Clock gene expression in the liver of streptozotocin-induced and spontaneous type 1 diabetic rats.

Several investigations have shown a relation between diabetes and alterations of the liver circadian clock. We investigated the diurnal expression of clock genes and clock-controlled genes (CCGs) in 3-hour intervals for a 24-h period in the livers of male streptozotocin (STZ)-treated rats, male spontaneous type 1 diabetic LEW.1AR1-iddm (Iddm) rats, and Iddm rats treated for 10 days with insulin. Hepatic mRNA was extracted, and the relative expression of clock genes (Per1, Per2, Bmal1, Clock, Cry1), as well as CCGs (Dbp, E4bp4, RevErbα, Rorα, Pparγ), was analyzed by reverse transcription followed by real-time polymerase chain reaction. Diabetic STZ and Iddm rats, as well as insulin-substituted Iddm rats, exhibited a significant diurnal expression pattern of clock genes as determined by Cosinor analysis; however, the MESOR (midline estimating statistic of rhythm) of Bmal1, Per2, and Clock transcript expression was altered in Iddm and insulin-substituted Iddm rats. The hepatic expression of the CCGs Dbp and RevErbα revealed a diurnal rhythm in all investigated groups. Insulin administration to Iddm rats normalized the enhanced MESOR in the expression of Dbp, RevErbα, and E4bp4 to the levels of normoglycemic controls. Cosinor analysis indicated no diurnal rhythm of Pparγ expression in the livers of diabetic STZ or Iddm rats or in those of insulin-substituted Iddm rats. Also, insulin substitution could not reverse the decreased MESOR of Pparγ expression in Iddm rats. In consequence of the diabetic disease, changes in the expression of clock genes and CCGs suggest alterations in the hepatic peripheral clock mechanism.

GLUT4 in the endocrine pancreas--indicating an impact in pancreatic islet cell physiology?

The glucose transporter GLUT4 is well known to facilitate the transport of blood glucose into insulin-sensitive muscle and adipose tissue. In this study, molecular, immunohistochemical, and Western blot investigations revealed evidence that GLUT4 is also located in the mouse, rat, and human endocrine pancreas. In addition, high glucose decreased and insulin elevated the GLUT4 expression in pancreatic α-cells. In contrast, high glucose increased GLUT4 expression, whereas insulin led to a reduced expression level of the glucose transporter in pancreatic ß-cells. In vivo experiments showed that in pancreatic tissue of type 2 diabetic rats as well as type 2 diabetic patients, the GLUT4 expression is significantly increased compared to the nondiabetic control group. Furthermore, type 1 diabetic rats exhibited reduced GLUT4 transcript levels in pancreatic tissue, whereas insulin treatment of type 1 diabetic animals enhanced the GLUT4 expression back to control levels. These data provide evidence for the existence of GLUT4 in the endocrine pancreas and indicate a physiological relevance of this glucose transporter as well as characteristic changes in diabetic disease.

Catecholamines are the key for explaining the biological relevance of insulin-melatonin antagonisms in type 1 and type 2 diabetes.

In this paper, we analyze the biological relevance of melatonin in diabetogenesis. As has recently been demonstrated, melatonin decreases insulin secretion via specific melatonin receptor isoforms (MT1 and MT2) in the pancreatic ß-cells. In addition, type 2 diabetic rats, as well as patients, exhibit decreased melatonin levels, whereas the levels in type 1 diabetic rats are increased. The latter effects were normalized by insulin substitution, which signifies that a specific receptor-mediated insulin-melatonin antagonism exists. These results are in agreement with several recent genome-wide association studies, which have identified a number of single nucleotide polymorphisms in the MTNR1B gene, encoding the MT2 receptor, that were closely associated with a higher prognostic risk of developing type 2 diabetes. We hypothesize that catecholamines, which decrease insulin levels and stimulate melatonin synthesis, control insulin-melatonin interactions. The present results support this assertion as we show that catecholamines are increased in type 1 but are diminished in type 2 diabetes. Another important line of inquiry involves the fact that melatonin protects the ß-cells against functional overcharge and, consequently, hinders the development of type 2 diabetes. In this context, it is striking that at advanced ages, melatonin levels are reduced and the incidence of type 2 diabetes is increased. Thus, melatonin appears to have a protective biological role. Here, we strongly repudiate misconceptions, resulting from observations that melatonin reduces the plasma insulin level, that the blockage of melatonin receptors would be of benefit in the treatment of type 2 diabetes.

