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Biomedical subjects

R Urrutia

Publications and source records attributed to R Urrutia.

At least 19 recordsLinked to original sources
PubMed2005

Environmental evidence of fossil fuel pollution in Laguna Chica de San Pedro lake sediments (Central Chile).

This paper describes lake sediment spheroidal carbonaceous particle (SCP) profiles from Laguna Chica San Pedro, located in the Biobío Region, Chile (36 degrees 51' S, 73 degrees 05' W). The earliest presence of SCPs was found at 16 cm depth, corresponding to the 1915-1937 period, at the very onset of industrial activities in the study area. No SCPs were found at lower depths. SCP concentrations in Laguna Chica San Pedro lake sediments were directly related to local industrial activities. Moreover, no SCPs were found in Galletué lake (38 degrees 41' S, 71 degrees 17.5' W), a pristine high mountain water body used here as a reference site, suggesting that contribution from long distance atmospheric transport could be neglected, unlike published data from remote Northern Hemisphere lakes. These results are the first SCP sediment profiles from Chile, showing a direct relationship with fossil fuel consumption in the region. Cores were dated using the 210Pb technique.

Air Pollutants↗
PubMed2005

Effects of pulp and paper mill effluents on the microplankton and microbial self-purification capabilities of the Biobío River, Chile.

Most studies focus on the ecotoxicity of pulp and paper mill effluents, rather than on how they affect the physicochemical and biological structure and the intrinsic ecological capabilities of the receiving watercourses. We investigated the impact of such effluents on the water quality, microplankton system and microbial self-purification capacity (degradation of polymeric organic compounds via extracellular enzymes) of the Biobío River in Chile. The physicochemical impact on the water quality was indicated by raised conductivity, by the pollution of the water body with nitrate, nitrite and soluble reactive phosphorus, by the appearance of tannin and lignin, and by the steady accumulation of inorganic and organic suspended matter (SPM) along the river. From the biological structure of the microplankton system, very low and declining concentrations of chlorophyll a and heterotrophic flagellate densities were determined. The pulp and paper mill effluents introduced high bacterial abundances and biomass concentrations into the river water. This reflects the effective use made of the abundantly available inorganic and organic nutrients within this industrial and municipal process water by bacteria adapted to these extreme environments, additionally supported by concomitant low grazing pressure derivable from low heterotrophic flagellate abundances. Indeed, in one section of the river affected by a pulp mill, the plant was found to significantly contribute to the self-cleaning capacity of the river. However, this elevated degradation capacity was not enough to compensate for the additionally discharged organic material which, together with the toxic effects of the paper plant effluents, significantly interferes with the ecological status of the Biobío River.

Animals↗
PubMed2005

Use of Daphnia spp. for the ecotoxicological assessment of water quality in an agricultural watershed in South-Central Chile.

Because of the importance of surface waters from the Chillán River watershed (Chile) for recreation, agricultural irrigation, and the production of drinking water, local concern about river water quality has increased considerably during the last decade. Agricultural and forestry activities in the watershed, characterized by an intensive use of pesticides, are thought to play an important role in the generation of non-point-source pollution, whereas the discharge of urban wastewater from the city of Chillán constitutes a major point source of pollution. In the present investigation, acute and chronic laboratory bioassays using Daphnia spp. were conducted on surface water samples from 17 river stations located throughout the watershed. Sampling occurred on 6 occasions during a 16-month period (2000 to 2001) and included both high and low flow conditions. Almost all toxic effects observed in summer were directly related to the discharge of urban wastewater, whereas toxicity in rural areas was mainly detected during the winter period when rainfall and river flow are high. Toxicity test results were compared with measured physicochemical water-quality data. Mortality and alterations in reproductive success of Daphnia spp. were not consistently reflected in detected chemical pollution. With only one exception (atrazine), detected pesticide concentrations were below known toxicity levels. However, additive and synergistic effects of the presence of a mixture of pesticides could not be excluded as a possible cause of observed toxicity. At several stations, filtering of the water sample led to a strong decrease in toxicity, which suggests the presence of xenobiotics attached to the smaller sediment fraction. Inclusion of sediment chemical analysis and sediment toxicity testing in future work should therefore be encouraged. The presented approach provided information about the adverse effects of human activities on surface water quality in the watershed, not easily obtained from classical monitoring schemes. In specific cases, the approach may represent an economically attractive alternative to physicochemical analyses. Modifications to the proposed methodology should be introduced if the effects of intrastorm and interstorm variability of water quality are to be analyzed.

