A new simple temperature-controlled membrane oxygenator for the perfusion of isolated rat livers

A new temperature-controlled membrane oxygenator for perfusing isolated rat livers was assembled using a combination of heat-exchangeable rubber tubing and silicon rubber tubing. The apparatus supplied enough oxygen to satisfy the requirements of hemoglobin-free perfused livers.     

A simple membrane oxygenator for the isolated rat liver perfusion

Summary Silastic capillary tubing was used in isolated rat liver perfusions as a simple membrane oxygenator supplying sufficient oxygen to cover the requirements of the liver.     

A simple method for the collection of leucocytes from rat blood

Zusammenfassung Blutleukozyten von Ratten wurden mit menschlichem (normalem) Serum behandelt. Die Viabilität dieser Präparationen konnte dadurch demonstriert werden, dass die Zellen durch Phytohemagglutinin in «blast» Formen umgewandelt wurden.     

A new type of antiserum to thyrotropin-releasing hormone; detection of new TRH-immunoreactive cell groups in rat hypothalamus

Summary We produced a new type of antiserum to thyrotropin-releasing hormone (TRH) in rabbits. The immunogen is TRH-BSA, the production of which is based on the formation of an amide bond using carbodiimide (EDC). The specificity of the antiserum was assessed by enzyme immunoassay (EIA) and immunohistochemistry. When using the anti-TRH serum for immunohistochemistry in rat hypothalamus, new magnocellular groups were detected in the ventrolateral parts of the posterior hypothalamus and the dorsal parts of the third ventricle. Colchicine treatment was found not to be necessary to visualize perikarya containing TRH.     

A simple method for the verification of a successful cannulation of the rat cerebral ventricles

Summary A simple and inexpensive method for the cannulation of the cerebral ventricles of the rat is described. The required tools consist of a stereotaxic instrument, a standard steel injection cannula and a polyethylene tube.This work was done under a contract with the Association of Finnish Life Insurance Companies.     

A new dopaminergic terminal plexus in the ventral horn of the rat spinal cord. Immunohistochemical studies at the light and electron microscopic levels

Summary It has been thought that the ventral motor column in the rat spinal cord is virtually free of dopaminergic fibers. However, a new dopaminergic terminal plexus was visualized at all spinal levels in the ventral horn using electron as well as light microscopic immunohistochemistry.     

Fluid and protein clearance in the rat endometrium. Part II: Ultrastructural evidence for the presence of alternative,non-lymphatic clearance mechanisms in the rat endometrium

Summary In the rat endometrium, resident macrophages and exudate phagocytes ensure proteolysis by means of phagocytosis, macro-and micropinocytosis. Using exogenous tracer particles no ultrastructural evidence could be obtained for the occurrence of endometrial prelymphatics. It is suggested that the free tissue fluid may be drained via the fenestrated (probably venous) blood capillaries.Supported by a grant from the Nationaal Fonds voor Wetenschappelijk Onderzoek-Fonds voor Geneeskundig Wetenschappelijk Onderzoek (Belgium).     

Influence of bistramide A on the twitch tension in rat atrial heart muscle

Bistramide A, a new toxin isolated from the UrochordateLissoclinum bistratum Sluiter, was applied to rat auricular heart muscle bundles. At a stimulation frequency of 0.2 Hz, the toxin induces a dose-dependent reduction of the stimulated twitch tension force; it decreases and shortens the duration of the plateau and the slow repolarizing phase of the action potential. In the control solution, switching from a stimulation frequency of 0.2 Hz to 1 Hz decreases the force with which a positive potentiation develops either at a maintained high frequency or after switching from 1 Hz to 0.2 Hz. Bistramide A reduces both the force evoked at 1 Hz and the potentiation. The data suggest that Bistramide A blocks Na⁺ conductance; inhibits Ca⁺⁺ channels in a time-and frequency-dependent manner; reduces Na⁺–Ca⁺⁺ exchange activity; but does not modify the ability of the sarcoplasmic reticulum to be refilled although the rate of Ca⁺⁺ accumulation is decreased.     

Brain chopping: A new method for the rapid removal of newborn rat brain

Summary A new multi-bladed air-driven guillotine is described. This device removes newborn rat heads and slices them into several thin wafers, which facilitates freezing and permits easy regional dissection.Supp. in part by U. S. Public Health Service Grant. AA-01391.     

Evidence for no relationship of the pineal activity with the neonatal shrinkage of Leydig cells in the rat

Summary The collective volume of Leydig cells in prenatally pinealectomized newborn rats declined as sharply as in intact newborn rats. Also, the collective volume in pups born in the dark declined in a fashion similar to that in pups born in the light. The results indicate that neither the pineal gland nor the light is responsible for the neonatal shrinkage of Leydig cells.This study was supported in part by a Grant-in Aid for Scientific Research (No. 548067) from the Ministry of Education, Science and Culture of Japan.     

Morphological evidence for calcium storage in the chromatoid body of rat spermatids

Summary Morphological evidence for probable Ca²⁺ storage in the vesicular elements of the rat spermatid chromatoid body is documented using the K-pyroantimonate method, combined with EDTA chelation. Some vesicles are related to the microtubules associated with the chromatoid body. A possible involvement of Ca²⁺ in the intracellular movement and/or structural integrity of the chromatoid body is discussed.     

