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Biomedical subjects

G Reinhardt

Publications and source records attributed to G Reinhardt.

At least 55 records · Page 3Linked to original sources

In vitro exposure of the early mouse embryo to Herpes Simplex Virus-1 strain Wal.

The causes of early embryonic death are not clearly understood, but one of them may be viral infection. To study the interaction between the virus and the undifierentiated cell, early mouse embryos in morula and blastocyst stages were exposed to Herpes Simplex Virus-1 WAL (HSV-1 WAL). In one group of a total of 167 embryos, 108 were exposed to HSV-1 WAL; the rest were maintained as controls. After washing in culture medium, these embryos were cultured on a murine fetal fibroblast monolayer for viral isolation. None showed cytopathic effects in the susceptible monolayer. In a second group, 140 empryos were exposed and 106 were maintained as controls. These embryos were cultured without a monolayer or washing to permit continuous viral contact. Eighty-seven of the exposed embryos and 74 control empryos developed normally 2 to 3 d post hatching with no morpnological differences between the two groups. No statistical differences were observed when the proportion of natched and degenerated embryos was compared. Our results indicated that the cells of early mouse embryos are not susceptible to HSV-1 WAL. We concluded that possibly the susceptibility of empryonic cells to viral agents partially depends on stage of differentiation.

Journal Article↗

Parathyroid hormone secretion by brain and pituitary of sheep.

The secretion of immunoreactive PTH by different brain regions and pituitary of sheep was studied in vitro. Separate tissue samples of gyrus, internal capsule, basal ganglia, cerebellum, and pituitary were incubated in culture medium with low, normal, and high Calcium (Ca)(0.7 mM, 1.2 mM and 2.4 mM) concentrations. PTH release in medium containing low Ca were observed by all samples. The concentrations increased in the fifth and sixth hour from 100% (1st hour basis value) up to 300%. The PTH release showed an inverse relationship to the Ca concentration in the culture medium. To induce any significant decrease in medium concentrations of PTH, 1.25(OH)2D (100 ng/ml) was added to the culture medium. This effect could be reversed by DB-cAMP. Our results indicate that secretion of immunoreactive PTH by brain and pituitary of sheep may occur in vitro. The secretion depends on the content of Ca, 1.25(OH)2D, and DB-cAMP.

Animals↗

[Ala214,38]aprotinin: preparation by partial desulphurization of aprotinin by means of Raney nickel and comparison with other aprotinin derivatives.

Treatment of aprotinin with Raney nickel in the presence or absence of denaturants yielded [Ala2 14,38]aprotinin. Aprotinin and [Ala2 14,38]aprotinin were separated by ion exchange chromatography at pH 8 using CM-Sepharose, fast flow. [Ala2 14,38]aprotinin is a proteinase inhibitor, but it possesses lower affinities than aprotinin, for the enzymes trypsin, alpha-chymotrypsin, pancreatic kallikrein and plasmin as reflected by higher Ki values [Ala2 14,38]aprotinin is slowly degraded by trypsin. The optical activity of [Ala2 14,38]aprotinin in different solvents is quite similar to that of aprotinin, or that of its hydrolysis products, [seco-15/16]aprotinin or [di-seco-15/16,39/40]-aprotinin. This is taken as good evidence for analogous molecular conformations of all these substances.

Amino Acid Sequence↗

Structures of liposome membranes as models for similar features of cytoplasmic membranes of bacteria.

To characterize a special kind of membrane structure, visible in the cytoplasmic membranes of a Streptomyces hygroscopicus strain, liposome membranes were prepared from their extracted lipid mixture and from their lipid fractions (phospholipids, glycolipids, neutral lipids) and investigated by freeze-fracture electron microscopy. Liposome membranes made of the extracted lipid mixture reveal this special membrane structure, named wafer structure, from its regular pattern of bulges (30-40 nm in diameter). That is the proof that this membrane feature is a lipid structure. Liposome membranes prepared from the lipid fractions show the wafer structure if they are made of the phospholipid fraction only or in combination of this fraction with one or both of the other lipid fractions, indicating that wafer structure formation is primarily connected with the phospholipid content of the membranes. The glycolipid- and neutral lipid fractions amplify this phospholipid structure only. Additional to the wafer structure a raspberry structure with bulges of 55-65 nm in diameter is visible in some case. Obviously both structures are related.

