Evolutionary adaptation of an HP1-protein chromodomain integrates chromatin and DNA sequence signals.

Members of the diverse heterochromatin protein 1 (HP1) family play crucial roles in heterochromatin formation and maintenance. Despite the similar affinities of their chromodomains for di- and tri-methylated histone H3 lysine 9 (H3K9me2/3), different HP1 proteins exhibit distinct chromatin-binding patterns, likely due to interactions with various specificity factors. Previously, we showed that the chromatin-binding pattern of the HP1 protein Rhino, a crucial factor of the Drosophila PIWI-interacting RNA (piRNA) pathway, is largely defined by a DNA sequence-specific C2H2 zinc finger protein named Kipferl (Baumgartner et al., 2022). Here, we elucidate the molecular basis of the interaction between Rhino and its guidance factor Kipferl. Through phylogenetic analyses, structure prediction, and in vivo genetics, we identify a single amino acid change within Rhino's chromodomain, G31D, that does not affect H3K9me2/3 binding but disrupts the interaction between Rhino and Kipferl. Flies carrying the rhinoG31D mutation phenocopy kipferl mutant flies, with Rhino redistributing from piRNA clusters to satellite repeats, causing pronounced changes in the ovarian piRNA profile of rhinoG31D flies. Thus, Rhino's chromodomain functions as a dual-specificity module, facilitating interactions with both a histone mark and a DNA-binding protein.

Training intensity influences left ventricular dimensions in young competitive athletes.

Background: In young athletes, exercise causes changes in the heart that include growth in wall thickness and mass of the left ventricle and expansion of the heart's chambers. The heart's function is either preserved or enhanced, but this may change to the opposite over time. Objective: This study aimed to assess structural and functional cardiac adaptations in relation to exercise training time, intensity, and performance in young competitive athletes. Methods: A total of 404 children and adolescents (14.23 ± 2.0 years, 97 females) were enrolled in the Munich Cardiovascular Adaptations in Young Athletes Study (MuCAYA-Study). Eighty-five participants were examined two times a year. Two-dimensional echocardiography was performed to assess left ventricular structure and function. Training time and intensity was measured with the MoMo physical activity questionnaire, maximum aerobic capacity by cardiopulmonary exercise testing, and strength with the handgrip strength test. Results: Maximum aerobic capacity significantly influenced interventricular septal thickness in diastole. Training intensity significantly influenced left ventricular internal diameter in diastole and systole, and left ventricular mass indexed to body surface area. Within one year, interventricular wall thickness, relative wall thickness and left ventricular mass, indexed to body surface area and height, increased significantly. Training intensity and aerobic capacity contributed to cardiac adaptations in young competitive athletes, as represented by altered structural parameters but preserved cardiac function. Within a year, however, structural changes and a decline in diastolic performance were observed within the longitudinal sub-sample. Conclusion: Our results confirm the hypothesis that cardiac adaptations to exercise occur at a young age. Cardiac adaptation in our cohort was influenced by exercise intensity and maximum aerobic capacity.

The Drosophila ZAD zinc finger protein Kipferl guides Rhino to piRNA clusters.

RNA interference systems depend on the synthesis of small RNA precursors whose sequences define the target spectrum of these silencing pathways. The Drosophila Heterochromatin Protein 1 (HP1) variant Rhino permits transcription of PIWI-interacting RNA (piRNA) precursors within transposon-rich heterochromatic loci in germline cells. Current models propose that Rhino's specific chromatin occupancy at piRNA source loci is determined by histone marks and maternally inherited piRNAs, but also imply the existence of other, undiscovered specificity cues. Here, we identify a member of the diverse family of zinc finger associated domain (ZAD)-C2H2 zinc finger proteins, Kipferl, as critical Rhino cofactor in ovaries. By binding to guanosine-rich DNA motifs and interacting with the Rhino chromodomain, Kipferl recruits Rhino to specific loci and stabilizes it on chromatin. In kipferl mutant flies, Rhino is lost from most of its target chromatin loci and instead accumulates on pericentromeric Satellite arrays, resulting in decreased levels of transposon targeting piRNAs and impaired fertility. Our findings reveal that DNA sequence, in addition to the H3K9me3 mark, determines the identity of piRNA source loci and provide insight into how Rhino might be caught in the crossfire of genetic conflicts.