The insulin-melatonin antagonism: studies in the LEW.1AR1-iddm rat (an animal model of human type 1 diabetes mellitus).

AIMS/HYPOTHESIS: It is well documented that melatonin influences insulin secretion mediated by G-protein-coupled melatonin receptor isoforms MT1 and MT2, which are present in rat and human pancreatic islets, as well as in rat insulinoma cells. Recent investigations have proven that hyperinsulinaemic Goto-Kakizaki (GK) rats, which are a rat model of type 2 diabetic rats, and humans have decreased melatonin plasma levels, whereas a streptozotocin-induced rat model of diabetes developed reduced insulin levels combined with increased melatonin levels. METHODS: Plasma levels of glucose, insulin and melatonin as well as RNA expression of pineal Aanat, Hiomt (also known as Asmt), insulin receptor, adrenoceptor ß1 and the clock genes Per1 and Bmal1 (also known as Arntl) were determined in male and female LEW.1AR1-iddm rats as well as in insulin-substituted LEW.1AR1-iddm rats. RESULTS: Severe hypoinsulinaemia in diabetic LEW.1AR1-iddm rats was associated with decreased body weight and increased melatonin plasma levels combined with mainly elevated expression of Aanat, Hiomt, pineal insulin receptor and adrenoceptor ß1. The changes were normalised by insulin substitution. Diurnal profiles of plasma melatonin and of antagonistic clock genes Per1 and Bmal1 were maintained in diabetic and insulin-substituted rats. CONCLUSIONS/INTERPRETATION: The assumed causal relation between elevated melatonin and reduced insulin levels in LEW.1AR1-iddm rats is supported by the observation that insulin substitution normalised these changes. Further support for this interpretation comes from the observation that in GK rats an increase of plasma insulin was combined with a decrease of plasma noradrenaline (norepinephrine), the most important activator of melatonin synthesis. These relationships between the noradrenergic and insulin pathway support the existence of melatonin-insulin antagonism.

Transcripts of calcium/calmodulin-dependent kinases are changed after forskolin- or IBMX-induced insulin secretion due to melatonin treatment of rat insulinoma beta-cells (INS-1).

The objective of the present study was to examine the effects of melatonin on transcripts of isoforms of calcium/calmodulin-dependent protein kinases in rat insulinoma beta-cells INS-1. Investigations show that calcium/calmodulin-dependent kinase IV and calcium/calmodulin-dependent kinase 2d are expressed in human and rat pancreatic islets and INS-1 cells. By application of either forskolin or 3-isobutyl-1-methylxanthine for 6 hours, calcium spiking was evoked and the release of insulin was increased. The expression of the calcium/calmodulin-dependent kinase IV and calcium/calmodulin-dependent kinase 2d transcripts was significantly increased due to forskolin or 3-isobutyl-1-methylxanthine. Acute melatonin treatment (6 h) in the presence of either forskolin or 3-isobutyl-1-methylxanthine caused a significant decrease in insulin release and induced significant downregulation of calcium/calmodulin-dependent kinase IV and calcium/calmodulin-dependent kinase 2d transcripts in INS-1 batch cultures. The attenuating effect of melatonin on transcripts could be almost completely reversed by preincubation with the melatonin receptor antagonist luzindole. Thus, the insulin-inhibiting effect of melatonin in INS-1 cells is associated with significant changes in transcripts of calcium-signaling components suggesting that melatonin influences gene expression of components, which are known to be involved in insulin secretion or insulin gene expression.

Did the gradual loss of GLUT2 cause a shift to diabetic disorders in the New Zealand obese mouse (NZO/Hl)?

