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C Gutierrez

Publications and source records attributed to C Gutierrez.

At least 19 recordsLinked to original sources
PubMed2006

Potential involvement of galectin-3 and SNAP23 in Aeromonas hydrophila cytotoxic enterotoxin-induced host cell apoptosis.

We investigated the potential of the cytotoxic enterotoxin (Act) of Aeromonas hydrophila to bind to 1869 human and 4319 yeast proteins, using protein microarray technology. Act was capable of binding nine different human proteins, including the SNARE complex scaffolding protein synaptosomal-associated protein 23 (SNAP23), galectin-3, and guanylate kinase 1 (GUK-1). Act was also able to bind to four of the yeast proteins examined, which included the vesicle tethering protein Vsp52. We verified interaction of Act with murine and human SNAP23, galectin-3, and GUK-1 by sandwich Western blot analysis. In order to determine the physiological relevance of Act binding to these three proteins, we performed small interfering RNA (siRNA) gene knockdown experiments in RAW 264.7 cells, a murine macrophage cell line in which Act-induced signaling and cell death is well characterized. Based on real-time reverse transcriptase-polymerase chain reaction, siRNA transfection of RAW 264.7 cells with specific oligonucleotides reduced the expression of genes encoding SNAP23, galectin-3, and GUK-1 by 62, 63, and 99%, respectively. Knockdown of galectin-3 and SNAP23, but not GUK-1, significantly reduced Act-induced apoptosis of host cells, as determined by TUNEL (TdT-mediated dUTP nick end labeling) assay, lactate dehydrogenase release, Giemsa staining, and reduction in activation of caspase 3, compared to toxin-treated macrophages that were transfected with a random sequence control siRNA. We also performed these assays using a human intestinal epithelial cell line (HT-29) and observed a similar trend of galectin-3 and SNAP23 association with Act-induced apoptosis. This is the first report of putative protein binding partners for this toxin and potential mediators/regulators of Act-induced apoptosis.

Aeromonas hydrophila↗
PubMed2006

Pulmonary toxicity associated with sirolimus treatment in kidney transplantation.

INTRODUCTION: An important side effect of sirolimus, a drug often used in organ transplantation, is pulmonary toxicity. MATERIALS AND METHODS: We present five kidney transplant patients who developed this toxicity associated with sirolimus. All underwent chest radiography computed tomography, fiberoptic bronchoscopy with bronchoalveolar lavage (BAL), microbiological studies of the bronchial aspirate, blood, and sputum, and cytomegalovirus (CMV) polymerase chain reaction (PCR) in blood as well as two had transbronchial biopsies. RESULTS: All five were men of mean age 54.8 +/- 10.3 years. In two sirolimus formed part of de novo therapy, and three were converted from calcineurin inhibitors. The mean treatment time was 16.6 +/- 13.7 months, with trough levels of 11.3 +/- 3 ng/mL. The patients presented with fever, cough, dyspnea, anemia, and dyslipidemia. The radiological pattern was diffuse alveolointerstitial (n = 2), or bilateral basal interstitial (n = 2), or bilateral basal alveolar (n = 1). The cell count in the BAL was 95% to 99% macrophages. In two patients cultures for bacteria were positive: Hemophilus and Pseudomonas. Tests for fungi, mycobacteria, pneumocystis, and legionella, as well as PCR for CMV were all negative. Transbronchial biopsy yielded insufficient material in one patient and a deposit of fibrinoid material and nonnecrotizing granuloma in the other. Antibacterial therapy was started, three with cotrimoxazole and two with ganciclovir, with no response. The respiratory symptoms improved after withdrawal of sirolimus (mean, 2.4 +/- 1.5 days). The mean hospital stay was 19.8 +/- 14.1 days. CONCLUSION: Pulmonary toxicity due to sirolimus should be included in the differential diagnosis of kidney transplant patients who display signs of interstitial pneumonia. Its diagnosis is difficult requiring exclusion of other pulmonary diseases. Resolution of the symptoms was quick after suspension of the drug.

Adult↗
PubMed2006

Posttransplant lymphoproliferative disease: a series of 23 cases.

