Dual effect of 2,3-diphosphoglycerate on the Bohr effects of human blood

Summary The influence of the red cell concentration of 2,3-diphosphoglycerate (2,3-DPG, 0.5–26 moles/g erythrocytes) on the CO₂-Bohr effect (pH varied by CO₂ at constant base excess) and the fixed acid-Bohr effect (pH varied by fixed acid or base at constantP _CO2) was studied in human blood at plasma pH values ranging between pH 7.2 and pH 7.6.Elevation of red cell 2,3-DPG concentration leads to a numerical decrease of the CO₂-Bohr coefficient referring to plasma pH. The fixed acid-Bohr coefficients are numerically smaller than the corresponding CO₂-Bohr coefficients and exhibit a maximum at normal red cell 2,3-DPG concentrations. The Bohr coefficients referring to red cell pH are distinctly higher than those referring to plasma pH, especially at high 2,3-DPG levels. This is due on the one hand to the physico-chemical properties of the intact red cell membrane, and on the other hand to a 2,3-DPG-induced decrease in the ratio pH_cell/pH_plasma.From the results it is concluded that 2,3-DPG exerts a dual effect on the Bohr coefficients of whole blood which is mediated 1. by the direct effect of 2,3-DPG on the allosteric properties of hemoglobin (as reflected by changes of the Bohr coefficients referring to red cell pH), and 2. by the effect of 2,3-DPG on pH_cell/pH_plasma.     

Effect of histamine and stress on the gastric mucosal Ca2+ content during the development of gastric ulcers

The influence of histamine, its triazole derivative (3--aminoethyl-1,2,4-triazole) and immobilization stress on the gastric mucosal Ca²⁺ content during the development of gastric ulcers in guinea pigs and rats was investigated.A considerable fall in the concentration of Ca²⁺ in gastric tissues of guinea pigs after administration of histamine 0.25 mg/kg, down to 80% (8.0 mol/g), its derivative (1 mg/kg) to 72% (7.2 mol/g) and stress to 76% (7.6 mol/g) was recorded by atomic absorption-spectrophotometric techniques, while the calcium level in the controls stood at 9.9 mol/g. Similar changes (90-65%) were seen in the blood plasma.In rats, the Ca²⁺-decreasing effects of stress ran closely parallel with increasing ulceration, and depended on the duration of stress. The immobilization of rats evoked a slight rise in the Na⁺ content of the gastric tissues. After 4 h immobilization, the tissue concentration of Na⁺ was increased to 116% of control levels. Cimetidine (100 mol kg^–1 more than 50% inhibited the development of gastric ulcers and prevented the change in Ca²⁺ concentration ions. Thus, the data suggest that Ca²⁺ ions take part not only in the regulation of secretion, but also in stress tissue dystrophy.     

α4β1 and α5β1 Control Cell Migration on Fibronectin by Differentially Regulating Cell Speed and Motile Cell Phenotype

Computer-aided time lapse fluorescence videomicroscopy was used to study single cell migration behavior of human aortic endothelial cells on fibronectin coated substrates of varying protein surface density. The role of receptors _₅ _₁, __v _₃, and ₄₁ in mediating cell adhesion and migration on fibronectin was characterized using integrin specific monoclonal antibodies. Matrix density had a direct effect on controlling the proportion of migrating cells and the directional persistence of cell movement (p < 0.01). While there was relatively little influence of fibronectin surface density on absolute migration speed, the ability of endothelial cells to disperse over a surface, as measured by the dispersion coefficient, was biphasic with respect to the surface density of this matrix protein (p < 0.005). Both cell speed and the proportion of migrating cells was controlled by ₄₁ (p < 0.01). However, ₅₁ selectively regulated the transformation of stationary cells to those exhibiting motile behavior (p <0.05). Migratory responses on fibronectin were not influenced by blockade of the _v3 receptor. It is noteworthy that cell surface adhesive receptors which control commitment to a motile phenotype are not necessarily the same as those that control migration speed. © 1998 Biomedical Engineering Society. PAC98: 8722-q     

Down-regulation of tumor necrosis factor α activity by acute ethanol treatment in human peripheral blood monocytes

