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Biomedical subjects

J Schilling

Publications and source records attributed to J Schilling.

15 recordsLinked to original sources

Penetration of fleroxacin into human lung, muscle, and fat tissue.

Fleroxacin is a new fluoroquinolone with established potent antimicrobial and pharmacokinetic properties, and the aim of this study was to determine its rate of penetration into human lung, muscle, and fat tissues. In a total of eight patients undergoing lung surgery, plasma and tissue concentrations of unmetabolized fleroxacin were determined. This was done in subgroups of two patients each, receiving either 6, 8, 12 or 24 hours prior to surgery one single oral dose of 400 mg fleroxacin. It was found that fleroxacin penetrates well into muscle and lung tissues, but not into fat tissue. The levels in muscle and lung were 1.1 to 4.7 times higher than the according plasma concentrations.

Adipose Tissue

[The course of pregnancy with placenta praevia diagnosed in the 2nd and 3rd trimester].

Selected clinical features of the course of pregnancy in 118 gravidas with placenta praevia diagnosed ultrasonographically on the first breeding were compared. The clinical features significantly connected with bleeding and the prognostic features for the way of delivery were stated. The longest bleeding was observed in patients with partial or marginal placenta praevia. The most common anterior location of low placenta praevia was connected with the lowest risk as regards the cesarean section due to hemorrhage. The earlier occurrence of the bleeding was associated with its longer duration. The time of the occurrence of the first bleeding and its duration were associated with a duration of the pregnancy and a body mass of the newborn. The recurrence of bleeding was of no prognostic importance. The appearance of the first bleeding after the 28th week of pregnancy was significantly connected with the necessity of performing a cesarean section due to hemorrhage in case of central, partial or marginal placenta praevia. In case of low-lying placenta the way on delivery was significantly related to the time of appearance of the first bleeding and the distance between the placenta and the internal os. Fisher's linear discriminative function of the two features allowed for the determination of a mathematical formula for the prognosis of the way of delivery in case of a low-lying placenta.

Adolescent

Synthesis and characterization of the Kunitz protease-inhibitor domain of the beta-amyloid precursor protein.

To understand the pathological process by which amyloid is deposited in Alzheimer's disease, it is important to characterize the proteolytic processing events of the beta-amyloid precursor protein (beta-APP) from which the amyloid-forming fragment is excised. A potentially important component in beta-APP processing is the 57-amino acid (aa) Kunitz serine protease inhibitor (KPI) located within the extracellular domain of both the 751- and 770-aa isoforms of beta-APP. We have synthesized DNA encoding the 57-aa KPI domain as a necessary step in identifying the role of the protease inhibitor in beta-APP processing and amyloid formation. A bacterial secretion system directed by the alkaline phosphatase signal peptide of Escherichia coli linked to a synthetic gene encoding KPI was used to produce soluble, extracellular recombinant KPI (reKPI) protein. The reKPI protein was purified to homogeneity from bacterial supernatants and was biochemically and biologically characterized. Complete aa sequence analysis confirmed the fidelity of the reKPI, and fast-atom bombardment mass-spectral analysis was used to document that reKPI was of the predicted Mr. The reKPI is as active on a molar basis as the inhibitor-containing beta-APP when assayed for inhibition of trypsin activity. Together these data suggest that reKPI protein is properly folded and lacking in modified aa. Hence, this reKPI will be an important reagent in gaining a better understanding of the role of the KPI domain in beta-APP function and metabolism, as well as in the proteolytic events involved in beta-amyloid formation.

Alzheimer Disease

A synaptic vesicle specific GTP-binding protein from ray electric organ.

A cDNA encoding a synaptic vesicle associated GTP-binding protein was identified by screening a lambda gt11 expression library derived from the electric lobe of Discopyge ommata with polyclonal antibodies recognizing vesicle-specific proteins of Mr 25,000. Nucleotide sequence analysis defines an open reading frame of 218 amino acids. The protein belongs to the ras superfamily and shares about 75% amino acid identity with smg-25A, B and C identified in bovine brain and rab3A characterized in rat brain. Northern blot analysis revealed a 4.5 kb transcript present only in neural tissues, the highest level of expression being observed in electric lobe. Western blot analysis of total tissue homogenates derived from D. ommata detected the protein in electric organ, forebrain and to a lesser extent in electric lobe and spinal cord. No immunoreactivity was detected in non-neuronal tissues. Blotting of subcellular fractions derived from electric ray electric organ revealed that the GTP-binding protein co-purifies with synaptic vesicles. The neural specific expression and the localization to synaptic vesicles suggest a role of this protein in synaptic vesicle trafficking and targeting.

Amino Acid Sequence

Conformation of apolipoprotein B-100 in the low density lipoproteins of tangier disease. Identification of localized conformational response to triglyceride content.

The low density lipoproteins (LDL) from patients with Tangier disease are enriched in triglycerides, 27% of LDL mass versus 7% for normal LDL. To study whether this unique LDL core lipid composition affects the surface disposition of apolipoprotein (apo) B-100, we analyzed the LDL by protease digestion and in competitive radioimmunoassays. Limited proteolytic digestion of Tangier LDL by Staphylococcus aureus V8 protease generated a prominent fragment of 120 kDa (cleavage site at residue 1076), which was not visible in similarly digested normal LDL. In competitive radioimmunoassay, Tangier LDL bound weakly to the apoB-specific monoclonal antibody MB20, compared with control LDL. We localized the MB20 epitope between residues 1031 and 1084 of apoB-100, probably very near residue 1076. DNA sequencing of exon 21 of apoB genomic clones (coding for residues 1014-1084) from a Tangier patient revealed no difference from the normal DNA sequence, thus eliminating a protein polymorphism as a basis for the altered protease sensitivity and antibody binding. When the triglyceride contents of Tangier LDL were reduced to 10% of mass by incubation with normal high density lipoproteins, production of the 120-kDa fragment by proteolysis decreased and MB20 binding increased in affinity, implying a change toward normal conformation of apoB-100. Thus, using two independent techniques, proteolytic digestion and binding of monoclonal antibodies, we have demonstrated an alternative conformation of apoB-100 in the vicinity of residue 1076, which reflects the content of triglycerides in the LDL particle.