The New Zealand obese mouse (NZO/Hl) is characterised by hereditary obesity and type-2 diabetes, including insulin resistance, hyperinsulinaemia, and glucose intolerance. In other diabetic models, it has been revealed that the proper functioning of the glucose transporter isoform 2 (GLUT2) is essential for adequate secretion of insulin. The aim of this study was to compare the distribution of islet cells and GLUT2, as well as the expression of GLUT2-mRNA, in the pancreas of NZO mice and metabolically unimpaired NMRI (Naval Medical Research Institute) mice. Pancreas tissue was obtained from different stages of development. For molecular determination of the expression level of GLUT2-mRNA, total-RNA was extracted from the pancreas and analysed by quantitative real-time RT-PCR. All investigated NZO mice displayed increased weight, elevated hyperinsulinaemia, and slightly enhanced blood glucose levels compared with the NMRI control mice. By means of immunofluorescence microscopy drastically reduced insulin levels were detected, which might be compensated by the observed islet cell hyperplasia and hypertrophy. Furthermore, the normally peripheral localisation of the alpha-cells within islets was disturbed. By contrast, there were no changes in somatostatin cell distribution. However, considerable differences appeared with regard to GLUT2: whereas the beta-cells of NMRI mice showed dense immunostaining of the GLUT2 transporter on the cell surface, in all age groups of NZO mice, GLUT2 on the plasma membranes was reduced and dispersed in the cytoplasm. These findings agree with the molecular biological results, which displayed decreased mRNA-expression of GLUT2. In summary, the observed alteration of islet morphology and of GLUT2 expression in diabetic mice complements our previous results from a superfusion protocol and further clarifies the mechanisms of diabetogenesis in NZO mice.

[Corneal wound healing after hyperopic PRK and LASIK]. / Korneale Wundheilungsreaktionen nach hyperoper PRK und LASIK.

BACKGROUND: The purpose of this study was to investigate the influence of different postoperative treatments on the wound healing reaction in the anterior stroma after PRK and in the interface area after LASIK. METHODS: Seventy-two corneal buttons of refractively treated rabbit eyes underwent different postoperative eyedrop regimens with antibiotics and/or steroids or additional UV-B irradiation. Morphological and immunohistological investigations were performed 6 months postoperatively by light and transmission electron microscopy. RESULTS: PRK eyes showed interdigitations between the epithelia and the anterior stroma. LASIK-treated eyes showed only minor changes between epithelia and stroma in the incisional region. Only a slight increase in deposits of fibrillar extracellular matrix components were detectable in the interface region. CONCLUSIONS: The clinically important problem of haze after PRK is caused by the interdigitations between epithelia and anterior stroma. The delicate wound healing reactions in the interface region in LASIK eyes corresponded to the clinically visible minor changes in these corneas.

Indication of circadian oscillations in the rat pancreas.

The central circadian oscillator of the suprachiasmatic nucleus controls diurnal rhythmicity of the body with light as its dominant zeitgeber. Recently, peripheral oscillators have been detected in liver and heart, which follow as yet unidentified cues. In this study real-time reverse transcription-polymerase chain reaction (RT-PCR) was used in analysis of the expression of the major clock genes Per1, Per2, Bmal1, Cry1, Tim (timeless) and Clock, as well as of the output genes Dbp and Rev-erbalpha in the pancreatic tissue of rats. The results presented here indicate a robust circadian expression of clock genes (e.g. Per1 and Bmal1) and the probable existence of a peripheral oscillator in the pancreas. Whether this oscillator regulates the diverse functions of the islets of Langerhans remains to be elucidated.

Circannual morphometric investigations of the rat suprachiasmatic nucleus after pinealectomy, ganglionectomy and thyroidectomy.