The genes within our DNA encode the essentials of our body plan and how each task in the body is achieved. However, our genome also contains many repetitive regions of DNA that do not encode functional genes. Some of these regions are genetic parasites known as transposons that try to multiply and spread around the DNA of their host. To prevent transposon DNA from interfering with the way the body operates, humans and other animals have evolved elaborate defense mechanisms to identify transposons and prevent them from multiplying. In one such mechanism, known as the piRNA pathway, the host makes small molecules known as piRNAs that have sequences complementary to those of transposons, and act as guides to silence the transposons. The instructions to make these piRNAs are stored in the form of transposon fragments in dedicated regions of host DNA called piRNA clusters. These clusters thereby act as genetic memory, allowing the host to recognize and silence specific transposons in other locations within the host's genome. In fruit flies, a protein called Rhino binds to piRNA clusters that are densely packed to allow piRNAs to be made. However, it remained unclear how Rhino is able to identify and bind to piRNA clusters, but not to other similarly densely packed regions of DNA. Baumgartner et al. used a combination of genetic, genomic, and imaging approaches to study how Rhino finds its way in the fruit fly genome. They found that another protein called Kipferl interacts with Rhino and is required for Rhino to bind to nearly all piRNA clusters. Since Kipferl can by itself bind to the sequences that Rhino needs to find, the results suggest that Kipferl acts to recruit and initiate Rhino binding within densely packed piRNA clusters. Further experiments found that, in flies lacking Kipferl, Rhino binds to regions of DNA called Satellite repeats, hinting that these selfish sequences may compete for Rhino for their own benefit. The finding that Kipferl and Rhino work together to define the memory system of the piRNA pathway strongly advances our understanding of how a sequence-specific defense system based on small RNAs can be established.

Panoramix SUMOylation on chromatin connects the piRNA pathway to the cellular heterochromatin machinery.

Nuclear Argonaute proteins, guided by small RNAs, mediate sequence-specific heterochromatin formation. The molecular principles that link Argonaute-small RNA complexes to cellular heterochromatin effectors on binding to nascent target RNAs are poorly understood. Here, we explain the mechanism by which the PIWI-interacting RNA (piRNA) pathway connects to the heterochromatin machinery in Drosophila. We find that Panoramix, a corepressor required for piRNA-guided heterochromatin formation, is SUMOylated on chromatin in a Piwi-dependent manner. SUMOylation, together with an amphipathic LxxLL motif in Panoramix's intrinsically disordered repressor domain, are necessary and sufficient to recruit Small ovary (Sov), a multi-zinc-finger protein essential for general heterochromatin formation and viability. Structure-guided mutations that eliminate the Panoramix-Sov interaction or that prevent SUMOylation of Panoramix uncouple Sov from the piRNA pathway, resulting in viable but sterile flies in which Piwi-targeted transposons are derepressed. Thus, Piwi engages the heterochromatin machinery specifically at transposon loci by coupling recruitment of a corepressor to nascent transcripts with its SUMOylation.

Exercise Training Duration and Intensity Are Associated With Thicker Carotid Intima-Media Thickness but Improved Arterial Elasticity in Active Children and Adolescents.

Even though exercise generally has a positive effect on health, intensive exercise can have adverse effects on the vascular system of adults. This study aimed to investigate the association between training duration and intensity and vascular structure and function in 427 physically active children and adolescents (14.0 ± 1.94 years). In this study, we examined carotid intima-media thickness (cIMT), carotid diameter, and cIMT:diameter-ratio as parameters of carotid arterial structure and arterial compliance (AC), stiffness index ß (ß), elastic modulus (Ep), and carotid pulse wave velocity (PWVß) as parameters of carotid arterial function with high-resolution ultrasound. We collected central systolic blood pressure (cSBP) and aortic pulse wave velocity (aPWV) as parameters of central arterial stiffness with an oscillometric device. We used the MoMo Physical Activity Questionnaire to record training duration and intensity. Training duration (p = 0.022) and intensity (p = 0.024) were associated with higher cIMT. Further, training duration was associated with lower central arterial stiffness (cSBP: p = 0.001; aPWV: p = 0.033) and improved AC (p < 0.001). Higher training intensity was related to improved AC (p < 0.001) and larger carotid diameter (p = 0.040). Boys presented thicker cIMT (p = 0.010), improved AC (p = 0.006), and lower central arterial stiffness (cSBP: p < 0.001; aPWV: p = 0.016) associated with higher training duration. Girls presented improved AC (p = 0.023) and lower Ep (p = 0.038) but higher ß (p = 0.036) associated with higher training duration. Only boys demonstrated thicker cIMT (p = 0.016) and improved AC (p = 0.002) associated with higher training intensity. A quintile analyses of the training duration revealed thicker cIMT of children and adolescents in Q1 and Q5 than that in Q4 and Q5. Besides, Q1 showed lower cSBP compared to Q4 and Q5. Regarding training intensity, Q5 had thicker cIMT than Q2 and Q3. Although a higher training load is associated with thicker cIMT, the common carotid artery is also more elastic. This suggests that a higher training load leads to a functional adaptation of the carotid artery in youth.

