Structural and functional aspects of cardosins.
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Amino Acid Sequence↗
Biomedical subjects
J Costa
Publications and source records attributed to J Costa.
Crystallisation, structure solution, and initial refinement of plant cardosin-A.
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Amino Acid Sequence↗
Patterns of regular drug use in Spanish childbearing women: changes elicited by pregnancy.
OBJECTIVE: To determine drug use in Spanish women before pregnancy and from conception to the awareness of pregnancy (early period of pregnancy, EPP), as well as to analyse attitudinal changes when pregnancy was planned or known. METHODS: Trained gynaecologists used a structured questionnaire to collect demographic and obstetric characteristics, histories of regular drug taking before pregnancy, attitudes towards drug taking during pregnancy and current drug use in the EPP. Women were interviewed at their first antenatal visit during the first trimester. RESULTS: Two hundred and seventy-two women were included (mean age 29.3 years and 66.3 days of gestation). Before pregnancy, 24% regularly took drugs, 70% of them more than twice a week; a significantly higher frequency was found in those receiving public antenatal care and in those who had had less education. In 39% of women, awareness of pregnancy did not change their attitudes towards regular drug use. Among those who decided to suppress their regular drug intake, 58% did so when their pregnancy was confirmed and 42% when they planned it. In women who planned their pregnancy, 30.1% stopped when they tried to become pregnant. However, 62% of all women took drugs during the EPP. In private antenatal care significantly more drugs were taken per patient. By the 168 women 278 compounds were consumed during the EPP: 40% were analgesics (mainly paracetamol and acetylsalicylic acid) and 25% were digestive and metabolic drugs (mainly antacids and laxatives). Drugs were often used more than twice a week, particularly in women receiving public antenatal care and in those who had had less education. CONCLUSION: Drug taking is common in Spanish women of childbearing age, and many of those in our study did not decide to stop during the EPP. Few women avoid drugs when planning a pregnancy. Therefore, gynaecologists must advise against drug taking in patients who wish to become pregnant and suggest that unnecessary drug use be avoided when the pregnancy is already diagnosed.
Adult↗
Observation of deposits on disposable contact lenses by bio-, light and scanning electron microscopy.
There has been little published on the presence of deposits on disposable hydrogel contact lenses. The purpose of the present study was to determine the possible existence of such deposits. Seventy-two disposable lenses (polymacon; FDA group 1; non-ionic, low water contact lenses) were fitted to six patients and replaced weekly in the course of a total follow-up period of six weeks. By means of biomicroscopy, light microscopy and scanning electron microscopy, different types of deposits were observed on all the lenses studied. Our results also indicate that the accumulation rate of these deposits expressed as a percentage of the total area of the lens which they covered increased after the third replacement of the lenses over the 4th, 5th and 6th weeks. Finally, we observed that the amount of deposits varies in the different patients studied in the course of a 6 week follow-up period.
Journal Article↗
In vitro alpha1-3 or alpha1-4 fucosylation of type I and II oligosaccharides with secreted forms of recombinant human fucosyltransferases III and VI.
Transgalactosylation of chitobiose and chitotriose employing beta-galactosidase from bovine testes yielded mixtures with beta1-3 linked galactose (type I) and beta1-4 linked galactose (type II) in a final ratio of 1:1 for the tri- and 1:1.4 for the tetrasaccharide. After 24 h incubations of the two purified oligosaccharide mixtures with large amounts (20-fold increase compared with standard conditions) of human alpha1,3/4-fucosyltransferase III (FucT III), the type I tri-/tetrasaccharides were completely converted to the Lewis(a) structure, whereas approximately 10% fucosylation of the type II isomers to the Lewis(x) oligosaccharides was observed in long-term incubations. Employing large amounts of human alpha1,3-fucosyltransferase VI (FucT VI), the type I trisaccharide substrate was exclusively fucosylated at the proximal O-4 substituted N-acetylglucosamine (GlcNAc) (20%) whereas almost all of the type II isomers was converted to the corresponding Lewis(x) product. 45% of the type I tetrasaccharide was fucosylated at the second GlcNAc solely by FucT VI. The type II isomer was almost completely alpha1-3 fucosylated to yield the Lewisx derivative with traces of a structure that contained an additional fucose at the reducing GlcNAc. The results obtained in the present study employing high amounts of enzyme confirmed our previous results that FucT III acts preponderantly as a beta1-4 fucosyltransferase onto GlcNAc in vitro. Human FucT VI attaches fucose exclusively in an alpha1-3 linkage to 4-substituted GlcNAc in vitro and does not modify any 3-substituted GlcNAc to yield Lewis(a) oligosaccharides. With 8-methoxycarbonyloctyl glycoside acceptors used under standard conditions, FucT III acts exclusively on the type I and FucT VI only on the type II derivative. With lacto-N-tetraose, lacto-N-fucopentraose I, or LS-tetrasaccharide as substrates, FucT III modified the 3-substituted GlcNAc and the reducing glucose; FucT VI recognized only lacto-N-neotetraose as a substrate.
