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Biomedical subjects

A Duran

Publications and source records attributed to A Duran.

At least 37 records · Page 2Linked to original sources
PubMed1997

Characterization of CHS4 (CAL2), a gene of Saccharomyces cerevisiae involved in chitin biosynthesis and allelic to SKT5 and CSD4.

We have cloned CHS4, a gene that complements the resistance to Calcofluor of the Saccharomyces cerevisiae cal2 mutant. We show that CHS4 is allelic to the previously described SKT5 and CSD4 genes. CHS4 encodes a 696 amino acids protein with no potential transmembrane domain. chs4-null mutants are resistant to Calcofluor white and exhibit a considerable reduction in cell wall chitin and in chitin synthase III (CSIII) activity. Biochemical characterization of chitin synthase III from these null mutants indicates that the defect is due to a reduced V(max) of the enzyme. This defect can be overcome in vitro by trypsin treatment of the membrane preparations. Chs4p does not act as a transcriptional or translational regulator of CHS3, the gene coding for the catalytic subunit of CSIII activity, and we therefore propose that Chs4p would be an essential component of the CSIII complex, acting as a post-translational regulator of this activity. In addition to the chitin defect, the chs4 mutant shows a severe defect in mating.

Alleles↗
PubMed1997

CHS5, a gene involved in chitin synthesis and mating in Saccharomyces cerevisiae.

The CHS5 locus of Saccharomyces cerevisiae is important for wild-type levels of chitin synthase III activity. chs5 cells have reduced levels of this activity. To further understand the role of CHS5 in yeast, the CHS5 gene was cloned by complementation of the Calcofluor resistance phenotype of a chs5 mutant. Transformation of the mutant with a plasmid carrying CHS5 restored Calcofluor sensitivity, wild-type cell wall chitin levels, and chitin synthase III activity levels. DNA sequence analysis reveals that CHS5 encodes a unique polypeptide of 671 amino acids with a molecular mass of 73,642 Da. The predicted sequence shows a heptapeptide repeated 10 times, a carboxy-terminal lysine-rich tail, and some similarity to neurofilament proteins. The effects of deletion of CHS5 indicate that it is not essential for yeast cell growth; however, it is important for mating. Deletion of CHS3, the presumptive structural gene for chitin synthase III activity, results in a modest decrease in mating efficiency, whereas chs5delta cells exhibit a much stronger mating defect. However, chs5 cells produce more chitin than chs3 mutants, indicating that CHS5 plays a role in other processes besides chitin synthesis. Analysis of mating mixtures of chs5 cells reveals that cells agglutinate and make contact but fail to undergo cell fusion. The chs5 mating defect can be partially rescued by FUS1 and/or FUS2, two genes which have been implicated previously in cell fusion, but not by FUS3. In addition, mating efficiency is much lower in fus1 fus2 x chs5 than in fus1 fus2 x wild type crosses. Our results indicate that Chs5p plays an important role in the cell fusion step of mating.

Amino Acid Sequence↗
PubMed1997

Localisation of the Schizosaccharomyces pombe rho1p GTPase and its involvement in the organisation of the actin cytoskeleton.

The Schizosaccharomyces pombe rho1p GTPase directly activates the (1-3) beta-D-glucan synthase and participates in the regulation of cell wall growth and morphogenesis in this fission yeast. Indirect immunofluorescence experiments using rho1p tagged with hemagglutinin have revealed that rho1p was located at the growing tips during interphase and at the septum prior to cytokinesis, localising to the same areas as actin patches. In S. pombe cdc10-129 mutant cells, arrested in G1, HA-rho1p accumulates at one tip whereas in cdc25-22 mutants, arrested in G2, HA-rho1p accumulates at both tips. In tea1-1 and tea2-1 cdc11-119 mutant cells, HA-rho1p is localised to the new growing tips. Overexpression of different rho1 mutant alleles caused different effects on cortical actin patch distribution, (1-3) beta-D-glucan synthase activation, and sensitivity to cell wall specific antifungal drugs. These results indicate that multiple cellular components are activated by rho1p. Overexpression of the dominant negative rho1T20N allele was lethal as was the rho1+ deletion. Moreover, when rho1+ expression was repressed in actively growing S. pombe, cells died in about 10 to 12 hours. Under these conditions, normal cell morphology was maintained but the level of (1-3) beta-D-glucan synthase activity decreased and the actin patches disappeared. Most cells lysed after cytokinesis during the process of separation, and lysis was not prevented by an osmotic stabiliser. We conclude that rho1p localisation is restricted to growth areas and regulated during the cell cycle and that rho1p is involved in cell wall growth and actin cytoskeleton organisation in S. pombe.

