Protective effects of an extract of the freshwater microalga Scenedesmus rubescens on UV-irradiated skin cells.

BACKGROUND: Skin ageing results from intrinsic but also extrinsic factors of which UV irradiation is a main cause. It is hence of interest to have means to protect skin from UV irradiation-induced damage. We selected an extract of the freshwater microalga Scenedesmus rubescens and assessed its potential to protect skin from photoageing caused by UV irradiation. METHODS: Skin cells in vitro and ex vivo were analysed for markers of UV irradiation-induced photodamage such as decreased viability, decreased collagen content, hyperpigmentation and sunburn cells. RESULTS: We found that a dry extract of the microalga Scenedesmus rubescens was able to suppress cellular signs of ageing induced by UV irradiation. It enhanced dermal fibroblast viability, rescued dermal collagen content, inhibited the formation of sunburn cells and inhibited tyrosinase activity. CONCLUSION: An extract of Scenedesmus rubescens showed broad activity against markers of UV irradiation-induced cutaneous ageing. It may therefore be used as a preventive or regenerative agent for anti-ageing strategies.

Characterization of activating mutations of NOTCH3 in T-cell acute lymphoblastic leukemia and anti-leukemic activity of NOTCH3 inhibitory antibodies.

Notch receptors have been implicated as oncogenic drivers in several cancers, the most notable example being NOTCH1 in T-cell acute lymphoblastic leukemia (T-ALL). To characterize the role of activated NOTCH3 in cancer, we generated an antibody that detects the neo-epitope created upon gamma-secretase cleavage of NOTCH3 to release its intracellular domain (ICD3), and sequenced the negative regulatory region (NRR) and PEST (proline, glutamate, serine, threonine) domain coding regions of NOTCH3 in a panel of cell lines. We also characterize NOTCH3 tumor-associated mutations that result in activation of signaling and report new inhibitory antibodies. We determined the structural basis for receptor inhibition by obtaining the first co-crystal structure of a NOTCH3 antibody with the NRR protein and defined two distinct epitopes for NRR antibodies. The antibodies exhibit potent anti-leukemic activity in cell lines and tumor xenografts harboring NOTCH3 activating mutations. Screening of primary T-ALL samples reveals that 2 of 40 tumors examined show active NOTCH3 signaling. We also identified evidence of NOTCH3 activation in 12 of 24 patient-derived orthotopic xenograft models, 2 of which exhibit activation of NOTCH3 without activation of NOTCH1. Our studies provide additional insights into NOTCH3 activation and offer a path forward for identification of cancers that are likely to respond to therapy with NOTCH3 selective inhibitory antibodies.

Evidence for involvement of copper ions and redox state in regulation of butane monooxygenase in Pseudomonas butanovora.

Pseudomonas butanovora possesses an alcohol-inducible alkane monooxygenase, butane monooxygenase (BMO), that initiates growth on C(2)-C(9) alkanes. A lacZ transcriptional reporter strain, P. butanovora bmoX::lacZ, in which the BMO promoter controls the expression of beta-galactosidase activity, was used to show that 1-butanol induced the BMO promoter in the presence or absence of O(2) when lactate-grown, BMO-repressed cells were washed free of lactate and incubated in NH(4)Cl-KNa phosphate buffer. In contrast, when lactate-grown cells of the reporter strain were incubated in phosphate buffer containing the mineral salts of standard growth medium, 1-butanol-dependent induction was significantly repressed at low O(2) (1 to 2% [vol/vol]) and totally repressed under anoxic conditions. The repressive effect of the mineral salts was traced to its copper content. In cells exposed to 1% (vol/vol) O(2), CuSO(4) (0.5 microM) repressed 1-butanol-dependent induction of beta-galactosidase activity. Under oxic conditions (20% O(2) [vol/vol]), significantly higher concentrations of CuSO(4) (2 microM) were required for almost complete repression of induction in lactate-grown cells. A combination of the Cu(2+) reducing agent Na ascorbate (100 microM) and CuSO(4) (0.5 microM) repressed the induction of beta-galactosidase activity under oxic conditions to the same extent that 0.5 microM CuSO(4) alone repressed it under anoxic conditions. Under oxic conditions, 2 microM CuSO(4) repressed induction of the BMO promoter less effectively in butyrate-grown cells of the bmoX::lacZ strain and of an R8-bmoX::lacZ mutant reporter strain with a putative BMO regulator, BmoR, inactivated. Under anoxic conditions, CuSO(4) repression remained highly effective, regardless of the growth substrate, in both BmoR-positive and -negative reporter strains.

