Diurnal variation of corticotropin-releasing factor binding sites in the rat brain and pituitary

Summary 1. Corticotropin-releasing factor (CRF) is thought to be involved in the regulation of the diurnal activity of the hypothalamus-pituitary-adrenal (HPA) axis and to act as a neurotransmitter in the brain. To date it is unknown whether the binding sites of the central CRF system are subject to diurnal variations. 2. We measured the number of CRF binding sites over the course of a complete 24-hr light-dark cycle in the pituitary, amygdala, bed nucleus of the stria terminalis (BNST), cingulate cortex, visceral cortex, paraventricular nucleus of the hypothalamus, hippocampus, and locus ceruleus of rats byin vitro receptor autoradiography with iodinated ovine CRF. A 24-hr time course was also established for plasma CRF and corticosterone. 3. The diurnal pattern of plasma CRF does not correlate with the pattern of plasma corticosterone. Within the brain, CRF binding in the basolateral nucleus of the amygdala showed a U-shaped curve with maximum levels in the morning and a wide hallow between 1500 and 0100. A biphasic profile with a small depression in the afternoon and a more pronounced depression in the second half of the activity period is characteristic for the other brain areas and the pituitary. The profile for the pituitary correlates with those for the BNST and the area of the locus ceruleus. Furthermore, the diurnal pattern of CRF binding sites in the BNST correlates with that of the hippocampus, and the daytime pattern of the visceral cortex is similar to that of both the hippocampus and the BNST. 4. Since the CRF-binding profiles in the brain and the pituitary clearly differ from the profiles of both plasma CRF and corticosterone, one may assume that the diurnal pattern of central CRF binding sites is not directly coupled to the activity of the HPA axis.     

A new approach for rhizosphere research by X-ray microanalysis of microliter soil solutions

Lolium perenne growing with high root density on a fine nylon mesh (Kuchenbuch and Jungk, 1982) caused the development of element gradients in the rhizosphere below the mesh. Micro-liter soil solutions from 2-mg soil samples were sprayed onto Formvar-coated grids and analyzed by X-ray microanalysis in a transmission electron microscope. The results were comparable to those obtained by flame photometry and atomic absorption spectrometry (AAS) of conventional soil solutions from 1 g soil. X-ray microanalysis of micro-soil solutions allows the application of different extraction procedures to even small amounts of soil usually available from rhizosphere experiments. Information about soil buffering characteristics in the rhizosphere can thus be obtained. Aluminum accumulation in the rhizosphere of small segments of single Picea abies fine roots grown in undisturbed natural forest soil could be detected with this technique.     

Isolation and visualisation of alkali stable protein/DNA complexes

Distinct polypeptides, 54,000–68,000 daltons in size, are alkali-stably bound to eukaryotic DNA. DNA fragments several hundred base pairs in length associated with these polypeptides are preferentially retained on glass fibre filters from solutions containing 1 M sodium chloride. About 50 percent of the protein/DNA complexes present in total DNA are retained on filters together with about 2 percent of the DNA. This preferential binding is demonstrated (a) by the ratio of ³H and ³⁵S radioactivity retained on filters after filtration of DNA from [³H]thymidine and L-[³⁵S]methionine labelled cells, (b) radioiodination of the material retained on filters and passing filters respectively and (c) by electron microscopical visualisation of the polypeptide component in the complexes after chemical modification with dinitrofluorobenzene (DNFB) followed by incubation with dinitrophenyl (DNP) specific antibodies.     

Body movements and retinal pattern displacements while approaching a stationary object in the walking fly,Calliphora erythrocephala

When a walking fly approaches a stationary object two types of body movements are distinguishable. Type I body movements are characterized by low frequencies (0.4–1.3 Hz) and large amplitudes (28–65°). Superimposed on these movements are type II body movements which are characterized by high frequencies (7.3–10.6 Hz) and small amplitudes (5.9–8.2°) (Figs. 3–6; Table 1). Type II movements occur no matter whether the fly is fixating a pattern or orientating itself in homogeneous surroundings without any pattern. In contrast, only 72% of the flies with immobilized heads and 62% of the flies with movable heads make type I body movements. The amplitude of type I and type II body movements increases slightly after immobilization of the head. Binocular as well as monocular pattern projection occurs for the whole walking trajectory (Fig. 7–9). Monocular pattern projection seems to be more frequent in flies with immobilized heads than in those with movable heads. The degree of pattern fluctuations in the visual field of the flies increases slightly along the walking trajectory. Near the starting point in the centre of the arena it amounts to 5–7°, while at the end of the walking trajectory it amounts to 8–10° (Table 2). The following conclusions and hypothesis can be drawn from these experiments. 1. The graph BT for the direction of the fly's logitudinal axis can be approximated by the first derivative of the walking trajectory WT, that means, dWT(x)/dxBT(x) (Fig. 11). 2. The amplitudes of type II body movements are caused by the alternating movements of the legs during forward motion, while type I body movements are classified as exploring movements. During evolution of visually guided behaviour it is possible that blowflies have adapted their elementary movement detector system to type II body movements. 3. The types of pattern projection into the visual field of the fly while approaching an object can be explained by a simple neuronal network characterized by either inhibitory and/or excitatory influences of the visually activated neurones on the motor neurones generating the propulsive forces, that means the forward motion. In addition it is postulated that the large frontal and antero-lateral receptive fields of these neurones are not coupled with the motor centres on the same side of the body (Fig. 12).     

