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B Scharf

Publications and source records attributed to B Scharf.

At least 19 recordsLinked to original sources
PubMed2002

Microscale evaluation of the effects of grazing by invertebrates with contrasting feeding modes on river biofilm architecture and composition.

River biofilms are a valuable food resource for many invertebrates. In the present study biofilms were cultivated in a rotating annular bioreactor with river water as sole source of inoculum. The resulting biofilms were then presented to starved snails, ostracods, and mayflies as sole food source. The biofilms were then removed and microscopically examined to determine areas that had been grazed. The grazed and ungrazed areas were marked and analyzed for the effects of grazing using confocal laser scanning microscopy and image analyses. Samples were treated with fluorescent probes for nucleic acids to quantify bacterial biomass and fluor-conjugated lectins to quantify exopolymer, and far red autofluorescence was imaged to quantify algal or photosynthetic biomass. Grazing by snails significantly reduced algal biomass (1.1 +/- 0.6 micro m 3 micro m 2 to 0.02 +/- 0.04 micro m 3 micro m 2), exopolymer (5.3 +/- 3.4 micro m 3 micro m 2 to 0.18 +/- 0.18 micro m 3 micro m 2), and biofilm thickness (154 micro m +/- 50 to 11 micro m +/- 5.2; ANOVA, p < or= 0.05). Although bacterial biomass was influenced by grazing snails the impact was not statistically significant (p <or= 0.05). Ostracods had a significant (p <or= 0.05) impact on the algal biomass and exopolymer but not on bacteria. Mayfly grazing resulted in reduction of biofilm thickness to approximately 40 mm and reductions in all biofilm components with little evidence of selectivity. Thus grazing consistently resulted in a significant reduction in autotrophic biomass and exopolymer with a resultant increased importance of bacterial biomass within the grazed regions. Examination of grazed biofilms after a recovery period in the absence of grazing indicated that grazed regions remained predominantly bacterial after 28 days. A comparison of grazing in diclofop methyl and atrazine contaminated (1 ppb and 10 ppb) versus control biofilms indicated no significant influence of the contaminants on grazing patterns.

Animals↗
PubMed2001

Mutational analysis of the Rhizobium lupini H13-3 and Sinorhizobium meliloti flagellin genes: importance of flagellin A for flagellar filament structure and transcriptional regulation.

Complex flagellar filaments are unusual in their fine structure composed of flagellin dimers, in their right-handed helicity, and in their rigidity, which prevents a switch of handedness. The complex filaments of Rhizobium lupini H13-3 and those of Sinorhizobium meliloti are composed of three and four flagellin (Fla) subunits, respectively. The Fla-encoding genes, named flaA through flaD, are separately transcribed from sigma(28)-specific promoters. Mutational analysis of the fla genes revealed that, in both species, FlaA is the principal flagellin and that FlaB, FlaC, and FlaD are secondary. FlaA and at least one secondary Fla protein are required for assembling a functional flagellar filament. Western analysis revealed a ratio close to 1 of FlaA to the secondary Fla proteins (= FlaX) present in wild-type extracts, suggesting that the complex filament is assembled from FlaA-FlaX heterodimers. Whenever a given mutant combination of Fla prevented the assemblage of an intact filament, the biosynthesis of flagellin decreased dramatically. As shown in S. meliloti by reporter gene analysis, it is the transcription of flaA, but not of flaB, flaC, or flaD, that was down-regulated by such abortive combinations of Fla proteins. This autoregulation of flaA is unusual. We propose that any combination of Fla subunits incapable of assembling an intact filament jams the flagellar export channel and thus prevents the escape of an (as yet unidentified) anti-sigma(28) factor that antagonizes the sigma(28)-dependent transcription of flaA.

Amino Acid Sequence↗
PubMed2000

VisN and VisR are global regulators of chemotaxis, flagellar, and motility genes in Sinorhizobium (Rhizobium) meliloti.