Mechanisms of apoptosis after ischemia and reperfusion: role of the renin-angiotensin system.

BACKGROUND: Apoptosis plays a key role in the pathogenesis of cardiac diseases. We examined the influence of the renin-angiotensin system (RAS) on different regulators of apoptosis using an isolated hemoperfused working porcine heart model of acute ischemia (2 h), followed by reperfusion (4 h). METHODS AND RESULTS: 23 porcine hearts were randomized to 5 groups: hemoperfused non-infarcted hearts (C), infarcted hearts (MI: R. circumflexus), infarcted hearts treated with quinaprilat (Q), infarcted hearts treated with angiotensin-I (Ang I), and infarcted hearts treated with angiotensin-I and quinaprilat (QA). Fas, Bax, bcl-2 and p53 proteins were increased in MI hearts and further elevated by Ang I. Quinaprilat reduced Bax and p53. Bcl-2 was elevated in Q and reduced in QA. An early upregulation of caspase-3 gene and protein expression was detected in MI and Ang I hearts compared to C. Q reduced caspase-3 gene expression, but had no effect on caspase-3 and Fas protein. CONCLUSIONS: These data suggest that the RAS plays a pivotal role in cardiac apoptosis which is the early and predominant form of death in myocardial infarction. Ischemia/reperfusion induces programmed cell death via extrinsic and intrinsic pathways. Early treatment with quinaprilat attenuated cardiomyocyte apoptosis.

Influence of malonyl-CoA and palmitate concentration on rate of palmitate oxidation in rat muscle.

5-Aminoimidazole-4-carboxamide 1-beta-D-ribofuranoside (AICAR) is taken up by perfused skeletal muscle and phosphorylated to form 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuraosyl-5'-monopho sph ate (analog of 5'-AMP) with consequent activation of AMP-activated protein kinase, phosphorylation of acetyl-CoA carboxylase, decrease in malonyl-CoA, and increase in fatty acid oxidation. This study was designed to determine the effect of increasing levels of palmitate on the rate of fatty acid oxidation. Malonyl-CoA concentration was manipulated with AICAR at different palmitate concentrations. Rat hindlimbs were perfused with Krebs-Henseleit bicarbonate containing 4% bovine serum albumin, washed bovine red cells, 200 microU/ml insulin, 10 mM glucose, and different concentrations of palmitate (0. 1-1.0 mM) without or with AICAR (2.0 mM). Perfusion with medium containing AICAR was found to activate AMP-activated protein kinase in skeletal muscle, inactivate acetyl-CoA carboxylase, and decrease malonyl-CoA at all concentrations of palmitate. The rate of palmitate oxidation increased as a function of palmitate concentration in both the presence and absence of AICAR but was always higher in the presence of AICAR. These results provide additional evidence that malonyl-CoA is an important regulator of the rate of fatty acid oxidation at palmitate concentrations in the physiological range.

Evidence for 5' AMP-activated protein kinase mediation of the effect of muscle contraction on glucose transport.

The intracellular signaling proteins that lead to exercise-stimulated glucose transport in skeletal muscle have not been identified, although it is clear that there are separate signaling mechanisms for exercise- and insulin-stimulated glucose transport. We have hypothesized that the 5'AMP-activated protein kinase (AMPK) functions as a signaling intermediary in exercise-stimulated glucose uptake. This hypothesis was based on recent studies showing the following: 1) muscle contraction increases AMPK activity and 2) perfusion of rat hindlimb skeletal muscles with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR), a compound that results in increased AMPK activity, increased insulin-stimulated glucose uptake. In the current study, isolated rat epitrochlearis muscles were treated to contract in vitro (via electrical stimulation for 10 min) and/or incubated in the absence or presence of AICAR (2 mmol/l), insulin (1 micromol/l), or wortmannin (100 nmol/l). Both contraction and AICAR significantly increased AMPK activity, while the enzyme was not activated by insulin. AICAR, contraction, and insulin all increased 3-O-methylglucose (3MG) transport by threefold to fivefold above basal. The phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin completely blocked insulin-stimulated transport, but did not inhibit AICAR- or contraction-stimulated transport. The increase in glucose transport with the combination of maximal AICAR plus maximal insulin treatments was partially additive, suggesting that these stimuli increase glucose transport by different mechanisms. In contrast, there was no additive effect on glucose transport with the combination of AICAR plus contraction. These data suggest that AICAR and contraction stimulate glucose transport by a similar insulin-independent signaling mechanism and are consistent with the hypothesis that AMPK is involved in exercise-stimulated glucose uptake.

