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Biomedical subjects

B Fischer

Publications and source records attributed to B Fischer.

At least 19 recordsLinked to original sources
PubMed1992

Analysis of catalytic properties of hen egg white lysozyme during renaturation from denatured and reduced material.

Catalytic properties of hen egg white lysozyme were analyzed during the renaturation of the enzyme from completely reduced and denatured material. The formation of intermediate folding products and the generation of native lysozyme was monitored by acetic acid/urea electrophoresis. The results showed that during the beginning of renaturation almost all reduced and denatured lysozyme is converted to forms possessing lower compactness than native lysozyme, probably as a result of formation of only one or two disulfide bonds. Kinetic analysis of lysozyme during renaturation showed that the generation of lysozyme with four disulfide bonds was not necessarily equivalent to the formation lysozyme with native-like catalytic properties. It appeared that the formation rate of the structures of the structures of the substrate binding site and of the catalytic site were limited by the generation of four disulfide bonds containing lysozyme. The catalytic properties of intermediate folding products made it evident that the final structures of the substrate binding site and of the catalytic site were formed after the generation of all disulfide bonds.

Animals↗
PubMed1992

Frequency of recurrent atrial fibrillation after catheter ablation of overt accessory pathways.

The effect of successful catheter ablation of overt accessory pathways on the incidence of atrial fibrillation (AF) was studied in 129 symptomatic patients with (n = 75) or without (n = 54) previous documented AF. Fourteen had had ventricular fibrillation. Factors predictive of recurrence were examined, including electrophysiologic parameters. Atrial vulnerability was defined as induction of sustained AF (greater than 1 minute) using single, then double, atrial extrastimuli at 2 basic pacing cycle lengths. When compared to patients with only reciprocating tachycardia, patients with clinical AF included more men (77 vs 54%, p = 0.008) and were older (35 +/- 12 vs 29 +/- 12 years, p = 0.01). They had a significantly shorter cycle length leading to anterograde accessory pathway block (252 +/- 42 vs 298 +/- 83 ms, p less than 0.001), greater incidences of atrial vulnerability (89 vs 24%, p less than 0.001) and subsequent need for cardioversion (51 vs 15%, p less than 0.001). After discharge, the follow-up period was 35 +/- 12 months (range 18 to 76); 7 patients with previous spontaneous AF (9%) had recurrence at a mean of 10 months after ablation. Age, presence of structural heart disease accessory pathway location, atrial refractory periods and accessory pathway anterograde conduction parameters were not predictive of AF recurrence. Persistence of atrial vulnerability after ablation was the only factor associated with further recurrence of AF. Atrial vulnerability was observed after ablation in only 56% of patients with previous AF versus 89% before ablation. It is concluded that successful catheter ablation of accessory pathways prevents further recurrence of AF in 91% of patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1992

Characterization of human tissue-type plasminogen activator with monoclonal antibodies: mapping of epitopes and binding sites for fibrin and lysine.

The study defines interactions between human tissue-type plasminogen activator (t-PA) and 21 mouse monoclonal antibodies (mAb). Characterization includes epitope distribution, reactivity of different forms of t-PA with antibodies, and modification of t-PA function by antibody binding. Eighteen antibodies are directed against t-PA A-chain. These antibodies recognize four distinct epitopes (A, B, C, D) and one partially overlapping epitope (D'). The remaining three antibodies are directed against two different epitopes (E, F) on catalytically active t-PA B-chain. A-chain reactive antibodies do not bind to the reduced form of t-PA, while B-chain reactive antibodies bind to reduced and deglycosylated t-PA forms. The latter antibodies associate more tightly with sc t-PA than with tc t-PA and have a higher affinity for t-PA-PAI 1 complex as compared to free t-PA. The analysis of functional effects of antibodies reveals that antibodies directed against all above defined epitopes inhibit interactions between t-PA and fibrin: a) binding of t-PA to fibrin, b) fibrinolytic activity of t-PA, and c) fibrin activation of sc t-PA amidolytic activity. The observations support the assumption that several sites of t-PA are involved in fibrin binding and that fibrin-bound t-PA is closely surrounded by the fibrin mesh. Many antibodies quench also binding of t-PA to lysine-Sepharose. Experiments with free, non-fixed lysine confirm strong competition between lysine and mAb 16 and 18, directed against epitope A, and mAb 29, binding to epitope F.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1992

Oxygen tension in follicular fluid falls with follicle maturation.

