Influence of cosmetic formulations on the skin's circadian clock.

OBJECTIVE: The circadian rhythm was set into focus by awarding the Nobel Price of Physiology/Medicine to Jeffrey Hall, Michael Rosbash and Michael Young in late 2017. Numerous publications elucidated the molecular mechanisms driving the circadian biorhythms of our body, peripheral organs and each single cell. However, there is minor knowledge on the circadian rhythm of the skin, which has its own peripheral circadian clock in contact with cosmetic formulations. The skin's epidermal clock is excessively influenced by environmental factors like UV radiation or modern lifestyle, which may induce epidermal jetlag. Here, we give an overview on the current knowledge about the epidermal circadian clock and provide a cosmetic solution to protect and preserve the biorhythm of the skin. METHODS: Quantitative RT-PCR to analyse the gene expression of circadian clock genes and the downstream DNA repair gene OGG1 in keratinocytes irradiated with UV-B. In vivo study to determine skin parameters dependent on the circadian cycle and interference of cosmetic formulations to them by assessment of morning and evening values at each measurement day after 28, 56 and 84 days of the study. RESULTS: UV-B irradiation leads to a pronounced delay in circadian clock and downstream gene expression which interferes in the proper function of epidermal stem cells and as thus skin function. The use of a cosmetic active ingredient prevents cyclobutane pyrimidine dimer formation, protects epidermal stem cells and resets the circadian gene expression. It preserves the circadian changes in skin hydration, reduces daily fluctuations of skin redness and strengthens the skin barrier. CONCLUSION: The skin has its own circadian biorhythm to gain full functionality. Interruption of this oscillation will lead to functional impairments. Here we show a cosmetic solution to protect and preserve the skin's circadian rhythm. DNA protection, ROS elimination and stimulation of circadian gene expression seem to be crucial to keep the skin in balance.

OBJECTIF: Le rythme circadien a été mis en lumière par l'attribution du prix Nobel de physiologie/médecine à Jeffrey Hall, Michael Rosbash et Michael Young fin 2017. De nombreuses publications ont élucidé les mécanismes moléculaires qui régissent les biorythmes circadiens de notre corps, des organes périphériques et de chaque cellule. Cependant, il existe peu de connaissances sur le rythme circadien de la peau, qui possède sa propre horloge circadienne périphérique, avec les formules cosmétiques. L'horloge épidermique de la peau est excessivement influencée par des facteurs environnementaux comme les rayons UV ou le mode de vie moderne, qui peuvent induire un "décalage horaire de l'épiderme". Nous donnons ici un aperçu des connaissances actuelles sur l'horloge circadienne épidermique et proposons une solution cosmétique pour protéger et préserver le biorythme de la peau. MÉTHODES: Test RT-PCR quantitatif pour analyser l'expression des gènes de l'horloge circadienne et du gène de réparation de l'ADN OGG1 en aval dans des kératinocytes irradiés aux UV-B. Étude in vivo pour determiner les paramètres cutanés dépendant du cycle circadien et l'interférence des formules cosmétiques sur ces paramètres par évaluation des résultats du matin et du soir à chaque jour de mesure après à 28, 56 et 84 jours d'étude. RÉSULTATS: L'irradiation aux UV-B entraîne un retard prononcé de l'horloge circadienne et de l'expression des gènes en aval, ce qui interfère avec le bon fonctionnement des cellules souches épidermiques et donc en conséquence avec la fonction de la peau. L'utilisation d'un principe actif cosmétique empêche la formation de dimère de cyclobutane pyrimidine, protège les cellules souches épidermiques et rétablit l'expression génique circadienne. Il préserve les changements circadiens dans l'hydratation de la peau, réduit les fluctuations quotidiennes des rougeurs cutanées et renforce la barrière cutanée. CONCLUSION: La peau a son propre biorythme circadien pour acquérir une assurer une parfaite fonctionnalité. L'interruption de cette oscillation entraînera des troubles fonctionnels. Nous présentons ici une solution cosmétique pour protéger et préserver le rythme circadien de la peau. La protection de l'ADN, l'élimination des ROS et la stimulation de l'expression des gènes circadiens semblent être cruciales pour maintenir l'équilibre de la peau.