Posttransplant lymphoproliferative disease (PTLD) is a rare but clinically important disorder due to its increasing incidence and its impact on renal function and the life of the patient. Between 1979 and 2005, this center performed 1614 kidney transplants, and 23 patients had PTLD. We undertook a retrospective study, analyzing risk factors, presentation, and evolution of the disorder. The most common clinical presentation was fever and adenopathy. All cases except one received calcineurin inhibitors, and nine were treated with monoclonal or polyclonal antibodies. Serology for Epstein Barr virus (EBV) was negative in nine patients at the time of transplant, and in five it became positive on diagnosis of PTLD. The predominant disorder was non-Hodgkin's lymphoma (NHL), either polymorphous (n = 11) or monomorphous (n = 7). The base therapy consisted of reducing or suspending calcineurin inhibitors and the addition of sirolimus and antivirals. Three patients received rituximab, and five chemotherapy. The disease progressed in 36% of the polymorphous NHL, in 67% of the monomorphous, and in 100% of the Hodgkin's lymphoma, whereas 10 patients had remission. Renal function worsened on diagnosis in eight patients, and the graft was infiltrated in five (confirmed histologically). Five patients lost the graft and 12 died; six due to infection and five due to PTLD. Survival was worse in the patients aged over 55 years. We conclude that in most cases EBV is positive on diagnosis of the PTLD, an age older than 55 years affords a poor prognosis, and lymphocyte infiltration of the graft is common, as is worsening renal function.

Humans↗
PubMed2006

Anterior uveitis associated with treatment with intravenous cidofovir in kidney transplant patients with BK virus nephropathy.

BACKGROUND: Polyomavirus-associated nephropathy (PVAN) is an important cause of kidney dysfunction and graft loss. Different treatment regimens have been used, including low-dose intravenous cidofovir. Anterior uveitis, a complication of this treatment, has been reported after its use in patients with cytomegalovirus-associated retinitis. We analyzed the incidence and associated risk factors for this disorder in patients with PVAN. PATIENTS AND METHODS: The study included 14 kidney-transplant patients who had received low doses of cidofovir after being diagnosed with PVAN. RESULTS: Five (35%) patients developed an episode of anterior uveitis. The mean number of cidofovir doses given was 6.8 +/- 1.6 as compared with 9.1 +/- 2.1 in patients who did not develop the disease. Creatinine clearance at diagnosis of the nephropathy and after terminating treatment was lower in the uveitis patients, who had a graft survival of 40% versus 100% in the patients who did not develop eye involvement. Treatment was suspended in the affected patients, with complete resolution in 80% after the administration of topical corticoids and cycloplegics. CONCLUSIONS: Anterior uveitis secondary to low-dose treatment with cidofovir is a common complication in patients with PVAN and is associated with the degree of renal involvement. In the absence of larger studies, cidofovir should be used with caution in patients with creatinine clearance below 30 mL/min.

Adult↗
PubMed2006

Vertebral osteoma in a dromedary camel.

A case of vertebral osteoma observed in a female 4-year-old dromedary camel is presented. The patient showed a progressive ataxia, incoordination and finally a permanent recumbence position. Treatment was unfruitful and the animal was humanely euthanized. Postmortem examination revealed an expansive growth at vertebral body of the 10th thoracic vertebra, which was histologically classified as an osteoma. To the author's knowledge, this would be the first description of an osteoma in a camelid species.

Animals↗
PubMed2006

Impact of lung transplant operation on bronchiolitis obliterans syndrome in patients with chronic obstructive pulmonary disease.

Previous studies suggest that bilateral (BLT) compared with single lung transplantation (SLT) for patients with chronic obstructive pulmonary disease (COPD) results in improved long-term survival. The effect of transplant operation on bronchiolitis obliterans syndrome (BOS) is unknown. A retrospective study of all lung transplant recipients with pre-transplant diagnoses of COPD at the University of Toronto and at Duke University was performed. Data collected were age, gender, date and type of transplant, acute rejection, survival, presence and time of BOS. 221 (bilateral n = 101, single n = 120) patients met our criteria. Patients with BLT were younger (53.0 vs. 55.3 years; p = 0.034), more likely to be male (56.3% vs. 42.4%; p = 0.039) and more likely to be transplanted at the University of Toronto (79.6% vs. 16.1%; p < 0.001). Freedom from BOS was similar at 1 year post-transplant. However, BLT recipients were more commonly free from BOS 3 years (57.4% vs. 50.7%) and 5 years (44.5% vs. 17.9%) post-transplant (p = 0.024). Survival of BLT was better than SLT recipients at 3 and 5 years post-transplant (BLT vs. SLT: 67.5% vs. 61.1% and 60.7% vs. 34.1%, respectively; p = 0.018). Similar trends on survival were observed after development of BOS. BLT results in lower rates of BOS in patients with COPD that are eligible for both SLT and BLT.