We karyometrically investigated the nucleus suprachiasmaticus (SCN) which had been manipulated in several ways in order to analyze the functional importance of the pineal gland on the primary pacemaker of mammals in the course of the year. The manipulation modes were (i) pinealectomy (PX), and (ii) sympathetic denervation of the pineal by bilateral extirpation of the upper cervical ganglia (ganglionectomy, GX). Additionally, the influence of the inactivated pineal, obtained through hypothyroidism which was realized by (iii) subtotal thyroidectomy (TX), was also investigated. With respect to annual oscillations the results of our investigations were able to illustrate the following. (1) The SCN consists of at least two parts (ventrolateral and dorsomedial) each with different functions and relationships. The nuclei of the ventrolateral cells are bigger and there are many indications both in our own research and from literature that the neurons of this part are involved in the generation of rhythms. (2) The size of the cell nuclei of the ventrolateral part shows annual patterns. In the course of the year the maxima of the nuclear volume were registered in March and September (bimodal pattern, equinox, L:D = 12:12). PX, GX or TX only negligibly changed the bimodal annual pattern. However, in comparison the smallest cell nuclei were registered in the winter (short day). The much smaller cell nuclei of the dorsomedial part likewise show bimodal patterns but only in the experimental groups. The control group of this part shows an unimodal annual curve with a minimum at long-day conditions (June, L:D = 16:8). (3) All manipulations which inactivated the pineal or reduced the content of melatonin (PX, GX, and TX) were followed by an increase (activation) of cell nuclei of the SCN. In contrast to these effects, an increase of thyroxine (by exposure to cold), has an opposite effect (not documented here). In conclusion these results indicate, without a doubt, that a negative correlation exists, functionally, between the SCN and the pineal (in the same annual experiment the nuclear size of the pinealocytes was increased, under short-day conditions in December, and decreased under long-day circumstances in June). Additionally, it could be shown that the degree of negative correlation between the pineal and the SCN was seasonally dependent. The lowest effects of PX, GX and TX were registered at short-day conditions (December, L:D = 8:16).

Comparison between xanthine oxidases from buttermilk and microorganisms regarding their ability to generate reactive oxygen species.

Xanthine oxidase (XO) forms uric acid from xanthine. It is assumed that at the same time oxygen is reduced by the XO to reactive oxygen species (ROS), mainly to .O2- and to H2O2. Under certain conditions such ROS can be highly damaging to cellular structures. Therefore, XO was frequently used as a model system, in which the impact of ROS on cellular compounds and structures has been investigated. In this in vitro study xanthine oxidases from buttermilk and from microorganisms were compared regarding their ability to generate ROS. It could be shown that both enzymes are able to transform xanthine to uric acid but differ significantly in their reductive properties to oxygen. XO from buttermilk reduces oxygen to both .O2- and H2O2 whereas XO from microorganisms generates H2O2, but fails to form .O2-. Since .O2- are involved in maintaining transition metal-mediated formation of hydroxyl radicals (.OH) from H2O2, we conclude that XO from microorganisms is therefore largely unsuitable in studies investigating just the interaction of .O2- with other ROS on cellular compounds.

Postirradiated and nonirradiated gliosarcoma: immunophenotypical profile.

BACKGROUND: Thirty-one gliosarcomas (25 nonirradiated and 6 postirradiated tumors) were selected based on the presence of two distinctive areas: a malignant gliomatous and a sarcomatous component. In all cases, the sarcomatous component appears like fibrosarcoma or malignant fibrous histiocytoma. Two tumors showed additional areas consisting of osteochondroid differentiation. METHODS: All tumors were examined using antibodies against Ulex europaeus agglutinin I (UEA), glial fibrillary acidic protein (GFAP), vimentin (VM), epithelial membrane antigen (EMA), desmin, collagen IV, alpha-1-antitrypsin (alpha-1-AT) and smooth muscle actin (SMA). RESULTS: While the VM highlighted the sarcomatous areas of all tumors there were only scattered spindle cells that were GFAP-positive in the same areas. The alpha-1-AT was diffusely reactive in the sarcomatous areas in 20 cases. Focal immunoreactivity was seen with SMA (20 tumors), UEA (8 tumors), EMA (5 tumors), collagen IV (5 tumors) and desmin (4 tumors) in the nonvascular sarcomatous cells. CONCLUSIONS: The range of immunophenotypical expression is likely to be a reflection of the capacity of a multipotential progenitor to undergo divergent differentiation. There is a very little morphological difference between the postirradiated and nonirradiated tumors except that a higher proportion of postirradiated tumors are immunoreactive to SMA and desmin.

Pineal influence on annual nuclear volume changes in ventromedial hypothalamic nucleus (VMH) neurons of the male Wistar rat.