The known 41 flagellar, chemotaxis, and motility genes of Sinorhizobium (Rhizobium) meliloti contained in the "flagellar regulon" are organized as seven operons and six transcription units that map to a contiguous 45-kb chromosomal region. By probing gene expression on Western blots and with lacZ fusions, we have identified two master regulatory genes, visN and visR, contained in one operon. The gene products probably form a heterodimer, VisNR, acting as a global transcription activator of other flagellar genes. The related 27-kDa VisN and VisR proteins are LuxR-type proteins with typical ligand- and DNA-binding domains. The vis operon itself is constitutively transcribed; however, to activate flagellar genes, VisNR seemingly requires the binding of a yet-unknown effector. Gene expression in tester strains with known deficiencies revealed a hierarchy of three classes of flagellar genes: class I comprises visN and visR; class II, controlled by VisNR, comprises flagellar assembly (class IIA) and motor (class IIB) genes; and class III comprises flagellin and chemotaxis genes that require functional class I and class IIA genes for expression. In contrast to their enterobacterial counterparts, mot genes belong to class II without exerting control over class III genes. While the general hierarchy of gene expression resembles the enterobacterial scheme, the assignment of mot genes to class IIB and the global control by a LuxR-type VisNR activator are new features distinguishing the S. meliloti flagellar gene system.

Amino Acid Sequence↗
PubMed2000

[Risk stratification after myocardial infarct].

In industrialized countries the rate of sudden cardiac death remains unchanged. The most frequently encountered structural heart disease in these patients is coronary artery disease. Despite the era of thrombolytic therapy of acute myocardial infarction patients carry an increased risk of sudden cardiac arrhythmogenic death within a time period of one to two years following the acute event. Therefore, risk stratification post-MI before patient discharge is furthermore mandatory. The spectrum of non-invasive techniques for risk stratification includes the clinical risk profile, measurement of left ventricular global function (LV ejection fraction), the resting ECG (QT dispersion), an ECG stress test (detection and severity of myocardial ischemia), ambulatory ECG monitoring (number and type of ventricular arrhythmias), surface high resolution ECG (detection of ventricular late potentials), measurement of T wave alternans (TWA, alternans ratio), and measurements of the activity and balance of the autonomous nervous system (heart rate variability, baroreflex sensitivity = BRS). Programmed ventricular stimulation (PVS) serves as an invasive risk stratification technique (detection of an arrhythmogenic substrate). The prognostic power of the non-invasive techniques is limited; in general, the prognostic value of a negative test is reasonably high (90 to 100% depending on the test used), whereas the prognostic value of a positive test is rather low (4 to 42% depending on the test used). Combining several non-invasive tests may significantly improve the positive predictive value above 50%, but this goes along with a significant decreases of sensitivity below 50%. Therefore, a combination of several non-invasive tests (detection and exclusion of a large number of low-risk individuals) with the invasive method of PVS (detection of an arrhythmogenic substrate, i.e. a high-risk patient) seems reasonable, as has been convincingly shown by several smaller prognostic studies.

Arrhythmias, Cardiac↗
PubMed1997

Electron transfer proteins from the haloalkaliphilic archaeon Natronobacterium pharaonis: possible components of the respiratory chain include cytochrome bc and a terminal oxidase cytochrome ba3.

Natronobacterium pharaonis, an aerobic haloalkaliphilic archaebacterium, expresses high concentrations of redox proteins as do alkaliphilic eubacteria. The first redox protein characterized from N. pharaonis was halocyanin [Scharf, B., & Engelhard, M. (1993) Biochemistry 32, 12894-12900], a small blue copper protein. It is a peripheral membrane protein and is conjectured to function in a manner similar to plastocyanin. In the present work, the respiratory chain is further elucidated and the purification and characterization of the most abundant components cytochrome bc and cytochrome ba3 from the membrane fraction are described. The cytochrome bc complex consists of a 14 and an 18 kDa subunit in a 1:1 ratio, with heme c bound to the larger polypeptide. An Fe-S subunit similar to that found in eukaryotic bc complexes has not yet been identified. The second membrane complex carries two different heme groups of the ba3-type as well as copper. It contains two subunits of 36 and 40 kDa. This cytochrome ba3 binds carbon monoxide, a feature common to terminal oxidases. There is no spectroscopic evidence for a second terminal oxidase; hence, under the growth conditions chosen the respiratory chain of N. pharaonis appears to be unbranched. In addition to these cytochromes, a succinate dehydrogenase which is solubilized from the membrane by detergents was isolated. A cytochrome c which was isolated from the cytosol has an unusually high molecular weight and a redox potential of -142 mV. A second cytosolic protein, ferredoxin, was purified to homogeneity. A comparison of the redox potentials of the isolated proteins with those obtained from the native membrane allows the construction of a possible electron transfer chain.

