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Biomedical subjects

C Gutierrez

Publications and source records attributed to C Gutierrez.

At least 55 records · Page 3Linked to original sources

Relationship of oligomerization to DNA binding of Wheat dwarf virus RepA and Rep proteins.

Members of the genus Mastrevirus (family Geminiviridae) produce a complementary-sense (c-sense) transcription unit with the potential to encode two proteins, RepA and Rep. In the present work, we have studied the DNA-protein complexes formed by the Wheat dwarf virus (WDV) RepA protein within the WDV large intergenic region. WDV RepA forms large nucleoprotein complexes near the TATA boxes of the viral complementary-sense and virion-sense (v-sense) promoters (the RepA C- and V-complexes, respectively), a location similar to those of WDV Rep-DNA complexes but with distinct DNase I footprints. We have also studied the relationship of oligomerization of WDV RepA and Rep proteins to DNA-protein complex formation. Using chemical cross-linking, we have determined that both WDV proteins can form oligomers in solution. Interestingly, the pH is critical for the monomer-oligomer equilibrium and small changes produce a displacement in such a way that at pH </= 7.0, the predominant species is an octamer while at pH >/= 7.4 it is a monomer. Complex formation is also strongly affected by pH and occurs more efficiently at pH 7.0-7.4. We found that preformed oligomers interact very poorly with DNA. Thus, our data are consistent with a stepwise model for protein-DNA complex assembly in which monomers interact with DNA and then with other monomers to assemble an oligomeric structure on the DNA. These results may be relevant for studies on the DNA binding, replication, and transcription properties of geminivirus proteins.

Base Sequence↗

Camel trypanosomosis in the Canary Islands: assessment of seroprevalence and infection rates using the card agglutination test (CATT/T. evansi) and parasite detection tests.

Trypanosomosis due to Trypanosoma evansi (surra) is a major enzootic disease of the dromedary camel. Thus, the purpose of the present study was to assess seroprevalence and infection rates in the Canary Islands using antibody(-card agglutination test-CATT/T. evansi) and parasite detection tests (micro-Haematocrit Centrifugation technique, Giemsa stained blood smears, microscopic examination of lymph node aspirates and mouse inoculation). PCV was also determined. 745 dromedary camels (483 females and 262 males) were examined. Trypanosomes were detected in seven animals. 36 animals yielded CATT positive results while 709 animals were negative. All parasitologically positive animals were also CATT positive. Results showed a good correlation between CATT positive and low PCV and a higher seroprevalence in older animals. Trypanocidal drugs have not been registered in Spain and, consequently, if vigilance is not exercised the prevalence could be increased in the future.

Agglutination Tests↗

Neurochemical and connectional organization of the dorsal pulvinar complex in monkeys.

To investigate the organization of the dorsal pulvinar complex, patterns of neurochemical staining were correlated with cortico-pulvinar connections in macaques (Macaca mulatta). Three major neurochemical subdivisions of the dorsal pulvinar were identified by acetylcholinesterase (AChE) histochemistry, as well as immunostaining for calbindin-D(28K) and parvalbumin. The dorsal lateral pulvinar nucleus (PLd) was defined on histochemical criteria as a distinct AChE- and parvalbumin-dense, calbindin-poor wedge that was found to continue caudally along the dorsolateral edge of the pulvinar to within 1 mm of its caudal pole. The ventromedial border of neurochemical PLd with the rest of the dorsal pulvinar, termed the medial pulvinar (PM), was sharply defined. Overall, PM was lighter than PLd for AChE and parvalbumin and displayed lateral (PMl) and medial (PMm) histochemical divisions. PMm contained a central "oval" (PMm-c) that stained darker for AChE and parvalbumin than the surrounding region. The neurochemically defined PLd was labeled by tracer injections in the inferior parietal lobule (IPL) and dorsolateral prefrontal cortex but not the superior temporal gyrus (STG). Label within PMl was found after prefrontal and IPL and, to a lesser extent, after STG injections. The PMm was labeled after injections of the IPL and STG, but only sparsely following prefrontal injections. The histochemically distinct subregion or module of PMm, PMm-c, was labeled only by STG injections. Overlapping labeling was found in dorsal pulvinar divisions PMl and PLd following paired IPL/prefrontal, but not IPL/STG or these particular STG/prefrontal, injections. Thus, PLd may be a visuospatially related region whereas PM appears to contain several types of territories, some related to visual or auditory inputs, and others that receive directly converging input from posterior parietal and prefrontal cortex and may participate in a distributed cortical network concerned with visuospatial functions.

