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Biomedical subjects

K Schneider

Publications and source records attributed to K Schneider.

At least 19 recordsLinked to original sources

EPR spectroscopic characterization of an 'iron only' nitrogenase. S = 3/2 spectrum of component 1 isolated from Rhodobacter capsulatus.

The alternative nitrogenase of Rhodobacter capsulatus, isolated from a nifHDK deletion mutant, has been purified to near homogeneity and identified as an 'iron only' nitrogenase. The dithionite-reduced component 1 ('FeFe protein') of this enzyme showed an EPR spectrum consisting of two components: a minor S = 1/2 signal at g = 1.93 and a very characteristic S = 3/2 signal of near-stoichiometric intensity at g = 5.44. This resonance is very close to the highest possible g value (g = 5.46) for the coinciding two intradoublet subspectra of an S = 3/2 system of maximal rhombicity (E/D = 0.33). The deviation from axial symmetry (increasing E/D) correlates with the stability, activity and substrate selectivity of the different (Mo, V, Fe) nitrogenases.

Electron Spin Resonance Spectroscopy

A desiccation-related Elip-like gene from the resurrection plant Craterostigma plantagineum is regulated by light and ABA.

The resurrection plant Craterostigma plantagineum tolerates an extreme loss of cellular water. Therefore this plant is being studied as model system to analyse desiccation tolerance at the molecular level. Upon dehydration, new transcripts are abundantly expressed in different tissues of the plant. One such desiccation-related nuclear gene (dsp-22 for desiccation stress protein) encodes a mature 21 kDa protein which accumulates in the chloroplasts. Sequence analysis indicates that dsp-22 is closely related to early light inducible genes (Elip) of higher plants and to a carotene biosynthesis related gene (cbr) isolated from the green alga Dunaliella bardawil. In contrast to other desiccation-related genes, light is an essential positive factor regulating the expression of dsp-22: ABA-mediated gene activation leads to the accumulation of the transcript only in the presence of light. During the desiccation process, light acts at the transcriptional and post-transcriptional levels. The implications of these different controls and the possible role of the dsp-22 protein in the desiccation/rehydration process are discussed.

Abscisic Acid

Expression of desiccation-related proteins from the resurrection plant Craterostigma plantagineum in transgenic tobacco.

Three cDNAs encoding desiccation-induced proteins from the resurrection plant Craterostigma plantagineum were each ligated to a triplicated CaMV 35S promoter and a nopaline synthase 3'-flanking region in an Agrobacterium vector and introduced into tobacco. Transgenic plants expressed the encoded Craterostigma proteins at high levels. This did not lead to changes in the phenotype, in the growth habit or in basic photosynthetic parameters. In tobacco, one protein was targeted to the chloroplast stroma which is its normal location in Craterostigma. These desiccation-related proteins are not sufficient per se to increase drought tolerance as measured by ion-leakage tests.

Biological Transport

Mass, center of mass, and moment of inertia estimates for infant limb segments.

To quantify limb dynamics, accurate estimates are needed of anthropometric inertia parameters (mass, center-of-mass location, and moments of inertia). These estimates, however, are not available for human infants; therefore, the movement dynamics of infants have not been studied extensively. Here, regression equations for the masses, center-of-mass locations, and transverse moments of inertia of upper and lower limb segments (upper arm, forearm, and hand; thigh, leg, and foot) of 0.04 to 1.50 yr old infants are provided. A mathematical model of the human body was used to determine the anthropometric inertia parameters for upper limbs in 44 infants and for lower limbs in 70 infants. Stepwise linear regressions were used to fit the distributions of the anthropometric inertia parameters. The regression equations accounted for significant amounts of the variance (64-98%), and the R2-values compared favorably when our equations were cross-validated. Consequently, these regression equations can provide, for infants of similar ages, reasonable estimates of upper and lower limb anthropometric inertia parameters, suitable for equations of motion in the analysis of limb dynamics in human infants.