The ventromedial hypothalamic nucleus (VMH) regulates various autonomic, endocrine, and behavioral activities. These activities show annual changes, and the pineal gland is involved in their adjustment to environmental cues. Therefore, this study investigated whether the VMH belongs to the effector structures of the pineal gland. To abolish the rhythmic melatonin release, male Wistar rats were subjected to pinealectomy (PX) or ganglionectomy (sympathetic denervation of the pineal gland, GX) regularly at the beginning of any of the four seasons. Brains from animals of PX-, GX-, and sham-operated control groups were prepared 3 months later for measurement of the nuclear volume, which changes according to the general gene activity. At each of the four seasons, 2000 nuclei of VMH neurons stemming from 18 animals per group were measured to obtain both seasonal daily mean values and annual mean values, respectively, as well as to calculate annual curves of the nuclear volume using empirical regression and locally adjusted polynomial approximation. The major findings are the following. First, inactivation of the pineal function influences the nuclear activity of VMH neurons, (2) PX and GX mainly depress the nuclear activity, indicating that the pineal influence on the VMH may predominantly be a stimulatory one. Third, size and direction of the changes caused by PX and GX vary in a seasonally dependent manner. Fourth, the annual rhythm of the nuclear activity of the VMH is modified by PX and GX. To explain how the pineal effects on the VMH may be mediated, a possible inhibitory influence of the suprachiasmatic nucleus (SCN), which has been activated in the same animals following both PX and GX, is discussed. In conclusion, the results confirm that the nuclear activity of VMH neurons underlies pineal influences. This also indicates an involvement of the pineal gland in many VMH-regulated functions.

[Effect of ganglionectomy (superior cervical ganglia) at normal temperature and exposure to cold on the circadian rhythm of liver glycogen and blood glucose levels of the Wistar rat with regard to the pineal gland]. / Zum Einfluss von Gangliektomie (Ganglia cervicalia superiora) bei Normaltemperatur und Kälteexposition auf Leberglykogen- und Blutglukose-Tagesmuster der Wistar-Ratte unter Berücksichtigung der Epiphysis cerebri.

Liver glycogen content and blood glucose level of 80 male Wistar rats were measured 4 times a day, 30 d after following surgeries: sham-operation without exposure to cold, sham-operation with exposure to cold (283 K, 72 h before killing), extirpation of the ganglia cervicalia superiora (GX) without exposure to cold, extirpation of the ganglia cervicalia superiora with exposure to cold (283 K, 72 h before killing). Exposure to cold alone as well as GX with and without exposure to cold modify the unimodal daily patterns (curves were calculated by means of empirical regression) of liver glycogen content and blood glucose level: there are changes from contrary localization of the maxima (and minima) of both, liver glycogen content and blood glucose level (exposure to cold alone) to nearly equal localization (GX alone). Compared to control group, all surgeries reduce the liver glycogen content statistical-significantly and also enhance the blood glucose level. Combination of GX with exposure to cold shows the strongest effect. In this case, the influence of GX is visible, but the effect is not significantly different from this of exposure to cold alone, which causes a high activation of the metabolism. To the contrary, a statistically significant influence is to be seen after GX alone, but the effect is lower than this one of exposure to cold. Extirpation of the ganglia cervicalia superiora and the resulting sympathetic denervation of the pineal gland is answered by comparable reactions of the investigated parameters as the extirpation of the pineal gland itself (90 d post operationem). Diminuation of liver glycogen content and enhancement of blood glucose level characterize the influence of gangliectomy (30 d post operationem) in this way, that the ratio of effect of factors causing hypo- and hyperglycemia is changed in favour of the latter.

[Thyroid reactions in Wistar rats during the circadian rhythm after ganglionectomy at normal temperature and under cold exposure with regard to the influence of the epiphysis cerebri]. / Schilddrüsenreaktionen der Wistar-Ratte im Tagesgang nach Gangliektomie unter Normaltemperatur und Kälteexposition unter Berücksichtigung des Einflusses der Epiphysis cerebri.

