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Biomedical subjects

M T Gil

Publications and source records attributed to M T Gil.

At least 19 recordsLinked to original sources

[A long-term comparative study of calcitriol versus alphacalcidol in patients with secondary hyperparathyroidism on hemodialysis].

UNLABELLED: Calcitriol has traditionally been the most widely used treatment for secondary hyperparathyroidism (SHPT) in uremic patients. There are currently no crossover equivalence studies of alphacalcidol versus calcitriol establishing which of the two derivatives is more active and better tolerated. The objective of this study was to compare the long term effect on control of PTH of similar doses of alphacalcidol versus calcitriol in the treatment of SHPT in these patients. METHODS: We conducted a retrospective study on 21 hemodialysis patients with stable SHPT of varying severity treated with intravenous calcitriol. In July 2002, the pharmacy of the reference hospital decided to substitute calcitriol for alphacalcidol based on the similarity of the two drugs. The conversion was made substituting a similar amount of drug. Mean absolute serum levels and percentage change in PTH, calcium and phosphorus were compared between the two periods and at 0, 3, 6, 9, 12 and 15 months after starting treatment with alphacalcidol. Student's t-test for paired means was used to compare the values between the two periods. RESULTS: In the calcitriol period, mean PTH levels were 275.2 +/- 111.7 pg/ml. The mean dose of drug used was 1.7 +/- 0.8 mcg postdialysis, and serum calcium and phosphorus levels were 10.1 +/- 0,5 mg/dl and 5,2 +/- 0,9 mg/dl, respectively (p < 0.01). Mean dialysate calcium content was 2,9 +/- 0,3 mEq/l. In the alphacalcidol period, PTH increased (441.6 +/- 178.3 pg/ml) (p < 0.001) and the percentage of patients with PTH < 300 pg/ml decreased (24% at the end of the period), in spite of significantly increasing the mean drug dose (2,3 +/- 0,9 mcg postdialysis) (p < 0.05). Serum calcium levels did not show significant differences (10.2 +/- 0.7 mg/dl) (p = NS), but phosphorus control was improved (4,7 +/- 0,5 mg/dl) (p < 0.01). The percentage of patients with PTH < 300 pg/ml decreased progressively from the start of treatment with alphacalcidol from 75% to 24% at the end of follow-up. Our results seem to suggest that the dose of alphacalcidol and calcitriol are not equivalent and we need to increase the dose of alphacalcidol to obtain a similar result to calcitriol on suppression of PTH in uremic patients with SPTH.

Adult↗

[Functional dependency evaluation of hemodialysis patients: a multicentric study].

BACKGROUND: There has been a change in the hemodialysis population characteristics over the last years with a progressive increase in patient,s age and associated comorbility and mortality. This older hemodialysis population are more functionally and medically dependent increasing the time taken to perform nursing work. The objective of this study was to evaluate the degree of functional dependency on hemodialysis patients and the need of care by nursing workload. METHODS: A transversal descriptive study was done during 1 month (april 2005) on 586 patients from 10 HD Units in Spain. No exclusion criteria were used. The Delta Test, who was used to evaluate the patients dependency needs, is a workload measure instrument base on three subscales: dependency, Physical Deficiency and mental deficiency. The indicators are measure on a scale of one to three with each level representing an increasing demand on nursing time. The results obtained from the Delta Test were analyzed taking into account the following variables: age, average time in dialysis, Charlson comorbidity Index, geographic location and HD unit. RESULTS: 46% of the patients show some degree of dependency, of these 12.8% were moderated and 8.1% severe; the subscale analysis showed that 19.6% and 6.7% had, respectively, a moderate to severe physical and mental health deficiency. The dependency degree varied significantly between HD Units and geographic location with a range of 0% to 59.8%. The degree of dependency were statistically associated with age and CCI. The higher CCI (r: 0,21; p < 0.001) and age (r: 0,26; p < 0.001) the higher was the Delta Test Score for dependency level. Patients times of initiation on dialysis were not associated with an increase in the degree of dependency. The aspects evaluated by the Delta Test that showed a higher score were those related to patients mobility. The patients assistance requirements during the HD session are basically related to a lack of mobility due to musculoskeletal disease and to a lesser extent to behavior alterations. CONCLUSIONS: HD units are attending patients with an important degree of dependency which impose and added workload to the healthcare personnel. In occasions, it is very difficult to attend and give proper care with the current legally established nurse to patient ratio which. This suggests the need to implement a better staffing policy. The Delta Test provides an objective, adaptable and standardized instrument for measuring degree of dependency of HD patients.

