CBP is involved in the activity of several transcription factors that are nuclear endpoints of intracellular signal transduction pathways. In some instances, the transcription factors are phosphorylated upon cell activation, which induces their interaction with CBP. CBP itself is a phosphoprotein and can be phosphorylated by cycle-dependent kinases or by MAP kinases. Here we show that CBP phosphorylation by p44 MAP kinase/ERK1 results in the stimulation of its HAT enzymatic activity. The p44 MAP kinase/ERK1 phosphorylation sites are located in the C-terminal part of the protein, outside of the HAT domain. These sites are required for enzymatic stimulation, suggesting that phosphorylation by p44 MAP kinase/ERK1 induces a conformational change of the CBP molecule. Our data suggest that, in some instances, CBP itself might be a target for signal transduction pathways.

3T3 Cells↗
PubMed1999

Inflammatory bowel disease in horses: 11 cases (1988-1998).

OBJECTIVE: To determine clinical and surgical abnormalities in, and long-term outcome of, horses that undergo surgery because of colic secondary to inflammatory bowel disease (IBD). DESIGN: Retrospective study. ANIMALS: 11 horses. PROCEDURE: Medical records of horses that had undergone abdominal surgery and in which IBD had been diagnosed on the basis of histologic examination of intestinal biopsy specimens were reviewed. RESULTS: 5 horses were examined because of acute colic and 6 were examined because of chronic colic. At surgery, all 11 horses had edematous or hemorrhagic bowel segments suggestive of IBD. In addition, 6 horses had circumferential mural bands (CMB) causing constriction of the small (4 horses) or large (2) intestine. Intestinal resections were performed in 7 horses. All 11 horses survived surgery and were discharged from the hospital; 10 horses were still alive at the time of follow-up (1.5 to 7 years after surgery). CLINICAL IMPLICATIONS: Results suggest that IBD is an uncommon cause of colic in horses. Surgical resection of segments of intestine with constrictive CMB may relieve clinical signs of colic. Horses with IBD that had surgery had a good prognosis for long-term survival.

Acute Disease↗
PubMed1999

Ablation of the nephrosplenic space for treatment of recurring left dorsal displacement of the large colon in a racehorse.

Left dorsal displacement of the large colon was diagnosed in a 3-year-old Thoroughbred gelding. Conservative treatment that included lunging after administration of phenylephrine and rolling while under general anesthesia was not successful. The displaced segment of colon was reduced surgically through a ventral midline incision, but displacement recurred 4 days after surgery. Ablation of the nephrosplenic space by suturing the nephrosplenic ligament to the splenic capsule was performed through the left flank. At followup 1 year after surgery, the horse had not had further signs of colic and had returned to racing. Other methods to prevent large colon displacement, including colon resection and colopexy, are not recommended for athletic horses. Ablation of the nephrosplenic space, however, may be efficacious.

Animals↗
PubMed1999

Mitogenic up-regulation of the PRL-1 protein-tyrosine phosphatase gene by Egr-1. Egr-1 activation is an early event in liver regeneration.

The cellular signals that initiate cell growth are incompletely understood. Insight could be provided by understanding the signals regulating the transcriptional induction of immediate-early genes which occurs within minutes of the growth stimulus. The expression of the PRL-1 gene, which encodes a unique nuclear protein-tyrosine phosphatase, is rapidly induced in regenerating liver and mitogen-treated cells. Transcription of the PRL-1 gene increased in the rat liver remnant within a few minutes after partial hepatectomy and largely explained the increase in steady-state PRL-1 mRNA in the first few hours posthepatectomy. Egr-1 (early growth response factor) specifically bound a region of the proximal PRL-1 promoter P1 (-99). Egr-1 binding activity was more rapidly induced in regenerating liver than mitogen-treated H35 and NIH 3T3 cells, remained elevated through 4 h posthepatectomy, and appeared to be dependent not only on new Egr-1 protein synthesis but on post-translational regulation of Egr-1. Egr-1 efficiently transactivated a PRL-1 promoter reporter construct containing an intact not mutant Egr-1 site, and the Egr-1 site largely accounted for PRL-1 gene up-regulation in response to mitogen stimulation. These data predict that Egr-1 activation is an early event in liver regeneration and mitogen-activated cells that provides a regulatory stimulus for a subset of immediate-early genes.

3T3 Cells↗
PubMed1999

Oligomeric hydrolysable tannins from Tibouchina multiflora.

Two hydrolysable tannins, nobotanin O and nobotanin P, were isolated from the leaf extract of Tibouchina multiflora (Melastomataceae) and their dimeric and tetrameric structures elucidated on the basis of spectral data and chemical correlations with nobotanin B and K, respectively. Thirteen known hydrolysable tannins including nobotanins A, B, C and J, which are oligomers characteristic of the Melastomataceae, were also isolated.

