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Biomedical subjects

I Díaz

Publications and source records attributed to I Díaz.

18 recordsLinked to original sources

A constitutive cystatin-encoding gene from barley (Icy) responds differentially to abiotic stimuli.

A barley cDNA clone encoding a cysteine proteinase inhibitor was characterized. The deduced amino acid sequence of this barley cystatin (Hv-CPI) contains the motif QXVXG conserved among members of the cystatin superfamily. The gene (Icy), located on chromosome 2, was expressed in embryos, developing endosperms, leaves and roots as assessed by northern blot analysis. Western blot analysis detected a slightly retarded band in leaves that was not present in roots or seeds. In these two organs a more precise location of Hv-CPI was done by immuno-histochemical analysis, with polyclonal antibodies raised against the recombinant CPI protein expressed in Escherichia coli. This protein efficiently inhibited papain (Ki 2.0 x 10(-8) M) and ficin (Ki 2.2 x 10(-8) M) and, to a lesser extent, chymopapain (Ki 1.6 x 10(-7) M) and was inactive against bromelain. The Icy mRNA expression in vegetative tissues increased in response to anaerobiosis, dark and cold shock (6 degrees C).

Amino Acid Motifs↗

Removal of bowel aerobic gram-negative bacteria is more effective than immunosuppression with cyclophosphamide and steroids to decrease natural alpha-galactosyl IgG antibodies.

Natural alpha-Galactosyl (Gal) antibodies play an important role in the rejection of pig xenografts by humans and Old World monkeys. In this study we investigate the efficacy of two different strategies to reduce the serum level of natural anti-Gal antibodies. On the one hand, removal of aerobic gram-negative bacteria from the intestinal flora, because anti-Gal antibodies appear to be produced as a result of the continuous sensitization by these microorganisms. On the other hand, we studied the effect on these antibodies of an immunosuppressive regimen of cyclophosphamide and steroids. Ten baboons were treated for three months with norfloxacin (Nor Group; n=6) or cyclophosphamide and steroids (CyP Group; n=4). A further four baboons did not receive any treatment (Control Group). Aerobic gram-negative bacteria became negative in stools of the Nor Group after two weeks of treatment, and remained undetectable until week 7. Thereafter, a gradual increase on the fecal concentration of aerobic gram-negative bacteria was observed despite the norfloxacin treatment. The mean anti-Gal IgG in the Nor Group gradually declined from week 4 to 9 to a mean of 62.7 +/- 18% of the baseline level, and during this period were significantly lower than in the CyP (P<0.02) and the Control (P<0.05) groups. No differences were observed between the three groups during the 16 weeks of follow-up in serum levels of anti-Gal IgM, hemolytic anti-pig antibodies, total IgG, IgM and IgA. In conclusion, removal of normal aerobic gram-negative bacteria from the intestinal flora is more effective than immunosuppression with CyP and steroids in reducing the level of natural anti-Gal antibodies, although there is no discernible effect on IgM antibodies.

Animals↗

Impact of stage III-IV endometriosis on recipients of sibling oocytes: matched case-control study.

OBJECTIVE: To evaluate the impact of severe endometriosis on IVF-ET outcome in women receiving oocytes from the-same donor. DESIGN: A matched case-control study. SETTING: Oocyte donation program at the Instituto Valenciano de Infertilidad. PATIENT(S): Fifty-eight recipients were included in a matched case-control study of IVF-ET in our oocyte donation program. Twenty-five patients were diagnosed by laparoscopy with stage III-IV endometriosis (group I), while the remaining 33 were free of the disease (group II). On the day of retrieval, oocytes from a single donor were donated to recipients from both groups. Some of the donors supplied oocytes for more than 2 patients. Recipients received steroid replacement therapy for endometrial preparation. INTERVENTION(S): Ovarian stimulation and oocyte retrieval in donors. Uterine embryo transfer (ET) in recipients after appropriate exogenous hormone replacement therapy (HRT). MAIN OUTCOME MEASURE(S): Pregnancy, implantation, miscarriage, and live birth rates. RESULT(S): The number of oocytes donated and fertilized, as well as the number of available and transferred embryos, was not statistically different between the two groups. Pregnancy, implantation, and miscarriage rates were not affected by stage III-IV endometriosis when compared with the control group. The live birth rate was 28.0% in the group with endometriosis and 27.2% in the control group. CONCLUSION(S): These results show that implantation is not affected by stage III-IV endometriosis. Given the contemporary methods of endometrial preparation for transfer of embryos derived from donor oocytes, any potential negative effect of severe endometriosis on the uterine environment is undetectable.