As the most commonly used drug that can modulate both metabolic and immune pathways, ethanol is evaluated in this report as a regulator of tumor necrosis factor (TNF) production in human peripheral blood monocytes (M) in combination with a variety of stimuli. While acute ethanol treatment did not induce TNF in M, it was a potent down-regulator of M TNF production whether induced by the combination of interferon- plus muramyl dipeptide (MDP) (P<0.001), lipopolysaccharide (LPS) alone (P<0.01), or interferon- plus LPS. Down-regulation of M TNF by ethanol was dose dependent and statistically significant in the biologically relevant, 25–150 mM, ethanol concentration range. We also demonstrate that these ethanol concentrations did not affect M viability. TNF down-regulation by ethanol was most effective when ethanol was administered 4 hr prior to MDP stimulation; however, it was also effective—though to a lesser extent—if it was added at the time of MDP stimulation. Furthermore, ethanol also down-regulated TNF production of thein vivo preactivated M of trauma patients, which produce hyperelevated levels of TNF. We have previously shown that the majority of posttrauma elevated M TNF is produced by the M subpopulation expressing high-affinity type I Fc receptors (FcRI). When the FcRI cross-linking-stimulated M subpopulation was treated with acute ethanol, TNF production was suppressed again both inin vivo preactivated M of trauma patients and in M of normal controls. In experiments utilizing cyclooxygenase inhibitor, we also demonstrate that ethanol has a direct, prostaglandin E₂-independent, effect on M TNF production. These results demonstrate that acute ethanol exposure has the potential to down-regulate M production of TNF significantly regardless of the TNF-inducing stimulus. Decreased capacity of M to produce TNF might, therefore, contribute to the immunological and metabolic abnormalities described after ethanol uptake.     

A comparative immunohistological study of cerebellar enolases

Summary A comparative immunohistological study of the neurone-specific enolase and enolase, demonstrates the exclusive neuronal localization of enolase and its absence from glial cells. In contrast, enolase is located in astroglial cells. The validity of enolase as a neuronal marker and enolase as an astrocytic marker, is confirmed both by a double labelling technique, using antibodies to and to revealed with fluorescence or peroxidase in the same tissue sections, and by immunoelectronmicroscopy.     

Comparison of the properties of five pox virus strains isolated from monkeys

Summary Five strains of monkey pox viruses were compared with respect to their cultural characteristics in primary and continuous cell cultures and the lesions developed in embryonated eggs and in rabbit skin as well as to their hemagglutinating activity.Four strains (Copenhagen 65-31 65-32 and 7-61) appeared to be similar in their properties. The cytopathogenic effect (CPE) was identical to that induced by vaccinia virus. There was no detectable virus multiplication in an pig kidney cell line (PEK). All four strains produced small, white, compact, hemorrhagic pock-like lesions on the chorioallantoic membrane.The strain 64–7275, isolated from healthy monkeys kidneys, had all properties of variola virus. It multiplied in the PEK cell line with a CPE. The lesions on the CAM were more compact without hemorrhage. In rabbit skin no detectable reaction occurred after infection with this strain.     

Immunocytochemical characterization of porcine zona pellucida during follicular development

Sections of bovine ovaries fixed in Bouin's fluid or methanol-acetic acid and embedded in paraffin were incubated with chicken polyclonal antibodies to HPLC-purified zona glycoproteins ZP3 and ZP3. Oocytes of primordial follicles as well as of primary follicles showed weak labelling with anti-ZP3 and anti-ZP3. No immunostaining could be observed in the follicle cells. The ZP of primary follicles displayed distinct immunoreactivity for both ZP3 and ZP3. In secondary follicles, distinct labelling with anti-ZP3 and weak labelling with anti-ZP3 could be seen in the oocyte. The ZP showed immunoreactivity with antibodies to ZP3 and ZP3. Both antibodies labelled single follicle cells. In tertiary follicles, the oocytes were weakly labelled with anti-ZP3 and anti-ZP3. Some granulosa cells showed staining for ZP3 and ZP3. The ZP displayed strong immunoreactivity for ZP3 and ZP3. Cells of the corona radiata were strongly immunopositive for ZP3 and ZP3. Similar histotopography of immunoreactive cells could be seen in preovulatory follicles. The characteristic pattern observed for the distribution of ZP3 and ZP3 strongly suggests that in the porcine ovary both the oocyte and the follicle cells contribute to the synthesis of the ZP, perhaps in sequence.     