Carbohydrate Sequence↗
PubMed1999

HIV-1 Tat-mediated activation of glycogen synthase kinase-3beta contributes to Tat-mediated neurotoxicity.

Human immunodeficiency virus type 1 (HIV-1) Tat induces neuronal apoptosis. To examine the mechanism(s) that contribute to this process, we studied Tat's effects on glycogen synthase kinase-3beta (GSK-3beta), an enzyme that has been implicated in the regulation of apoptosis. Addition of Tat to rat cerebellar granule neurons resulted in an increase in GSK-3beta activity, which was not associated with a change in protein expression and could be abolished by the addition of an inhibitor of GSK-3beta (lithium). Lithium also enhanced neuronal survival following exposure to Tat. Coprecipitation experiments revealed that Tat can associate with GSK-3beta, but direct addition of Tat to purified GSK-3beta had no effect on enzyme activity, suggesting that Tat's effects might be mediated indirectly. As the activation of platelet activating factor (PAF) receptors is critical for the induction of neuronal death by several candidate HIV-1 neurotoxins, we determined whether PAF can also activate GSK-3beta. Application of PAF to neuronal cultures activated GSK-3beta, and coincubation with lithium ameliorated PAF-induced neuronal apoptosis. These findings are consistent with the existence of one or more pathways that can lead to GSK-3beta activation in neurons, and they suggest that the dysregulation of this enzyme could contribute to HIV-induced neuronal apoptosis.

Animals↗
PubMed1999

Neuromuscular electrical stimulation and dynamic bracing as a treatment for upper-extremity spasticity in children with cerebral palsy.

We have investigated a therapeutic regimen using neuromuscular electrical stimulation (NMES) and dynamic bracing to assess their effectiveness in reducing upper-extremity spasticity in children with cerebral palsy. Nineteen patients between 4 and 21 years of age with documented diagnoses of spastic cerebral palsy were treated. The patients included in the study followed a regimen of two 30-minute sessions of NMES of the antagonist extensors combined with dynamic orthotic traction during the day. A static brace was used at night. Spasticity of the wrist and fingers was assessed periodically using the Zancolli classification. Treatment ranged from 3 to 43 months. After treatment with electrical stimulation and dynamic bracing, all the patients moved up 1 to 3 levels in the Zancolli classification and showed a marked improvement in upper-extremity function. These results show that combining NMES and dynamic orthotic traction dramatically decreases spasticity of the upper extremity in young patients with cerebral palsy.

Adolescent↗
PubMed1999

Can dialysis adequacy be achieved by tailoring the dialysis prescription in an Asian pediatric population on nightly intermittent peritoneal dialysis?

This study was undertaken to determine whether tailoring the dialysis prescription in Asian children on nightly intermittent peritoneal dialysis (NIPD), without adding high-dose therapy for cost-savings purposes, was able to achieve dialysis adequacy and improvement in nutrition. Eight children (age range: 5.5-20 years) on NIPD for a mean of 2.1 +/- 0.6 years, were studied at baseline and at 3 months and 9 months after their dialysis dose was tailored. Dialysis adequacy was measured by weekly Kt/Vurea and creatinine clearance (CCr). Fat-free mass (FFM) and percent body fat (%FAT) as measured by bioelectrical impedance, together with anthropometric measurements, serum total protein, and albumin, were used as indicators of nutrition. After the dialysis prescription was tailored, the mean weekly Kt/Vurea increased from 1.89 +/- 0.35 to 2.12 +/- 0.54 at 9 months, and total CCr increased from 36.4 +/- 11.51 L/1.73 m2 to 48.30 +/- 14.30 L/1.73 m2. The increase occurred despite a decline in residual renal function and was attributable to significant improvements in the peritoneal clearances of urea and creatinine (p < 0.05). The mid arm muscle circumference (MAMC) increased significantly (p = 0.006), while FFM increased from 25.68 +/- 7.92 kg to 26.95 +/- 9.83 kg, and %FAT decreased from 21.56% +/- 8.41% to 18.66% +/- 8.16%. The increase in FFM correlated significantly with a decrease in serum creatinine (r = -0.94, p = 0.005). In conclusion, tailoring the dialysis prescription in NIPD, without adding high-dose therapy, resulted in a trend of improvement in dialysis adequacy and some nutritional parameters.

Adolescent↗
PubMed1998

Histone acetyltransferase activity of CBP is controlled by cycle-dependent kinases and oncoprotein E1A.