Actins↗
PubMed1996

Characterization of cwl1+, a gene from Schizosaccharomyces pombe whose overexpression causes cell lysis.

From a Schizosaccharomyces pombe genomic library we have isolated the gene cwl1+ that causes cell lysis when it is overexpressed in the absence of an osmotic stabilizer. Southern hybridization showed that cwl1+ exists as a single copy in the S. pombe genome. The cwl1+ gene nucleotide sequence revealed a putative open reading frame of 924 bp encoding a polypeptide of 308 amino acids with a calculated Mt of 27000. The cwl1+ DNA hybridizes to a major RNA transcript of 1.5 kb whose 5' end maps at a position 452 bp upstream from the predicted translation start. Comparison of the amino acid sequence with those included in the current databases, showed no significant similarity to any known sequences. Cells overexpressing the cwl1+ gene under the control of the S. pombe nmt inducible promoter displayed a reduced cell wall content, were unable to separate after division and lysed drastically in the absence of osmotic stabilizer. Disruption of the cwl1+ gene caused no noticeable phenotype.

Amino Acid Sequence↗
PubMed1996

New device for artificial increasing intraocular pressure.

BACKGROUND: For certain special investigations, particularly in glaucoma diagnostics, intraocular pressure has to be increased. Mostly, a suction cup apparatus is used for that purpose. These devices, however, have some disadvantages. They are at least uncomfortable, sometimes risky for the patient and induce a strong astigmatism which can influence the results of the corresponding investigation. METHOD: A very simple mechanical device has been constructed and tested, by means of which the intraocular pressure can be increased for diagnostic purposes. A ring on a lever, which is moved towards the eye by small weights, compresses the eye through the lids without the need of even local anesthetics. The device can be easily mounted to any head fixation which is normally used in combination with ophthalmological equipment. e.g. slit lamp, visual field analyzer or fundus camera. Tests have been performed in four different groups: one group of 19 healthy volunteers, one of 21 patients with Graves' disease, one of 21 patients with high myopia and one of 20 patients with open-angle glaucoma. RESULTS: Pressure elevations could be achieved in all patients with very good reproducibility with respect to the applied forces. However, in patients with exophthalmos (Graves' disease), the elevation effect was somewhat higher than in normal eyes. The induced astigmatism was only about 0.5 dpt for pressure elevations of about 20 mmHg. CONCLUSION: The tests performed showed that the new device is easy to handle and comfortable for the patients. If very high accuracy is needed concerning the pressure values achieved, an individual calibration should be performed.

Adult↗
PubMed1995

99mTc direct labeling of anti-CEA monoclonal antibodies: quality control and preclinical studies.

The anti-carcinoembryonic B2C114 monoclonal antibody was radiolabeled with 99mTc by a direct method and quality control tested in vitro by instant thin layer chromatography, gel column scanning and cellulose acetate electrophoresis and assessed in vivo for radioimmunodetection on a murine spontaneous mammary carcinoma. The optimal results of percent 99mTc bound to protein were obtained at a dithiothreitol:antibody molar ratio ranging from 800:1 to 1000:1 and at a methylene diphosphonate:stannous fluoride weight ratio of 4.3:1. Although cysteine removed up to 18% of the label during the first 4 h, the stability of the tracer appeared to be excellent in human serum at 37 degrees C and when challenged with DTPA. 99mTc-labeled B2C114 demonstrated good and specific in vivo tumor targeting.

Animals↗
PubMed1995

Papulacandin B resistance in budding and fission yeasts: isolation and characterization of a gene involved in (1,3)beta-D-glucan synthesis in Saccharomyces cerevisiae.

Papulacandin B, an antifungal agent that interferes with the synthesis of yeast cell wall (1,3)beta-D-glucan, was used to isolate resistant mutants in Schizosaccharomyces pombe and Saccharomyces cerevisiae. The resistance to papulacandin B always segregated as a recessive character that defines a single complementation group in both yeasts (pbr1+ and PBR1, respectively). Determination of several kinetic parameters of (1,3)beta-D-glucan synthase activity revealed no differences between S. pombe wild-type and pbr1 mutant strains except in the 50% inhibitory concentration for papulacandin B of the synthases (about a 50-fold increase in mutant activity). Inactivation of the synthase activity of both yeasts after in vivo treatment with the antifungal agent showed that mutant synthases were more resistant than the corresponding wild-type ones. Detergent dissociation of the S. pombe synthase into soluble and particulate fractions and subsequent reconstitution indicated that the resistance character of pbr1 mutants resides in the particulate fraction of the enzyme. Cloning and sequencing of PBR1 from S. cerevisiae revealed a gene identical to others recently reported (FKS1, ETG1, CWH53, and CND1). Its disruption leads to reduced levels of both (1,3)beta-D-glucan synthase activity and the alkali-insoluble cell wall fraction. Transformants containing the PBR1 gene reverse the defect in (1,3)beta-D-glucan synthase. It is concluded that Pbr1p is probably part of the (1,3)beta-D-glucan synthase complex.