Testosterone does not affect agrin cleavage in mobility-limited older men despite improvement in physical function.

In a subset of men, sarcopenia and physical dysfunction occur due to destabilization of the neuromuscular junction (NMJ), which is manifested by elevated serum concentrations of C-terminal agrin fragment (CAF). Testosterone administration improves physical function in some studies; however, its effects on serum circulating CAF concentrations remain unknown. Here we evaluate the effects of testosterone administration on circulating CAF levels in mobility-limited men with low testosterone aged 65 or older participating in the Testosterone in Older Men with Mobility Limitations (TOM) Trial. We analyzed the difference in change in serum CAF levels between testosterone and placebo groups, as well as its association with muscle strength and physical function. Association of change in serum CAF levels with serum total (TT) and free testosterone (FT) was also evaluated. Men randomized to testosterone experienced significant improvement in muscle strength and physical function (assessed by loaded stair-climbing power). However; testosterone administration was not associated with a reduction in serum CAF levels (effect size = -50.3 pm; 95% CI = -162.1 to 61.5 pm; p = 0.374); there was no association between changes in CAF levels with changes in TT (p = 0.670) or FT (p = 0.747). There was no association between changes in serum CAF levels with improvement in either muscle strength or stair-climbing power. In conclusion, testosterone treatment in mobility-limited older men with low to low-normal testosterone levels did not reduce serum CAF levels. Additionally, testosterone-induced improvements in muscle strength and physical function were not associated with changes in serum CAF concentrations. These findings suggest that improvement in physical function with testosterone replacement in older men with mobility limitations and elevated CAF levels is mediated by mechanisms other than stabilization of the NMJ.

Grifolin derivatives from Albatrellus ovinus as TRPV1 receptor blockers for cosmetic applications.

OBJECTIVE: Blocking the TRPV1 receptor is an interesting approach for the treatment of sensitive skin. Here we investigated the potential of grifolin derivatives from Albatrellus ovinus to act as TRPV1 receptor blockers and their potential to serve as cosmetic active ingredients. METHODS: Binding characteristics of grifolin derivatives from Albatrellus ovinus were determined in competitive and functional in vitro assays to achieve IC50 values. The TRPV1 receptor was activated in vivo with capsaicin and noxious heat to investigate skin reddening, microcirculation, skin sensations and heat pain thresholds. RESULTS: Grifolin derivatives extracted from Albatrellus ovinus proved to inhibit the TRPV1 receptor in vitro and in vivo. Besides suppression of the TRPV1 receptor activity upon chemical stimulation with capsaicin, thermal activation was shown to be inhibited as well by application of cosmetic formulations containing 3% Albatrellus ovinus extract. The reduction of stinging and burning sensations as well as reduction of reddening and microcirculation upon irritation with capsaicin or thermal stress proved efficacy in vivo. CONCLUSION: Grifolin derivatives from Albatrellus ovinus are able to serve as fungal-derived TRPV1 receptor blockers with capability to serve as a cosmetic active ingredient on sensitive skin.

Effects of a One-Year Physical Activity Program on Serum C-Terminal Agrin Fragment (CAF) Concentrations among Mobility-Limited Older Adults.