Anthropometry

Characterization of the murine cytomegalovirus genes encoding the major DNA binding protein and the ICP18.5 homolog.

In several herpesviruses the genes for the major DNA binding protein (MDBP), a putative assembly protein, the glycoprotein B (gB), and the viral DNA polymerase (pol) collocate. In murine cytomegalovirus (MCMV), two members of this gene block, pol (Elliott, Clark, Jaquish, and Spector, 1991, Virology 185, 169-186) and gB (Rapp, Messerle, Bühler, Tannheimer, Keil, and Koszinowski, 1992, J. Virol., 66, 4399-4406) have been characterized. Here the two other MCMV genes are characterized, the gene encoding the MDBP and the ICP18.5 homolog encoding a putative assembly protein. Like in human cytomegalovirus (HCMV) the genes order is pol, gB, ICP18.5, and MDBP. The 4.2-kb MDBP mRNA is expressed first in the early phase, whereas the 3.0-kb ICP18.5 mRNA is a late transcript. The open reading frame of the MDBP gene has the capacity of encoding a protein of 1191 amino acids with a predicted molecular mass of 131.7 kDa. The MCMV ICP18.5 ORF is translated into a polypeptide of 798 amino acids with a calculated molecular mass of 89.1 kDa. Comparison of the amino acid sequences of the predicted proteins of MCMV with the respective proteins of HCMV, Epstein-Barr virus (EBV), and herpes simplex virus type-1 (HSV-1) reveals a striking homology ranging from 72% (HCMV), 50% (EBV), to 45% (HSV-1) for the MDBP sequence and from 74% (HCMV), 51% (EBV), to 49% (HSV-1) for the ICP18.5 sequence. These results establish the close relationship of the two cytomegaloviruses, and underline the usefulness of the murine model for studies on the biology of the CMV infection.

Amino Acid Sequence

Risk factors for early intraventricular hemorrhage in low birth weight infants.

Because earlier studies suggested that preterm infants with germinal matrix hemorrhage or intraventricular hemorrhage or both (GMH/IVH) present within the first 12 postnatal hours are at greatest risk for the development of high-grade hemorrhage and neurodevelopmental disability, we examined the risk factors for this insult among 229 neonates of 600 to 1250 gm birth weight in a multicenter study. All had echoencephalography (ECHO) within the first 11 hours and serially for the next 20 days; risk factor data were collected prospectively. Forty-three infants had GMH/IVH within the first 5 to 11 hours (mean age at ECHO 7.7 hours): 18 GMH and 21 grade II, 1 grade III, and 3 grade IV IVH. One hundred eighty-six infants did not have GMH/IVH at a mean age of 7.9 hours. Both groups of infants were similar in birth weight, gestational age, maternal risk factors, cord pH values, and surfactant therapy before ECHO. The group with early IVH had more vertex presentations than the group without early IVH (79% vs 55%, p = 0.043), less maternal tocolytic use (42% vs 60%, p = 0.029), and more vaginal deliveries (67% vs 44%, p = 0.005). In the first 21 days, severe IVH developed in 12 infants with early IVH and in 6 infants without early IVH (p < 0.001). There were more neonatal deaths (16% vs 6%, p = 0.035) and more deaths at any time during the primary hospitalization (23% vs 9%, p = 0.010) among the early IVH group than among the group without early IVH. Multivariate analysis indicated that the mode of delivery, fetal presentation, and birth weight were important and independent prognostic indicators of IVH.

Birth Weight

[The fetal brain in vaginal sonography].

Transvaginal sonographic imaging of the fetal brain performed in the second and third trimester of pregnancy is presented. By this technique coronal and sagittal sections of the central nervous system can be obtained which are similar to those performed via cranial sonography postnatally.

Echoencephalography

Long-term results after partial and unilateral nephrectomy in childhood.