Investigation of the “valency” of antibodies by means of azoproteins

Summary The valency of antibodies was studied by the method of exhaustion of antisera against mono-and diazoproteins, and subsequent cross reactions both with the antibodies left over in the supernatant fluid of the serum and with the precipitating and nonprecipitating antibodies isolated from the precipitate.It was proved that the antibodies interact with the antigens as multivalent compounds.The valency determined with regard to the azoproteins is dependent upon the number of groups introduced.Thus, bivalent antibodies correspond to monoazoproteins and trivalent ones to diazoproteins.The valency of antibodies is, evidently, determined by the structural similarity of the heterologous and the immunizing antigens as well as by the less complete specific conformity between the individual structural peculiarities of the antigen and its antibody.From the Tashkent Pharmaceutical Institute (Director-Docent M. A. Azizov)Presented by Active Member AMN SSSR N. N. Zhukov-Verezhnikov     

Response to therapy of a type III hyperlipoproteinemic subject with the rare apolipoprotein E1 (Gly127 → Asp,Arg158 → Cys) variant

Summary In a preceding paper, we described the molecular biological defects in a patient with a severe form of the familial lipoprotein disorder type III hyperlipoproteinemia (HLP) and an unusual apolipoprotein (apo) El phenotype and 1/null genotype. The index case was a 60-year-old white male of German ancestry who suffered from a myocardial infarction at age 50 years. He had distinctly elevated levels of plasma lipids (triglycerides 551 mg/dl and cholesterol 747 mg/dl, respectively) and typical clinical signs of this inborn error of lipoprotein metabolism. His mutant apo E1 was shown to be identical to a rare (already described) apo E1 (Gly₁₂₇Asp, Arg₁₅₈Cys) variant. A second independent defect at the molecular level was a nucleotide deletion of a guanosine (G) in the codon for amino acid 31 of the proband's apo 3 allele. This single base deletion (not described before) changed his apo 3 allele to a nonfunctional null allele devoid of a stable gene product. Here we describe the response to combined dietary and medical treatment of the patient with this unusual form of type III HLP. His response to therapy was excellent, similar to patients with classical type III HLP and homozygosity for apo E2. However, the correct diagnosis of this familial lipoprotein disorder seems to be necessary, even in patients without the expected apo E2/2 phenotype, in terms of the prompt and beneficial response to therapeutic interventions.Abbreviations Apo Apolipoprotein - Chol cholesterol - HDL high density lipoproteins - HDL-C HDL-cholesterol - HLP hyperlipoproteinemia - IDL intermediate density lipoproteins - IEF isoelectric focusing - LDL low density lipoproteins - LDL-C LDL-cholesterol - TG triglycerides - VLDL very low density lipoproteins - VLDL-C VLDL-cholesterol Dedicated to Prof. Dr. G. Schettler on the occasion of his 75th birthday on April 13, 1992     

Kombinationsformen der Arteriitis

Zusammenfassung Ausgehend von den UntersuchungenRössles zum Formenkreis der rheumatischen Gewebsveränderungen, wird an Hand von 4 eigenen Fällen die Problematik der morphologischen Differentialdiagnose zwischen rheumatischer Arteriitis, Periarteriitis (Panangiitis) nodosa und Thrombangitis obliterans unter Berücksichtigung der Literatur aufgezeigt. Mit der morphologischen Definition der verwendeten Begriffe fibrinöse Entzündung und proliferierende und granulierende Angiitis wird versucht, die drei Formen der Gefäßwandentzündung morphologisch abzugrenzen. Die überschneidungen der morphologischen Symptome und die Kombinationsformen werden aus Beschreibung und Abbildung einschlägiger veröffentlichter Fälle aufgezeigt. Aus dem Nachweis dieses Schrifttumsvergleichs und dem der Differentialdiagnose der 4 eigenen Fälle wird gefolgert, daß bei solchen nicht-klassischen Gefäßentzündungen die Annahme von Kombinationsformen richtiger ist als die immer stärkere Erweiterung des sog. rheumatischen Formenkreises.