Bronchiolitis Obliterans↗
PubMed2006

Pre-implantation multiple cytokine mRNA expression analysis of donor lung grafts predicts survival after lung transplantation in humans.

While current donor selection with clinical findings is generally effective, the imprecise nature of the assessment forces clinicians to remain on the conservative side. A reliable biological marker would assist donor selection and would improve donor organ utilization. We collected biopsies from 169 donor lungs before implantation. Expression levels of IL-6, IL-8, IL-10, TNF-alpha, IFN-gamma and IL-1beta were measured by quantitative real-time RT-PCR (qRT-PCR). Seventeen cases died within 30 days after transplantation. No donor factor was significantly associated with 30-day mortality. Univariate analysis of the 84 cases for development of the prediction model showed that IL-6, IL-8, TNF-alpha and IL-1beta were risk factors for mortality and IL-10 and IFN-gamma were protective factors. We analyzed the cytokine expression ratios of risk to protective cytokines. A stepwise logistic regression for 30-day mortality demonstrated that a model containing the ratio of IL-6/IL-10 was the most predictive (p = 0.0013). When applied to the remaining 85 cases for validation, the test of model fit was significant (p = 0.039). Using the cytokine ratio, we were able to define three risk groups with striking differences in survival (p = 0.0003). Multi-cytokine analysis of the donor lung graft with qRT-PCR shows significant promise as a strategy to biologically evaluate the donor lung prior to implantation.

Biopsy↗
PubMed2006

The effect of reflux and bile acid aspiration on the lung allograft and its surfactant and innate immunity molecules SP-A and SP-D.

Gastro-esophageal reflux and related pulmonary bile acid aspiration were prospectively investigated as possible contributors to postlung transplant bronchiolitis obliterans syndrome (BOS). We also studied the impact of aspiration on pulmonary surfactant collectin proteins SP-A and SP-D and on surfactant phospholipids--all important components of innate immunity in the lung. Proximal and distal esophageal 24-h pH testing and broncho-alveolar lavage fluid (BALF) bile acid assays were performed prospectively at 3-month posttransplant in 50 patients. BALF was also assayed for SP-A, SP-D and phospholipids expressed as ratio to total lipids: phosphatidylcholine; dipalmitoylphosphatidylcholine; phosphatidylglycerol (PG); phosphatidylinositol; sphingomyelin (SM) and lysophosphatidylcholine. Actuarial freedom from BOS was assessed. Freedom from BOS was reduced in patients with abnormal (proximal and/or distal) esophageal pH findings or BALF bile acids (Log-rank Mantel-Cox p < 0.05). Abnormal pH findings were observed in 72% (8 of 11) of patients with bile acids detected within the BALF. BALF with high levels of bile acids also had significantly lower SP-A, SP-D, dipalmitoylphosphatidylcholine; PG and higher SM levels (Mann-Whitney, p < 0.05). Duodeno-gastro-esophageal reflux and consequent aspiration is a risk factor for the development of BOS postlung transplant. Bile acid aspiration is associated with impaired lung allograft innate immunity manifest by reduced surfactant collectins and altered phospholipids.

Bile Acids and Salts↗
PubMed2005

An outbreak of abortions and high neonatal mortality associated with Trypanosoma evansi infection in dromedary camels in the Canary Islands.

Trypanosoma evansi was diagnosed for the first time in the Canary Islands (Spain) in 1998 in a dromedary camel. Seroprevalences of 4.8% up to 9% have been observed using different diagnostic methods. Affected animals have been treated but the dissemination of the disease is unknown. This article presents an outbreak of abortions and high neonatal mortality attributable to T. evansi infection in camels as well as the clinical assessment of the affected animals. The patients were diagnosed by routine checking (three pregnant animals), after abortion (five dams), or after delivered premature or weak calves (eight dams). At clinical examination, 2 out of 16 affected animals showed moderate signs of chronic form, particularly hyporexia and intolerance to exercise. The aborted fetuses were aged 6-8 months of gestation, approximately. The main laboratorial findings were regenerative anemia (haemolytic anemia), lymphocytic and monocytic leukocytosis, hyperproteinemia, hyperglobulinemia, hypoglycaemia, serum urea increased and serum iron decreased. Treatment using trypanocidal drug (Cymelarsan) resulted highly effective. Massive treatment would be recommended in the entire camel population in the Canary Islands (less than 2000 animals), as therapeutic or preventive measure, in order to control and to achieve an eventual eradication of the disease.