Bacterial Outer Membrane Proteins↗
PubMed1997

[Cardiovascular diseases caused by unemployment? An analysis from the viewpoint of social medicine].

Unemployment has become a sociopolitical problem of great importance in the Western industrialised countries. Although negative effects on social life and psyche resulting from unemployment are regarded as scientifically accepted today, a possible causal relationship between job loss and somatic illnesses is still a matter of controversy. A possible target organ is the cardiovascular system. The aim of this study was, therefore, to check by means of extensive literature analysis to what extent unemployment can be seen to influence cardiovascular morbidity. Particular attention was paid to the method used and the clinical relevance of the results. Person-related epidemiological studies published since 1980 which investigated changes in cardiovascular risk factors associated with unemployment or prevalence rates of manifest disease influenced by unemployment, were included in the final evaluation. In some cases statistically significant associations were found between unemployment and the increase in cholesterol levels or systolic/diastolic blood pressure, but the clinical relevance of such slight changes is questionable. To consider unemployment as an independent, social, cardiovascular risk factor is at present unwarranted. An increase in the prevalence rates of coronary heart disease or arterial hypertonia causally linked in some studies with unemployment is scientifically questionable due to severe methodological shortcomings. On the basis of the currently available methodologically acceptable studies, the question of a qualitative contribution of unemployment to cardiovascular disease cannot be answered conclusively.

Adult↗
PubMed1997

On the role of the olivocochlear bundle in hearing: 16 case studies.

Earlier we presented data (Scharf et al. (1994) Hear. Res. 75, 11-26) from a young patient (S.B.) who had undergone a vestibular neurotomy, during which the olivocochlear bundle (OCB) was severed. Those data are complemented by measurements on 15 other patients-some like S.B. with normal audiometric thresholds, none with a loss greater than 35 dB at experimental frequencies. Comparisons of performance for the same ear before and after surgery or between the operated and healthy ears do not provide evidence that the lack of OCB input impairs the following psychoacoustical functions: (1) detection of tonal signals, (2) intensity discrimination, (3) frequency selectivity, (4) loudness adaptation, (5) frequency discrimination within a tonal series, (6) in-head lateralization. Data on single-tone frequency discrimination are equivocal. These mostly negative results apply to listening both in the quiet and, where relevant, in noise. The only clear change in hearing after a vestibular neurotomy is that most patients detect signals at unexpected frequencies better than before. This change suggests an impaired ability to focus attention in the frequency domain. Although limited in scope, our finding that human hearing without OCB input is essentially normal agrees with much of the relevant literature on animal behavior and with the patients' self-reports.

Acoustic Stimulation↗
PubMed1997

Loudness adaptation and excitation patterns: effects of frequency and level.