Improved Carotid Elasticity but Altered Central Hemodynamics and Carotid Structure in Young Athletes.

Young athletes most often exceed the physical activity recommendations of the World Health Organization. Therefore, they are of special interest for investigating cardiovascular adaptions to exercise. This study aimed to examine the arterial structure and function of young athletes 12-17 years old and compare these parameters to reference values of healthy cohorts. Carotid intima-media thickness (cIMT), carotid diameter, cIMT÷carotid diameter-ratio (cIDR), arterial compliance (AC), elastic modulus (Ep), ß stiffness index (ß), and carotid pulse wave velocity (PWVß) were determined using ultrasound in 331 young athletes (77 girls; mean age, 14.6 ± 1.30 years). Central systolic blood pressure (cSBP) and aortic PWV (aPWV) were measured using the oscillometric device Mobil-O-Graph. Standard deviation scores (SDS) of all parameters were calculated according to German reference values. The 75th and 90th percentiles were defined as the threshold for elevated cIMT and arterial stiffness, respectively. Activity behavior was assessed with the MoMo physical activity questionnaire, and maximum power output with a standard cardiopulmonary exercise test. One-sample t-tests were performed to investigate the significant deviations in SDS values compared to the value "0". All subjects participated in competitive sports for at least 6 h per week (565.6 ± 206.0 min/week). Of the 331 young athletes, 135 (40.2%) had cIMT >75th percentile, 71 (21.5%) had cSBP >90th percentile, and 94 (28.4%) had aPWV>90th percentile. We observed higher cIMT SDS (p < 0.001), cIDR SDS (p = 0.009), and AC SDS (p < 0.001) but lower ß SDS (p < 0.001), Ep SDS (p < 0.001), and PWVß SDS (p < 0.001) compared to the reference cohort. The cSBP SDS (p < 0.001) and aPWV SDS (p < 0.001) were elevated. In conclusion, cIMT and cIDR were higher in young athletes than in a reference cohort. Furthermore, young athletes presented better carotid elasticity and lower arterial stiffness of the carotid artery. However, central arterial stiffness was higher compared to the reference cohort. The thickening of the carotid intima-media complex in combination with a reduction in arterial stiffness indicates a physiological adaptation to exercise in youth.

Vascular diameter and intima-media thickness to diameter ratio values of the carotid artery in 642 healthy children.

In children, arterial alterations occur with increased intima-media thickness as well as vascular diameter enlargement. Both conditions correlate with higher cardiovascular risk in adults, and both the array and proportion of these alterations are important hemodynamic parameters. In terms of functional adaptation processes, they influence several arterial wall properties as for example the shear and tensile stress of the vessel. There are no reference values for the vascular diameter and intima-media thickness/diameter ratio of the carotid artery in children. Therefore, this study aimed to assess vascular diameter, intima-media thickness/diameter ratio and related tensile stress values in children and to further investigate the influence of sex, age, body mass index, and blood pressure. The parameters were measured with high-resolution semi-automated ultrasound. Sex- and age-dependent values were calculated with the LMS method for a cross-sectional sample of 642 healthy, non-obese children aged 8-17 years. The mean vascular diameter was 5.45 ± 0.46 mm; the median intima-media thickness/diameter ratio was 0.085 (0.079-0.092); the median tensile stress was 105.4 (95.2-116.4) kPa. The vascular diameter and the tensile stress were higher, and the intima-media thickness/diameter ratio was lower in boys than in girls. In comparison to the normal weight study population the excludedobese children had a significantly higher diameter, a lower intima-media thickness/diameter ratio, and a higher tensile stress. In multiple regression analyses of diameter, intima-media thickness/diameter ratio, and tensile stress, all parameters were influenced by sex and body mass index. Furthermore, systolic and diastolic blood pressure significantly influenced the vascular diameter, and systolic blood pressure significantly influenced the intima-media thickness/diameter ratio. Conclusion: This study is the first to report values for the diameter, the intima-media thickness/diameter ratio of the carotid artery, and the related tensile stress allowing a more differentiated view of cardiovascular adaptations as it combines structural and functional vascular parameters. What is known: • Intima-media thickness and vascular diameter are related to a higher cardiovascular risk in adults • The intima-media thickness/diameter ratio gives information about hemodynamic and functional vessel adaptation What is new: • Values for vascular diameter, intima-media thickness/diameter ratio, and tensile stress of the carotid artery in children are presented in this study • Intima-media thickness as a surrogate marker for arterial health in children should be complemented by intima-media thickness/diameter ratio measurement.