Agriculture↗
PubMed2004

Association of distinct alpha(2) adrenoceptor and serotonin transporter polymorphisms with constipation and somatic symptoms in functional gastrointestinal disorders.

BACKGROUND: The role of genetics in the phenotypic manifestations of irritable bowel syndrome (IBS) is unclear. Our aims were: (1) to compare the prevalence of polymorphisms of alpha 2 (alpha(2)) adrenoceptors, norepinephrine transporter, and serotonin transporter protein (soluble carrier protein member 4 (SLC6A4)) promoter in patients with lower functional gastrointestinal disorders (FGID) and in healthy controls; and (2) to test associations of these genetic variations with symptoms of IBS and high somatic symptom scores. METHODS: Validated bowel and somatic symptom questionnaires characterised the phenotype: 90 with IBS constipation (IBS-C), 128 IBS diarrhoea, 38 IBS alternating bowel function, and 20 chronic abdominal pain. Logistic regression analyses assessed associations of different polymorphisms for alpha(2) adrenoceptor and SLC6A4 with IBS or chronic abdominal pain phenotypes and high somatic score. RESULTS: Two distinct polymorphisms independently appeared to be associated with the phenotype IBS-C: alpha(2C) Del 322-325 (odds ratio (OR) 2.48 (95% confidence interval (CI) 0.98, 6.28); p = 0.05) and alpha(2A) -1291 (C-->G) (OR 1.66 (95% CI 0.94, 2.92); p = 0.08) relative to wild-type. Overall, the alpha(2C) Del 322-325 polymorphism (alone or combined with other polymorphisms) was also significantly associated with a high somatic symptom score (OR 2.2 (95% CI 1.06, 4.64); p = 0.03). Combinations of polymorphisms were also associated with high somatic scores. CONCLUSION: Functionally distinct alpha(2A) and alpha(2C) adrenoceptor and serotonin transporter polymorphisms are associated with constipation and high somatic symptoms in patients with lower functional gastrointestinal disorders, although the strength of the genetic contribution to the phenotype is unclear.

Adolescent↗
PubMed2001

The Sp1-like protein BTEB3 inhibits transcription via the basic transcription element box by interacting with mSin3A and HDAC-1 co-repressors and competing with Sp1.

Sp1-like proteins are characterized by three conserved C-terminal zinc finger motifs that bind GC-rich sequences found in promoters of numerous genes essential for mammalian cell homeostasis. These proteins behave as transcriptional activators or repressors. Although significant information has been reported on the molecular mechanisms by which Sp1-like activators function, relatively little is known about mechanisms for repressor proteins. Here we report the functional characterization of BTEB3, a ubiquitously expressed Sp1-like transcriptional repressor. GAL4 assays show that the N terminus of BTEB3 contains regions that can act as direct repressor domains. Immunoprecipitation assays reveal that BTEB3 interacts with the co-repressor mSin3A and the histone deacetylase protein HDAC-1. Gel shift assays demonstrate that BTEB3 specifically binds the BTE site, a well characterized GC-rich DNA element, with an affinity similar to that of Sp1. Reporter and gel shift assays in Chinese hamster ovary cells show that BTEB3 can also mediate repression by competing with Sp1 for BTE binding. Thus, the characterization of this protein expands the repertoire of BTEB-like members of the Sp1 family involved in transcriptional repression. Furthermore, our results suggest a mechanism of repression for BTEB3 involving direct repression by the N terminus via interaction with mSin3A and HDAC-1 and competition with Sp1 via the DNA-binding domain.

Amino Acid Motifs↗
PubMed2001

First report on chlorinated pesticide deposition in a sediment core from a small lake in central Chile.

This paper presents a first report on chlorinated pesticide deposition analyzed through sedimentary records in a small mesotrophic lake (Chica de San Pedro) in central Chile. The sediment core was sliced and dated using 210Pb, 137Cs and pollen analyses. Organochlorine pesticides were analyzed by gas chromatography with electron capture detection (GC-ECD). From these results, pesticide deposition over the last 50 years was estimated. No pesticides were detected below the 1940 slice of the core. Concentrations were in the range 0.640-1.4 ng/g d.w. for total DDTs, 0.046-0.362 ng/g d.w. for lindane and 0.015-0.310 ng/g d.w. for alpha-hexachlorohexane. Highest concentrations of pp'DDT were found in 1993-1996 and higher concentrations of pp'-DDE and pp'-DDD were found in the seventies (1972-1978). Total organic carbon (TOC) normalized data were used for statistical analysis. Although significant correlation was observed between concentrations of DDE and DDD, no correlation was found for DDT, suggesting that it had a different source. Factorial analysis grouped DDE together with DDD, while DDT was grouped together with gamma- and alpha-HCH. Total DDT fluxes were highest during the 1970s, while those for HCHs have been increasing in the 1990s. In Chile, organochlorine compounds were banned in 1985, and the historical deposition patterns seem to indicate that such measures have been effective. On the other hand, results point out a relatively new occurrence of pp'-DDT in the watershed, but the source remains unknown.