The increase in follicular diameter during maturation is accompanied by an increase in vascular supply. The oxygen tension (PO2) in the follicular fluid of the growing follicle should therefore give an insight whether the oxygen delivery during this pertinent process is sufficient. Follicular fluid of 20 patients subjected to in vitro fertilization and embryo transfer were sampled in heparinized glass capillaries which were placed between the puncturing needle and the collecting reservoir. All patients were treated with human menopausal gonadotropin (HMG) and follicle-stimulating hormone (FSH). The PO2, PCO2 and pH of the follicular fluid were measured with an automatic gas analyser (Ciba Corning). Simultaneously, the same parameters were estimated in maternal capillary blood taken from the arterialized well perfused earlobe. Follicular size was measured by ultrasound. With growing follicular size (FS), the oxygen partial pressure (PO2) fell from 80 to 50-60 mmHg (PO2 = 114.4-3.3.FS; r = 0.79) while the carbondioxide partial pressure (PCO2) in the follicle increased from 35 to 50 mmHg (PCO2 = 15.1 +/- 1.9.FS; r = 0.71). In correlation to these results, the pH in follicular fluid fell from 7.40 to 7.30 (pH = 7.6-0.019.FS; r = 0.62). The correlation between the PCO2 and the pH demonstrates a deviation from the regression line if no metabolic alterations take place (delta log PCO2/delta pH = -0.69). The increase in ovarian blood flow during follicular maturation is obviously not sufficient to bridge the growing diffusion distance between the capillary network surrounding the follicle and its center.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbon Dioxide↗
PubMed1992

Characteristics of "anti" saccades in man.

Four subjects - all made large numbers of Express saccades in the normal gap task - were instructed to make saccades in the direction opposite to the side where a visual stimulus appeared ("anti" task). Gap and overlap trials were used. Saccadic reaction time (SRT), velocity and amplitude of the corresponding eye movements were analysed and compared to those of saccades made in the normal task. The velocity of "anti saccades" was found to be slightly (up to 15%) but significantly slower in two subjects. The distributions of SRTs in normal gap tasks show a small group of anticipatory saccades (with SRT below 80 ms and slower velocities) followed by a group of saccades with fast reaction times between 80 ms and 120 ms (Express saccades) followed by another large group ranging up to 180 ms (regular saccades). In the gap anti task there are anticipatory saccades and saccades with SRTs above 100 ms; Express saccades are missing. The distribution of SRTs obtained in the overlap anti task was unimodal with a mean value of 231 ms as compared to 216 ms in the normal task. The introduction of the gap therefore clearly decreases the reaction times of the anti saccades. Control experiments show that the delay of anti saccades is not due to an interhemispheric transfer time but must be attributed to the saccade generating system taking more time to program a saccade to a position where no visual stimulus appears. These data are discussed as providing further evidence for the existence of a reflex-like pathway connecting the retina to the oculomotor nuclei mediating the Express saccade.

Humans↗
PubMed1992

Dead zone for express saccades.