Acetylcholinesterase↗

DNA replication and cell cycle in plants: learning from geminiviruses.

Plant cell growth and development depend on continuous cell proliferation which is restricted to small regions of the plant called meristems. Infection by geminiviruses, small DNA viruses whose replicative cycle relies on host cell factors, is excluded from those proliferating areas. Since most of the replicative factors are present, almost exclusively, in proliferating cells, geminivirus infection is believed to induce a cellular state permissive for viral DNA replication, e.g. S-phase or, at least, some specific S-phase functions. The molecular basis for this effect seems to be the interference that certain geminivirus proteins exert on the retinoblastoma-related (RBR) pathway, which analogously to that of animal cells, regulates plant cell cycle activation and G(1)-S transition. In some cases, geminiviruses induce cell proliferation and abnormal growth. Mechanisms other than sequestering plant RBR probably contribute to the multiple effects of geminivirus proteins on cellular gene expression, cell growth control and cellular DNA replication. Current efforts to understand the coupling of geminivirus DNA replication to cell cycle and growth control as well as the directions in which future research is aiming are reviewed.

Cell Cycle↗

Juvenile papillomatosis of the breast in male infants: two case reports.

Juvenile papillomatosis of the breast ("Swiss cheese disease") is a benign localized proliferative condition of the breast which occurs almost exclusively in young adult women. Patients with this lesion often have a family history of breast carcinoma, and rarely carcinoma may coexist with the lesion at the time of diagnosis. We present two cases of male infants with juvenile papillomatosis of the breast. The pathology and clinical management of this novel lesion is discussed.

Breast Neoplasms, Male↗

Geminiviruses and the plant cell cycle.

Geminiviruses are plant DNA viruses with a small genome that infect a large variety of plant species. Viral proteins regulate viral DNA replication and transcription. Also they appear to have an impact on cellular gene expression. Cellular proteins directly involved in DNA replication, such as PCNA, have long been known to accumulate in cells expressing Rep tomato golden mosaic geminivirus. This effect can be a direct effect of the viral protein and/or be mediated by interference with the G1/S transition control, namely the pathway controlled by the retinoblastoma-related (RBR) protein, analogous to the human retinoblastoma (RB) tumour suppressor protein. Different geminiviruses seem to have evolved two mechanisms to interact with plant RBR proteins. One is dependent on a LxCxE amino acid motif present in proteins, such as RepA, encoded by members of the Mastrevirus genus, and another seems to be mediated by the viral Rep protein, which lacks the LxCxE motif, encoded by members of the Begomovirus, and perhaps the Curtovirus genus. These and other aspects of the relationships between geminivirus replication and cell cycle control pathways will be discussed.

Amino Acid Motifs↗

Involvement of differential efficiency of transcription by esigmas and esigma70 RNA polymerase holoenzymes in growth phase regulation of the Escherichia coli osmE promoter.

Transcription of the gene osmE of Escherichia coli is inducible by elevated osmotic pressure and during the decelerating phase of growth. osmE expression is directed by a single promoter, osmEp. Decelerating phase induction of osmEp is dependent on the sigmas (RpoS) factor, whereas its osmotic induction is independent of sigmas. Purified Esigmas and Esigma70 were both able to transcribe osmEp in vitro on supercoiled templates. In the presence of rpoD800, a mutation resulting in a thermosensitive sigma70 factor, a shift to non-permissive temperature abolished induction of osmEp after an osmotic shock during exponential phase, but did not affect the decelerating phase induction. Point mutations affecting osmEp activity were isolated. Down-promoter mutations decreased transcription in both the presence and the absence of sigmas, indicating that the two forms of RNA polymerase holoenzyme recognize very similar sequence determinants on the osmE promoter. Three up-promoter mutations brought osmEp closer to the consensus of Esigma70-dependent promoters. The two variant promoters exhibiting the highest efficiency became essentially independent of sigmas in vivo. Our data suggest that Esigmas transcribes wild-type osmEp with a higher efficiency than Esigma70. A model in which an intrinsic differential recognition contributes to growth phase-dependent regulation is proposed. Generalization of this model to other sigmas-dependent promoters is discussed.