Serum thyroxin (T4), serum TSH, and pituitary TSH were measured by radioimmunoassay (RIA) and serum cholesterol by Liebermann-Burchard reaction in rats 4 times a day (light-dark cycle: 14 L: 10 D) after gangliectomy (bilateral extirpation of the Ganglia cervicalia superiora) at cold and normal temperature conditions. 80 male Wistar rats were divided into 4 groups: sham-operated group, 24 degrees C (297 K); sham-operated group, 10 degrees C (283 K); gangliectomy, 24 degrees C (297 K), and gangliectomy, 10 degrees C (283 K). We have sacrificed the rats 30 d after operations at the following day-times: middle light, middle darkness, 1 h after light "on" and 1 h after light "off" (they were exposed to cold 72 h before killing). It was found that gangliectomy significantly depressed blood level of thyroxin. On the other hand, it enhanced the serum cholesterol and TSH levels as well as the pituitary TSH content. Exposure to cold increased thyroxin, serum TSH and pituitary TSH. The cholesterol level, however, was significantly decreased. Gangliectomy causes a reduction of the cold-induced stimulation of thyroxin (significant), serum TSH, and pituitary TSH content (significant). The cholesterol (in relation to the cold-exposure alone) was significantly increased under these conditions. We have found similar results in another long-time experiment (90 d exposure) after gangliectomy as well as after pinealectomy. There also appears a lowered thyroxin and an increased cholesterol level (in dependency on the seasons). Gangliectomy induced a decrease of the pineal weight and a compensatory thyroid growth. Exposure to cold induced an increase of pituitary and pineal weights. Gangliectomy provokes a reduction of the cold-induced augmentation of the pineal weight. The results indicate that gangliectomy diminishes the total levels of circulating T4 in the presence of an intact pineal gland and reduces the cold-induced increase of T4 in long-time experiments (30 and 90 d post operationem). Both gangliectomy and cold condition led to an enhancement of serum TSH and pituitary TSH content. The exposure to cold was found to have a more severe influence. In the present study, we also have discussed the sympathetic denervation effect of the gangliectomy in relation to the thyroid and pineal gland. Due to certain contradictory data in the literature, we have also discussed the TRH-5-hydroxytryptamine (respectively melatonin) antagonism, though we were not able to determine whether peripheral and/or central mechanisms play the more important role in their regulation.

Circadian changes of ether-a-go-go-related-gene (Erg) potassium channel transcripts in the rat pancreas and beta-cell.

Evidence has previously been presented that circadian rhythms play a role in islet hormone secretion. Here, RT-PCR was used to monitor the circadian expression of ether-a-go-go-related gene (Erg) potassium channel isoforms and Erg1 splice variants. Immunohistochemistry was used to identify the pancreatic distribution patterns of ERG1a and ERG1b, as well as ERG2 and ERG3. The influence of ERG on insulin secretion was monitored by perfusion of rat INS-1 beta-cells with the blockers E-4031 and rBeKm-1. We identified Erg1a, Erg1b, Erg2 and Erg3 transcripts in islets and INS-1 cells. Immunohistochemistry showed differential expression of ERG isoforms in the islet. Ca(2+) imaging and electrophysiological recordings of INS-1 cells during ERG blocking by E-4031 indicated functional ERG channels. Serum shock treatment of INS-1 cells elicited a time-dependent expression response for Erg transcripts. These results add to the current understanding of the function of ERG channels in beta-cells and the circadian secretion processes of insulin.

Circadian and age-dependent expression patterns of GLUT2 and glucokinase in the pancreatic beta-cell of diabetic and nondiabetic rats.