OBJECTIVES: C-terminal Agrin Fragment (CAF) has been proposed as a potential circulating biomarker for predicting changes in physical function among older adults. To determine the effect of a one-year PA intervention on changes in CAF concentrations and to evaluate baseline and longitudinal associations between CAF concentrations and indices of physical function. DESIGN: Ancillary study to the Lifestyle Interventions and Independence for Elders Pilot (LIFE-P), a multi-site randomized clinical trial designed to evaluate the effects of chronic exercise on the physical function of older adults at risk for mobility disability. SETTING: Four academic research centers within the U.S. PARTICIPANTS: Three hundred thirty three older adults aged 70 to 89 with mild to moderate impairments in physical function. INTERVENTION: A 12-month intervention of either structured physical activity (PA) or health education promoting successful aging (SA). MEASUREMENTS: Serum CAF concentrations and objectives measures of physical function - i.e. gait speed and performance on the Short Physical Performance Battery (SPPB). RESULTS: The group*time interaction was not significant for serum CAF concentrations (p=0.265), indicating that the PA intervention did not significantly reduce serum CAF levels compared to SA. Baseline gait speed was significantly correlated with baseline CAF level (r = -0.151, p= 0.006), however the association between CAF and SPPB was not significant. Additionally, neither baseline nor the change in CAF concentrations strongly predicted the change in either performance measure following the PA intervention. CONCLUSION: In summary, the present study shows that a one-year structured PA program did not reduce serum CAF levels among mobility-limited older adults. However, further study is needed to definitively determine the utility of CAF as a biomarker of physical function.

Outcome prediction using clinical scores and biomarkers in patients with presumed severe infection in the emergency department.

BACKGROUND: Severe infections play an important role in the emergency department (ED) and early risk stratification is essential. We compared the prognostic value of APACHE II, SOFA, and MEDS scores, and the biomarkers C-reactive protein (CRP), procalcitonin (PCT), and interleukin 6 (IL-6). METHODS: We performed a prospective observational study. Patients aged 18 years or older with a severe infection, from whom blood cultures were taken, were included. RESULTS: Two hundred and eleven patients were included. The 30-day mortality rate was 8.5%. All scores and biomarkers showed significant area under the curve (AUC) values of receiver operating characteristic curve analysis for death within 30 days: 0.801 for APACHE II, 0.785 for MEDS, 0.708 for SOFA, 0.693 for CRP, 0.651 for PCT, and 0.716 for IL-6. For treatment in an ICU and need for mechanical ventilation, these parameters had significant AUC values, too. For renal replacement therapy, only APACHE II, SOFA, and PCT showed significant AUC values. According to the trend observed, the AUC values were highest for the APACHE II score. CONCLUSIONS: All investigated parameters have a predictive value in patients with an infection in the ED. According to the trend observed, the APACHE II score seems to have the best discriminative power. Use of the APACHE II score already at the time of admission to the ED may be useful for stratifying patients at risk for ICU treatment, thereby using the same score in the ED and the ICU.

Microbial diagnostics in patients with presumed severe infection in the emergency department.

INTRODUCTION: Sepsis in the early stage is a common disease in emergency medicine, and rapid diagnosis is essential. Our aim was to compare pathogen diagnosis using blood cultures (BC) and the multiplex polymerase chain reaction (PCR) test.Methods. At total of 211 patients admitted to the multidisciplinary emergency department of our university hospital between 2006 and 2009 with suspected severe infection from any origin were studied. Blood samples for BC (aerobic and anaerobic) and multiplex PCR were taken for identification of infectious microorganisms immediately after hospital admission. Results of the BC and PCR correlated with procalcitonin concentration (PCT) and clinical diagnosis of sepsis (≥2 positive SIRS criteria) as well as with severity of disease at admission and with clinical outcome measures. RESULTS: Results of the BC were available in 200 patients (94.8%) and PCR were available in 119 patients (56.3%), respectively. In total, 87 BC (43.5%) were positive and identified 94 pathogens. In 45 positive PCRs, 47 pathogens (37.8%) were found. Identical results were obtained in 81.4%. In addition, BC identified 9 Gram-positive and 3 Gram-negative bacteria, while PCR added 5 Gram-negative pathogens. Coagulase-negative staphylococci were detected in blood cultures only (n=20, 21.3%), whereas PCR identified significantly more Gram-negative bacteria than BC. In patients with positive PCR results, the PCT level was significantly higher than in patients with negative PCR (15.0±23.3 vs. 8.8±32.8 ng/ml, p<0.001). This difference was not observed for BC (10.6±25.7 vs. 11.6±44.9 ng/ml, p=0.075). The APACHE II score correlated with PCR (19.2±9.1 vs. 15.8±8.9, p<0.05) and was also higher in positive BC (18.7±8.7 vs. 14.4±8.0, p<0.01). Positive PCR and BC were correlated with negative clinical outcomes (e.g., transfer to ICU, mechanical ventilation, renal replacement therapy, death). CONCLUSION: In patients admitted with suspected severe infection, a high percentage of positive BC and PCR were observed. Positive findings in the PCR correlate with elevated levels of PCT and high APACHE II scores.