Activities of Daily Living↗

[Evaluation of health-related quality of life (HRQL) based on information from Coop/Wonca sheets in hemodialysis patients].

UNLABELLED: There has been increasing interest to find instruments to assess the HRQL quickly and easily in daily clinical practice. The main objective of this study is to measure the HRQL inhemodialysis patients using the Coop-Wonca Charts and to analyse whether they can be a useful instrument to assess the patient functional status and the HRQL in this population. METHODS: A descriptive cross-sectional study was performed between March 2003 and May 2003 in 163 hemodialysis patients (106 males and 57 females). The HRQL has been measured according to the validated Spanish version of the Coop-Wonca Charts. The questionnaire has nine different scales with a single-item. The answers score from 1 to 5 with the higher scores indicating the worst health or functional problems. RESULTS: The filling middle time of Coop-Wonca charts was less than five minutes. The charts were easily understood for the patients and can be self-administered. The dimensions of "Physical fitness" (3.66 +/- 0.8) and "Overall health" (3.43 +/- 0.8) showed the higher score, and the item aSocial activities) showed the lower scores (better quality of life) (1.98 +/- 1.3). The factors related to the worse health state are the variables: sex (females), the comorbility (diabetes and/or hepatopathy), the labour status (retired) and the way of transport (taxi/ambu lance). The time in hemodialysis was an independent variable and showed the worse score in the "general quality of life" (p < 0.05) and in the "perception of pain" (p < 0.01). The way of transport used (ambulances) was also an independent variable with worse scores for the dimensions "Physical fitness" (p < 0.01), "Daily activities" (p < 0.05) and "Health status" (p < 0.05). The older patients showed a worse score in the dimension "Physical fitness" (p < 0.01) and the patients with more than prescribed medicines scored worse in "Social activities" (p < 0.01). Males (p < 0.05), widows and single (p < 0.05) scored worse in social support. CONCLUSIONS: The Coop-Wonca Charts could be a good instrument to measure of HRQL in the HD patients. Their application is easy, fast (less than 5 minutes), easily understood, and can be self-administered.

Activities of Daily Living↗

Secretion of the rotavirus VP8* protein in Lactococcus lactis.

Secretion of the VP8* subunit of the VP4 capsid protein of rotavirus by Lactococcus lactis has been achieved. For this purpose, a secretion vector has been constructed with the lactococcal signal sequence AL9 and the VP8*-encoding gene fragment. The amount of VP8* secreted by L. lactis in the culture supernatant was quantified and visualised by Western blot. Furthermore, it was shown to retain its hemagglutination capability, indicating that the conformation of the secreted peptide may be retaining its biological activity.

Amino Acid Sequence↗

Alterations in sperm protein phosphorylation in male infertility.

Protein phosphorylation is involved in sperm capacitation, so the effect of protein phosphatase inhibitors on the capacitation of spermatozoa of males with unexplained infertility was investigated. d-mannose ligand specific receptor expression in fresh, living spermatozoa, capacitated or treated with calyculin A (an inhibitor of protein phosphatases 1 and 2A), was studied in three groups of men: pre-vasectomy (fertile) males, males in couples with male infertility, and males in couples with infertility of unknown aetiology. Flow cytometry showed significant differences between infertile couples with a male factor and fertile couples (P < 0.05), both after capacitation and after treatment with calyculin A. In the group of couples with infertility of unknown aetiology (n = 15), d-mannose receptor expression was diminished in six cases after classical capacitation. However, when the spermatozoa of these six men were treated with calyculin A, five showed an increased specific d-mannose receptor expression. From these results it is suggested that in vitro treatment of spermatozoa with inhibitors of protein phosphatases may be of great value in some cases of unexplained infertility.

Flow Cytometry↗

[Distribution of hepatitis C virus genotypes among the hemodialysis population in the province of Alicante].