Aborted Fetus↗
PubMed2005

Congenital duplication of the caudal region (monocephalus dipygus) in a kid goat.

A case of congenital duplication (monocephalus dipygus) in a goat is described. Two pelvis and four pelvic limbs were observed in the kid. Conjoined or fused symmetric twins were diagnosed. Associated abnormalities were cleft palate and anal atresia. Most of the classically recognized teratogens were ruled out by history and serology. However, progenitors were related in the second degree. Thus, genetic factors could be suspected in this case. To the authors' best knowledge, this is the first report of monocephalus dipygus in a goat.

Animals↗
PubMed2005

Microarray and proteomics analyses of human intestinal epithelial cells treated with the Aeromonas hydrophila cytotoxic enterotoxin.

We performed microarray analyses on RNA from human intestinal epithelial (HT-29) cells treated with the cytotoxic enterotoxin (Act) of Aeromonas hydrophila to examine global cellular transcriptional responses. Based on three independent experiments, Act upregulated the expression of 34 genes involved in cell growth, adhesion, signaling, immune responses (including interleukin-8 [IL-8] production), and apoptosis. We verified the upregulation of 14 genes by real-time reverse transcriptase-PCR and confirmed Act-induced production of IL-8 by enzyme-linked immunosorbent assay on supernatants from nonpolarized and polarized HT-29 cells. Maximal production of IL-8 in response to Act required the presence of intracellular calcium, since chelation of calcium with BAPTA-AM significantly reduced Act-induced IL-8 production in HT-29 cells. We also examined activation of mitogen-activated protein kinases and, as demonstrated by Western blot analysis of apical side-treated polarized HT-29 cells, Act induced phosphorylation of p38, c-Jun NH(2)-terminal kinase, and extracellular signal-regulated kinase 1/2. In addition, KinetWorks proteomics screening of whole-cell lysates revealed Act-induced phosphorylation of cyclic AMP-response element binding protein (CREB), c-Jun, adducin, protein kinase C, and signal transducer and activator of transcription 3 (STAT3) and decreased phosphorylation of protein kinase Balpha, v-raf-1 murine leukemia viral oncogene homolog 1 (i.e., Raf1), and STAT1. We verified activation of CREB and activator protein 1 in polarized cells by gel shift assay. This is the first description of human intestinal epithelial cell transcriptional alterations, phosphorylation or activation of signaling molecules, cytokine production, and calcium mobilization in response to this toxin.

Aeromonas hydrophila↗
PubMed2003

Validation of egg yolk antibody testing as a method to determine influenza status in white leghorn hens.

Determination of the avian influenza (AI) status of a flock has traditionally been done by detection of serum antibodies. However, for many diseases, detection of antibodies in egg yolk has been effective in monitoring the disease status of laying flocks. This study compared the utility of egg yolk vs. serum for determining AI status in laying hen flocks. Specific-pathogen-free white leghorn hens were inoculated via the respiratory tract with a low-pathogenic H7N2 AI virus or sterile allantoic fluid or subcutaneously with an inactivated oil emulsion vaccine produced from the same AI virus or normal allantoic fluid. Antibody levels were determined by the agar gel immunodiffusion (AGID) test, the hemagglutination-inhibition (HI) test, and the enzyme-linked immunosorbent assay (ELISA). Anti-influenza antibodies were detected in sera of all live virus-inoculated hens by day 7 postinoculation (PI) (AGID and ELISA tests), but detection of antibodies in egg yolk was delayed by a few days, with all being positive by day 14 PI. Sera from all vaccinated hens were positive by day 14 PI (AGID and ELISA tests), and egg yolk was positive by day 18 PI. The HI test was less sensitive than the ELISA and AGID tests in detecting anti-influenza antibodies in both sera and yolk. Serum and yolk from all control birds remained negative throughout the study. These studies show that currently used serologic tests can detect antibodies in serum and yolk samples from hens exposed to live AI virus or from those that have been vaccinated. Antibody is detected earlier in the serum than in the yolk and antibody is detected earlier from birds exposed to a live infection compared to birds vaccinated with an inactivated oil emulsion vaccine.

Animals↗
PubMed2002

Pharmacologic restoration of the early insulin response in pre-diabetic monkeys controls mealtime glucose excursions without peripheral hyperinsulinaemia.