Carbohydrate Sequence↗
Genetic confirmation of the specific status of Triatoma petrochii (Hemiptera: Reduviidae: Triatominae).
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Alleles↗
Synergistic sedation with propofol and midazolam in intensive care patients after coronary artery bypass grafting.
OBJECTIVES: To evaluate and compare the clinical efficacy, impact on hemodynamics, safety profiles, and cost of combined administration of propofol and midazolam (synergistic sedation) vs. midazolam and propofol administered as sole agents, for sedation of mechanically ventilated patients after coronary artery bypass grafting. DESIGN: Prospective, controlled, randomized, double-blind clinical trial. SETTING: Intensive care unit of SCIAS-Hospital de Barcelona. PATIENTS: Seventy-five mechanically ventilated patients who underwent coronary artery bypass graft surgery under low-dose opioid anesthesia. INTERVENTIONS: According to the double-blind method, patients were randomly assigned to receive propofol (n = 25), midazolam (n = 25), or propofol combined with midazolam (n = 25). Infusion rates were adjusted to stay between 8 and 11 points on Glasgow Coma Score modified by Cook and Palma. MEASUREMENTS AND MAIN RESULTS: Mean +/- SD duration of sedation was 14.4 +/- 1.5 hrs, 14.1 +/- 1.1 hrs, and 14.7 +/- 1.9 hrs for the propofol, midazolam, and synergistic groups, respectively. The induction dose was 0.55 +/- 0.05 mg/kg for propofol as sole agent, 0.05 +/- 0.01 mg/kg for midazolam as sole agent, and 0.22 +/- 0.03 mg/kg for propofol administered in combination with 0.02 +/- 0.00 mg/kg of midazolam (p = .001). The maintenance dose was 1.20 +/- 0.03 mg/kg/hr for propofol as sole agent, 0.08 +/- 0.01 mg/kg/hr for midazolam as sole agent, and 0.50 +/- 0.09 mg/kg/hr for propofol administered in combination with 0.03 +/- 0.01 mg/kg/hr of midazolam (p < .001). All sedative regimens achieved similar efficacy in percentage of hours of adequate sedation (93% for propofol, 88% for midazolam, and 90% for the synergistic group, respectively). After induction, both propofol and midazolam groups had significant decreases in systolic blood pressure, diastolic blood pressure, left atrial pressure, and heart rate. Patients in the synergistic group had significant bradycardia throughout the study, without impairment in other hemodynamic parameters. Patients sedated with propofol or synergistic regimen awoke sooner and could be extubated before those patients sedated with midazolam (0.9 +/- 0.3 hrs and 1.2 +/- 0.6 hrs vs. 2.3 +/- 0.8 hrs, respectively, p = .01). Synergistic sedation produced cost savings of 28% with respect to midazolam and 68% with respect to propofol. CONCLUSIONS: In the study conditions, the new synergistic treatment with propofol and midazolam administered together is an effective and safe alternative for sedation, with some advantages over the conventional regimen with propofol or midazolam administered as sole agents, such as absence of hemodynamic impairment, >68% reduction in maintenance dose, and lower pharmaceutical cost.
Adult↗
Morphometrical features of the corneal epithelium in mammals.
The study has been conducted on 40 species of mammals included in different taxonomic orders: 10 carnivores (felines and canides), 13 primates and 17 herbivores (artiodactyles, perisodactyles and proboscideans). Domestic, experimental and wild animals have been included in the sample. The purpose of the study is to offer new data regarding morphometric aspects of the epithelium of the cornea of mammals. We also aim to establish the relationships and differences between the different epithelia studied and to determine the morphometric characteristics which best define the epithelium of the different species under study. The results obtained have demonstrated the existence of correlations between thicknesses, number of cellular layers and number of the epithelial cells in the different groups. It has also been established that the variants in thickness and number of layers define and characterize the epithelia. Finally, the study reveals that morphometric characteristics of the epithelium of primates and carnivores have more similarities than those of herbivores.