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Summary With reference to the investigations ofRössle On the Classification of the Rheumatic Tissue Changes, the problems that are associated with the differential diagnosis of rheumatic arteritis, periarteritis (panangiitis) nodosa, and thromboangiitis obliterans are discussed, exemplified by the report of four cases, and by a review of the literature. By applying the morphological criteria of fibrinous inflammation, proliferative and granulomatous angiitis, an attempt is made histologically to differentiate these three forms of inflammation of the vascular wall. The overlapping of the morphological criteria and the combined forms that mav occur are illustrated in the four cases reported. As shown by comparing the cases in the literature, and as exemplified by the differential diagnosis of the four cases, the inference is made, that with such types of non-classical vascular inflammation, it is better to refer to combined forms than to the more diffuse so-called rheumatic classification.

     

Der gegenwärtige Stand der Mastzell-Forschung

Zusammenfassung Seit der Erstbeschreibung der MZ durchEhrlich (1877) ist die Kenntnis über die biologische Rolle dieser Zellen wesentlich erweitert worden; diese, mit metachromatischen Granula erfüllten Zellen haben nicht nur die Eigenschaft, Substanzen zu speichern (Mastzellen), sie haben auch die Fähigkeit, biologisch aktive Substanzen zu bilden, diese an das Erfolgsgewebe abzugeben (sekretorische Zellen, Heparinocyten, Histaminocyten) und sind sowohl in morphologischer als auch in funktioneller Hinsicht in den Bauplan der vegetativen nervösen Peripherie mit eingeschlossen (neurohormonale Zellen).     

Tuning properties of auditory cortex cells in the awake squirrel monkey

Summary Pure tone bursts elicited in primary auditory cortex (AI) cells of the awake squirrel monkey a wide range of response patterns which consisted of one or more excitatory or inhibitory temporal response components. In almost 60% of these cells, response patterns were frequency and/or intensity dependent. Response components such as early and late onset excitation, offset excitation and on-off excitation; as well as tonic excitation or inhibition often varied independently with changes in these stimulus parameters. Individual cells were therefore considered as multiple bandpass filters, and each discrete response component was analyzed separately for its tuning properties. A correlation between best frequencies of the various excitatory components (BEF), and between BEFs and best frequencies of inhibitory components (BIF), in cells which responded with more than one discrete response component, disclosed a significantly higher correlation between BEF/BIF pairs compared with BEF/BEF pairs, presumably reflecting certain lateral inhibition like processes. Applying Q_10dB factor, and Hf-Lf bandwidth at 10 dB above threshold, as measures of the sharpness of response areas, revealed that approximately 65% of all response areas could be defined as narrow by either one of these 2 measures, with no distinction, in that regard, between excitatory and inhibitory components. The average response bandwidths of the narrowly and the broadly tuned components, at 10 dB above threshold, were 0.4±0.18 and 1.42±0.68 octaves respectively. A comparison with the medial geniculate body (MGB) of the squirrel monkey, applying the Hf-Lf measure of sharpness of tuning, showed a significantly higher proportion of narrow response areas in the AI. Narrow response areas in both these regions were equally narrow, whereas the broad response areas of MGB cells were significantly broader. These results suggest a sharpening of response areas throughout the geniculo-cortical transformation.     

Identification and quantitation of cortisol metabolites in rat liver homogenate and -slices

Summary Cortisol-1, 2-H³ was incubated with rat liver homogenate and/or rat liver slices in the presence of a NADPH-generating system. The following metabolites could be identified in adult male rats: -cortol, allo--cortol, 3-allo--cortol, 20-hydroxy-cortisol, 11, 17, 20, 21-tetrahydroxy-5-pregnan-3-one, 3-allotetrahydrocortisol, tetrahydrocortisol, trace amounts of allotetrahydrocortisol and two highly polar metabolites only partly identified. In female rats only tetrahydrocortisol, allotetrahydrocortisol and allodihydrocortisol could be detected in significant amounts.The radioactive metabolites mentioned above were localized and quantitated on paper chromatograms by a 4-radiochromatogram scanner. A nearly perfect correlation was found between these results so obtained and those given by liquid-scintillation counting of each metabolite after its elution from the paper.Part of this work was supported by grant n° 695 of the National Fonds voor Wetenschappelijk Geneeskundig Onderzoek.Stagiair of the Nationaal Fonds voor Wetenschappelijk Onderzoek.     