Alterations in glucose sensing are well-known in both humans and animal models of non-insulin-dependent diabetes mellitus. However, the circadian- and age-dependent expression of glucose-sensing genes has not previously been investigated in vivo. In the present paper, we show a progressive loss of beta-cell GLUT2-mRNA and, by immunocytochemistry, a gain of soluble, cytoplasmic GLUT2-protein in Goto-Kakizaki rat islets. We report that GLUT2-mRNA shows significant diurnal variation, which is stronger in metabolically healthy rats. We also demonstrate the significant diurnal variation of glucokinase-mRNA, with higher levels in the pancreas of 6-week-old Goto-Kakizaki rats than in Wistar rats. This leads to a maximum glucose phosphorylation capacity in-phase with food intake, enhanced glucose-stimulated insulin secretion, and prevents postprandial hyperglycemia. Perfusion experiments showed a reduction in glucose-stimulated insulin secretion in Goto-Kakizaki rat islets with an impaired first phase. Hyperglycemia and hypoinsulinemia in newborn and up to 3-week-old Goto-Kakizaki rats are thus probably due to reduced pancreatic beta-cell content, reduced beta-cell insulin content and impaired glucose sensing. The de-compensation of the metabolic situation in 42-week-old Goto-Kakizaki rats is likely to be caused by beta-cell destruction accompanied by negligible accumulation of GLUT2 in the cell membrane and further reduction of glucokinase expression.

A reticular variant of elastolytic giant cell granuloma.

An 80-year-old man presented with a distinctive reticular form of elastolytic giant cell granuloma (EGCG). This represents a rare subset of EGCG, thus belonging to a larger group of granulomatous skin diseases with poorly understood pathogenesis. The patient exhibited numerous erythematous to brownish, sharply demarcated patches of reticulate appearance with partial central atrophy and nonelevated margins, which involved the trunk and especially chest, shoulders, upper back and arms. Skin biopsy revealed a granulomatous infiltrate with multinucleate histiocytes containing remnants of elastic fibres in conjunction with a significant loss of elastic fibres throughout the dermis. There is little published information on treatment, which remains unsatisfactory.

Influence of oxygen concentration on redox cycling of alloxan and dialuric acid.

Alloxan, a chemical diabetogen, decays in the absence of reductants into alloxanic acid. In the presence of glutathione, it is reduced via the alloxan radical into dialuric acid, which autoxidizes back to alloxan. During this redox cycling process, reactive oxygen species are formed that destroy beta-cells in islets of Langerhans. Previous experiments were conducted with oxygen concentrations about ten times as high as within cells. The aim of our in vitro study was to evaluate the impact of different oxygen concentrations (0, 25, 250 micromol/l) at a given initial ratio of glutathione and alloxan on this redox cycling. Reduction of alloxan, oxidation of glutathione, and the formation of glutathiol (GSSG) were continuously recorded by HPLC for 90 minutes at 25 degrees C in air, calibration gas, or argon. In the absence of reductants, alloxan irreversibly decomposed into alloxanic acid regardless of oxygen presence. When the reaction system contained glutathione, decomposition was significantly retarded and therefore influenced by oxygen. In argon, decay could not be observed due to its reduction and the absence of oxygen. Increasing oxygen concentration enabled a redox cycling and therefore an ongoing decay. The highest decomposition along with the highest consumption of glutathione occurred at 250 micromol/l oxygen. The lower the oxygen, the more dialuric acid could be detected. After calculation, about 33 redox cycles per hour generates an amount of reactive oxygen species sufficient to damage pancreatic beta cells and induce insulin deficiency.

[Technical possibilities of experimentation, registration and surgery for solving chronobiological problems in animal experiments]. / Versuchs-, registrier- und operationstechnische Möglichkeiten zur Lösung chronobiologischer Fragestellungen im Tierversuch.

Common possibilities of keeping and registering in chronobiological experiments and special techniques of surgical operations (pinealectomy, thyroidectomy, and exstirpation of the ganglia cervicalia superiora) are presented with a special chronoendocrinological conception for an experiment to investigate the role of the pineal gland.

[Morphological investigations of the rat thyroid under lead intoxication and experimental hypothyroid (author's transl)]. / Morphologische Untersuchungen an der Rattenschilddrüse nach Bleiintoxikation und epxerimenteller Hypothreose.

Male white Wistar-rats are treated alone and in combination with methylthiouracil and lead acetate. The histologic-cytological and the caryometrical results and the weights of the thyroids show an increase thyroid activity following as well methylthiouracil as lead application. The thyroxin-iodine blood level and the cholesterol blood level are altered. The strongest enhancement of the activity is obtained in the combination group methylthiouracil plus lead acetate. More slow activations are present in the groups with singular application methylthiouracil or lead acetate application. The findings are caused by encymatic inhibition of the applicated substances.