[Septic circulatory shock and septic cardiomyopathy]. / Septischer Kreislaufschock und septische Kardiomyopathie.

Septic shock is not only a circulatory shock but is also a cardiac shock, the consequence of a potentially reversible heart impairment known as septic cardiomyopathy. Disturbances of macrocirculation as well as microcirculation, an individually heterogeneous reduction of cardiac function, and an extensive impairment of demand-oriented regulation of heart function characterize the septic shock state. Bacterial toxins, inflammatory mediators, and a disseminated intravasal coagulopathy are responsible for these disturbances; for the impairment of cardiac regulation, the interaction of endotoxin with the cardiac pacemaker current I(f) also plays a role. Circulatory shock as well as septic cardiomyopathy should be quantified: The lowering of systemic vascular resistance characterizes the extent of circulatory shock, and the reduction of relative cardiac output in relation to afterload characterizes the extent of septic cardiomyopathy. The intensity of circulatory as well as of cardiac impairment correlates with an unfavorable prognosis. Treatment of septic circulatory shock and of septic cardiomyopathy is predominantly symptomatic; first causal approaches are under investigation.

[Specific aspects in septic patients: initial phase in the emergency department, age, sex and post-ICU-care]. / Spezielle Aspekte bei Sepsispatienten : Initiale Phase auf der Notaufnahme, Lebensalter, Geschlecht, Postintensivphase.

This article deals with specific aspects of the patient with sepsis and his treatment. With adequate therapy (antibiotics started within the first hour, early goal-directed therapy) initiated as early as possible, the patient with community-aquired severe sepsis in the emergency department has a considerable better chance to survive than the patient with prolonged severe sepsis in the ICU. The average age of patients with severe sepsis and septic shock treated at the ICU is rising, with consequences like oligosymptomatic presentation, altered pharmakokinetics according to older-age-induced reduced organ functions and prolonged stay at the ICU due to comorbidity. Due to sexual dimorphisms of the immune system, women have a better prognosis in sepsis than men. In an animal sepsis model activation of the estrogen receptor beta improves prognosis. Within six months after having survived severe sepsis, morbidity and mortality is still increased. Taking care for the patient in a post-ICU outpatient department during this time will help to recognize these problems and to effectively treat the patient as soon as possible.

Isolated diastolic dysfunction--diagnostic value of tissue Doppler imaging, colour M-mode and N-terminal pro B-type natriuretic peptide.

INTRODUCTION: For diagnosis of diastolic dysfunction of the left ventricle (DDF), measurement of relaxation velocity (V(R)) by tissue Doppler imaging (V(R)), flow propagation velocity of transmitral inflow (v(p)) as well as the measurement of serum levels of N-terminal pro b-type natriuretic peptide (NT-proBNP) compete with the standard echocardiographic DDF-measures because of several disadvantages of the latter. METHODS: We examined the diagnostic value of method 1, 2 and NT-proBNP in 120 patients with echocardiographic-proven DDF and in 20 patients without. Patients were classified according to the DDF-stage by standard echocardiographic parameters (transmitral E/A-ratio, deceleration time, isovolumetric relaxation time) into stage I, II and III and furthermore subdivided by the presence of dyspnoea. RESULTS: V(R) and v(p) were significantly lower in patients with DDF than in patients without DDF, with no difference between the various DDF stages. Symptomatic patients showed a trend to a lower V(R). NT-proBNP was elevated in patients with DDF: Symptomatic patients with a DDF at stage I and patients with a DDF at stage II and III independent of the presence of symptoms had elevated NT-proBNP levels. CONCLUSION: All three methods tested identified patients with DDF. NT-proBNP and v(p) were able to discriminate between symptomatic and asymptomatic patients.