Adenosine increases interleukin 6 release and decreases tumour necrosis factor release from rat adrenal zona glomerulosa cells, ovarian cells, anterior pituitary cells, and peritoneal macrophages.

Adenosine modifies interleukin 6 (IL-6) and tumour necrosis factor (TNF) release from immune tissues. Because adenosine alters endocrine function and endocrine cells secrete cytokines, its effects on IL-6 and TNF release from rat adrenals, ovaries, and anterior pituitaries were compared with its effects on cytokine release from rat peritoneal macrophages. Adenosine increased basal IL-6 release and decreased basal TNF release from adrenal zona glomerulosa and zona fasciculata/reticularis cells. IL-6 and TNF release from zona glomerulosa cells was greater (20x) than that of other adrenal cells. An A2 agonist modified adrenal IL-6 and TNF release at lower concentrations than an A1 agonist. Adenosine augmented adrenal IL-6 release stimulated by endotoxin (LPS), interleukin 1 beta (IL-1 beta), adrenocorticotrophic hormone, and angiotensin II. LPS- and IL-1 beta-stimulated adrenal TNF release was inhibited by adenosine. Adenosine increased IL-6 release and inhibited TNF release from ovarian cells. Anterior pituitary cells released IL-6, but no detectable TNF. Adenosine, via A2 receptors, stimulated IL-6 secretion from these cells. Peritoneal macrophage IL-6 release was increased and TNF release decreased by adenosine. Thus, in immune and endocrine tissues, adenosine increases IL-6 release, but inhibits TNF release.

AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle.

5-Aminoimidazole-4-carboxamide ribonucleoside (AICAR) has previously been reported to be taken up into cells and phosphorylated to form ZMP, an analog of 5'-AMP. This study was designed to determine whether AICAR can activate AMP-activated protein kinase (AMPK) in skeletal muscle with consequent phosphorylation of acetyl-CoA carboxylase (ACC), decrease in malonyl-CoA, and increase in fatty acid oxidation. Rat hindlimbs were perfused with Krebs-Henseleit bicarbonate containing 4% bovine serum albumin, washed bovine red blood cells, 200 microU/ml insulin, and 10 mM glucose with or without AICAR (0.5-2.0 mM). Perfusion with medium containing AICAR was found to activate AMPK in skeletal muscle, inactivate ACC, and decrease malonyl-CoA. Hindlimbs perfused with 2 mM AICAR for 45 min exhibited a 2.8-fold increase in fatty acid oxidation and a significant increase in glucose uptake. No difference was observed in oxygen uptake in AICAR vs. control hindlimb. These results provide evidence that decreases in muscle content of malonyl-CoA can increase the rate of fatty acid oxidation.

[Insulin injection aid. Experiences with its use in Type II diabetes patients]. / Insulin-Injektionshilfe. Erfahrungen mit dem Einsatz bei Typ-II-Diabetikern.

This article reports on the experience gained with an insulin injection aid for use by type II diabetics. The baseline data provided by a questionnaire survey are discussed in detail. The "pen" proved simple to use and was readily accepted. A number of shortcomings were identified and improvements suggested. The significantly superior metabolic control observed in patients using such aids is briefly discussed. In addition, detailed consideration is given to the models presently available and desirable future prospects are identified.

[Attitude to the special education student with reference to social integration in early adulthood--studies in a rural district]. / Einstellungen zum Hilfsschüler und Aspekte der gesellschaftlichen Integration im jungen Erwachsenenalter--Untersuchungen in einem Landkreis.

This paper deals with cognition of attitudes to pupils from a special school for mentally retarded children in a rural district on the base of using polarity profiles. Although children from Secondary Schools and adults have little understanding of the retarded children's handicaps, they do not appear to hold one-sided stereotyped opinions and in the areas of social behavior and ways of working positive attributes of the personality are emphasized. In comparison with this results recorded self-assessments of former pupils of special schools, together with the assessments of others, in real working situations show a satisfactory level of integration, although after initial difficulties. Opportunities for advice and care are considered necessary, even after pupils have left school.

Effects of NaCl and CaCl(2) on Cell Enlargement and Cell Production in Cotton Roots.