Chile↗
PubMed2001

TGFbeta-mediated signaling and transcriptional regulation in pancreatic development and cancer.

Transforming growth factor-beta (TGFbeta) plays a critical role in pancreatic development and cell proliferation. Binding of TGFbeta to its membrane receptor kinases activates the Smad signaling proteins, allowing them to translocate to the nucleus and participate in the transcriptional control of TGFbeta target genes. In addition, there is an increasing number of cellular mechanisms affecting the final response of a cell to TGFbeta. This includes crosstalk with other signaling pathways and the induction of TGFbeta early response genes, such as the TGFbeta-inducible early response gene (TIEG) family of transcription factors. Like the Smads, TIEGs behave as downstream effector proteins in TGFbeta-mediated pancreatic growth control. The discovery of the Smads and TIEGs has provided new insights into TGFbeta-regulated functions. Their significance in pancreatic development and cancer is discussed in this review.

Journal Article↗
PubMed2001

A conserved alpha-helical motif mediates the interaction of Sp1-like transcriptional repressors with the corepressor mSin3A.

Sp1-like proteins are defined by three highly homologous C(2)H(2) zinc finger motifs that bind GC-rich sequences found in the promoters of a large number of genes essential for mammalian cell homeostasis. Here we report that TIEG2, a transforming growth factor beta-inducible Sp1-like protein with antiproliferative functions, represses transcription through recruitment of the mSin3A-histone deacetylase complex. The interaction of TIEG2 with mSin3A is mediated by an alpha-helical repression motif (alpha-HRM) located within the repression domain (R1) of TIEG2. This alpha-HRM specifically associates with the second paired amphipathic helix (PAH2) domain of mSin3A. Mutations in the TIEG2 alpha-HRM domain that disrupt its helical structure abolish its ability to both bind mSin3A and repress transcription. Interestingly, the alpha-HRM is conserved in both the TIEG (TIEG1 and TIEG2) and BTEB (BTEB1, BTEB3, and BTEB4) subfamilies of Sp1-like proteins. The alpha-HRM from these proteins also mediates direct interaction with mSin3A and represses transcription. Surprisingly, we found that the alpha-HRM of the Sp1-like proteins characterized here exhibits structural and functional resemblance to the Sin3A-interacting domain previously described for the basic helix-loop-helix protein Mad1. Thus, our study defines a mechanism of transcriptional repression via the interactions of the alpha-HRM with the Sin3-histone deacetylase complex that is utilized by at least five Sp1-like transcriptional factors. More importantly, we demonstrate that a helical repression motif which mediates Sin3 interaction is not an exclusive structural and functional characteristic of the Mad1 subfamily but rather has a wider functional impact on transcriptional repression than previously demonstrated.

Amino Acid Motifs↗
PubMed2001

Sequence-specific transcriptional repression by KS1, a multiple-zinc-finger-Krüppel-associated box protein.

The vertebrate genome contains a large number of Krüppel-associated box-zinc finger genes that encode 10 or more C(2)-H(2) zinc finger motifs. Members of this gene family have been proposed to function as transcription factors by binding DNA through their zinc finger region and repressing gene expression via the KRAB domain. To date, however, no Krüppel-associated box-zinc finger protein (KRAB-ZFP) and few proteins with 10 or more zinc finger motifs have been shown to bind DNA in a sequence-specific manner. Our laboratory has recently identified KS1, a member of the KRAB-ZFP family that contains 10 different C(2)-H(2) zinc finger motifs, 9 clustered at the C terminus with an additional zinc finger separated by a short linker region. In this study, we used a random oligonucleotide binding assay to identify a 27-bp KS1 binding element (KBE). Reporter assays demonstrate that KS1 represses the expression of promoters containing this DNA sequence. Deletion and site-directed mutagenesis reveal that KS1 requires nine C-terminal zinc fingers and the KRAB domain for transcriptional repression through the KBE site, whereas the isolated zinc finger and linker region are dispensable for this function. Additional biochemical assays demonstrate that the KS1 KRAB domain interacts with the KAP-1 corepressor, and mutations that abolish this interaction alleviate KS1-mediated transcriptional repression. Thus, this study provides the first direct evidence that a KRAB-ZFP binds DNA to regulate gene expression and provides insight into the mechanisms used by multiple-zinc-finger proteins to recognize DNA sequences.