Dissection of a (betaalpha)8-barrel enzyme into two folded halves.

The (betaalpha)8-barrel, which is the most frequently encountered protein fold, is generally considered to consist of a single structural domain. However, the X-ray structure of the imidazoleglycerol phosphate synthase (HisF) from Thermotoga maritima has identified it as a (betaalpha) 8-barrel made up of two superimposable subdomains (HisF-N and HisF-C). HisF-N consists of the four N-terminal (betaalpha) units and HisF-C of the four C-terminal (betaalpha) units. It has been postulated, therefore, that HisF evolved by tandem duplication and fusion from an ancestral half-barrel. To test this hypothesis, HisF-N and HisF-C were produced in Escherichia coli, purified and characterized. Separately, HisF-N and HisF-C are folded proteins, but are catalytically inactive. Upon co-expression in vivo or joint refolding in vitro, HisF-N and HisF-C assemble to the stoichiometric and catalytically fully active HisF-NC complex. These findings support the hypothesis that the (betaalpha)8-barrel of HisF evolved from an ancestral half-barrel and have implications for the folding mechanism of the members of this large protein family.

Directed evolution of a (beta alpha)8-barrel enzyme to catalyze related reactions in two different metabolic pathways.

Enzymes participating in different metabolic pathways often have similar catalytic mechanisms and structures, suggesting their evolution from a common ancestral precursor enzyme. We sought to create a precursor-like enzyme for N'-[(5'-phosphoribosyl)formimino]-5-aminoimidazole-4-carboxamide ribonucleotide (ProFAR) isomerase (HisA; EC ) and phosphoribosylanthranilate (PRA) isomerase (TrpF; EC ), which catalyze similar reactions in the biosynthesis of the amino acids histidine and tryptophan and have a similar (betaalpha)(8)-barrel structure. Using random mutagenesis and selection, we generated several HisA variants that catalyze the TrpF reaction both in vivo and in vitro, and one of these variants retained significant HisA activity. A more detailed analysis revealed that a single amino acid exchange could establish TrpF activity on the HisA scaffold. These findings suggest that HisA and TrpF may have evolved from an ancestral enzyme of broader substrate specificity and underscore that (betaalpha)(8)-barrel enzymes are very suitable for the design of new catalytic activities.

MPH1, a yeast gene encoding a DEAH protein, plays a role in protection of the genome from spontaneous and chemically induced damage.

We have characterized the MPH1 gene from Saccharomyces cerevisiae. mph1 mutants display a spontaneous mutator phenotype. Homologs were found in archaea and in the EST libraries of Drosophila, mouse, and man. Mph1 carries the signature motifs of the DEAH family of helicases. Selected motifs were shown to be necessary for MPH1 function by introducing missense mutations. Possible indirect effects on translation and splicing were excluded by demonstrating nuclear localization of the protein and splicing proficiency of the mutant. A mutation spectrum did not show any conspicuous deviations from wild type except for an underrepresentation of frameshift mutations. The mutator phenotype was dependent on REV3 and RAD6. The mutant was sensitive to MMS, EMS, 4-NQO, and camptothecin, but not to UV light and X rays. Epistasis analyses were carried out with representative mutants from various repair pathways (msh6, mag1, apn1, rad14, rad52, rad6, mms2, and rev3). No epistatic interactions were found, either for the spontaneous mutator phenotype or for MMS, EMS, and 4-NQO sensitivity. mph1 slightly increased the UV sensitivity of mms2, rad6, and rad14 mutants, but no effect on X-ray sensitivity was observed. These data suggest that MPH1 is not part of a hitherto known repair pathway. Possible functions are discussed.