UNLABELLED: Hepatitis C virus (HCV) genotypes are irregularly distributed among the different geographic area and groups at risk. OBJECTIVE: To study the different HCV genotypes and subtypes of hemodialyzed patients from Alicante. METHODS: We studied 640 patients on haemodialysis (HD) and we determined the RNA-HCV and the genotypes in the 120 patients with antibodies against HCV (HCV-Ab). We compared the results with the genotypes of 1,370 patients from other groups at risk in the same geographic area. RESULTS: RNA-HCV was not found in the serum in 15% (18/120) of the patients on HD who were HCV-Ab positive. Prevalence of the different genotypes in the 102 patients with positive viral RNA was the following: 1b: 56.8% (58/102), 1a: 19.6% (20/102), 3: 17% (17/102), 2a-2c: 1.9 (2/102), 2b: 0.9% (1/102) 4: 2.9 (3/102), 5: 0.9% (1/102). In conclusion, the genotype 1b was the most frequent in the patients studied in all these areas, and was the same as in the rest of the country. This genotype has been associated with the most severe hepatic disease and poor response to treatment, affecting the prognosis of these patients. The most frequent genotypes in HD in Alicante were 1b, 3 and 1a. HCV genotypes distribution among the HD units was not uniform in the different geographic areas. HCV genotypes distribution in the HD population is similar to other groups at risk from the same geographic area.

Adult↗

VP7 and VP4 genotypes among rotavirus strains recovered from children with gastroenteritis over a 3-year period in Valencia, Spain.

Between September 1996 and May 1999, the incidence and distribution of the main human rotavirus G genotypes (VP7 associated: G1-G4) and P genotypes (VP4 associated: P[8], P[4], P[6] and P[9]) among children with rotavirus gastroenteritis were determined using reverse transcription and polymerase chain reaction (RT-PCR)-based genotyping methods. From a total of 145 rotavirus strains examined, we identified the G type in 131 (90.3%) and the P type in 127 (87.5%) of the samples. An overall predominance of genotypes P[8] G1 (42.7%) and P[8] G4 (32.4%) was found during the period of study, with much lower incidence of genotypes P[4] G2 (5.5%) and P[8] G3 (2%). P[6] and P[9] types were not detected, neither were unusual combinations of P and G types. A significant genotypic shift was observed: whereas P[8] G4 was the most prevalent genotype during the first year of the study (60%), the genotype P[8] G1 gradually increased to account for 62.3% of the strains analysed in the following winter season. Mixed G types revealing dual infections G1/G4 and G3/G4 were found at low frequency (2%).

Acute Disease↗

Homotypic protection against rotavirus-induced diarrhea in infant mice breast-fed by dams immunized with the recombinant VP8* subunit of the VP4 capsid protein.

The outer capsid proteins VP4 and VP7 induce neutralizing antibody against rotavirus. We have investigated in a mouse model the protection mediated by immunization with VP8*, the amino-terminal tryptic fragment of VP4. BALB/c female mice immunized with simian rotavirus SA11 VP6 and VP8* proteins expressed in Escherichia coli were mated with seronegative males. Litters were orally challenged with the SA11 strain (P5B[2], G3) or with the murine rotavirus strain EDIM (P10[16], G3) to verify the degree of protection against diarrhea induced in the newborns. Only those pups born to dams immunized with VP8* did not develop diarrhea after having been orally challenged with the SA11 strain. Pups born to naive dams but foster nursed by VP8*-immunized dams did not develop diarrhea after having been orally infected with the SA11 strain, but they suffered diarrhea when challenged with the EDIM strain. These results support the concepts that (1) VP8* is a highly immunogenic polypeptide that induces effective homotypic protection against disease in pups born to dams immunized with this antigen and (2) in newborn mice the protection against disease is mediated by neutralizing secretory antibodies present in the milk rather than by serum antibodies transferred through the placenta to the offspring.

Animals↗

Decreased sperm function of patients with myotonic muscular dystrophy.

Myotonic muscular dystrophy (MMD) is a genetic disease caused by a defective enzyme, myotoninkinase. Male patients with MMD are reported to have reduced fertility. The purpose of this work was to study sperm capacitation and acrosome reaction in the ejaculates of sterile males with MMD and of healthy males (control group). The expression of the specific D-mannose receptors was explored by microscopic examination and by flow cytometry analysis. In addition, the binding patterns of Pisum sativum (PSA) lectin to acrosome content and outer acrosomal membrane in the spermatozoa of each group were analysed. Both the capacitation and the acrosome reaction in the spermatozoa of the MMD group were deficient and these findings strongly suggest that these anomalies may account for the sterility of these patients.

Acrosome Reaction↗

Rotavirus-specific cytotoxic T lymphocytes recognize overlapping epitopes in the amino-terminal region of the VP7 glycoprotein.