Can School-Based Physical Activity Projects Such as Skipping Hearts Have a Long-Term Impact on Health and Health Behavior?

Low physical activity, limited motor skills, and an increased number of overweight or obese children are major public health problems. Numerous school-based programs try to improve physical activity and health behavior in children but investigations on sustainable effects of these programs are rare. Therefore, we examined the long-term effects of the Skipping Hearts health promotion project. 486 children (57.7% female, 9.0 ± 0.6 years at baseline) participated in this non-randomized controlled longitudinal trial within a follow-up period of 3.5 years. Of these, 286 subjects received a one-time 90-min workshop in rope skipping (Basic-Workshop) and 140 additionally received 10 lessons in rope skipping (Champion-Program), 78 students served as controls. Anthropometrics, blood pressure, motor skills, screen-based media use, self-assessment of physical fitness, and physical activity were collected at both measurement points; endurance capacity and health-related quality of life only at follow-up. Standard deviation scores of body-mass-index (η2 = 0.005) and systolic blood pressure (η2 = 0.006) decreased, while diastolic blood pressure (η2 = 0.004), motor performance (η2 < 0.001), physical fitness, subjective physical activity (η2 = 0.008), and screen-based media use (η2 = 0.001) increased without significant difference in development between groups (all p > 0.05). At follow-up, groups did not differ in endurance capacity (η2 = 0.010) and health-related quality of life (η2 < 0.001). Skipping Hearts does not affect the long-term improvement of health status, motor performance, or health behavior. To improve the effects, the project should be implemented as a daily routine in schools to force the transfer of health behavior-related knowledge. Nevertheless, the project offers a physical activity that can be performed in children's everyday life without high costs.

Sleep disorders in migrants and refugees: a systematic review with implications for personalized medical approach.