Aminoglycosides↗
PubMed1994

Cisplatin-5-fluorouracil in small cell lung cancer. A phase II study in 109 patients.

One hundred nine previously untreated patients with small cell lung cancer were treated for five consecutive days with 20 mg/m2/day of cisplatin and 600 mg/m2/day of 5-fluorouracil. One cycle of chemotherapy was administered every three weeks. The patients received a median number of three cycles. Then they were transferred to CAE chemotherapy. A 77% overall response rate (95% confidence interval of 0.70-0.85) was observed after initial cisplatin-5FU treatment. Twenty-three complete responses (21%) and 62 partial responses (56%) were obtained. In cerebral metastases the response rate was high at 91% (21 out of 23), with 43% complete responses. In the limited forms, statistical survival at 1 year was 25%. A Grade 3-4 thrombocytopaenia was observed in 10 patients (9%) and a Grade 2-3 leukopaenia in four patients. Three patients suffered from a Grade 2 cardiac toxicity. The cisplatin-5-fluorouracil combination demonstrates promising initial response rate in small cell lung cancers. Its main interest is in its important action on cerebral metastases and its moderate haematological toxicity.

Adult↗
PubMed1994

Synchronized intermittent mandatory ventilation with and without pressure support ventilation in weaning patients with COPD from mechanical ventilation.

This prospective study compared two weaning modalities in COPD patients requiring mechanical ventilation (MV) for acute respiratory failure. Nineteen patients with COPD were studied when their precipitating illness was controlled. Although they satisfied the conventional bedside weaning criteria, they could not tolerate any reduction in the respirator rate below 10 cycles/min. At this time, patients were randomized into two groups receiving either synchronized intermittent mandatory ventilation (SIMV) with pressure support ventilation (PSV) (group 1) or SIMV alone (group 2). The volumetric support of ventilation (SIMV rate) was progressively decreased in both groups according to the patient's tolerance with a concurrent decrease in the barometric support of ventilation (PSV levels from 15 cm H2O to 6 cm H2O). At each step of SIMV rate, we found no difference between group 1 and group 2 in arterial blood gases, blood pressure, heart rate, airway occlusion pressure, maximal inspiratory pressure, and oxygen cost of breathing (OCB). At each step, however, group 1 patients showed significantly higher spontaneous tidal volume and lower spontaneous breathing frequency than did group 2 patients. We found a slight but not significant tendency to a shorter weaning period with than without PSV, but no difference in the weaning success. We concluded that (1) conventional weaning criteria might be inaccurate in COPD patients, (2) SIMV appeared very useful in weaning COPD patients from MV, (3) PSV marginally reduced the weaning period when added to SIMV, and (4) the OCB was not significantly improved with PSV.

Aged↗
PubMed1993

The Schizosaccharomyces pombe cwg2+ gene codes for the beta subunit of a geranylgeranyltransferase type I required for beta-glucan synthesis.