Simple loudness adaptation for pure tones was measured at frequencies from 0.125 to 16 kHz and at sensation levels from 5 to 60 dB. Sixteen young listeners with normal hearing participated in four experiments. Most of the loudness measurements were obtained by the method of successive magnitude estimation; some were also obtained by loudness matching. The two indices of loudness adaptation gave similar results. At all frequencies, loudness adaptation increased as sensation level decreased. After 6 min, average loudness declined at most frequencies by about 20% at 40-dB sensation level (SL) and by between 70% and almost 100% at 5-dB SL. Adaptation also increased with increasing frequency, and was especially marked at 16 kHz, where loudness declined more than 60% at a sensation level as high as 40 dB. Most of the adaptation occurred usually within the first 3 min of exposure, but loudness continued to diminish at a slower rate up to around 6 min. The dependence of loudness adaptation on frequency and level can be largely accounted for by the restricted-excitation-pattern hypothesis. Adaptation is assumed to take place when excitation is restricted to a narrow region of the cochlea. This hypothesis is supported by a quantitative analysis based on excitation patterns derived from measurements of masking.

Adolescent↗
PubMed1996

Protonation changes during the photocycle of sensory rhodopsin II from Natronobacterium pharaonis.

The Fourier Transform Infrared (FTIR) spectra of photocycle intermediates of sensory rhodopsin II (pSRII) from Natronobacterium pharaonis were measured. The results of the FTIR experiments indicate considerable conformational movements of pSRII already at the stage of the early K-like intermediate which persist at least during the lifetime of the long lived intermediate. These changes in the amide bond region are more intense than those described for sensory rhodopsin I (SRI) and are quite similar to those observed for rhodopsin. Concomitantly with the deprotonation of the Schiff base a carboxyl group located in a hydrophobic environment is protonated. In analogy to bacteriorhodopsin, this carboxyl group might arise from Asp-75 which probably serves as counter ion to the Schiff base. The protonation reaction differs from the situation observed in SRI where the protonation is pH independent over the range of pH 5-8.

Archaea↗
PubMed1996

Midline granuloma presenting as orbital cellulitis.

BACKGROUND: Lethal midline granuloma usually presents with rhinorrhoea and redness of the skin above the nose. Early ocular symptoms are very rare. We here describe a patient who presented with acute orbital cellulitis. PATIENT: A 73-year-old woman had a 24-h history of severe pain around her left eye. We saw the typical clinical picture of orbital cellulitis. A CT scan revealed a diffuse infiltration of the left upper and lower lid, the anterior orbit and the ethmoidal sinuses. RESULT: On surgical exploration we found a granular, partly necrotic tumour. Histological examination revealed an angiocentric nasal T-cell lymphoma (midline granuloma). CONCLUSION: Midline granuloma should be included in the differential diagnosis of acute orbital cellulitis.

Acute Disease↗
PubMed1996

Expectancy of line segment orientation.

The question was asked whether briefly flashed line segments are easier to detect when presented at an expected, rather than an unexpected, orientation. Detection rates were measured in a two-interval forced choice (2IFC) paradigm that did not require the subject to identify the orientation of the line segment, only to detect its presence. The 2IFC paradigm was used to rule out bias or criterion effects. Subjects were led to expect lines in a particular or primary orientation by being presented lines with that orientation as cues before every trial, and by being tested with only that orientation during practice. Lines of the orthogonal, probe orientation replaced the primary on 25% of experimental trials. When the stimulus location was known in advance, lines of the primary orientation were detected more accurately than were probe lines, but when stimulus location was not known, detection rates were equal. Detection rates were also equal when subjects were informed of the probe at the end of the practice period, so that both orientations were expected; hence the subjects' expectations, not the probability of stimulus occurrence, are necessary for the effect to occur. Thus expecting a line of a particular orientation at a particular location facilities its detection.

Form Perception↗
PubMed1995

Ligand binding and protein dynamics in cupredoxins.