The saccadic eye movements of three humans and one non-human primate (a male rhesus monkey) have been measured for target eccentricities between 0.3 and 15 deg. With a gap task (fixation point offset precedes target onset by 200 ms) and a target at 4 deg, all subjects produced reasonable amounts of express saccades as indicated by a clear peak in the distribution of their saccadic reaction times (SRT): about 100 ms in human subjects and 70 ms in the monkey. This peak disappeared with decreasing target eccentricity below 2 deg, but saccades of longer (regular) reaction times were still present. Thus it was found that there exists a dead zone for express saccades. In addition, small saccades have a much stronger tendency to overshoot the target and their velocity falls above the main sequence as defined by the least square fit of an exponential v = vo(1-exp(-a/ao)) to the maximal velocity (v) versus amplitude (a) relationship (vo and ao are constants fitted). It is concluded that for small saccades the express way is blocked functionally or does not exist anatomically.

Animals↗
PubMed1992

Regulated inactivation of homologous gene expression in transgenic Nicotiana sylvestris plants containing a defense-related tobacco chitinase gene.

The class I chitinases are vacuolar proteins implicated in the defense of plants against pathogens. Leaves of transgenic Nicotiana sylvestris plants homozygous for a chimeric tobacco (Nicotiana tabacum) chitinase gene with Cauliflower Mosaic Virus (CaMV) 35S RNA expression signals usually accumulate high levels of chitinase relative to comparable leaves of non-transformed plants. Unexpectedly, some transgenic plants accumulated lower levels of chitinase than nontransformed plants. We call this phenomenon silencing. The incidence of silencing depends on the early rearing conditions of the plants. When grown to maturity in a greenhouse, approximately 25% of plants raised as seedlings in closed culture vessels were of the silent type; none of the plants raised from seed in a greenhouse showed this phenotype. Silencing is also developmentally regulated. Plants showed three patterns of chitinase expression: uniformly high levels of expression in different leaves, uniformly low levels of expression in different leaves, and position-dependent silencing in which expression was uniform within individual leaves but varied in different leaves on the same plant. Heritability of the silent phenotype was examined in plants homozygous for the transgene. Some direct descendants exhibited a high-silent-high sequence of activity phenotypes in successive sexual generations, which cannot be explained by simple Mendelian inheritance. Taken together, the results indicate that silencing results from stable but potentially reversible states of gene expression that are not meiotically transmitted. Gene-specific measurements of chitinase and chitinase mRNA showed that silencing results from co-suppression, i.e. the inactivation of both host and transgene expression in trans. The silent state was not correlated with cytosine methylation of the transgene at the five restriction sites investigated.

Base Sequence↗
PubMed1992

Ultrastructural and autoradiographic study of preimplantation rabbit embryos grown in conventional or uterine flushing-supplemented culture media.

Rabbit morulae and blastocysts were cultured in conventional culture media [Ham's F10 or BSM II supplemented with bovine serum albumin (BSA) or serum] or in Ham's medium supplemented with synchronous or asynchronous uterine flushings, mostly for 2 days, and afterwards investigated by light and electron microscopy and by autoradiography. Ultrastructure and cell proliferation differed considerably between cultured embryos and noncultured controls. Cultured embryos displayed more dead cells. They were developmentally retarded (predominance of smooth endoplasmic reticulum rather than the age-specific rough endoplasmic reticulum, mitochondria still round to ovoid shaped) and showed nonspecific signs of cells damage (swollen mitochondria and Golgi complex vesicles, increased number of lysosomes). All these features were also present in embryos grown in uterine flushing-supplemented media, but were less pronounced. Cell damage and impaired cell proliferation had affected trophoblast cells more than embryoblast cells. Endoderm could be differentiated only if culture had been started with blastocysts--not with morulae--and seems to require uterine secretions. No significant ultrastructural differences were observed between embryos cultured in synchronous or in asynchronous uterine flushings. Present results indicate that cultured preimplantation rabbit embryos deviate clearly from those grown in vivo and maintain, for some time, a better cellular structure--and probably function--in the presence of uterine flushings than in conventional culture media. Specific abnormal morphologic features related to a particular medium could not be identified.

Animals↗
PubMed1992

Sense and antisense transcripts of human papillomavirus type 16 in cervical cancers.