Genes for C4b-binding protein α- and β-chains (C4BPA andC4BPB) are located on chromosome 1, band 1q32, in humans and on chromosome 13 in rats

C4b-binding protein is involved in the regulation of the complement system. It is a multimeric protein composed of seven identical -chains and a single copy of a unique -chain. The latter was identified only recently and its structure determined by cDNA cloning. Both subunits in C4b-binding protein belong to the same superfamily of proteins composed predominantly of tandemly arranged short consensus repeats (SCR) approximately 60 amino acid residues in length. The gene for the human -chain is known to be located in a gene cluster on chromosome 1, band 1q32, which is called the regulators of complement activation (RCA) gene cluster. We have used cDNA probes for both - and -chains of human C4b-binding protein to localize their genes with an in situ hybridization technique. We find the genes for both chains to be located on chromosome 1, band 1q32, in the human. This suggests that the -chain gene is also a member of the RCA gene cluster and that the - and -chain genes are located close to each other. The cDNA probes for the - and -chains also were used to screen mouse-rat somatic cell hybrids using Southern blotting to localize their genes in the rat. Both the - and -chain genes were shown to be located on chromosome 13 in the rat. These are the second and third genes to be located on rat chromosome 13, and the results suggest that the genes for the - and the -chains together with the gene for coagulation factor V represent a conserved chromosomal region in rat and man.     

Hemoglobin heterogeneity under conditions of abnormal erythropoiesis

Experiments on rats showed that changes in the hemoglobin profile during hypoxia are determined by switching of erythropoiesis from the basic to emergency mode. The basic mode of erythropoiesis is typical of the mature organism under normal conditions. The reserve or emergency mode is realized in fetuses, senile animals, and during hypoxia. This mode is characterized by production of large erythrocytes with high content of fetal hemoglobin.     

Interferon-γ for treatment of severe atopic eczema in two children

Two 4- and 5-year-old children suffering from refractory atopic dermatitis were treated with recombinant interferon- (rIFN-). rIFN- was injected at 50 g subcutaneously three times a week in the first child for 3 weeks, followed by three times 25 g in week 4. In the other child two treatment courses of 4 weeks were given after a break of 2 weeks. Therapy was well tolerated. In child one reductions in eczematous body surface and severity of lesions were observed, while no beneficial effect was seen in the other. Clinical chemistry data remained unchanged. Immunological studies performed in parallel showed a decrease in total serum IgE of 50% in child 1, a decrease in spontaneous in vitro IgE production, an increase in in vitro production of interleukin-6, and a normalization of previously decreased in vitro lymphocyte responses to several mitogens. While marked immunological changes were noted during IFN- treatment, clinical benefits were not encouraging. Diminished IFN- production has been claimed to be a major pathogenic factor in atopic eczema. Our results indicate that the pathogenesis is more complex. Clinically, we were unable to confirm previous observations in adults. Further studies are needed before IFN- can be recommended for therapy of pediatric atopic eczema.Abbreviations IFN- interferon- - IL interleukin     

Characterization and localization of α-connectin (titin 1): An elastic protein isolated from rabbit skeletal muscle

Summary A simplified procedure to isolate-connectin (titin 1, TI), a gigantic elastic protein, from rabbit skeletal muscle is described. A rapid column chromatography step to concentrate-connectin is introduced. Separation of-connectin from-connectin is introduced. Separation of-connectin from-connectin (titin 2, TII) in the presence of 4 M urea at pH 7.0 did not cause any change in the secondary structure of-connectin as judged by circular dichroic spectra. Ultraviolet absorption spectra and the amino acid composition of-connectin (MW, approximately 3×10⁶) were similar to those of its proteolytic product,-connectin (MW, approximately 2×10⁶). Circular dichroic spectra suggested that both- and-connectin consist of 60%-sheet and 30%-turn. It thus appears that the whole elastic filament of connectin has a folded-strand structure. Proteolysis of-connectin by calpain resulted in formation of-connectin and smaller peptides. The-connectin interacted with both myosin and actin filaments similarly to-connectin. Polyclonal antibodies raised against 1200 kDa peptides obtained from aged rabbit skeletal myofibrils reacted with-connectin (titin 1, TI) but only weakly with-connectin (titin 2, TII) in rabbit skeletal muscle. Immunoelectron microscopy and indirect immunofluorescence microscopy revealed that the antibodies bound at the Z-line and at the epitope regions in the I-band near the binding site of a monoclonal antibody SMI whose position depends on sarcomere length. It thus appears that-connectin extends from the edge of M-line to the above epitope region in the I-band.     