Rotavirus-specific cytotoxic T lymphocytes (CTL) play an important role in the resolution of rotavirus infection. The outer capsid glycoprotein, VP7, elicits a class I MHC-restricted CTL response. Vaccinia virus recombinants expressing the VP7 genes from simian rotavirus SA11 (serotype G3) and from the RF strain of bovine rotavirus (serotype G6) were used to analyze the CTL activity to this antigen in BALB/c (H-2(d)) and C57BL/6 (H-2(b)) mice neonatally infected with homologous and heterologous rotaviruses. A vaccinia virus recombinant expressing the first amino-terminal 88 amino acids of VP7 was constructed and used to search for cross-reactive CTL against this region of the protein. By using synthetic Kb, Db, and Kd motif-fitting peptides two overlapping CTL epitopes have been identified located in the first hydrophobic domain (H1) of VP7. Splenocytes obtained from rotavirus SA11-infected C57BL/6 mice induced the strongest CTL response against target cells sensitized with a peptide containing a Kb-restricted CTL epitope (amino acids 8-16). A second Kd-restricted epitope (residues 5-13) was recognized by splenocytes derived from rotavirus-infected BALB/c mice. These findings reveal the existence of CTL epitopes in the H1 signal sequence of the VP7 glycoprotein that coexist with a CTL epitope (residues 31-40) previously described within the H2 region.

Animals↗

Viral proteins VP2, VP6, and NSP2 are strongly precipitated by serum and fecal antibodies from children with rotavirus symptomatic infection.

Rotavirus-specific IgA has been correlated with immune protection against rotavirus reinfection and symptomatic disease. Systemic and mucosal antibody responses were determined by an enzyme-linked immunosorbent assay in 11 infants with severe rotavirus gastroenteritis. Geometric mean titers of antirotavirus serum IgG and IgA antibodies were significantly higher during the convalescence of the disease (P < 0.001 vs. acute-phase titers). Rotavirus-specific fecal sIgA antibodies increased 4 times during the convalescence in 9 (81.8%) children (P < 0.001). The serum IgG and IgA antibody and fecal sIgA antibody responses to individual rotavirus polypeptides were characterized by radioimmunoprecipitation assay (RIPA) using Staphylococcus aureus protein A and the lectin jacalin to precipitate IgG- and IgA-immune complexes, respectively. The main IgG response was directed toward the structural viral proteins VP2, VP4, and VP6 and toward the nonstructural protein NSP2. Serum IgA reactivity was detected by RIPA in all serum samples, with major responses to VP2, VP6, and NSP2. Interestingly, fecal sIgA in convalescent samples reacted strongly toward NSP2 and VP6. These data reinforce the antigenic importance of rotaviral proteins other than VP4 and VP7, such as VP2, VP6, and NSP2, as main targets in the immune response to rotavirus.

Antibodies, Viral↗

[Virus-specific serum and fecal antibodies response in children with acute rotavirus gastroenteritis].

BACKGROUND: The analysis of the immune response to rotavirus infection and the characterization of the viral antigens recognized by specific antibodies are of great concern in evaluating the protection against rotavirus. MATERIAL AND METHODS: The levels of rotavirus-specific fecal (sIgA) and serum antibodies (IgM, IgG and IgA) were evaluated by ELISA in 25 children with acute gastroenteritis for rotavirus, in 11 of them during the acute and convalescent phases. The specificity of serum antibodies to viral polypeptides was characterized by immunoblotting. RESULTS: Serum IgM antibodies with a geometric mean titer (GMT) of 1/3,973 were the predominant antibodies detected during the acute phase. In comparison, IgG and IgA serum antibodies and sIgA coproantibodies levels were higher in the convalescent phase (GMT = 1/5,799, 1/257 and 1/137 respectively). Significant differences were observed for all the isotypes of immunoglobulins evaluated during the infection and in the convalescence (p < 0.01). Rotavirus-specific serum antibodies recognized mainly the structural VP6, VP7 and VP3/VP4 proteins. Other polypeptides also detected were VP1, VP5 and the non-structural NS34 protein. CONCLUSIONS: Rotavirus infection produce an intense humoral immune response both in serum and in the gut. Specific antibodies react against structural proteins of the internal (VP6) and external (VP7) capsids of rotavirus.

Antibodies, Viral↗