In a clinical follow-up, 47 patients after unilateral or partial nephrectomy were examined. The average age at operation was 5.8 years (range 3 days-17.1 years), the average follow-up period after surgery was 11.9 years (range 8.2-18.3 years). Main indications for surgery were hydronephrosis (n = 27), other congenital malformations (n = 6) and neoplasia (n = 9). In one patient, we found hypertension, two patients presented with moderately diminished creatinine clearance, two further patients suffered from unilateral hydronephrosis after partial nephrectomy. We found by ultrasonography marked compensatory hypertrophy in cases of solitary kidneys (225.6 +/- 49.3%) as well as contralateral kidneys after partial nephrectomy (158.7 +/- 55.6%).

Adolescent

Mood-congruent and mood-incongruent learning.

We report two experiments that investigate the effect of an induced mood on the incidental learning of emotionally toned words. Subjects were put in a happy or sad mood by means of a suggestion technique and rated the emotional valence of a list of words. Later on, they were asked to recall the words in a neutral mood. For words with a strong emotional valence, mood-congruent learning was observed: strongly unpleasant words were recalled better by sad subjects and strongly pleasant words were recalled better by happy subjects. The reverse was true for slightly toned words: here, mood-incongruent learning was observed. Both effects are predicted by a two-component processing model that specifies the effect of the mood on the cognitive processes during learning. Further evidence for the model is given by rating times measured in Experiment 2.

Affect

Selective removal of molybdenum traces from growth media of N2-fixing bacteria.

A new method for the selective removal of traces of molybdenum from growth media of N2-fixing bacteria (Rhodobacter capsulatus and Klebsiella pneumoniae) was developed. This method is based on the filtration of nutrient solutions through a layer of activated carbon (pulverized charcoal). The adsorption of Mo (molybdate) to activated carbon was optimal if a charcoal suspension (50 g/liter) was degassed by boiling before use and if the pH of the solutions, which had to be purified, was adjusted to values between 1.5 and 4. In this pH region no or only negligible amounts of other metal ions were adsorbed. The activated carbon method was compared with other Mo-eliminating procedures, including 8-hydroxyquinoline/dichloromethane extraction, Chelex 100 chromatography, and treatment with Mo-starved Azotobacter vinelandii cells. The activated carbon filtration appeared to be the most effective, specific, and rapid method. Whereas the untreated Rhodobacter growth medium was contaminated with 1.2 ppb Mo, as analyzed by inductively coupled plasma mass spectrometry (ICP-MS), the activated carbon-treated medium was below the ICP-MS detection limit (less than 0.05 ppb). A similarly effective removal of Mo impurities was obtained by the Azotobacter treatment. Even at low optical densities (2-5 at 436 nm) Mo traces were removed very rapidly within 10-15 min. However, because the Mo uptake/Mo adsorption capacity of A. vinelandii depended on freshly cultivated cells and on the growth phase at which the cells were harvested, this microbiological method was generally more time-consuming and less reproducible than the activated carbon method.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption

Demonstration of a molybdenum- and vanadium-independent nitrogenase in a nifHDK-deletion mutant of Rhodobacter capsulatus.