The product of the Schizosaccharomyces pombe cwg2+ gene is involved in the biosynthesis of beta-D-glucan. When grown at the non-permissive temperature, cwg2-1 mutant cells lyse in the absence of an osmotic stabilizer and display a reduced (1-3) beta-D-glucan content and (1-3) beta-D-glucan synthase activity. The cwg2+ gene was cloned by the rescue of the cwg2-1 mutant phenotype using an S. pombe genomic library and subsequently verified by integration of the appropriate insert into the S. pombe genome. Determination of the nucleotide sequence of this gene revealed a putative open reading frame of 1065 bp encoding a polypeptide of 355 amino acids with a calculated M(r) of 40,019. The cwg2+ DNA hybridizes to a main transcript, the 5' end of which maps to a position 469 bp upstream of the predicted start of translation. The sequence between the transcription and the translation start sites is unusually long and has several short open reading frames which suggest a translational control of the gene expression. Comparative analysis of the predicted amino acid sequence shows that it possesses significant similarity to three Saccharomyces cerevisiae proteins, encoded by the DPR1/RAM1, CDC43/CAL1 and ORF2/BET2 genes respectively, which are beta subunits of different prenyltransferases. When grown at 37 degrees C, cwg2-1 mutant extracts were specifically deficient in geranylgeranyltransferase type I activity, as measured in vitro. Multiple copies of the CDC43 gene can partially suppress the growth and (1-3) beta-D-glucan synthase defect of the cwg2-1 mutant at the restrictive temperature. In a similar manner, the cwg2+ gene can partially suppress the cdc43-2 growth defect. These results indicate that cwg2+ is the structural gene for the beta subunit of geranylgeranyltransferase type I in S. pombe and that this enzyme is required for (1-3) beta-D-glucan synthase activity. The functional homology of Cwg2 with Cdc43, which has been implicated in the control of cell polarity, suggests a link between two morphogenetic events such as establishment of cell polarity and cell wall biosynthesis.

Amino Acid Sequence↗
PubMed1993

A shared pathway in atrioventricular nodal reentrant tachycardia and atrial flutter: implications for pathophysiology and therapy.

Atrioventricular (AV) nodal reentrant tachycardia and atrial flutter are considered 2 distinct supraventricular tachycardias. Recent clinical and experimental data suggest that both these tachycardias include an area in the lower right atrial septum in their reentrant pathways. This study was designed to test the hypothesis that there is an association between the mechanisms of AV nodal reentrant tachycardia and atrial flutter because of a shared pathway of reentry. Consecutive patients referred for evaluation and management of supraventricular tachycardia, thought to be due to AV nodal reentry, underwent electrophysiologic testing protocols designed to induce both AV nodal reentrant tachycardia and atrial flutter, if present. Fifteen of 29 patients (52%) had both AV nodal reentrant tachycardia and atrial flutter induced during electrophysiologic testing. Seven of these 15 patients (47%) underwent transcatheter radiofrequency current application (mean power 34 +/- 4 W) against the tricuspid annulus above the coronary sinus. In each patient, neither AV nodal reentrant tachycardia nor atrial flutter could be induced after the procedure. Repeat study after successful ablation (mean 6 days) showed no inducible supraventricular arrhythmia of either type at baseline study or during isoproterenol infusion. Atrial flutter occurs frequently (15 of 29 patients; 52%) in patients with AV nodal reentrant tachycardia, because of a shared pathway in their reentry circuits. Because of this shared pathway, both arrhythmias can be ablated at the same site. These observations promote new insights into the mechanism and therapeutics of supraventricular tachycardias.

Adult↗
PubMed1993

Effect of pulse frequency and amplitude of D-Trp6-luteinizing hormone-releasing hormone on the pulsatile secretion of prolactin and LH.

This work analyzes the effect of the pulse amplitude and frequency of a potent LHRH analog, D-Trp6-LHRH, in a perfusion system of isolated rat pituitary cells. To this purpose, we studied the LH and PRL secretion in different conditions: basal secretion, secretion after increasing concentrations of D-Trp6-LHRL (0.001, 0.01, 0.1 and 1 nM) secretion in function of the pulses frequency (2,3, and 4 pulses per h) and amplitude (0.1, 1 and 10 nM). The principal findings were: 1. The basal LH and PRL secretions was pulsatile; 2. The stimulation of LH by the analog was not dose-dependent; 3. When more than 2 pulses per h were administered, a rapid desensitization of gonadotroph to release LH (at 20-30 min) occurred; 4. There was a loss of pulsatility of PRL secretion with an increase in the pulse frequency and amplitude of the D-Trp6-LHRH, which was produced parallelly to the desensitization of the gonadotroph to release LH. In summary, these findings suggest that a rapid loss of pulsatility of the PRL when the D-Trp6-LHRH pulse frequency and amplitude is increased might be due to the early desensitization of the gonadotroph to the analog.

Animals↗
PubMed1992

[Loeffer's syndrome: is Taenia responsible?].

Taenia saginata is a parasite of the gastrointestinal tract which is often encountered in France. Its responsibility in the occurrence of pulmonary eosinophilic infiltrates is only exceptionally suspected. We report a case of Loeffler's syndrome most probably caused by Taenia saginata. This case illustrates the diagnostic problems encountered to determine the responsibility of this cestode.

Adult↗
PubMed1991

A method for analysis of the local atrial evoked response for determination of atrial capture in permanent pacing systems.