Bacterial Proteins↗

Syncope associated with hypertrophic cardiomyopathy in a dromedary camel.

A case of hypertrophic cardiomyopathy in a dromedary camel (Camelus dromedarius) is described for the first time. The patient, a castrated 9-year-old animal, presented with a 6-month history of several episodes of syncope after moderate exercise and later at rest. The syncope had a sudden onset and a duration of 30 to 45 seconds. After clinical, electrocardiographical and echocardiographical examination, a tentative diagnosis of hypertrophic cardiomyopathy was made. At necropsy, the heart had a globose shape and was firm on palpation. The left ventricular free wall and the interventricular septum were thickened in cross-section and the left ventricular lumen was small. The clinical diagnosis was confirmed by histologic examination of heart tissue that demonstrated hypertrophy of myocardial fibres with vesicular nuclei and the presence of diffuse interstitial fibrosis.

Animals↗

Osteoporosis and lung transplantation: a prospective study.

STUDY OBJECTIVE: Osteoporosis is a well-recognized complication of lung transplantation that may significantly impair the quality of life of transplant recipients. We performed a prospective study of bone mineral density (BMD) before and after transplantation to determine the degree of bone mass loss associated with lung transplantation Patients and design: We conducted a prospective study of BMD in 28 patients with various end-stage respiratory diseases pretransplantation and 6 to 12 months posttransplantation. The BMD of the lumbar spine (LS) and femoral neck (FN) were measured. All 28 patients were treated only with vitamin D and calcium supplementation posttransplant. The primary endpoint was the percentage change in BMD. The secondary endpoint was the incidence of fractures posttransplant. A univariate analysis was conducted to determine the various risk factors associated with bone mass loss pretransplant and posttransplant. RESULTS: Prior to transplantation, moderate to severe bone disease was evident. The mean (+/- SD) pretransplant T score (the number of SDs from the peak bone mass) and Z score (the number of SDs from the age-matched mean) for the LS were -1.72 +/- 1.37 and -1.44 +/- 1.31, respectively. The mean pretransplant T score and Z score for the FN were -2.65 +/- 1.01 and -1.5 +/- 1.43, respectively. Within 6 to 12 months posttransplant, the mean BMD for the LS decreased by 4.76% (p < 0.001), while the mean BMD for the FN decreased by 5.3% (p < 0.001). Five of the 28 patients (18%) suffered osteoporotic fractures posttransplant, while no fractures were documented pretransplant. The cumulative steroid dose posttransplant was associated with a drop in BMD for the LS and FN (r = 0.39, p = 0.039 and r = 0.63, p < 0.001, respectively), while a negative association was found between cumulative steroid use pretransplant and baseline LS and FN T scores (r = -0.4, p = 0. 02 and r = -0.43, p = 0.023, respectively). CONCLUSION: Within 6 to 12 months after lung transplantation, there is a significant decrease in BMD at both the LS and FN levels (approximately 5%) despite vitamin D and calcium supplementation. This drop in BMD is associated with a relatively high incidence of osteoporotic fractures posttransplant.

Absorptiometry, Photon↗

Applying a prediction rule to identify low-risk patients with community-acquired pneumonia.