Amino Acid Sequence

Protease-specificity of Kunitz inhibitor domain of Alzheimer's disease amyloid protein precursor.

The putative inhibitor domain of Alzheimer's disease amyloid protein precursor was purified from E. coli containing a synthetic gene encoding the Kunitz domain. The purified protein (A4 inhibitor) inhibited the activity of trypsin, forming a 1:1 molar complex with the enzyme. It also strongly inhibited plasmin (Ki = 7.5 x 10(-11) M) from human serum and tryptase (Ki = 2.2 x 10(-10) M) from rat mast cells (tryptase M). In addition, it inhibited rat pancreatic trypsin, alpha-chymotrypsin and kallikrein and human serum kallikrein, but did not inhibit rat chymase, pancreatic elastase, alpha-thrombin, urokinase, papain or cathepsin B.

Amino Acid Sequence

Sequence of histone 2B of Drosophila melanogaster.

The complete sequence of histone 2B of Drosophila has been determined by using an improved Beckman sequenator. Comparing these data with those previously published by other investigators on the histone 2B of calf [Iwai, K., Hayashi, H., & Ishikawa, K. (1972) J. Biochem. (Tokyo) 72, 357--367], trout [Koostra, A., & Bailey, G. S. (1978) Biochemistry 17, 2504--2510], and Patella (a limpet) [van Helden, P. D., Strickland, W. N., Brandt, W. F., & von Holt, C. (1979) Eur. J. Biochem. 93, 71--78], it is possible to assess the evolutionary stability of this protein. There is little conservation of sequence in the N-terminal portion of the molecule (residues 1--26 numbering according to calf H2B), while the remainder of the protein, which we designate the C-terminal portion, is highly conserved. In the region of 27--125 residues, there are 9 substitutions in the composite data among the 98 positions, 8 of them conservative. These data indicate that very different selective pressures operate on the two different portions of the H2B molecule, implying the existence of two well-defined regions. Studies on the structure of the nucleosome by others have suggested that the C-terminal portion of H2B is involved in histone-histone interactions while the N-terminal portion is a relatively free "tail" binding to DNA. The sequence data indicate that the function of the C-terminal region of H2B requires considerable sequence specificity while that of the N-terminal region does not.

Amino Acid Sequence

Amino acid sequence of a mouse immunoglobulin mu chain.

The complete amino acid sequence of the mouse mu chain from the BALB/c myeloma tumor MOPC 104E is reported. The C mu region contains four consecutive homology regions of approximately 110 residues and a COOH-terminal region of 19 residues. A comparison of this mu chain from mouse with a complete mu sequence from human (Ou) and a partial mu chain sequence from dog (Moo) reveals a striking gradient of increasing homology from the NH2-terminal to the COOH-terminal portion of these mu chains, with the former being the least and the latter the most highly conserved. Four of the five sites of carbohydrate attachment appear to be at identical residue positions when the constant regions of the mouse and human mu chains are compared. The mu chain of MOPC 104E has a carbohydrate moiety attached in the second hypervariable region. This is particularly interesting in view of the fact that MOPC 104E binds alpha-(1 leads to 3)-dextran, a simple carbohydrate. The structural and functional constraints imposed by these comparative sequence analyses are discussed.

Amino Acid Sequence

Analysis of the diversity of murine antibodies to dextran B1355: N-terminal amino acid sequences of heavy chains from serum antibody.

The N-terminal amino acid sequences of two gamma and two mu chains from normally induced serum antibodies to dextran in BALB/c mice are presented. These heavy chains are derived from antibodies with three distinguishable idiotypes. These variable region (VH) sequences are all identical as far as they have been analyzed (27 to 53 residues). The light chains from these antibodies are all of the lambda type and are identical by isoelectric focusing analysis. Accordingly, the diversity of dextran antibodies appears to reside primarily in the heavy chains. The implications of these observations for antibody diversity are discussed.

Amino Acid Sequence

Rearrangement of genetic information may produce immunoglobulin diversity.

The nearly complete amino-acid sequences of 22 closely related immunoglobulin kappa variable (Vkappa) regions from the inbred NZB mouse are presented. This group of Vkappa regions is encoded by at least six germline Vkappa genes. These data also suggest that the mouse kappa gene is divided into three segments termed V or variable (residues 1 to 98 or 99), J or joining (residues 99 or 100 to 112) and C or constant (residues 113--219). Tonegawa et la. have recently described a similar J segment for mouse lambda chains. Inbred mice contain multiple Vkappa and Jkappa gene segments. Therefore, different combinations of V and J gene segments may be joined at the DNA level during the differentiation of individual lymphocytes to contribute to antibody diversity.

Amino Acid Sequence

[Methods for diagnosing placenta praevia].

All methods of placenta praevia diagnosis such as: amniography, cystography, arteriography soft X-ray, displacement, isotope technique, thermography, Doppler's technique, and NMR have been surveyed and compared. Historical background of each technique respective modifications, interpretation of the results, complications, contraindications and safety are discussed.

Diagnostic Imaging