Abortion, Spontaneous↗

A QTL on pig chromosome 4 affects fatty acid metabolism: evidence from an Iberian by Landrace intercross.

Three Iberian boars were bred to 31 Landrace sows to produce 79 F1 pigs. Six F1 boars were mated to 73 F1 sows. The F2 progeny from 33 full-sib families (250 individuals) were genotyped for seven microsatellites spanning the length of chromosome 4. Least squares procedures for interval mapping were used to detect quantitative trait loci (QTL). A permutation test was used to establish nominal significance levels associated with QTL effects, and resulting probability levels were corrected to a genomewide basis. Observed QTL effects were (genomewide significance, position of maximum significance in centimorgans): percentage of linoleic acid in subcutaneous adipose tissue (< 0.01, 81); backfat thickness (< 0.01, 83); backfat weight (< 0.01, 80); longissimus muscle area (0.02, 83); live weight (0.19, 88); and percentage of oleic acid in subcutaneous adipose tissue (0.25, 81). Gene action was primarily additive. The Iberian genotypes were fatter, slower growing, and had lower linoleic and higher oleic acid contents than Landrace genotypes. The interval from 80 to 83 cM contains the FAT1 and A-FABP loci that have been shown previously to affect fat deposition in pigs. This is the first report of a QTL affecting fatty acid composition of subcutaneous adipose tissue in pigs and provides a guide for the metabolic pathways affected by candidate genes described in this region of chromosome 4.

Adipose Tissue↗

Molecular cloning of a zinc finger autoantigen transiently associated with interphase nucleolus and mitotic centromeres and midbodies. Orthologous proteins with nine CXXC motifs highly conserved from nematodes to humans.

We have cloned a novel human autoimmune antigen in a patient suffering from rheumatoid arthritis with high levels of antibodies to the nucleolus organizer regions. Initially the human autoimmune serum was used to select a cDNA of 317 amino acids from a hamster expression library. Using the hamster DNA as a probe, we isolated the human homologous cDNA of 320 amino acids. Human and hamster polypeptides share a 95% amino acid homology. The deduced 36-kDa protein contains a putative amino-terminal NLS signal, nine cysteine-X-X-cysteine motifs highly conserved, and a carboxyl-terminal poly acidic region. Several homologous expressed sequence tags have been identified in data bases suggesting that orthologous proteins are present throughout evolution from worms to humans. A Drosophila expressed sequence tag was further completely sequenced for a full-length protein with 60% amino acid identity to the human homologue. Northern blot analysis revealed that this novel protein is widely distributed in human tissues with significantly higher expression levels in heart and skeletal muscle. Specific antibodies to the recombinant protein and transfection experiments demonstrated by immunofluorescence the localization of the protein predominantly but not exclusively to the nucleolus of interphase mammalian cells. In actinomycin D-treated cells the protein remains associated with the nucleolus but is not segregated, like other ribosomal factors such as upstream binding factor. In mitosis the protein was found to be associated with centromeres and concentrated at the midbody in cytokinesis. Transient distribution of this evolutionarily conserved zinc finger nucleolar autoantigen to the mitotic centromeres may provide the means for several aspects of cell cycle control and transcriptional regulation.

Amino Acid Sequence↗

Barley BLZ2, a seed-specific bZIP protein that interacts with BLZ1 in vivo and activates transcription from the GCN4-like motif of B-hordein promoters in barley endosperm.

A barley endosperm cDNA, encoding a DNA-binding protein of the bZIP class of transcription factors, BLZ2, has been characterized. The Blz2 mRNA expression is restricted to the endosperm, where it precedes that of the hordein genes. BLZ2, expressed in bacteria, binds specifically to the GCN4-like motif (GLM; 5'-GTGAGTCAT-3') in a 43-base pair oligonucleotide derived from the promoter region of a Hor-2 gene (B1-hordein). This oligonucleotide also includes the prolamin box (PB; 5'-TGTAAAG-3'). Binding by BLZ2 is prevented when the GLM is mutated to 5'-GTGctTCtc-3' but not when mutations affect the PB. The BLZ2 protein is a potent transcriptional activator in a yeast two-hybrid system where it dimerizes with BLZ1, a barley bZIP protein encoded by the ubiquitously expressed Blz1 gene. Transient expression experiments in co-bombarded developing barley endosperms demonstrate that BLZ2 transactivates transcription from the GLM of the Hor-2 gene promoter and that this activation is also partially dependent on the presence of an intact PB. A drastic decrease in GUS activity is observed in co-bombarded barley endosperms when using as effectors equimolar mixtures of Blz2 and Blz1 in antisense constructs. These results strongly implicate the endosperm-specific BLZ2 protein from barley, either as a homodimer or as a heterodimer with BLZ1, as an important transcriptional activator of seed storage protein genes containing the GLM in their promoters.