STUDY OBJECTIVES: To study the validity of a recently developed community-acquired pneumonia (CAP) severity prediction rule in estimating mortality, to determine its utility in decision making regarding hospitalization, and to assess factors influencing this decision. DESIGN: Retrospective chart review. SETTING: Two sites of the University Health Network, the Toronto General and Toronto Western Hospitals, tertiary-care teaching institutions with a sizable primary-care and secondary-care source of referrals, and a total of 900 beds. PATIENTS: Consecutive patients with CAP admitted between February and June 1996. MEASUREMENTS AND RESULTS: A single trained medical records extractor assembled data to compare our population to that used in developing the CAP prediction rule, in terms of mortality and to assess reasons for hospitalization. Two hundred fifty-five eligible patients were admitted, and 244 charts (96%) were available. Our patients tended to be older, with nearly four times as many residents of chronic care institutions (39% compared with 10%), and had a higher risk class distribution than the published cohort. Risk class-specific mortality was similar in four of five classes. Of the 71 patients in the low-risk classes, 67 had additional reasons for admission; 18 of which were psychosocial (homelessness, substance abuse, or inadequate home supports). CONCLUSIONS: The CAP severity prediction rule estimates mortality well. Admission of low-risk patients was linked to psychosocial and other medical reasons not captured by this rule. The rule can be very useful in assessing the need for hospitalization; however, there remains a significant percentage of patients with a low severity score who may require hospitalization for psychosocial and economic considerations.

Adult↗

Plasma soluble tumor necrosis factor-alpha receptors circulate in proportion to leptin levels during the menstrual cycle in lean but not in obese women.

OBJECTIVE: In recent studies serum leptin levels were significantly higher in the luteal phase than in the follicular phase, but the mechanism of changing leptin levels are unknown. Several research lines indicate a potential role for tumor necrosis factor (TNF-alpha) in ovulation and reproductive events. As TNF-alpha appears to regulate leptin secretion, we speculated that TNF-alpha might be involved in leptin variations during the menstrual cycle. DESIGN AND METHODS: Nine healthy never obese and ten overweight normally cycling women were studied. TNF-alpha action - through the plasma levels of the soluble fraction of the tumor necrosis factor receptors 1 and 2 (sTNFR1 and sTNFR2) - and leptin concentrations were measured in the follicular (F), peri-ovulatory (PO) and luteal phases (L) of their menstrual cycles. RESULTS: Circulating leptin levels were significantly associated with the stage of the menstrual cycle (P<0.001), being higher in PO and L phases. However, only three of ten overweight subjects vs eight of nine lean women (Chi square P=0.014 after Fisher's exact test) showed significantly higher leptin levels in the PO and L than in the F phase (95% confidence interval (95% CI) of the differences, 3.7 to 10.2 ng/ml, paired t-test P=0.001). In these women (group 1), the changes in leptin levels parallelled the variations observed in plasma sTNFR1 (2.50+/-0.1 vs 2.11+/-0.05 ng/ml, P<0.0001, 95% CI, 0.21 to 0.56) and sTNFR2 levels (5.19+/-0.28 vs 4.55+/-0.25 ng/ml, P<0.0001, 95% CI, 0. 47 to 0.81). In the remaining women (group 2), leptin (95% CI, -1 to 9.2 ng/ml, P=not significant (NS)), sTNFR1 (95% CI, -0.3 to 0.14 ng/ml, P=NS) and sTNFR2 levels (95% CI, -0.95 to 0.39 ng/ml, P=NS) were essentially unaltered throughout the menstrual cycle. Group 2 women were similar in age (36.1+/-2.9 vs 37.3+/-1.4 years) and significantly overweight (body mass index 31+/-2.9 vs 23.9+/-1. 2 kg/m(2)) compared with group 1 women. A negative correlation was observed between leptin levels in the follicular phase and the change in plasma leptin from F to L phase in all subjects (r=-0.67, P=0.002). CONCLUSIONS: Circulating leptin and sTNFRs levels change significantly during the menstrual cycle of most lean women. In contrast, the levels of these molecules remain essentially unaltered during the F, PO and L phases in the majority of overweight women. Obesity might be associated not only with blunted diurnal excursions and dampened pulsatility, but also with blunted excursions during the menstrual cycle.

Antigens, CD↗

Caseous lymphadenitis caused by Corynebacterium ulcerans in the dromedary camel.