Human papillomavirus type 16 (HPV-16) transcription was analysed in one squamous cervical carcinoma by cDNA cloning and DNA sequencing, and in eight additional squamous cervical carcinomas and 11 precancerous lesions by RNA-RNA in situ hybridization. The nucleotide sequences of the cDNA clones revealed structures of early HPV-16 mRNAs (E6*-E7-E1 E4-E5) in agreement with data reported for other premalignant and malignant tumours. cDNA clones possibly representing viral RNA of antisense orientation were also detected. These RNAs included sequences of the upstream regulatory region, part of the early and the late region of the genome. In three of eight squamous cervical carcinomas examined by in situ hybridization, signals specific for viral antisense RNA were also found. The antisense RNAs had a predominantly nuclear localization. Viral antisense RNA could not be detected in any of 11 HPV-16-positive premalignant lesions. The expression of HPV antisense RNA is likely to be linked to viral integration into the host genome. The possible effects of viral antisense transcription with regard to tumour progression remain to be determined.

Female↗
PubMed1992

Electrogram patterns predictive of successful radiofrequency catheter ablation of accessory pathways.

UNLABELLED: We assessed anterograde conduction electrogram parameters at successful ablation sites according to accessory pathway (AP) location and compared them to the most favorable electrograms at unsuccessful sites. A median of three radiofrequency energy impulses was applied to ablate 97% of 136 APs versus four impulses to ablate 90% of 65 concealed APs. Electrograms at successful sites showed variable A/V ratio (0.04-7), and a QS pattern of unipolar ventricular wave in 90%. Electrograms were different in right versus left AP: AV time 29 +/- 7 versus 38 +/- 10; and timing of ventricular deflection: 17 +/- 9 versus 2 +/- 9 msec, respectively. Analysis in each patient of the mapping parameters at successful versus "most favorable" unsuccessful sites showed an improvement in at least one parameter in 55%, no apparent change in all parameter values in 30%, and even less favorable parameters in 15% of patients. In patients with manifest AP, overall comparison of electrograms at successful versus unsuccessful sites showed no difference in A/V ratio (1.3 +/- 1.5 vs 1.2 +/- 1.6), unipolar pattern distribution, and AV time (34 +/- 10 vs 35 +/- 9), but earlier bipolar main ventricular potential (-4 +/- 12 vs -1.5 +/- 10 msec) and unipolar intrinsic deflection timings (-5.2 +/- 11 vs -1.8 +/- 10 msec). In patients with concealed AP, a retrograde continuous electrical activity was recorded in 72% of successful versus 38% of unsuccessful sites (P = 0.03). CONCLUSIONS: Electrogram characteristics at successful radiofrequency ablation are different in right and left manifest AP.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1992

Elimination of atrioventricular nodal reentrant tachycardia using discrete slow potentials to guide application of radiofrequency energy.

BACKGROUND: Ablation of the slow pathway has been performed to eliminate atrioventricular (AV) nodal reentrant tachycardia (AVNRT) either by a surgical approach or by using radiofrequency catheter technique guided by retrograde slow pathway activation mapping. From previous experience of midseptal and posteroseptal mapping, we were aware of the existence of peculiar slow potentials in most humans. Postulating their role in AVNRT, we studied these potentials and the effects of radiofrequency energy. METHODS AND RESULTS: Sixty-four patients (mean age, 48 +/- 19 years) with the usual form of AVNRT were studied. Slow, low-amplitude potentials were recorded when using the anterograde AV conducting system. Slow potentials occupied all (giving a continuum of electrograms) or some of the time between the atrial and ventricular electrograms. Their most specific patterns were their progressive response to increasing atrial rates, which resulted in a dramatic decline in amplitude and slope, a corresponding increase in duration, and a separation from preceding atrial potentials until the disappearance of any consistent activity. Slow potentials were recorded along a vertical band at the mid or posterior part of the septum near the tricuspid annulus. Radiofrequency energy applied at the slow potential site resulted in interruption of induced tachycardia within a few seconds and rendered tachycardia noninducible in all patients. A median of two impulses was delivered to each patient. In 69% of patients, postablation atrial stimulation cannot achieve a long atrial-His interval, which previously was critical for tachycardia induction or maintenance. No patient had AVNRT over a follow-up period of 1-16 months, and all had preserved AV conduction. In all except two patients, the PR interval was unchanged. In 47 patients, long-term electrophysiological studies confirmed the efficacy of ablation and the nonreversibility of results by isoproterenol; however, echo beats remained inducible in 40% of patients. CONCLUSIONS: An area showing slow potentials is present at the perinodal region in humans. In patients with AVNRT, application of radiofrequency energy renders tachycardia noninducible through the preferential modification of the anterograde slow pathway. With present clinical methods, the exact origin and significance of these physiological potentials cannot be specified.