In many crop species, supplemental Ca(2+) alleviates the inhibition of growth typical of exposure to salt stress. In hydroponically grown cotton seedlings (Gossypium hirsutum L. cv Acala SJ-2), both length and weight of the primary root were enhanced by moderate salinities (25 to 100 millimolar NaCl) in the presence of 10 millimolar Ca(2+), but the roots became thinner. Anatomical analysis showed that the cortical cells of these roots were longer and narrower than those of the control plants, while cortical cells of roots grown at the same salinities but in the presence of only 0.4 millimolar Ca(2+) became shorter and more nearly isodiametrical. Cell volume, however, was not affected by salinities up to 200 millimolar NaCl at either 0.4 or 10 millimolar Ca(2+). Our observations suggest Ca(2+)-dependent effects of salinity on the cytoskeleton. The rate of cell production declined with increasing salinity at 0.4 millimolar Ca(2+) but at 10 millimolar Ca(2+) was not affected by salinities up to 150 millimolar NaCl.

Research in mental retardation in the German Democratic Republic (GDR).

Research in mental retardation in the GDR has been influenced primarily by the brain damage--development theory, which assumes biological as well as social determinants in human development. The theory was based on the results of brain damage (encephalopathy) research, which distinguishes between primary and secondary symptoms of mental retardation. The importance of social factors such as family conditions for development was demonstrated in a variety of studies, including research with adopted children. Positive results have been achieved with early education programming for developmentally disabled children, as well as with medical treatment for some genetically induced metabolic disorders. Results from longitudinal studies suggest that intelligence continues to develop in educable mentally retarded adolescents. New diagnostic methods have been developed; test of motor skills, visuomotor coordination, and a developmental test for young infants. Further developments of traditional school entrance diagnosis for special education classes is reported. Several studies show that there has been some success in the vocational and social integration of mentally retarded persons in the GDR.

[Condition analysis of behavior disorders in children and adolescents in residential treatment in a district town]. / Zur Bedingungsanalyse der Verhaltensstörungen bei heimeingewiesenen Kindern und Jugendlichen einer Bezirksstadt.

The psychological-statistical analysis of the resolutions concerning accommodation in a home by the Youth Welfare Services of the town of Rostock in 1973 includes 75 children and young people with (dissocial) behaviour disorders, their average age at that time being 14.0 years. The main phenomena were playing truant or idling, running away as well as difficulties regarding discipline. A remarkable fact is the large percentage of parents with lacking vocational training and the frequency of families with more than four children (10 times as high as the average of the population) with the rate of divorces being at the same time high (39%). 55 per cent of the persons with behaviour disorders did not have a normal school development, for example the percentage of the children attending schools for backward children was 14 times as high as the average of the other school children. As a consequence, the necessity of making an early use of advisory and educational measures is pointed out, especially in cases of such families with many children who are lacking the basic conditions for an education.

[Disorders of psychological performance in mentally normal school children]. / Psychische Leistungsstörungen bei geistig normalentwickelten Schulkindern.

The authors deal with such psychic factors as weaknesses in concentration, reading and spelling which are to a major degree responsible for failure at school. These factors often occur in combination and may indicate early brain damage, in complex interaction with milieu conditions. Ample evidence of this is given in a literature survey. Principles of concentration training and a diagnostic verification procedure are then outlined, which have been derived from an analysis of attention requirements made at school. A therapy for reading and spelling weaknesses is dealt with, followed by a presentation of the results of training and catamneses. Using the method of Kossow mistakes were reduced by 78 percent and spelling marks improved considerably, after the children had returned to their original schools. In conclusion information is given on future projects for better assistance and prevention in the case of reading and spelling weaknesses at the elementary stage.

[On connection between music therapy and music pedagogics during childhood (author's transl)]. / Zur Beziehung zwischen Musiktherapie und Musikpädagogik im Kindesalter.

In this article is explained, that, in spite of existing communities between music therapy and cooperating remedial education on the one hand and music education on the other, we can separate with regard to the method music therapy as one form of psychotherapy from educational influence. But in practice at therapy of children with psychical disturbances we always must take into considerations the education object in view.

[Possibilities of behavior therapy in childhood]. / Möglichkeiten der Verhaltenstherapie im Kindesalter.

The author, after discussing methods of conditioning and their theoretical substantiation, describes the behavior disorders, forms of malconduct, and development disorders where methods of behavior therapy have so far been made successful use of. Possibilities of, and technical problems associated with, the treatment of nocturnal enuresis are discussed in addition to methods of treatment of autistic, electively mutistic, and stuttering children. Also discussed in this paper are methods of behavior therapy of states of anxiety, phobias, and hypermotor conditions as well as the instruction of educators in the use methods developed on the lines of behavior therapy. Behavior therapy is characterized as a form of psychotherapy which cannot replace general pedagogic influences.