BACKGROUND: Sleep disorders are very common in migrants and refugees, often as a comorbid disorder to different somatic or psychiatric diagnoses and psychological disturbances such as metabolic syndrome, post-traumatic stress disorder, depression, and anxiety disorders. OBJECTIVES: To review published prevalence rates as well as possible predictors for sleep disturbances in these vulnerable groups, including pre-migration stress, acculturation, and trauma before, during, and after migration, integration, and lifestyle in the host country with implications for predictive, preventive, and personalized medical approach (3PM). DATA SOURCES: Electronic databases PubMed, PsycInfo, and Web of Knowledge were searched using (combined) search terms "migrant," "asylum seeker," "refugee," "sleep disturbances," "sleep disorder," "insomnia," and "sleep wake disorder." STUDY ELIGIBILITY CRITERIA: Peer-reviewed studies from 2000 to 2018 reporting data on prevalence and/or predictors of any measure of sleep disturbance were included. PARTICIPANTS: Studies on international migrants and refugees, as well as internally displaced populations, were included. METHODS: We conducted a systematic review on the topic of sleep disorders in migrant and refugee populations. Only published articles and reviews in peer-reviewed journals were included. RESULTS: We analyzed five studies on sleep disorders in migrants, five studies on adult refugees, and three on refugee children and adolescents. Prevalence of sleep disorders in migrants and refugees ranges between 39 and 99%. In migrant workers, stress related to integration and adaptation to the host society is connected to higher risks of snoring, metabolic diseases, and insomnia. Sleep disturbances in refugees are predicted by past war experience. Sleep difficulties in adult and child refugees are strongly correlated to trauma. Torture of parents and grandparents can predict sleep disorders in refugee children, while being accompanied by parents to the host country has a protective effect on children's sleep. CONCLUSIONS AND IMPLICATIONS: Considering the differences in risk factors, vulnerability, and traumatic life events for different migrant populations, origins of sleep difficulties vary, depending on the migrant populations. Effects on sleep disturbances and sleep quality may be a result of integration in the host country, including changes of lifestyle, such as diet and working hours with implication for OSAS (obstructive sleep apnea) and insomnia. Compared with migrant populations, sleep disturbances in refugee populations are more correlated with mental health symptoms and disorders, especially PTSD (post-traumatic stress disorder), than with psychosocial problems. In juvenile refugee populations, psychological problems and disturbed sleep are associated with traumatic experiences during their journey to the host country. Findings highlight the need for expert recommendations for development of 3P approach stratified in the following: (1) prediction, including structured exploration of predisposing and precipitating factors that may trigger acute insomnia, screening of the according sleep disorders by validated translated questionnaires and sleep diaries, and a face-to-face or virtual setting and screening of OSAS; (2) target prevention by sleep health education for female and male refugees and migrant workers, including shift workers; and (3) personalized medical approach, including translated cognitive behavioral treatment for insomnia (CBT-I) and imagery rehearsal therapy for refugees and telehealth programs for improved CPAP adherence in migrants, with the goal to enable better sleep health quality and improved health economy.

Vascular Structure and Function in Children and Adolescents: What Impact Do Physical Activity, Health-Related Physical Fitness, and Exercise Have?

A physically active lifestyle can prevent cardiovascular disease. Exercise intervention studies in children and adolescents that aim to increase physical activity have resulted in reduced vascular wall thickening and improve cardiovascular function. Here we review the literature that explores the correlations between physical activity, health-related physical fitness, and exercise interventions with various measures of vascular structure and function in children and adolescents. While several of these studies identified improvements in vascular structure in response to physical activity, these associations were limited to studies that relied on questionnaires. Of concern, these findings were not replicated in studies featuring quantitative assessment of physical activity with accelerometers. Half of the studies reviewed reported improved vascular function with increased physical activity, with the type of vascular measurement and the way physical activity was assessed having an influence on the reported relationships. Similary, most of the studies identified in the literature report a beneficial association of health-related physical fitness with vascular structure and function. Overall, it was difficult to compare the results of these studies to one another as different methodologies were used to measure both, health-related physical fitness and vascular function. Likewise, exercise interventions may reduce both arterial wall thickness and increased vascular stiffness in pediatric populations at risk, but the impact clearly depends on the duration of the intervention and varies depending on the target groups. We identified only one study that examined vascular structure and function in young athletes, a group of particular interest with respect to understanding of cardiovascular adaptation to exercise. In conclusion, future studies will be needed that address the use of wall:diameter or wall:lumen-ratio as part of the evaluation of arterial wall thickness. Furthermore, it will be critical to introduce specific and quantitative measurements of physical activity, as intensity and duration of participation likely influence the effectiveness of exercise interventions.

Influence of Vigorous Physical Activity on Structure and Function of the Cardiovascular System in Young Athletes-The MuCAYA-Study.