We developed a method for detection and analysis of the local atrial evoked response. Eleven patients undergoing permanent dual chamber pacemaker implantations for standard clinical indications were included in the study. Using a pacing system emulator, charge balancing using a variable triphasic stimulus waveform to reduce polarization artifact amplitude was performed first. This could not be completed in one patient because of a poor signal-to-noise ratio. Subsequent analysis of the local atrial evoked response in the remaining ten patients showed the typical signal to be a biphasic waveform with an initial negative deflection followed by a positive deflection nearly equal in amplitude. The mean amplitude of the atrial evoked response was 3.1 +/- 1.4 mV, while the intrinsic P wave amplitude in these same patients averaged 5.6 +/- 3.0 mV. The summed evoked response, a parameter that is directly proportional to the area of the signal, was used to determine atrial pacing threshold. The median atrial pacing threshold determined by the algorithm was 1.00 V. There was no instance of failure to detect loss of capture, nor was loss of capture inaccurately determined when there was still successful atrial pacing. Atrial capture in permanent pacing systems can thus be determined using an algorithm to record and analyze the local atrial evoked response. This method could potentially be useful in the automatic determination of atrial pacing threshold.

Algorithms↗
PubMed1991

A new algorithm for minimizing pacemaker polarization artifact: universally applicable in permanent pacing systems.

Polarization artifacts that result from pacing may interfere with analysis of paced evoked responses during, e.g., automatic threshold tracking. We have developed a method for reduction of such artifacts that relies on the introduction of pacing stimuli during the refractory period of unipolar or bipolar paced captured beats after previous identification of a refractory period "template" or baseline. The refractory pacing stimuli cannot capture the heart, and thus any deviation from the template is due to polarization artifact alone. The artifact amplitude is measured and the precharge duration of the triphasic stimulus waveform is changed each time until artifact is minimized, as detected by repeated reversals in the polarity of the polarization artifact. In a series of 11 patients with unipolar and bipolar permanent pacing leads, mean initial artifact before balancing was 1.44 +/- 0.84 mV, which was reduced to 0.44 +/- 0.30 mV after balancing (P = 0.001). Initial precharge duration was 3.2 msec by design; mean final precharge duration was 3.30 +/- 0.34 msec. This algorithm is universally applicable in permanent pacing systems, as it is valid in unipolar and bipolar pacing and it does not require an intrinsic cardiac rhythm.

Algorithms↗
PubMed1991

Isolation and characterization of Schizosaccharomyces pombe mutants defective in cell wall (1-3)beta-D-glucan.

Schizosaccharomyces pombe thermosensitive mutants requiring the presence of an osmotic stabilizer to survive and grow at a nonpermissive temperature were isolated. The mutants were genetically and biochemically characterized. In all of them, the phenotype segregated in Mendelian fashion as a single gene which coded for a recessive character. Fourteen loci were defined by complementation analysis. Studies of cell wall composition showed a reduction in the amount of cell wall beta-glucan in three strains (JCR1, JCR5, and JCR10) when growing at 37 degrees C. Galactomannan was diminished in two others. Strains JCR1 and JCR5, with mutant alleles cwg1-1 and cwg2-1, respectively, were further studied. The cwg1 locus was mapped on the right arm of chromosome III, 18.06 centimorgans (cM) to the left of the ade5 marker; cwg2 was located on the left arm of chromosome I, 34.6 cM away from the aro5 marker. (1-3)beta-D-Glucan synthase activities from cwg1-1 and cwg2-1 mutant strains grown at 37 degrees C were diminished, as measured in vitro, compared with the wild-type strain; however, Km values and activation by GTP were similar to the wild-type values. Mutant synthases behaved like the wild-type enzyme in terms of thermostability. Analyses of round shape, lytic behavior, and low (1-3)beta-D-glucan synthase activity in cultures derived from ascospores of the same tetrad showed cosegregation of all these characters. Detergent dissociation of (1-3)beta-D-glucan synthase into soluble and particulate fractions and subsequent reconstitution demonstrated that the cwg1-1 mutant was affected in the particulate fraction of the enzymatic activity while cwg2-1 was affected in the soluble component. The antifungal agents Papulacandin B and Aculeacin A had similar effects on the enzymatic activities of the wild type and the cwg2-1 mutant strain, whereas the cwg1-1 mutant, when growing at 37 degrees C, had a more inhibitor-resistant (1,3)beta-D-glucan synthase. It is concluded that the cwg1+ and cwg2+ genes are related to (1,3)beta-D-glucan biosynthesis.

Aminoglycosides↗