Animals↗
PubMed2000

Rho GTPases mediate the regulation of cochlear outer hair cell motility by acetylcholine.

Outer hair cells are the mechanical effectors of the cochlear amplifier, an active process that improves the sensitivity and frequency discrimination of the mammalian ear. In vivo, the gain of the cochlear amplifier is regulated by the efferent neurotransmitter acetylcholine through the modulation of outer hair cell motility. Little is known, however, regarding the molecular mechanisms activated by acetylcholine. In this study, intracellular signaling pathways involving the small GTPases RhoA, Rac1, and Cdc42 have been identified as regulators of outer hair cell motility. Changes in cell length (slow motility) and in the amplitude of electrically induced movement (fast motility) were measured in isolated outer hair cells patch clamped in whole-cell mode, internally perfused through the patch pipette with different inhibitors and activators of these small GTPases while being externally stimulated with acetylcholine. We found that acetylcholine induces outer hair cell shortening and a simultaneous increase in the amplitude of fast motility through Rac1 and Cdc42 activation. In contrast, a RhoA- and Rac1-mediated signaling pathway induces outer hair cell elongation and decreases fast motility amplitude. These two opposing processes provide the basis for a regulatory mechanism of outer hair cell motility.

Acetylcholine↗
PubMed2000

TIEG proteins join the Smads as TGF-beta-regulated transcription factors that control pancreatic cell growth.

The control of epithelial cell proliferation, differentiation, and apoptosis requires a balance between signaling and transcriptional regulation. Recent developments in pancreatic cell research have revealed that transforming growth factor-beta (TGF-beta) signaling is important for the regulation of each of these phenomena. More importantly, perturbations in this pathway are associated with pancreatic cancer. A chief example of these alterations is the mutation in the TGF-beta-regulated transcription factor Smad4/DPC4 that is found in a large percentage of pancreatic tumors. Surprisingly, studies on transcription factors have remained an underrepresented area of pancreatic research. However, the discovery of Smad4/DPC4 as a transcription factor fueled further studies aimed at characterizing transcription factors involved in normal and neoplastic pancreatic cell growth. Our laboratory recently described the existence of a novel family of zinc finger transcription factors, TGF-beta-inducible early-response gene (TIEG)1 and TIEG2, from the exocrine pancreas that, similarly to Smads, participate in the TGF-beta response and inhibit epithelial cell proliferation. This review therefore focuses on describing the structure and function of these two families of transcription factor proteins that are becoming key players in the regulation of pancreatic cell growth.

Animals↗
PubMed1999

Three conserved transcriptional repressor domains are a defining feature of the TIEG subfamily of Sp1-like zinc finger proteins.

Sp1-like transcription factors are characterized by three highly homologous C-terminal zinc finger motifs that bind GC-rich sequences. These proteins behave as either activators or repressors and have begun to be classified into different subfamilies based upon the presence of conserved motifs outside the zinc finger domain. This classification predicts that different Sp1-like subfamilies share certain functional properties. TIEG1 and TIEG2 constitute a new subfamily of transforming growth factor-beta-inducible Sp1-like proteins whose zinc finger motifs also bind GC-rich sequences. However, regions outside of the DNA-binding domain that differ in structure from other Sp1-like family members remain poorly characterized. Here, we have used extensive mutagenesis and GAL4-based transcriptional assays to identify three repression domains within TIEG1 and TIEG2 that we call R1, R2, and R3. R1 is 10 amino acids, R2 is 12 amino acids, and R3 is approximately 80 amino acids long. None of these domains share homology with previously described transcriptional regulatory motifs, but they share strong sequence homology and are functionally conserved between TIEG1 and TIEG2. Together, these data demonstrate that TIEG proteins are capable of repressing transcription, define domains critical for this function, and further support the idea that different subfamilies of Sp1-like proteins have evolved to mediate distinct transcriptional functions.

Amino Acid Sequence↗
PubMed1999

Sp1 and its likes: biochemical and functional predictions for a growing family of zinc finger transcription factors.