Amino Acid Sequence↗

Microcomputer decision support system for intensive care.

This article describes a computer system to support decisions in Intensive Medicine developed for a multipurpose Intensive Care Unit (13 beds and about 700 yearly patients). The system is used in a microcomputer (486DX4). Works done in this area are mentioned; and the different programs which integrate the system, the advantages of its use and the results accomplished are described. Some of the most important results are the quality increment of the medical care given, physicians' possibilities of dedicating more time to their patients, cost reductions, and improvements in the development of teaching and in the conditions to carry out research work in the Intensive Care Unit (ICU). An analysis is made of the system requirements that guarantee its integration to the Unit work, and the easy and safe use of it by the physicians and nurses.

Critical Care↗

[Diagnostic biopsy with paranasal sinus endoscopy].

Rigid endoscopy allows clear visualization of the nasal fossa and paranasal sinuses and permits biopsies to be obtained under direct vision from relatively inaccessible sites. In most cases the biopsy can be made using local anesthesia, but selected patients require general anesthesia. We report the results of 31 patients who underwent biopsy with rigid nasal endoscopy under general anesthesia. Of the 31 patients, a diagnostic biopsy was achieved in 30 (96.7%). We define a diagnostic biopsy as one which yielded a histopathological diagnosis that did not change as a result of clinical findings, examinations or further biopsies performed during follow-up. We recommend the use of endoscopic nasal biopsy of nasal and paranasal neoplasms as an easy, safe and reliable technique.

Adenocarcinoma↗

Antinociceptive effects of S(+)-ketoprofen and other analgesic drugs in a rat model of pain induced by uric acid.

We investigated the antinociceptive properties of dexketoprofen trometamol [S(+)-ketoprofen tromethamine salt; SKP], a new analgesic, antiinflammatory drug, using the pain-induced functional impairment model in the rat (PIFIR), an animal model of arthritic pain. SKP was compared with racemic ketoprofen tromethamine salt (rac-KP), R(-)-ketoprofen tromethamine salt (RKP), ketorolac (KET), and morphine (MOR). We also assessed the effects of flurbiprofen (rac-FB) and its enantiomers (SFB and RFB) in the same model. Groups of six rats received either vehicle or analgesic drug and antinociception was evaluated by evaluating the dose-response curves over time. SKP was an effective antinociceptive drug in this model and was almost equally potent by either oral or intracerebroventricular administration. The oral potency of SKP was similar to that of oral KET and greater than that of oral MOR. No significant differences were observed between racemic ketoprofen and its enantiomers when administered orally. In the rat, significant bioinversion of RKP to SKP occurs when RKP is given orally. After oral administration of RKP, SKP was detectable in 30 min and surpassed the concentration of RKP after 3 h. Nevertheless, when the compounds were given intracerebroventricularly, some stereoselectivity in favor of SKP was observed. Stereoselectivity was observed with flurbiprofen, an analogue of ketoprofen that does not undergo significant metabolic inversion. Whereas SFB was an effective antinociceptive, RFB had no antinociceptive effect at the doses tested when given either orally or intracerebroventricularly.

Administration, Oral↗

Determination of neutral lipids from subcutaneous fat of cured ham by capillary gas chromatography and liquid chromatography.

The determination of neutral lipids in fat of cured ham is reported. Fat samples were extracted with chloroform-methanol (2:1) and neutral lipids and free fatty acids were separated on an aminopropyl minicolumn, the first fraction with chloroform-2-propanol (neutral lipids) and the second fraction with 2% acetic acid in diethyl ether (free fatty acids). Neutral lipids were fractionated with minicolumns, with aminopropyl and silica stationary phases. Two fractions were obtained with the first column: (A) triglyceride and cholesteryl esters and (B) cholesterol and mono- and diglycerides. Fraction A was applied to the silica column to obtain two new fractions: (C) cholesteryl esters and (D) triglycerides. Fractions B and C were analysed by capillary gas chromatography (cGC) and fraction D by cGC and HPLC. The R.S.D.s obtained were below 5% except for the monoglycerides (8%). Cholesteryl esters were determined by cGC in 5 min with R.S.D. 5%. The main triglycerides identified were PPO, POS, POO, POL and OOO (P = palmitic acid, O = oleic acid, L = linoleic acid; S = stearic). Monoglyceride and diglycerides having 18, 34 and 36 carbon atoms were the most abundant. The determination of triglycerides by HPLC was more difficult than by cGC because the linearity with HPLC was concentration dependent. The procedure allowed the determination of neutral lipid classes without derivatization of mono- and diglycerides.