In Rhodobacter capsulatus there exists, in addition to a conventional Mo-containing nitrogenase, a second, Mo-indendent nitrogenase which was demonstrated in wild-type cells as well as in cells of a nifHDK- mutant. To construct this R. capsulatus mutant, a 4-kb BglII-HindIII fragment encompassing nifK, nifD and most of the nifH coding region was substituted by an interposon coding for kanamycin resistance. The alternative nitrogenase is repressed by molybdenum. Mo concentration greater than 1 ppb in the growth medium prevented diazotrophic growth of nifHDK- cells and the expression of nitrogenase activity. The Mo-independent nitrogenase was maximally derepressed in activated carbon-treated media which contained less than 0.05 ppb Mo, high concentrations of iron (1 mM ferric citrate) and serine as N source. Under N2-fixing and optimal Mo-deficient conditions, nifHDK- cells grew with a doubling time of 9 h. The highest activity achieved with whole cells was 1.2 nmol ethylene.min-1.mg protein-1. Vanadium neither stimulated nor inhibited growth and activity. The alternative nitrogenase reduced acetylene to both ethylene and ethane. With whole cells (nifHDK-) the proportion of ethane varied over 2-5% depending on the amount of residual traces of Mo in the medium. The addition of Mo to a growing, nitrogenase-active culture resulted in a slow decrease of total activity but also in a simultaneous increase of ethane production up to 40%. In contrast, cell-free extracts and the purified enzyme did not show any or only very little ethane formation (0-0.4%). Both enzyme components appeared to be very labile proteins. Component 2 lost almost all its activity during cell breakage. With component 1 in crude extracts, if complemented with the stable component 2 of the Mo-nitrogenase from Xanthobacter autotrophicus, a recovery of 50% of the original whole cell activity could be achieved. During purification, component 1 (from the nifHDK- mutant) remained remarkably stable. The partially purified component 1 had a pH optimum (acetylene reduction) of 7.8-8.0, relatively high affinity to acetylene (Km = 0.055 mM) and was analyzed to contain 20 mol Fe atoms/mol protein, 0.2 mol Mo atoms and negligible amounts of V, W and Re. The dithionite-reduced dinitrogenase appeared to be ESR-silent. The results indicate that the alternative nitrogenase of R. capsulatus is not a vanadium enzyme but rather a heterometal-free Fe-nitrogenase or a nitrogenase with an as-yet-unidentified heterometal atom.

Chromatography, Gel

An ABA and GA modulated gene expressed in the barley embryo encodes an aldose reductase related protein.

In most higher plants a period of desiccation is the terminal event in embryogenesis. Excised barley embryos acquire desiccation tolerance at a precise developmental stage and cDNA clones have been isolated which are temporally linked with desiccation tolerance. One such clone (pG22-69) with a putative gene product of 34 kd displays high structural homology to mammalian genes encoding an NADPH dependent aldose reductase involved in the synthesis of sorbitol. This first aldose reductase gene of plants is expressed constitutively during embryo maturation and is modulated by the plant hormones abscisic acid (ABA) and gibberellic acid (GA). Immunohistochemistry showed that the protein is preferentially expressed in tissues formed at early stages in embryogenesis. Measurements of enzymatic activity indicate that pG22-69 encodes an active aldose reductase. The finding of this reductase activity and the cloning of the corresponding gene supports the existence of a metabolic pathway in plants playing a role in the synthesis of osmolytes like sorbitol. The significance of this work is that genes of related structure and functions are being used in diverse organisms to fulfil stress related biological requirements.

Abscisic Acid

Sonographic imaging of the thyroid in children.

High-resolution sonographic imaging of thyroid disorders in paediatrics has become an extremely accurate method and is being more frequently used. The need for scintigraphy has therefore dramatically decreased. The anatomy and sonographic morphology of the thyroid gland (normal findings, variants) in infants and children are presented, as are patterns of thyroid disorders and respective algorithms of diagnostic imaging.

Adolescent

[Nuclear magnetic resonance tomography of children's thorax].

Magnetic Resonance Imaging of the pediatric chest was performed in 53 patients. All examinations were performed using a 1.0 and 1.5 Tesla superconducting MR imager. 80% of the children were examined using anesthesia with a ventilation of 20% inspiration and 80% expiration. After performing the MR examination in 55% of the cases an operation was performed. In 29 patients anomalies of the brachiocephalic trunc were found. In four patients MR detected a doubled aortic arch, in four patients we found a dilated arch. In eight children a dilated pulmonary artery was diagnosed. In combination with eight tracheobronchoscopy, MR imaging proved to be the diagnostic method of choice for the evaluation of anomalies of the mediastinum. New developments like fast imaging techniques and 3D acquisition promise a further diagnostic improvement in the mediastinum.