Objective: Moderate physical activity (PA) is associated with a reduced risk to develop cardiovascular disease. However, junior athletes exercise between 10 and 20 h a week with intensities exceeding moderate levels by far. In this regard, the cardiovascular system has to increase its work five to six times compared to moderate intensities. This may result in potentially pathological adaptations of the cardiovascular system. The underlying process of vascular adaptations to exercise is yet not fully understood and hardly investigated in junior athletes. An increased blood pressure and pulse wave velocity, ventricular hypertrophy, arrhythmia, and even sudden cardiac death (SCD) has been reported in adult athletes. Studies, examining the cardiovascular system in children, its association to intensity and type of exercise, are rare. Therefore, we present the study protocol of a prospective cross-sectional study that investigates the influence of PA on the cardiovascular system in young athletes. Methods and Design: Children and adolescents, 7-18 years, presenting for their annual pre-participation screening at the Institute of Preventive Pediatrics, Faculty of Sports and Health Sciences, Technical University of Munich (TUM), are examined in this prospective cross-sectional study. Vascular parameters measured by ultrasound are carotid intima-media thickness (cIMT), vascular stiffness (AC, Ep, ß, PWV ß), and the vascular diameter (D) to calculate the IMT:Diameter-Ratio (IDR). Cardiac function is evaluated by a 12-lead ECG, and echocardiographic parameters (end-diastolic left ventricular diameter, left ventricular diastolic posterior wall thickness, diastolic septal thickness, left ventricular mass and relative wall thickness, ejection fraction, and shortening fraction). A cardiopulmonary exercise test is performed on a bicycle ergometer, muscular strength is assessed with the handgrip test, and physical activity with the MoMo questionnaire. Discussion: It is essential to follow young athletes over the course of their career in order to detect pathophysiological changes in the myocardium as soon as possible. If these changes are preceded or followed by changes in vascular structure and function is not known yet. Therefore, we present the study protocol of the Munich Cardiovascular adaptations in young athletes study (MuCAYA-Study) which investigates the association between vascular and cardiac adaptation to intensive exercise in junior athletes.

Remyelination promoting therapies in multiple sclerosis animal models: a systematic review and meta-analysis.

An unmet but urgent medical need is the development of myelin repair promoting therapies for Multiple Sclerosis (MS). Many such therapies have been pre-clinically tested using different models of toxic demyelination such as cuprizone, ethidium bromide, or lysolecithin and some of the therapies already entered clinical trials. However, keeping track on all these possible new therapies and their efficacy has become difficult with the increasing number of studies. In this study, we aimed at summarizing the current evidence on such therapies through a systematic review and at providing an estimate of the effects of tested interventions by a meta-analysis. We show that 88 different therapies have been pre-clinically tested for remyelination. 25 of them (28%) entered clinical trials. Our meta-analysis also identifies 16 promising therapies which did not enter a clinical trial for MS so far, among them Pigment epithelium-derived factor, Plateled derived growth factor, and Tocopherol derivate TFA-12.We also show that failure in bench to bedside translation from certain therapies may in part be attributable to poor study quality. By addressing these problems, clinical translation might be smoother and possibly animal numbers could be reduced.

Honokiol: a non-adipogenic PPARγ agonist from nature.

BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPARγ) agonists are clinically used to counteract hyperglycemia. However, so far experienced unwanted side effects, such as weight gain, promote the search for new PPARγ activators. METHODS: We used a combination of in silico, in vitro, cell-based and in vivo models to identify and validate natural products as promising leads for partial novel PPARγ agonists. RESULTS: The natural product honokiol from the traditional Chinese herbal drug Magnolia bark was in silico predicted to bind into the PPARγ ligand binding pocket as dimer. Honokiol indeed directly bound to purified PPARγ ligand-binding domain (LBD) and acted as partial agonist in a PPARγ-mediated luciferase reporter assay. Honokiol was then directly compared to the clinically used full agonist pioglitazone with regard to stimulation of glucose uptake in adipocytes as well as adipogenic differentiation in 3T3-L1 pre-adipocytes and mouse embryonic fibroblasts. While honokiol stimulated basal glucose uptake to a similar extent as pioglitazone, it did not induce adipogenesis in contrast to pioglitazone. In diabetic KKAy mice oral application of honokiol prevented hyperglycemia and suppressed weight gain. CONCLUSION: We identified honokiol as a partial non-adipogenic PPARγ agonist in vitro which prevented hyperglycemia and weight gain in vivo. GENERAL SIGNIFICANCE: This observed activity profile suggests honokiol as promising new pharmaceutical lead or dietary supplement to combat metabolic disease, and provides a molecular explanation for the use of Magnolia in traditional medicine.

2-(2,4-dihydroxyphenyl)-5-(E)-propenylbenzofuran promotes endothelial nitric oxide synthase activity in human endothelial cells.