The discovery and functional characterization of Sp1 as a GC-rich binding zinc finger protein provided a useful paradigm for understanding mechanisms mediating transcriptional activation in eukaryotic cells. This early paradigm suggested that promoters carrying GC-rich sequences are activated by Sp1 through its interaction with proteins from the basal transcriptional machinery to upregulate gene expression. Since the time of this seminal work, studies from several laboratories have led to the discovery of many Sp1-like transcription factors containing highly homologous DNA binding motifs that bind to similar sequences. Consequently, this knowledge poses many important questions regarding whether these related proteins have similar or antagonistic biochemical and functional properties to Sp1. The goal of this article is to use available database information and recent experimental evidence to describe the current repertoire of Sp1-like zinc finger transcription factors in mammalian cells. Furthermore, we discuss structural and functional studies that reveal that these proteins may share a role in morphogenetic pathways. Altogether, this information is aimed at better understanding how this growing family of transcription factors work to regulate gene expression and morphogenesis.

Amino Acid Sequence↗
PubMed1999

The transforming growth factor beta(1)-inducible transcription factor TIEG1, mediates apoptosis through oxidative stress.

Transforming growth factor beta(1) (TGF-beta(1))-inducible transcription factors have recently elicited interest because of their critical role in the regulation of cell proliferation, differentiation, and apoptosis. We have previously reported that the TGF-beta(1)-inducible transcription factor, TIEG1, induces apoptosis in a pancreas-derived cell line. However, the mechanisms underlying the apoptotic effects of this transcription factor remain to be defined. In this study, using the TGF-beta(1)-sensitive Hep 3B cell line, we have defined the mechanistic sequence of events that characterize TIEG1-mediated apoptosis and compared these events with the changes observed during TGF-beta(1)-induced apoptosis. Both TGF-beta(1)- and TIEG1-induced cell death were accompanied by an increase in the generation of reactive oxygen species and a loss of the mitochondrial membrane potential preceding the morphological changes of apoptosis. In contrast, increases in caspase 3-like activity and glutathione (GSH) depletion occurred later in the apoptotic process, concurrent with the morphological features of apoptosis. The antioxidant, trolox, decreased the formation of reactive oxygen species and apoptosis. These results demonstrate that similar to TGF-beta(1), TIEG1 induces apoptosis by a mechanism involving the formation of reactive oxygen species.

Antioxidants↗
PubMed1998

KRAB-independent suppression of neoplastic cell growth by the novel zinc finger transcription factor KS1.

The study of zinc finger proteins has revealed their potential to act as oncogenes or tumor suppressors. Here we report the molecular, biochemical, and functional characterization of KS1 (KRAB/zinc finger suppressor protein 1), a novel, ubiquitously expressed zinc finger gene initially isolated from a rat pancreas library. KS1 contains 10 C2H2 zinc fingers, a KRAB-A/B motif, and an ID sequence that has been shown previously to participate in growth factor-regulated gene expression. Northern blot analysis using pancreatic cell lines demonstrates that KS1 mRNA is inducible by serum and epidermal growth factor, suggesting a role for this gene in cell growth regulation. Biochemical analysis reveals that KS1 is a nuclear protein containing two transcriptional repressor domains, R1 and R2. R1 corresponds to the KRAB-A motif, whereas R2 represents a novel sequence. Transformation assays using NIH3T3 cells demonstrate that KS1 suppresses transformation by the potent oncogenes Ha-ras, Galpha12, and Galpha13. Deletion of the R1/ KRAB-A domain does not modify the transformation suppressive activity of KS1, whereas deletion of R2 abolishes this function. Thus, KS1 is a novel growth factor-inducible zinc finger transcriptional repressor protein with the potential to protect against neoplastic transformation induced by several oncogenes.

3T3 Cells↗
PubMed1998

Identification and chromosomal localization of CTNNAL1, a novel protein homologous to alpha-catenin.

Sodium butyrate (NaB) was shown to induce growth inhibition and apoptosis in a human pancreatic cancer cell line (AsPC-1). To identify the genes that are differentially regulated by NaB, we generated subtracted EST libraries highly enriched for up- or down-regulated transcripts using the suppression subtraction hybridization technique. One of the ESTs identified in the down-regulated library showed significant homology to human alpha-catenin. A cDNA of 2.45 kb that codes for a putative protein of 734 aa was cloned. The cloned cDNA was used as a template in an in vitro transcription-translation reaction. This produced a protein product of about 82 kDa in size. The gene designated as CTNNAL1 (catenin (cadherin-associated protein), alpha-like 1) was found to be ubiquitously expressed in many tissues including pancreas, heart, and skeletal muscle. A human BAC clone containing the gene was isolated and used as a probe for fluorescence in situ hybridization (FISH). Both radiation hybrid and FISH analysis mapped the gene to chromosome band 9q31.2, a region where frequent abnormalities have been observed in bladder carcinoma, esophageal cancers, and several other tumors.

Amino Acid Sequence↗