Aorta, Thoracic

[Standards for preclinical resuscitation--requirements for efficient therapy and scientific analysis. A prospective study using as an example the combined emergency medical service of the Munich administrative district and capital].

A standardized treatment protocol is essential for scientific evaluation of parameters influencing the outcome and survival of patients who have suffered cardiac arrest in a non-hospital situation. In addition, a standardized therapy algorithm permits effective, time-saving interaction of all members of the emergency team who work together to perform cardiopulmonary resuscitation (CPR) in any emergency outside the hospital. This paper gives the results obtained in 50 patients in whom on-the-spot resuscitation was performed by a specially trained team [emergency medical team (EMT) + on-scene physician] using an ACLS (advanced cardiac life support) protocol modified from the AHA (American Heart Association) standard. Two different algorithms were used one for ventricular fibrillation (VF) and pulseless ventricular tachycardia and one for asystole and pulseless bradycardia. When indicated, countershocks were first administered at a continuous energy level of 360 J, up to three times one after the other. All patients then received epinephrine intratracheally, 2 mg, immediately after intubation. In the case or persisting asystole a further 2-mg dose of epinephrine and then one 5-mg dose were given i.v. in keeping with the ACLS protocol. In the case of persisting VF or pulseless tachycardia we gave one 100-mg dose of lidocaine i.v. and then performed the next countershock at the same energy level. The time the team members actually needed for the single steps of the ACLS protocol was meticulously documented with the aid of a stop watch.(ABSTRACT TRUNCATED AT 250 WORDS)

Algorithms

Amino acid sequences of two proline-rich bactenecins. Antimicrobial peptides of bovine neutrophils.

Bactenecins are highly cationic polypeptides of the large granules of bovine neutrophils, exerting in vitro a potent antimicrobial activity. Two bactenecins, with an approximate molecular weight of 7000 and 5000, called Bac7 and Bac5, are characterized by a high content of proline (greater than 45%) and arginine (greater than 23%) residues. Their complete amino acid sequences were determined by automated Edman degradation combined, in the case of Bac5, with plasma desorption mass spectrometry. Bac7 comprises 59 residues and includes three tandem repeats of a tetradecamer characterized by several Pro-Arg-Pro triplets spaced by single hydrophobic amino acids. Resolution of the primary structure of Bac5 required fragmentation with N-bromosuccinimide as well as digestion of the obtained C-terminal fragment with carboxypeptidases P and Y directly in the mass spectrometer. Bac5 comprises 42 amino acid residues with a repeated motif of Arg-Pro-Pro triplets also alternating with single apolar residues.

Amino Acid Sequence

Identification of distinct NAD-linked hydrogenase protein species in mutants and nickel-deficient wild-type cells of Alcaligenes eutrophus H16.

By crossed immunoelectrophoresis with antibodies against the NAD-linked hydrogenase the presence of three hydrogenase protein species was demonstrated in crude extracts of Alcaligenes eutrophus H16. Protein 1 (antigen 1) exhibited NAD-reducing activity and was shown to be identical with the native heterotetrameric enzyme. Protein 2 (antigen 2) was catalytically inactive in the antibody-precipitated form and corresponded to the beta subunit (56 kDa) of the holoenzyme. Protein 3 (antigen 3) was serologically distinct from antigen 2 and catalyzed NADH-oxidizing (diaphorase) activity, suggesting that it either consists of the alpha peptide or of the alpha and gamma subunits of the diaphorase dimer. Tandem immunoelectrophoresis revealed that antigen 2 was the predominant protein species in cells cultivated under nickel deficiency. Low concentrations of the diaphorase-active antigen 3 were also detected under these conditions. Extracts from mutants defective in the catalytic activity of NAD-reducing hydrogenase still contained the four polypeptides. This was shown by immunodiffusion and immunoblotting with antibodies raised against the individual subunits. However, as observed with nickel-deficient cells, no complete tetrameric protein could be identified, and the dominant subunit species (70-80%) was the beta peptide.

Alcaligenes