Cardiac Pacing, Artificial↗
PubMed1992

Radiofrequency catheter ablation of left lateral accessory pathways via the coronary sinus.

BACKGROUND: The purpose of this study was to describe a new technique for catheter ablation of left lateral accessory pathways (APs) by radiofrequency energy applied at the epicardium through the coronary sinus wall using a unipolar configuration. METHODS AND RESULTS: In an overall group of 212 patients with left lateral APs, multiple endocardial ablation attempts of the AP were unsuccessful in eight patients. The mean +/- SD cumulative duration of previous attempts was 12 +/- 9 hours, using DC shocks and/or radiofrequency energy applied both at the atrial and/or ventricular AP insertions. Epicardial AP insertion was determined by bipolar and unipolar unfiltered distal electrograms by scanning the coronary sinus with a steerable 6F or 7F catheter with a 4-mm distal electrode. The local atrial to ventricular electrogram amplitude ratio was 0.3-1.6. At the ablation site, the catheter tip was slightly deflected toward the annulus to increase both the ventricular component of electrograms and contact with the epicardium. In four patients, epicardial electrogram timings were earlier than endocardial ones. The AP was ablated in seven of the eight patients with 20-30 W applied for 10-60 seconds. No complications occurred except a marked nonspecific pain during radiofrequency energy application; however, the catheter remained adherent to the coronary sinus wall, and its withdrawal was performed during a new radiofrequency application to decrease the risk of coronary sinus rupture. After ablation, echocardiograms, coronary artery angiograms, and levophase coronary sinus angiograms showed no abnormality in all patients except two who had a probable mural thrombus in the coronary sinus. AP conduction remained abolished for 1-10 months of follow-up in seven patients. CONCLUSIONS: Radiofrequency catheter ablation of left lateral APs can be achieved effectively and relatively safely via the mid or distal coronary sinus when endocardial approaches are unsuccessful.

Catheter Ablation↗
PubMed1992

Isolation and renaturation of bio-active proteins expressed in Escherichia coli as inclusion bodies.

Over-expression of recombinant proteins in Escherichia coli often results in the formation of insoluble and inactive material as inclusion bodies. Within the last years specific methods and strategies have been developed to produce bio-active proteins from these inclusion bodies. These methods include (i) isolation and purification of inclusion bodies, (ii) solubilization and reduction of the insoluble material, and (iii) renaturation of the proteins including formation of native disulphide bonds. Inclusion bodies are isolated from E. coli cells after desintegration of cells by mechanic forces. Due to their stability, inclusion bodies may be purified by washing with detergent solutions or low concentrations of denaturant. High concentrations of urea and guanidine hydrochloride are used in combination with reducing reagents such as 2-mercaptoethanol or dithiothreitol to reduce and solubilize proteins from inclusion bodies. After purification of solubilized materials, proteins are renatured and native disulphide bonds are formed by either air oxidation or glutathione reoxidation starting from reduced material, or by disulphide interchange starting from mixed disulphides containing peptides. Final yield of renatured proteins is raised by low concentrations of proteins and by adding low concentrations of denaturant during renaturation.