Animals↗
Detection of thyroglobulin, thyroid peroxidase, and RET/PTC1 mRNA transcripts in the peripheral blood of patients with thyroid disease.
PURPOSE: Detection of mRNA transcripts for thyroglobulin (TG), thyroid peroxidase (TPO) and RET/PTC1 in the peripheral blood of patients with thyroid disease. PATIENTS AND METHODS: TG, TPO, and RET/PTC1 mRNA were analyzed in 52 peripheral-blood samples from 44 patients diagnosed with thyroid carcinoma (24 patients), adenoma (five patients), and nodular hyperplasia (15 patients) by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: TG and TPO were identified in 13 patients (54.2%) with thyroid carcinoma, which includes five of eight patients with no clinical evidence of disease at the time of blood collection. Four of 5 patients had cervical lymph node metastases and/or extrathyroid extension at the time of the initial surgery. RET/PTC1 mRNA was detected in the peripheral blood of only one patient with papillary thyroid carcinoma. This sample was also positive for TG and TPO. TG and TPO were detected in two patients (10%) with benign thyroid nodules. All positive samples from patients with benign thyroid lesions were collected before surgery, whereas all samples collected after surgery were negative. RET/PTC1 mRNA was not detected in any of the patients with benign thyroid nodules. RT-PCR positivity for TG and TPO mRNA was higher in patients with carcinoma than in patients with benign lesions (P = .002). CONCLUSION: TG, TPO, and RET/PTC1 mRNA are detectable in the peripheral blood of patients with thyroid disease, which correlates with a diagnosis of carcinoma.
Adolescent↗
The molecular biology of breast cancer: accelerating clinical applications.
Recent advances in basic science have led to a better understanding of the molecular events important in the pathogenesis of breast cancer. Very little of this new knowledge, however, has had a significant impact on improving the diagnosis and therapy of breast cancer. We review many of the molecular events important in the pathogenesis of breast cancer, including inherited abnormalities in BRCA-1 and BRCA-2, p53, ATM, and PTEN and sporadic alterations in growth factors and their receptors, signal transduction, cell cycle control, DNA repair, cell death, angiogenesis, and invasion and metastasis. We suggest ways to speed up clinical applications of the new molecular knowledge base through the use of preclinical disease models, development of high throughput sample analysis and infrastructure programs to facilitate translational research, implementation of practice guidelines, and development of regional oncology networks. Only through the implementation of such a deliberate, multifaceted strategy will the gap between the research laboratory and the clinic be closed.
Animals↗
Rigid gas permeable contact lenses surface roughness examined by interferential shifting phase and scanning electron microscopies.
The anterior surface roughness of seven factory new rigid gas permeable (RGP) contact lenses has been studied by interferential shifting phase microscopy (ISPM) and scanning electron microscopy (SEM). Five lenses were fluorsilicone acrylate and two lenses were silicone acrylate. Their material Dk ranged from 14 to 210. ISPM is shown to be a reliable and non-destructive method to observe and measure the relief of the contact lens surface. Moreover, profile and contour data are easily stored for further quantitative studies. ISPM contour patterns of the studied lenses are qualitatively compared with those obtained by SEM for the same lenses. Results point out that ISPM gives similar accuracy but it is non-destructive and cheaper than SEM. Moreover, the quantitative study of surface roughness suggests that there is a relationship between surface roughness and Dk of the lens material: surface roughness increases with Dk and allows to distinguish between lenses with low, medium and high Dk.
Contact Lenses↗
Identification of the human Lewis(a) carbohydrate motif in a secretory peroxidase from a plant cell suspension culture (Vaccinium myrtillus L.).
This paper reports for the first time the presence of the human Lewis(a) type determinant in glycoproteins secreted by plant cells. A single glycopeptide was identified in the tryptic hydrolysis of the peroxidase VMPxC1 from Vaccinium myrtillus L. by HPLC/ESI-MS. The oligosaccharide structures were elucidated by ESI-MS-MS and by methylation analysis before and after removal of fucose by mild acid hydrolysis. The major structure determined is of the biantennary plant complex type containing the outer chain motif Lewis(a) [structure in text]. A corresponding fucosyltransferase activity catalyzing the formation of Lewis(a) type structures in vitro was identified in cellular extracts of the suspension cultures.
Amino Acid Sequence↗
Effects of p53 mutations on apoptosis in mouse intestinal and human colonic adenomas.