Endothelial nitric oxide synthase (eNOS) mediates important vaso-protective and immunomodulatory effects. Aim of this study was to examine whether lignan derivatives isolated from the roots of the anti-inflammatory medicinal plant Krameria lappacea influence eNOS activity and endothelial nitric oxide (NO) release. The study was performed using cultured human umbilical vein endothelial cells (HUVECs) and HUVEC-derived EA.hy926 cells. Among the eleven isolated compounds only 2-(2,4-dihydroxyphenyl)-5-(E)-propenylbenzofuran (DPPB) was able to increase eNOS enzyme activity. DPPB (1-10 µM) treatment for 24 h induced a significant and dose-dependent increase in eNOS activity as determined by the [(14)C]L-arginine/[(14)C]L-citrulline conversion assay. Immunoblotting studies further revealed a time-dependent DPPB-induced increase in eNOS-Ser(1177) and decrease in eNOS-Thr(495) phosphorylation, as well as increased AMPK phosphorylation at Thr(172), whereas Akt phosphorylation at Ser(473) was not affected. Si-RNA-mediated knockdown of AMPK and inhibition of CaMKKß by STO 609, as well as intracellular Ca(2+) chelation by Bapta AM abolished the stimulating effect of DPPB on eNOS-Ser(1177) and AMPK-Thr(172) phosphorylation. Furthermore, we could show that DPPB increases intracellular Ca(2+) concentrations assessed with the fluorescent dye Fluo-3-AM. DPPB enhances eNOS activity and endothelial NO release by raising intracellular Ca(2+) levels and increases signaling through a CaMKKß-AMPK dependent pathway.

Ratanhiaphenol III from Ratanhiae radix is a PTP1B inhibitor.

The inhibition of protein tyrosine phosphatase 1B (PTP1B) is considered a valid strategy to combat insulin resistance and type II diabetes. We show here that a dichloromethane extract of Ratanhiae radix ( RR_EX) dose-dependently inhibits human recombinant PTP1B in vitro and enhances insulin-stimulated glucose uptake in murine myocytes. By determination of the PTP1B inhibiting potential of 11 recently isolated lignan derivatives from RR_EX, the observed activity of the extract could be partly assigned to ratanhiaphenol III. This compound inhibited PTP1B in vitro with an IC (50) of 20.2 µM and dose-dependently increased insulin receptor phosphorylation as well as insulin-stimulated glucose uptake in cultured myotubes. This is the first report to reveal an antidiabetic potential for a constituent of rhatany root, traditionally used against inflammatory disorders, by showing its capability of inhibiting PTP1B.

Lignan derivatives from Krameria lappacea roots inhibit acute inflammation in vivo and pro-inflammatory mediators in vitro.

The roots of Krameria lappacea are used traditionally against oropharyngeal inflammation. So far, the astringent and antimicrobial properties of its proanthocyanidin constituents are considered to account for the anti-inflammatory effect. The aim of the present study was to characterize pharmacologically a lipophilic extract of K. lappacea roots and several isolated lignan derivatives (1-11) in terms of their putative anti-inflammatory activity. The dichloromethane extract (ID50 77 µg/cm²) as well compounds 1-11 (ID50 0.31-0.60 µmol/cm²) exhibited topical antiedematous properties comparable to those of indomethacin (ID50 0.29 µmol/cm²) in a mouse ear in vivo model. Two of the most potent compounds, 2-(2-hydroxy-4-methoxyphenyl)-5-(3-hydroxypropyl)benzofuran (5) and (+)-conocarpan (7), were studied regarding their time-dependent edema development and leukocyte infiltration up to 48 h after croton oil-induced dermatitis induction, and they showed activity profiles similar to that of hydrocortisone. In vitro studies of the isolated lignan derivatives demonstrated the inhibition of NF-κB, cyclooxygenase-1 and -2, 5-lipoxygenase, and microsomal prostaglandin E2 synthase-1 as well as antioxidant properties, as mechanisms possibly contributing to the observed in vivo effects. The present findings not only support the ethnopharmacological use of K. lappacea roots but also reveal that the isolated lignan derivatives contribute strongly to the anti-inflammatory activity of this herbal drug.

Quantitative analysis of anti-inflammatory lignan derivatives in Ratanhiae radix and its tincture by HPLC-PDA and HPLC-MS.