Animals↗
PubMed1992

[Focal junctional tachycardia in adults. Anatomo-electrophysiological aspects apropos of a case].

The authors report the case of a 65 year old patient with focal junctional tachycardia complicating infectious myocarditis which had a fatal outcome. The ECG recordings showed episodes of tachycardia alternating with junctional rhythm at 90/min. There were signs of retrograde conduction. The anatomopathological findings were typical of acquired myocarditic lesions in the lower part of the atrioventricular node with congenital abnormalities, in particular a bifid node and His bundle with an accessory paraseptal atrioventricular bundle. This is the first description, to the best of the authors's knowledge, of junctional tachycardia associated with a latent pre-excitation.

Aged↗
PubMed1992

[Radiofrequency treatment of junctional tachycardia].

Atrioventricular nodal tachycardias due to intranodal reentry or to an accessory pathway are accessible to radical cure with radiofrequency currents applied either at the site of recording of characteristic slow potentials or at the tricuspid or mitral atrioventricular rings. One hundred and six patients with atrioventricular nodal reentry were treated by modification of the slow anterograde reentrant pathway with a 100% success rate and without any serious complications (especially atrioventricular block). One hundred and eighty six patients had one or more overt or latent accessory pathways and were treated by the same method. The ablation site was decided on indirect criteria and not by the recording of the specific activity of the accessory pathway. The success rate was 97%, also with no significant complications. The duration of the treatment was 41 +/- 22 min for the accessory pathways. The exposure time to ionising radiation was 14 +/- 14 min and 31 +/- 34 min respectively. These results suggest that the indications of radiofrequency current ablation could be extended.

Adolescent↗
PubMed1991

The dynamic structure of rabbit blastocyst coverings. III. Transformation of coverings under non-physiological developmental conditions.

Under physiological conditions the zona pellucida disappears in the rabbit between Day 3 and early Day 4 post coitum (p.c.) and is replaced by a new layer, the neozona. The dissolution of the zona pellucida and the formation of the neozona was investigated in three different experimental approaches, all of them characterized by non-physiological developmental conditions for the embryo: Prevention of embryo migration from the oviduct into the uterus by postcoital (48 h p.c.) tubal ligation, in vitro culture, and asynchronous embryo transfer into uteri of recipient rabbits. Embryos of age 2 1/2, 3, 4 and 4 1/2 days p.c. were cultured for 12 to 72 h. The media used for in vitro culture were supplemented with BSA, serum or with uterine secretions that were collected either synchronously or asynchronously to the developmental stage of the cultured embryos. Three-day-old embryos were transferred into uteri of pseudopregnant foster rabbits of either synchronous (Day 3) or asynchronous stages (Day 0, 2, 4, 5, 6) and were recovered 24 to 72 h after transfer. The transformation of the coverings was evaluated by light and transmission electron microscopy. The dissolution of the zona pellucida was greatly disturbed in tube-locked embryos, and in cultured embryos if standard protein supplements (BSA or serum) had been used for in vitro culture. In many cases the zona was still completely preserved after 2 or 3 days in culture, at a time when it normally would have already been replaced by the neozona in vivo. The dissolution in vitro, however, progressed incomparably better if the culture medium had been substituted with synchronous or asynchronous uterine secretions. The formation of the neozona could not be verified in cultured blastocysts. After embryo transfer, the dissolution of the zona pellucida was completed in most cases by 2 days after transfer, irrespective of the recipients' progestational stage. Present results indicate that uterine components are essential for the dissolution of the rabbit zona pellucida. These components appear to be present in the uterine cavity constitutively, i.e. independently of the uterine progestational transformation, and need not be in synchrony with the embryo's developmental stage for dissolution of the zona. Normal formation of the neozona does not take place under the non-physiological developmental conditions of in vitro culture.

Animals↗