Derivative chromosomal structures from a ring chromosome 4

Summary This report describes the results from cultured lymphocytes studied at metaphase, anaphase, and interphase from an individual with a ring chromosome 4. A ring was present in 90.1% of metaphases. Special attention was directed towards the occurrence of derivative chromosomal structures, such as partially duplicated and triplicated rings, tricentric rings, chains of 3 interlocked rings, rod-shaped chromosomes, pulverized rings, and others.The clinical features of the individual (small stature and impaired mental development, hypoplastic thumbs, ptosis palpebrae hypoplastic external male genitalia, abnormal dermatoglyphic pattern) did not conform to a specific phenotype.

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Zusammenfassung Die Lymphocyten eines Patienten mit Ringchromosom 4 wurden in der Meta-, Ana-und Interphase beobachtet. 90,1% der Metaphasen wiesen einen Ring auf. Besondere Aufmerksamkeit wurde abnormen, aus dem Ring entstandenen Chromosomen zugewandt. Es fanden sich u. a.: partielle Doppel-und Dreifachringe, Ringe mit 3 Zentromeren, Ringe aus 3 miteinander verketteten Ringen, stabförmige Chromosomen und pulverisierte Ringe.Die Symptome des Patienten (Minderwuchs, Schwachsinn, Hypoplasie der Daumen, Ptosis palpebrae, Hypoplasie des äußeren genitale, abnorme Dermatoglyphen) waren zum größten Teil unspezifish.

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This work was supported in part by grants from the Deutsche Forschungsgemeinschaft and is part of a thesis by R.N. for a medical doctorate.     

Systems biology and its potential role in radiobiology

About a century ago, Conrad Röentgen discovered X-rays, and Henri Becquerel discovered a new phenomenon, which Marie and Pierre Curie later coined as radio-activity. Since their seminal work, we have learned much about the physical properties of radiation and its effects on living matter. Alas, the more we discover, the more we appreciate the complexity of the biological processes that are triggered by radiation exposure and eventually lead (or do not lead) to disease. Equipped with modern biological methods of high-throughput experimentation, imaging, and vastly increased computational prowess, we are now entering an era where we can piece some of the multifold aspects of radiation exposure and its sequelae together, and develop a more systemic understanding of radiogenic effects such as radio-carcinogenesis than has been possible in the past. It is evident from the complexity of even the known processes that such an understanding can only be gained if it is supported by mathematical models. At this point, the construction of comprehensive models is hampered both by technical inadequacies and a paucity of appropriate data. Nonetheless, some initial steps have been taken already and the generally increased interest in systems biology may be expected to speed up future progress. In this context, we discuss in this article examples of relatively small, yet very useful models that elucidate selected aspects of the effects of exposure to ionizing radiation and may shine a light on the path before us.     

Dipeptidyl peptidase IV inhibits the polymerization of fibrin monomers

A highly purified dipeptidyl peptidase IV from human placenta cleaves glycylproline from the N-terminal end of the fibrin α chain and inhibits the clotting of fibrin monomers. This result underlines the importance of the amino terminus of the fibrin α chain as an aggregation site masked by fibrinopeptide A. We speculate that the peptidase may hinder blood coagulation in intact vessels in vivo, because it is located on the surface of the capillary endothelium.     

The impact of mtDNA analysis between positions nt8306 and nt9021 for forensic casework

Identification in forensic casework by mtDNA sequencing is predominantly done by sequencing the non-coding control region (HVI and HVII). In an attempt to further increase the discriminatory power of mtDNA analysis, we sequenced a coding region between nt8306 and nt9021 to identify additional polymorphisms in a group of 61 unrelated German individuals who had mtDNA profiles that occurred more than two times each, as well as a control group of 119 unrelated Germans whose profiles occurred one or two times each. Within these 180 individuals, 38 different polymorphisms in this region were observed; 64.4% of these individuals displayed the Cambridge reference sequence profile plus A8860G. For 28 individuals with the two most common profiles, A263G-315.1insC (N = 18) and A263G-309.1insC-315.1insC (N = 10), additional polymorphisms in this coding region permitted further discrimination of 56 and 40% of the individuals, respectively.