We have examined the effects of inactivation of the p53 tumor suppressor gene on the incidence of apoptotic cell death in two stages of the adenoma-to-carcinoma progression in the intestine: in early adenomas where p53 mutations are rare and in highly dysplastic adenomas where loss of p53 occurs frequently. Homozygosity for an inactivating germ-line mutation of p53 had no effect on the incidence or the rate of progression of ApcMin/+-induced adenomas in mice and also did not affect the frequency of apoptosis in the cells of these adenomas. To examine the effect of p53 loss on apoptosis in late-stage adenomas, we compared the incidence of apoptotic cell death before and after the appearance of highly dysplastic cells in human colonic adenomas. The appearance of highly dysplastic cells, which usually coincides during colon tumor progression with loss of heterozygosity at the p53 locus, did not correlate with a reduction in the incidence of apoptosis. These studies suggest that p53 is only one of the genes that determine the incidence of apoptotic in colon carcinomas and that wild-type p53 retards the progression of many benign colonic adenoma to malignant carcinomas by mechanism(s) other than the promotion of apoptosis.
Adenoma↗
Stable expression of the Golgi form and secretory variants of human fucosyltransferase III from BHK-21 cells. Purification and characterization of an engineered truncated form from the culture medium.
Stable BHK-21 cell lines were constructed expressing the Golgi membrane-bound form and two secretory forms of the human alpha1, 3/4-fucosyltransferase (amino acids 35-361 and 46-361). It was found that 40% of the enzyme activity synthesized by cells transfected with the Golgi form of the fucosyltransferase was constitutively secreted into the medium. The corresponding enzyme detected by Western blot had an apparent molecular mass similar to those of the truncated secretory forms. The secretory variant (amino acids 46-361) was purified by a single affinity-chromatography step on GDP-Fractogel resin with a 20% final recovery. The purified enzyme had a unique NH2 terminus and contained N-linked endo H sensitive carbohydrate chains at its two glycosylation sites. The fucosyltransferase transferred fucose to the O-4 position of GlcNAc in small oligosaccharides, glycolipids, glycopeptides, and glycoproteins containing the type I Galbeta1-3GlcNAc motif. The acceptor oligosaccharide in bovine asialofetuin was identified as the Man-3 branched triantennary isomer with one Galbeta1-3GlcNAc. The type II motif Galbeta1-4GlcNAc in bi-, tri-, or tetraantennary neutral or alpha2-3/alpha2-6 sialylated oligosaccharides with or without N-acetyllactosamine repeats and in native glycoproteins were not modified. The soluble forms of fucosyltransferase III secreted by stably transfected cells may be used for in vitro synthesis of the Lewisa determinant on carbohydrates and glycoproteins, whereas Lewisx and sialyl-Lewisx structures cannot be synthesized.
Animals↗
The glycosylation of the aspartic proteinases from barley (Hordeum vulgare L.) and cardoon (Cynara cardunculus L.).
Plant aspartic proteinases characterised at the molecular level contain one or more consensus N-glycosylation sites [Runeberg-Roos, P., Tŏrmäkangas, K. & Ostman, A. (1991) Eur. J. Biochem. 202, 1021-1027; Asakura, T., Watanabe, H., Abe, K. & Arai, S. (1995) Eur. J. Biochem, 232, 77-83; Veríssimo, P., Faro, C., Moir, A. J. G., Lin, Y., Tang, J. & Pires, E. (1996) Eur. J. Biochem. 235, 762-768]. We found that the glycosylation sites are occupied for the barley (Hordeum vulgare L.) aspartic proteinase (Asn333) and the cardoon (Cynara cardunculus L.) aspartic proteinase, cardosin A (Asn70 and Asn363). The oligosaccharides from each site were released from peptide pools by enzymatic hydrolysis with peptide-N-glycanase A or by hydrazinolysis and their structures were determined by exoglycosidase sequencing combined with matrix-assisted laser desorption/ionization time of flight mass spectrometry. It was observed that 6% of the oligosaccharides from the first glycosylation site of cardosin A are of the oligomannose type. Modified type glycans with proximal Fuc and without Xyl account for about 82%, 14% and 3% of the total oligosaccharides from the first and the second glycosylation sites of cardosin A and from H. vulgare aspartic proteinase, respectively. Oligosaccharides with Xyl but without proximal Fuc were only detected in the latter proteinase (4%). Glycans with proximal Fuc and Xyl account for 6%, 86% and 92% of total oligosaccharides from the first and second glycosylation sites of cardosin A and from H. vulgare aspartic proteinase, respectively.