Interferon β1a Treatment Modulates TH1 Expression in γδ + T Cells from Relapsing-Remitting Multiple Sclerosis Patients

A paradigm exists that multiple sclerosis is causally related to dysregulation of T_H1 inflammatory cytokines and T_H2 antiinflammatory cytokines. The cytokine source(s) that initiate the imbalances are unknown. In this study, , CD4, and CD8 T cell receptor-positive (TCR+) cells were isolated from the blood of 26 definitive relapsing-remitting multiple sclerosis patients prior to interferon -1a (IFN1a) therapy and following 8–10 weeks of this therapy. The bioactivities of interferon (IFN), interleukin 10 (IL10), and interleukin 12 (IL12) were determined. The concentrations of IFN, IL10, and IL12 from each cell type did not change significantly with IFN1a treatment. The IL10 secreted by TCR+ cells strongly correlated with the IL12 secreted by the same TCR+ cells, supporting the paradigm. Furthermore, IFN1a therapy decreased the TCR+ cell secretion of T_H1 cytokines after 8–10 weeks of therapy.     

Trennung der intestinalen β-Galactosidasen beim lactose-toleranten Erwachsenen durch Ultrazentrifugation im Dichtegradienten

Zusammenfassung Die intestinalen-Galactosidasen von 4 lactose-toleranten, erwachsenen Mitteleuropäern wurden im Saugbiopsie-Gewebe nach Solubilisierung mit Triton X-100 in einem linearen Mannitol-Gradienten (5–20%) auf der Ultrazentrifuge bei 4°C und 44000 U/min getrennt. Bei 12stündiger Zentrifugation fanden sich 3 Fraktionen, von denen die beiden schneller sedimenticrenden Lactose spalten. Alle 3 Fraktionen hydrolysieren p-Nitrophenyl--Galactosid. Die 3 isolierten-Galactosidasen entsprechen wahrscheinlich der neutralen Bürstensaum-Lactase, der sauren lysosomalen Lactase und einer cytoplasmatischen Hetero--Galactosidase.     

Biotin-rich intranuclear inclusions in morule-lacking adenocarcinoma of the gallbladder: a new category of “neoplastic/non-morular” lesions

Biotin-rich intranuclear inclusions, also called optically clear nuclei, are observed in various neoplastic and non-neoplastic lesions, including pregnancy-related endometrium and benign and malignant neoplasms with morular structures. A recent study reported that lesions with biotin-rich intranuclear inclusions can be classified as (non-neoplastic) pregnancy-related endometrial and as (neoplastic) morular category. In the present report, we describe two cases of well-differentiated adenocarcinoma of the gallbladder in which biotin-rich intranuclear inclusions were found without morular structures. Immunohistochemically, as reported previously, the intranuclear inclusions were positive for biotin and two biotin-binding enzymes (pyruvic acid carboxylase and propionyl CoA carboxylase). Intranuclear expression of -catenin was also observed in neoplastic cells with and without intranuclear inclusion. We also detected a frame shift mutation of APC gene in one case but no mutation of -catenin gene in both cases. Although intranuclear expression of -catenin by mutation of APC gene might contribute to carcinogenesis in our cases, the relationships among intranuclear expressions of -catenin, biotin, biotin-binding enzymes and intranuclear inclusions remain unclear. Our cases are the first neoplastic lesions with biotin-rich intranuclear inclusions that lacked morular structures. We propose a new neoplastic/non-morular category for lesions with biotin-rich intranuclear inclusions.