Caseous lymphadenitis that affected the dorsal and ventral superficial lymph nodes in the left cervicothoracic region of a young dromedary camel is described. The agent isolated was Corynebacterium ulcerans. To our knowledge, this is the first description of purulent lymphadenitis caused by C. ulcerans in a species belonging to the Camelidae.

Animals↗

Characterization of OpuA, a glycine-betaine uptake system of Lactococcus lactis.

A Lactococcus lactis glycine-betaine transport system was identified by functional complementation of an Escherichia coli proP proU mutant with a gene library from L. lactis sbsp. cremoris. The cloned locus forms an operon highly homologous to opuA, encoding a glycine-betaine uptake system of Bacillus subtilis. Disruption of opuA in L. lactis abolished protection by glycine-betaine against elevated osmolarity. OpuA belongs to the so-called "ABC transporters" family, which comprise an extracellularly localized substrate-binding protein. In B. subtilis OpuA system, this binding protein is a lipoprotein, attached to the external face of the cytoplasmic membrane by its lipidic moiety. In contrast, in the L. lactis opuA operon, and in other gram-positive homologues as well, a fusion between the gene encoding the integral membrane protein and the substrate-binding protein components gave rise to a hybrid protein presumably attaching the substrate-binding protein to the surface of the cell via its covalent link to the integral membrane component. Mapping of L. lactis opuA transcription start identified one mRNA, more abundant in cells grown at elevated osmolarity. Construction of an opuA-gusA fusion confirmed that opuA transcription is directed by a promoter osmotically inducible in L. lactis. When recombined upstream from a lac transcriptional fusion in the chromosome of E. coli, the opuA promoter appeared as very strong, and only poorly stimulated by elevated osmotic pressure, suggesting the existence of a specific machinery involved in the osmotic signal transduction in L. lactis.

ATP-Binding Cassette Transporters↗

Geminivirus DNA replication.

Geminiviruses are DNA viruses which infect plants. They have a small genome and encode only a few proteins. Therefore, their DNA replication cycle relies largely on the use of cellular DNA replication proteins. The strategy used by geminiviruses to replicate their single-stranded DNA (ssDNA) genome consists of a first stage of conversion of ssDNA into double-stranded DNA (dsDNA) intermediates and, then, the use of dsDNA as a template to amplify viral dsDNA and to produce mature ssDNA genomes by a rolling-circle replication mechanism. In addition, the accumulating evidence indicates that viral DNA replication is somehow coupled to the cell cycle regulatory network of the infected cell. For these reasons, geminiviruses are excellent model systems to understand the regulation of DNA replication and cell cycle in plant cells. Recent years have witnessed significant progress in the identification of cis-acting signals and their interaction with trans-acting factors that contribute to geminivirus origin function. These and other aspects of the geminivirus DNA replication cycle will be reviewed.

DNA Replication↗

The cloning of plant E2F, a retinoblastoma-binding protein, reveals unique and conserved features with animal G(1)/S regulators.

Association of the retinoblastoma (Rb) protein with E2F transcription factors is central to cell cycle-specific gene expression and growth in animal cells. Whether Rb-E2F complexes are also involved in plant cell growth and differentiation is still unknown since E2F proteins have not yet been identified in plants. Here we report the isolation and characterisation of a wheat E2F (TmE2F) cDNA clone. Interestingly, the overall domain organisation of plant E2F is related to the human E2F-1/2/3 subset but its primary sequence is slightly more related to the E2F-4/5 subset. TmE2F-Rb binding depends on residues, located at the C-terminus, which are different from those of animal E2Fs. However, the acidic or hydrophobic nature of certain residues is maintained, strongly suggesting that they may have a crucial role in E2F activities. Plant E2F is expressed in proliferating cultured cells and in differentiated tissues and is up-regulated early in S phase. Our studies reinforce the idea that G(1)/S regulators in plants are unrelated to those of yeast cells but similar to those of animal cells and provide new tools to analyse the links between cell cycle regulators, plant growth and developmental signals.

Amino Acid Sequence↗