Cell Membrane↗

Distribution and concentration of immunoreactive parathyroid hormone in brain and pituitary of sheep.

We have studied the presence of immunoreactive parathyroid hormone (PTH) in the central nervous system and pituitary of sheep. The PTH concentrations were measured radioimmunologically by two different region-specific antibodies. We could demonstrate PTH in various areas of the brain, whole pituitary, parathyroid glands and plasma of 21 sheep. Measurable concentrations of the two different parathyroid regions (35-84 and 44-68 amino acids fragments) were found in all samples.

Animals↗

Occurrence of squalene and dehydrosqualene in streptomycetes.

Squalene, dehydrosqualene and related hydrocarbons were found to constitute an essential part of the neutral lipid fraction extracted from mycelia and membranes of S. hygroscopicus, S. griseus and S. noursei. In comparison with the fraction of the triglycerides, these terpenoid compounds failed to incorporate (U-14C)-acetic acid throughout pulse labelling experiments. This suggested that the pertinent precursors were formed via alternative routes, for instance by catabolising branched-chain amino acids.

Acetates↗

Modification by genetic changes of the pleiotropic interference of butyrolactone-type autoregulators with differentiation of Streptomyces griseus.

Two series of aerial-mycelium-negative (Amy-), anthracycline-nonproducing (Ant-) mutants were obtained from ancestral Amy+Ant+ strains of S. griseus: a) derivatives represented by the met- strain 39 which could not differentiate although they were still producing both the butyrolactone-type autoregulator 1 and NADP-glycohydrolase, and b) mutants whose incapability to form spores and anthracycline pigments was apparently caused by the loss of autoregulator production. These latter mutants responded to the addition of 1 or the naturally occurring dihydro derivative 2 with complete or at least partial reconstitution of differentiation-associated functions. All of the b)-type mutant strains exhibited similar biochemical alterations in the presence of 1 or 2 regardless of the presence of additional genetic changes in the primary metabolism. Two mutants, however, displayed an altered pattern of secondary product formation. In submerged cultures the major biochemical changes observed in presence of 1 (or 2) were an increase of the lipid level in the mycelium, an alteration of the lipid composition, and a stimulation of neutral proteinase production. All of the blocked autoregulator-negative mutants were discernible from the ancestral strains and strain 39 by their lack of NADP-glycohydrolase production. This suggested the existance of a common genetic locus or a common pleiotropic regulator gene controling both gene functions. Present ideas concerning the role of butyrolactone-type autoregulator 1 as a pleiotropic effector molecule interacting with development of S. griseus are summarized in a hypothetical scheme.

4-Butyrolactone↗

Pleiotropic effects of a butyrolactone-type autoregulator on mutants of Streptomyces griseus blocked in cytodifferentiation.

Mutants of Streptomyces griseus blocked in cytodifferentiation regained their capacity to form differentiated mycelia and/or anthracycline pigments in the presence of butyrolactone-type autoregulatory effectors such as trans-2-(6'-methylheptanol-1'-yl)-3-hydroxymethyl-4-butanolide+ ++. In the pertinent indicator strains, the effect has been correlated with the increase of lipid synthesis, with changes in the composition of lipid fraction and with the restoration of the production of neutral proteinases. The results suggest that autoregulatory butyrolactones from streptomycetes stimulate cytodifferentiation of their producers at an early stage of development.

4-Butyrolactone↗

Effect of the autoregulator from Streptomyces griseus JA 5142 on surface cultures of blocked mutant ZIMET 43682.

Zero time addition of the autoregulator (L-factor) from S. griseus (Lkm+Amy+) to surface cultures of its bald mutant ZIMET 43 682 (Amy-Lkm-) restored the capacity to form both anthracycline-type antibiotic leukaemomycin and aerial mycelium. The pertinent mycelia displayed the same growth rate and cellular levels of nucleic acids as the asporogeneous phenotype but the composition of fatty acids and phospholipids as well as the ratio of cytochromes b and c were altered. These differences indicate alterations in the cellular architecture of substrate and aerial hyphae. The results suggest that the autoregulator triggers the onset of a complex programme of differentiation at a very early stage.

4-Butyrolactone↗