Adipose Tissue↗

Mapping helper virus functions for cucumber mosaic virus satellite RNA with pseudorecombinants derived from cucumber mosaic and tomato aspermy viruses.

P-TAV is a strain of tomato aspermy virus (TAV) able to efficiently support the systemic accumulation of some (i.e., B2-satRNA) but not of other (i.e., Ix-satRNA) strains of the satellite RNA (satRNA) of cucumber mosaic virus (CMV) in both tobacco and in tomato. As reported for V-TAV, the failure to support the systemic accumulation of Ix-satRNA seems to be due to an inefficient support of its systemic movement. Pseudorecombinants obtained by the exchange of RNAs 1 + 2 between P-TAV and Trk7-CMV, an efficient helper for the systemic accumulation of Ix-satRNA, were assayed for their ability to support the accumulation of CMV-satRNAs in tobacco plants and protoplasts. Pseudorecombinants having RNAs 1 + 2 from CMV supported the systemic movement and accumulation of CMV-satRNA as efficiently as CMV, whereas pseudorecombinants having RNAs 1 + 2 from TAV supported the CMV-satRNA very poorly. Thus, the ability to support the systemic movement and accumulation of CMV-satRNA is determined primarily by RNAs 1 + 2 and not by RNA 3, which is presumed to encode movement functions in the cucumoviruses and only has a minor, modulating effect on the systemic accumulation of satRNA. This suggests that for systemic movement CMV-satRNA has to interact with (the gene products of) RNAs 1 and/or 2 or that these viral RNAs compete with the satRNA for interaction with the coat or other movement proteins.

Cucumovirus↗

ms2i6A deficiency enhances proofreading in translation.

The hypermodified base 2-methylthio-N6-isopentenyladenosine (ms2i6A) at position 37 occurs frequently in tRNAs that read codons starting with uridine. Here we have studied how ms2i6A affects the accuracy of poly(U) translation in vitro. Deficiency leads to a higher rejection rate of tRNA4(Leu) by more aggressive proofreading on the wild-type ribosome, but with the initial selection step unchanged. Our data indicate that ms2i6A has no effect on codon-anticodon interactions on wild-type ribosomes as long as aminoacyl-tRNA is in ternary complex with EF-Tu and GTP. ms2i6A deficiency in the cognate poly(U) reader tRNA(Phe) leads to increased misreading when the near-cognate competitor tRNA4(Leu) is wild-type. ms2i6A deficiency in tRNA4(Leu) gives a decreased error level in competition with wild-type tRNA(Phe).

Codon↗

[Treatment of bladder endometriosis using the Nd-Yag laser].

The good results achieved in the treatment of gynecological endometriosis via laser laparoscopy prompted us to perform endoscopic Nd:Yag laser photocoagulation in the present case of symptomatic vesical endometriosis described herein. To our knowledge, this is the first reported case in the world literature.

Adult↗

[Our experience in the treatment of superficial cancer of the bladder with BCG].

We report on our experience with 21 patients with superficial bladder cancer who were treated with intravesical BCG instillation. The BCG vaccine was effective in the treatment of carcinoma in situ. Tumor regression was achieved in 83.3% at 3 months, and 61.1% at 24 months. It is advisable to follow patients closely because of the possibility of tumor progression.

Adult↗

Nuclear magnetic resonance in advanced prostate cancer (stage D).

Nuclear Magnetic Resonance (NMR), a recently developed diagnostic method, has proved to be useful in the study of patients with prostatic disease. The authors report their experience with this technique in 10 cases of advanced prostate cancer (stage D) treated with LH-RH agonists (HOE 766-Buserelin). NMR provided information on the size of the prostate, the invasion of the vesical floor and seminal vesicles and the involvement of the pelvic and retroperitoneal nodes. It was also useful for ruling out metastases in the pelvis and spinal column, and showed an excellent correlation with the bone scan. In 2 untreated cases, NMR was sufficient by itself to diagnose metastatic prostate cancer. However, given the current lack of experience, it is not yet possible to draw conclusions concerning the superiority of this technique over other methods in the diagnosis and staging of prostatic carcinoma.

Adenocarcinoma↗