AIMS/HYPOTHESIS: This study sought first to compare the pharmacodynamics and pharmocokinetics of two rapid-onset, rapidly-reversible insulinotropic agents, nateglinide and repaglinide, in pre-diabetic Cynomolgus monkeys and second to use these agents to assess the metabolic effects of early insulin secretion on prandial glucose control. METHODS: First, equipotent doses of nateglinide (20 mg/kg) and repaglinide (0.1 mg/kg) or vehicle were given intragastrically to overnight-fasted ketamine-anesthetized pre-diabetic Cynomolgus monkeys and samples were obtained for measurement of plasma glucose, insulin, glucagon, NEFA and drug concentrations. Second, nateglinide, repaglinide or vehicle were administered 10 min before a glucose-supplemented liquid meal and prandial glucose and insulin profiles were compared. RESULTS: Although oral administration of nateglinide and repaglinide elicited similar maximum increments of plasma insulin (+403 and +448 pmol/l, respectively), the effects of nateglinide were more rapidly manifest and less prolonged. With nateglinide, insulin increased within 10 min and returned to baseline within 50 min. After repaglinide, the first increase occurred at 30 min and insulin concentrations remained increased for 3.5 h post-dose. When given 10 min before a meal, nateglinide increased early, but not total insulin release (AUC(0-210)=108 vs 150 nmol/l min for nateglinide and vehicle, respectively) and reduced prandial glucose excursions by 78%. Repaglinide increased total insulin release (AUC(0-210)=298 nmol/l min) and reduced glucose excursions by 53%. CONCLUSION/INTERPRETATION: Nateglinide is more rapid-acting and rapidly-reversible than is repaglinide. By restoring a more physiologic insulin profile, nateglinide is more effective than repaglinide in controlling prandial glucose excursions with less hyperinsulinaemia.

Animals↗
PubMed2002

Transient repressor effect of Fis on the growth phase-regulated osmE promoter of Escherichia coli K12.

Transcription of the gene osmE of Escherichia coli is osmotically inducible and regulated by the growth phase. Expression of osmE is directed by a single promoter, osmE (p), which is recognized by Esigma(70) and Esigma(s), two forms of RNA polymerase using, respectively, the sigma factors sigma(70) and sigma(s). Esigma(s) transcribes osmE (p) during entry into stationary phase. Esigma(70) is responsible for osmotic induction of osmE (p) during the exponential growth phase. In a search for proteins that can modulate osmE (p) expression in trans, we performed electrophoretic mobility shift experiments using a DNA fragment carrying osmE (p) and crude extracts from E. coli. One major retarded band was observed in these experiments. The Fis protein is responsible for this retarded band, and binds to several sites upstream and downstream of, and overlapping, the promoter region of osmE. In a fis mutant background, the kinetics of in vivo transcription of osmE (p) during growth demonstrated that Fis is not responsible for the repression of the promoter seen during early exponential phase. In contrast, expression of osmE (p) at elevated osmolarity during the mid-exponential growth phase is increased in the absence of Fis, demonstrating that Fis is able to act as a repressor in vivo at a particular stage of growth.

Bacterial Proteins↗
PubMed2002

A new evolutionary scenario for the Mycobacterium tuberculosis complex.

The distribution of 20 variable regions resulting from insertion-deletion events in the genomes of the tubercle bacilli has been evaluated in a total of 100 strains of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canettii, Mycobacterium microti, and Mycobacterium bovis. This approach showed that the majority of these polymorphisms did not occur independently in the different strains of the M. tuberculosis complex but, rather, resulted from ancient, irreversible genetic events in common progenitor strains. Based on the presence or absence of an M. tuberculosis specific deletion (TbD1), M. tuberculosis strains can be divided into ancestral and "modern" strains, the latter comprising representatives of major epidemics like the Beijing, Haarlem, and African M. tuberculosis clusters. Furthermore, successive loss of DNA, reflected by region of difference 9 and other subsequent deletions, was identified for an evolutionary lineage represented by M. africanum, M. microti, and M. bovis that diverged from the progenitor of the present M. tuberculosis strains before TbD1 occurred. These findings contradict the often-presented hypothesis that M. tuberculosis, the etiological agent of human tuberculosis evolved from M. bovis, the agent of bovine disease. M. canettii and ancestral M. tuberculosis strains lack none of these deleted regions, and, therefore, seem to be direct descendants of tubercle bacilli that existed before the M. africanum-->M. bovis lineage separated from the M. tuberculosis lineage. This observation suggests that the common ancestor of the tubercle bacilli resembled M. tuberculosis or M. canettii and could well have been a human pathogen already.

Base Sequence↗