Transforming viral proteins such as E1A force cells through the restriction point of the cell cycle into S phase by forming complexes with two cellular proteins: the retinoblastoma protein (Rb), a transcriptional co-repressor, and CBP/p300, a transcriptional co-activator. These two proteins locally influence chromatin structure: Rb recruits a histone deacetylase, whereas CBP is a histone acetyltransferase. Progression through the restriction point is triggered by phosphorylation of Rb, leading to disruption of Rb-associated repressive complexes and allowing the activation of S-phase genes. Here we show that CBP, like Rb, is controlled by phosphorylation at the G1/S boundary, increasing its histone acetyltransferase activity. This enzymatic activation is mimicked by E1A.

3T3 Cells↗
PubMed1998

Abnormal regulation of the Ke 6 gene, a new 17beta-hydroxysteroid dehydrogenase in the cpk mouse kidney.

The function encoded by the Ke 6 gene has been recently determined to be 17beta-hydroxysteroid dehydrogenase. Previously, the abnormal expression of the Ke 6 gene has been intimately associated with development of recessive polycystic kidney disease. The Ke 6 gene is normally expressed at very high levels in the kidney and liver and is severely down regulated in all recessive murine models of polycystic kidney disease that have been examined to date. Here, we report a detailed examination of the promoter region of the Ke 6 gene in normal mouse kidney cells (CTA) and in cells derived from mouse kidneys homozygous for the cpk (congenital polycystic kidney) mutation, using transfection analysis and DNA-protein gel shift assays. The minimal promoter region, P1 (+1 to -96), and a putative enhancer site, P3 (-165 to -256), within the Ke 6 gene 5' flanking sequence have been identified. We have also identified another region, P2 (-97 to -165), that may be responsible for the lower promoter activity of the Ke 6 gene in cpk cells. Furthermore, absence of binding of a 38 kDa nuclear protein to a 16 bp sequence element (P1A) within the minimal promoter of the Ke 6 gene suggests that the P1A element could be responsible for the overall reduction in promoter function in cpk cells.

17-Hydroxysteroid Dehydrogenases↗
PubMed1998

A rapid and sensitive assay for histone acetyl-transferase activity.

Histone acetyl-transferases (HATs) seem to be key elements in the regulation of transcription. We have designed an enzymatic assay to quantify HAT enzymatic activity. In this assay, the substrate is a peptide corresponding to the 24 first amino acids of histone H4 which is coupled to biotin. After acetylation using [14C]acetyl-CoA, the peptide is purified on streptavidin beads and the associated radioactivity is measured. This assay is sensitive, rapid and convenient.

3T3 Cells↗
PubMed1998

Renal medullary rim sign in 2 adult quarter horses.

This report describes a renal ultrasonographic abnormality (medullary rim sign), which was identified in 2 separate cases of spontaneously occurring disease associated with chronic and acute overdosage of phenylbutazone therapy. In horses, medullary rim sign has only been documented in neonatal foals experimentally administered large doses of phenylbutazone.

Animals↗
PubMed1997

The CREB-binding protein (CBP) cooperates with the serum response factor for transactivation of the c-fos serum response element.

The serum response element is one of the major promoter elements of the immediate early response to extracellular signals. The serum response element includes two main binding sites for proteins: the Ets box, which binds p62(TCF), and the CArG box, which binds p67(SRF). These two proteins are direct targets for signal transduction pathways; p62(TCF) is a nuclear end point of the Ras/mitogen-activated protein kinase pathway, and p67(SRF) is targeted by the Rho/Rac small G-proteins. The mechanism by which the signal is further transduced from the transcription factors to the basal transcriptional machinery is poorly understood. Recent data have suggested that the cAMP-responsive element-binding protein (CREB)-binding protein, a transcriptional adaptor involved in the transactivation through a wide variety of enhancer elements, participates in p62(TCF) activity. We here show that the CREB-binding protein also cooperates in the process of transactivation by p67(SRF). Cotransfections of expression vectors for the CREB-binding protein increased the expression, in response to serum, of reporters under the control of the c-fos serum response element. Interestingly, the C-terminal moiety of the CREB-binding protein was not necessary to observe this effect. The cooperation did not require the Ets box in the serum response element, and the CArG box was sufficient, indicating that the CREB-binding protein is able to cooperate with p67(SRF) in the absence of an Ets protein. Co-immunoprecipitation experiments using cell extracts showed that p67(SRF) could be retained with antibodies directed against the CREB-binding protein, suggesting that the two proteins form a multimolecular complex in live cells. The physical interaction between p67(SRF) and the CREB-binding protein was further confirmed by two-hybrid assays in mammalian cells. Our results indicate that the CREB-binding protein cooperates with p67(SRF) and, thus, suggest that the serum response element is regulated by a multimolecular complex, which includes the CREB-binding protein, p67(SRF), and p62(TCF), with multiple interactions between the components of the complex.

3T3 Cells↗