Type 1 copper sites bind nitric oxide (NO) in a photolabile complex. We have studied the NO binding properties of the type 1 copper sites in two cupredoxins, azurin and halocyanin, by measuring the temperature dependence of the ligand binding equilibria and the kinetics of the association reaction after photodissociation over a wide range of temperature (80-280 K) and time (10(-6)-10(2) s). In both proteins, we find nonexponential kinetics below 200 K that do not depend on the NO concentration. Consequently, this process is interpreted as geminate recombination. In azurin, the rebinding can be modeled with the Arrhenius law using a single pre-exponential factor of 10(8.3) s-1 and a Gaussian distribution of enthalpy barriers centered at 22 kJ/mol with a width [full width at half-maximum (FWHM)] of 11 kJ/mol. In halocyanin, a more complex behavior is observed. About 97% of the rebinding population can also be characterized by a Gaussian distribution of enthalpy barriers at 12 kJ/mol with a width of 6.0 kJ/mol (FWHM). The pre-exponential of this population is 1.6 x 10(12) s-1 at 100 K. After the majority population has rebound, a power-law phase that can be modeled with a gamma-distribution of enthalpy barriers is observed. Between 120 and 180 K, an additional feature that can be interpreted as a relaxation of the barrier distribution toward higher barriers shows up in the kinetics. Above 200 K, a slower, exponential rebinding appears in both cupredoxins. Since the kinetics depend on the NO concentration, this process is identified as bimolecular rebinding.(ABSTRACT TRUNCATED AT 250 WORDS)

Azurin↗
PubMed1995

The primary structure of sensory rhodopsin II: a member of an additional retinal protein subgroup is coexpressed with its transducer, the halobacterial transducer of rhodopsin II.

The blue-light receptor genes (sopII) of sensory rhodopsin (SR) II were cloned from two species, the halophilic bacteria Haloarcula vallismortis (vSR-II) and Natronobacterium pharaonis (pSR-II). Upstream of both sopII gene loci, sequences corresponding to the halobacterial transducer of rhodopsin (Htr) II were recognized. In N. pharaonis, psopII and phtrII are transcribed as a single transcript. Comparison of the amino acid sequences of vHtr-II and pHtr-II with Htr-I and the chemotactic methyl-accepting proteins from Escherichia coli revealed considerable identities in the signal domain and methyl-accepting sites. Similarities with Htr-I in Halobacterium salinarium suggest a common principle in the phototaxis of extreme halophiles. Alignment of all known retinal protein sequences from Archaea identifies both SR-IIs as an additional subgroup of the family. Positions defining the retinal binding site are usually identical with the exception of Met-118 (numbering is according to the bacteriorhodopsin sequence), which might explain the typical blue color shift of SR-II to approximately 490 nm. In archaeal retinal proteins, the function can be deduced from amino acids in positions 85 and 96. Proton pumps are characterized by Asp-85 and Asp-96; chloride pumps by Thr-85 and Ala-96; and sensors by Asp-85 and Tyr-96 or Phe-96.

Amino Acid Sequence↗
PubMed1994

Raman spectroscopic study of the blue copper protein halocyanin from Natronobacterium pharaonis.

Halocyanin, a blue copper protein from Natronobacterium pharaonis was studied by Raman spectroscopic techniques. Near-infrared Fourier transform Raman spectra, which for the first time have been employed to study copper proteins in both oxidation states, display the Raman bands of the protein and, in the oxidized state, also the preresonance-enhanced bands of the copper center. The frequency of the amide I band at 1676 cm-1 indicates a predominant beta-sheet protein structure, which is typical for small blue copper proteins. The Raman spectra provide no evidence for major redox-linked changes of the secondary structure. Resonance Raman spectra of the oxidized protein obtained upon excitation close to the 600-nm absorption band were measured in the pH range between 7.7 and 4.5. The vibrational band pattern in the Cu-cysteine stretching region is closely related to that of azurin, indicating far-reaching similarities of the coordination geometry of the copper center in both proteins. Significantly lower frequencies, however, are noted for the Cu-histidine stretches, which appear as a closely spaced doublet at ca. 260 cm-1. Lowering the pH to 4.5 leads to an increase of this band splitting with one component shifting down to 247 cm-1. This downshift is attributed to the rupture of a hydrogen bond between one of the histidine ligands and a nearby carboxyl group, which becomes protonated at such a low pH. On the other hand, no major changes in the Cu-cysteine stretching region are noted at pH 4.5, implying that the coordination geometry remains largely unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Azurin↗