A new twist to the No Miracles Argument for the success of science

J. D. Trout has recently developed a new defense of scientific realism, a new version of the No Miracles Argument. I critically evaluate Trout's novel defense of realism. I argue that Trout's argument for scientific realism and the related explanation for the success of science are self-defeating. In the process of arguing against the traditional realist strategies for explaining the success of science, he inadvertently undermines his own argument.     

Membrane protein folding on the example of outer membrane protein A of <Emphasis Type="Italic">Escherichia coli</Emphasis>

The biophysical principles and mechanisms by which membrane proteins insert and fold into a biomembrane have mostly been studied with bacteriorhodopsin and outer membrane protein A (OmpA). This review describes the assembly process of the monomeric outer membrane proteins of Gram-negative bacteria, for which OmpA has served as an example. OmpA is a two-domain outer membrane protein composed of a 171-residue eight-stranded -barrel transmembrane domain and a 154-residue periplasmic domain. OmpA is translocated in an unstructured form across the cytoplasmic membrane into the periplasm. In the periplasm, unfolded OmpA is kept in solution in complex with the molecular chaperone Skp. After binding of periplasmic lipopolysaccharide, OmpA insertion and folding occur spontaneously upon interaction of the complex with the phospholipid bilayer. Insertion and folding of the -barrel transmembrane domain into the lipid bilayer are highly synchronized, i.e. the formation of large amounts of -sheet secondary structure and -barrel tertiary structure take place in parallel with the same rate constants, while OmpA inserts into the hydrophobic core of the membrane. In vitro, OmpA can successfully fold into a range of model membranes of very different phospholipid compositions, i.e. into bilayers of lipids of different headgroup structures and hydrophobic chain lengths. Three membrane-bound folding intermediates of OmpA were discovered in folding studies with dioleoylphosphatidylcholine bilayers. Their formation was monitored by time-resolved distance determinations by fluorescence quenching, and they were structurally distinguished by the relative positions of the five tryptophan residues of OmpA in projection to the membrane normal. Recent studies indicate a chaperone-assisted, highly synchronized mechanism of secondary and tertiary structure formation upon membrane insertion of -barrel membrane proteins such as OmpA that involves at least three structurally distinct folding intermediates.     

A relevant in vitro rat model for the evaluation of blood-brain barrier translocation of nanoparticles

Poly(MePEG₂₀₀₀cyanoacrylate-co-hexadecylcyanoacrylate) (PEG-PHDCA) nanoparticles have demonstrated their capacity to reach the rat central nervous system after intravenous injection. For insight into the transport of colloidal systems across the blood-brain barrier (BBB), we developed a relevant in vitro rat BBB model consisting of a coculture of rat brain endothelial cells (RBECs) and rat astrocytes. The RBECs used in our model displayed and retained structural characteristics of brain endothelial cells, such as expression of P-glycoprotein, occludin and ZO-1, and immunofluorescence studies showed the specific localization of occludin and ZO1. The high values of transendothelial electrical resistance and low permeability coefficients of marker molecules demonstrated the functionality of this model. The comparative passage of polyhexadecylcyanoacrylate and PEG-PHDCA nanoparticles through this model was investigated, showing a higher passage of PEGylated nanoparticles, presumably by endocytosis. This result was confirmed by confocal microscopy. Thanks to a good in vitro/in vivo correlation, this rat BBB model will help in understanding the mechanisms of nanoparticle translocation and in designing new types of colloidal carriers as brain delivery systems.Received 4 March 2005; accepted 14 April 2005     

Serotonin is localized in endothelial cells of coronary arteries and released during hypoxia: A possible new mechanism for hypoxia-induced vasodilatation of the rat heart

Summary In this report we demonstrate the immunocytochemical localization of serotonin in endothelial cells of rat coronary vessels and a significant increase in the release of serotonin into the perfusate of Langendorff rat heart preparations during hypoxia. It is suggested that serotonin, localized in endothelial cells, is released during hypoxia and could provide part of a pathophysiological mechanism for vasodilatation to protect the heart from damage due to hypoxia.This research was supported by the British Heart Foundation and the excellent technical assistance of Jon Bokor is gratefully acknowledged.     

A simple method for the preparation of hydra chromosome spreads: introducing chromosome counts into hydra taxonomy

Summary A simple method to prepare chromosome spreads of hydra is described. Chromosome counts for a non-symbiotic brown hydra,Hydra vulgaris attenuata, and the Swiss strain of a symbiotic green hydra indicated a diploid number of about 2n=30 in each case. It is suggested that chromosome number may be used to define hydra species more precisely.This work was done while A. R. and M. R. were on leave in Oxford. We thank Prof. D. C. Smith F. R. S. for providing laboratory facilities. We wish to thank Prof. Smith and Dr A. E. Douglas for reading earlier drafts of this paper.     

Molecular evidence for increased hematopoietic proliferation in the spleen of the b/b laboratory rat

The splenomegaly and the appearance of a significant number of CFU-E (erythroid colony-forming units) and BFU-E¹ (erythroid burst-forming units) in the Belgrade laboratory rat (b/b) spleen prompted us to analyse further the molecular evidence for increased hematopoietic proliferation in the b/b spleen. Messenger RNAs (mRNAs) specific for globins, proteins for iron transport and deposition and the band 3 protein were used in rat erythropoietic tissues as markers for proliferation and erythroid differentiation. In the b/b spleen, all mRNAs analysed display an erythroid-specific pattern of expression. This analysis also revealed an enhanced level of mRNA for ferritin in the +/b spleen, whereas erythrocyte-specific mRNA production was normal.