Aspartic Acid Endopeptidases↗
Hepatitis C virus (HCV) genotypes in Spanish patients with HCV infection: relationship between HCV genotype 1b, cirrhosis and hepatocellular carcinoma.
BACKGROUND/AIMS: The influence of the infecting virus genotype on the progression of the underlying liver disease in patients with chronic hepatitis C virus (HCV) infection remains controversial. The aim of this study was to investigate the prevalence of HCV genotypes in Spanish patients with chronic HCV infection and to elucidate the relationship between the infecting genotype and the severity of the disease. METHODS: A cross-sectional, retrospective analysis of frequency distribution of HCV genotypes was carried out in 414 Spanish patients with chronic HCV infection, including 243 patients with asymptomatic or minimally symptomatic chronic hepatitis, 112 patients with cirrhosis and hepatocellular carcinoma and 59 patients with decompensated cirrhosis. HCV genotype was determined by restriction fragment length polymorphisms of the 5' non-coding region. RESULTS: Infection with HCV genotype 1b was found in 72% of patients with chronic hepatitis and in more than 90% of patients with cirrhosis, with or without hepatocellular carcinoma. Older age, infection with genotype 1b and absence of overt parenteral exposure as a possible source of infection were associated with cirrhosis and hepatocellular carcinoma by univariate analysis and this association was confirmed by regression analysis. CONCLUSIONS: HCV genotype 1b is associated with advanced liver disease in our geographical area. However, this may be related to a cohort-effect caused by over-representation of genotype 1b in older patients with more advanced disease, because, in our country, this HCV genotype appeared earlier in time and is therefore associated with more prolonged periods of infection.
Adult↗
Hepatitis G virus infection in chronic hepatitis C: frequency, features and response to interferon therapy.
BACKGROUND/AIMS: The pathogenic relevance of the hepatitis G virus (HGV) and its sensitivity to interferon are currently under investigation. This study aimed to investigate the prevalence of HGV infection in patients with chronic hepatitis C and to elucidate if HGV co-infection modifies the clinical course and the response to interferon therapy in this disease. METHODS: HGV-RNA was investigated by reverse transcription-polymerase chain reaction in serum from 143 consecutive patients who received interferon alpha-2b (3 MU t.i.w.) for 24 weeks. Baseline features and response to therapy in HGV-infected and non-infected patients were compared. To assess the antiviral effect of interferon, serial quantitative measurement of HCV-RNA and HGV-RNA in serum was performed in patients co-infected with HCV and HGV. RESULTS: Eight patients (5.6%) presented HGV-RNA sequences in serum. No significant differences were found between HGV-infected and non-infected patients in relation to age, sex, source of infection, liver function tests, liver histology and HCV genotype, nor in the biochemical response to interferon, which was sustained in 12% and 15%, transient in 37% and 30% and absent in 50% and 55% of HGV-infected and non-infected patients, respectively. HGV-RNA became negative in all treated patients, but sustained viral inhibition was observed only in those with low viral load. CONCLUSIONS: The prevalence of HGV infection in HCV-infected patients is relatively low in our geographical area. HGV co-infection does not appear to modify the clinical presentation nor the response to interferon in chronic hepatitis C. HGV is sensitive to interferon, particularly if pre-treatment viral load is low.
Adult↗
Comparison of cytospat and pipelle endometrial biopsy instruments.
OBJECTIVE: To compare two endometrial biopsy techniques, based on the quality of material obtained, cost effectiveness, and pain during the procedure. STUDY DESIGN: A single blind, randomized prospective study. RESULTS: A total of 365 women with peri/postmenopausal bleeding were randomized into two groups: 174 biopsied cytospat (C) and 191 biopsied with pipelle (P). Both procedures were then followed by either dilatation and curettage (D&C) or hysterectomy. Pain was evaluated in 379 patients, with the result being better tolerance for P vs. C (3.55 vs. 4.06, P = 0.07). With C, the sensitivity in histological evaluation was 82% for benign endometrium, 60% for endometrial hyperplasia and 60% for corpus uterine neoplasia as compared to the D&C and hysterectomy material. With P the sensitivity was 84, 71 and 60%, respectively for the three diagnoses, as compared to the D&C and hysterectomy material. Insufficient tissue for pathologic evaluation was present in 24% of C, 25% of P and 9.8% of D&C samples. CONCLUSIONS: Cost effectiveness for pipelle is slightly higher than for Cytospat.
Adult↗