Root preparations of Krameria lappacea (Dombey) Burdet et Simpson are traditionally used against oropharyngeal inflammation. Besides antimicrobial and astringent procyanidines, lignan derivatives, including ratanhiaphenol I, II, III and (+)-conocarpan, contribute to the activity of Ratanhiae radix, exerting a significant topical anti-inflammatory activity in vivo, and in vitro by inhibiting NF-κB and the formation of inflammatory prostaglandins and leukotrienes. Besides gravimetrical analysis of the ratanhiaphenols I, II and III, the content of these compounds in the herbal drug has never been determined. The developed HPLC method enables the quantification of twelve active lignan derivatives in the roots, and is also suitable for the determination of the constituents in Tinctura Ratanhiae. Separation was achieved on a phenyl-hexyl column material using a solvent gradient consisting of 0.02% aqueous TFA and a mixture of acetonitrile/methanol (75:25, v/v). Sensitivity, accuracy (recovery rates were between 95% and 105.6%), repeatability (RSD ≤ 4.6%), and precision (intra-day precision ≤ 4.8%; inter-day precision ≤ 3.4%) of the method were determined. HPLC-MS experiments in positive and negative electrospray ionization mode confirmed identity and peak purity of analytes. The analysis of several root and tincture samples revealed that (+)-conocarpan and ratanhiaphenol II dominated with contents of 0.49-0.71% and 0.51-0.53% in the roots and 0.66-0.68 mg/ml and 0.70-0.71 mg/ml in the commercial tinctures, respectively.

Computer-aided discovery, validation, and mechanistic characterization of novel neolignan activators of peroxisome proliferator-activated receptor gamma.

Peroxisome proliferator-activated receptor gamma (PPAR gamma) agonists are used for the treatment of type 2 diabetes and metabolic syndrome. However, the currently used PPAR gamma agonists display serious side effects, which has led to a great interest in the discovery of novel ligands with favorable properties. The aim of our study was to identify new PPARgamma agonists by a PPAR gamma pharmacophore-based virtual screening of 3D natural product libraries. This in silico approach led to the identification of several neolignans predicted to bind the receptor ligand binding domain (LBD). To confirm this prediction, the neolignans dieugenol, tetrahydrodieugenol, and magnolol were isolated from the respective natural source or synthesized and subsequently tested for PPAR gamma receptor binding. The neolignans bound to the PPAR gamma LBD with EC(50) values in the nanomolar range, exhibiting a binding pattern highly similar to the clinically used agonist pioglitazone. In intact cells, dieugenol and tetrahydrodieugenol selectively activated human PPAR gamma-mediated, but not human PPAR alpha- or -beta/delta-mediated luciferase reporter expression, with a pattern suggesting partial PPAR gamma agonism. The coactivator recruitment study also demonstrated partial agonism of the tested neolignans. Dieugenol, tetrahydrodieugenol, and magnolol but not the structurally related eugenol induced 3T3-L1 preadipocyte differentiation, confirming effectiveness in a cell model with endogenous PPAR gamma expression. In conclusion, we identified neolignans as novel ligands for PPAR gamma, which exhibited interesting activation profiles, recommending them as potential pharmaceutical leads or dietary supplements.

Accepting being poz: the incorporation of the HIV identity into the self.

In a previous study, the first author of the current study examined how people who were diagnosed with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) when it was considered a terminal illness incorporated their HIV/AIDS identity into the self over time. In the qualitative study reported here, we examined HIV identity incorporation in participants diagnosed with HIV after 1996, when it was considered a chronic illness in the United States. We uncovered a three-step process: diagnosis, a postdiagnosis turning point, and integration. We compared and contrasted the results to those from the previous study and studies of other chronic illnesses. The findings advance our understanding of HIV/AIDS, chronic illness, and identity. Practical implications for HIV/AIDS educators are also discussed.

Inhibition of 11beta-hydroxysteroid dehydrogenase type 1 by plant extracts used as traditional antidiabetic medicines.

Elevated glucocorticoids are a key risk factor for metabolic diseases, and the glucocorticoid-activating enzyme 11beta-hydroxysteroid dehydrogenase 1 (11beta-HSD1) represents a promising therapeutic target. We measured the potential of six traditional antidiabetic medicinal plants extracts to inhibit 11beta-HSD1 activity and glucocorticoid receptor (GR) activation in transfected HEK-293 cells. Leave extracts of Eriobotrya japonica preferentially inhibited 11beta-HSD1 over 11beta-HSD2. Extracts of roasted but not native coffee beans preferentially inhibited 11beta-HSD1 over 11beta-HSD2, emphasizing the importance of sample preparation. Thus, natural compounds inhibiting 11beta-HSD1 may contribute to the antidiabetic effect of the investigated plant extracts.