Androgens and corticosteroids increase in response to mirror images and interacting conspecifics in males of the Siamese fighting fish Betta splendens.

The role of hormones as modulators of aggressive behavior in fish remains poorly understood. Androgens and corticosteroids, in particular, have been associated with aggressive behavior in fish but it is still not clear if animals adjust the secretion of these hormones to regulate behavior during ongoing fights, in response to fight outcomes in order to adjust aggressive behavior in subsequent fights, or both. With its stereotyped displays and high aggression levels, the Siamese fighting fish Betta splendens is an excellent model to investigate this question. Here, we compared the behavioral and endocrine response of male B. splendens to fights where there is no winner or loser by presenting them with a size-matched live interacting conspecific behind a transparent partition or with a mirror image. The aggressive response started with threat displays that were overall similar in frequency and duration towards both types of stimuli. Fights transitioned to overt attacks and interacting with a live conspecific elicited a higher frequency of attempted bites and head hits, as compared with the mirror image. There was a pronounced increase in plasma androgens (11-ketotestosterone and testosterone) and corticosteroids (cortisol) levels in response to the aggression challenge, independent of stimulus type. Post-fight intra-group levels of these hormones did not correlate with measures of physical activity or aggressive behavior. A linear discriminant analysis including all behavioral and endocrine data was a poor classifier of fish from the conspecific and mirror trials, showing that overall the behavioral and endocrine response to mirror images and conspecifics was similar. The results show that fight resolution is not necessary to induce an evident increase in peripheral levels of androgens and corticosteroids in B. splendens. However, the function of these hormones during present and future aggressive contests remains to be clarified.

Marginal accuracy of nickel chromium copings fabricated by conventional and accelerated casting procedures, produced with ringless and metal ring investment procedures: A comparative in vitro study.

BACKGROUND: Conventional investing and casting techniques are time-consuming and usually requires 2-4 h for completion. Accelerated nonstandard, casting techniques have been reported to achieve similar quality results in significantly less time, namely, in 30-40 min. During casting, it is essential to achieve compensation for the shrinkage of solidifying alloy by investment expansion. The metal casting ring restricts the thermal expansion of investment because the thermal expansion of the ring is lesser than that of the investment. The use of casting ring was challenged with the introduction of the ringless technique. MATERIALS AND METHODS: A total of 40 test samples of nickel chromium (Ni-Cr) cast copings were obtained from the patterns fabricated using inlay casting wax. The 20 wax patterns were invested using metal ring and 20 wax patterns were invested using the ringless investment system. Of both the groups, 10 samples underwent conventional casting, and the other 10 underwent accelerated casting. The patterns were casted using the induction casting technique. All the test samples of cast copings were evaluated for vertical marginal gaps at four points on the die employing a stereo optical microscope. RESULTS: The vertical marginal discrepancy data obtained were tabulated. Mean and standard deviations were obtained. Vertical discrepancies were analyzed using analysis of variance and Tukey honestly significantly different. The data obtained were found to be very highly significant (P < 0.001). Mean vertical gap was the maximum for Group II (53.64 µm) followed by Group IV (47.62 µm), Group I (44.83 µm) and Group III (35.35 µm). CONCLUSION: The Ni-Cr cast copings fabricated with the conventional casting using ringless investment system showed significantly better marginal fit than that of cast copings fabricated from conventional and accelerated casting with metal ring investment and accelerated casting using ringless investment since those copings had shown the least vertical marginal discrepancies among the four methods evaluated in this study.

Assessing feasibility of conducting medication review with follow-up among older adults at community pharmacy: a pilot randomised controlled trial.

BACKGROUND: Medication review with follow-up is essential for optimising medication utilisation among the older adult population in primary healthcare. AIM: This study aimed to evaluate the feasibility of implementing medication reviews with follow-up for older adults in community pharmacies and examined potential outcomes on medication use. METHOD: A pilot randomised controlled trial was conducted with 4 cluster-randomised community pharmacies to assess the feasibility of the intervention. Two community pharmacies served as intervention and control groups. Both groups recruited older adults over 60 who were followed over 6 months. The translated Medication use Questionnaire (MedUseQ) was administered at baseline and 6 months for both groups. The outcomes were to assess the feasibility of conducting medication review with follow-up and the probable medication use outcomes from the intervention. RESULTS: The intervention and control groups comprised 14 and 13 older adults. A total of 35 recommendations were made by pharmacists in the intervention group and 8 in the control group. MedUseQ was easily administered, providing some evidence the feasibility of the intervention. However, there were feasibility challenges such as a lack of pharmacists, collaborative practice, difficulties with the tool language, time constraints, and limited funds. Questionnaire results provided a signal of improvement in medication administration, adherence, and polypharmacy among intervention participants. The incidence of drug related problems was significantly higher in the control group (median = 1) after 6 months, U = 15, z = - 2.98, p = 0.01. CONCLUSION: Medication review with follow-up is potentialy practical in community pharmacies, but there are feasibility issues. While these challenges can be addressed, it is essential to study larger sample sizes to establish more robust evidence regarding outcomes. CLINICAL TRIAL REGISTRY: ClinicalTrials.Gov NCT05297461.

Behavioral and endocrine responses to noninteractive live and video conspecifics in males of the Siamese fighting fish.

The physiological mechanisms underlying variation in aggression in fish remain poorly understood. One possibly confounding variable is the lack of standardization in the type of stimuli used to elicit aggression. The presentation of controlled stimuli in videos, a.k.a. video playback, can provide better control of the fight components. However, this technique has produced conflicting results in animal behavior studies and needs to be carefully validated. For this, a similar response to the video and an equivalent live stimulus needs to be demonstrated. Further, different physiological responses may be triggered by live and video stimuli, and it is important to demonstrate that video images elicit appropriate physiological reactions. Here, the behavioral and endocrine responses of male Siamese fighting fish Betta splendens to a matched-for-size conspecific fighting behind a one-way mirror, presented live or through video playback, were compared. The video playback and live stimulus elicited a strong and similar aggressive response by the focal fish, with a fight structure that started with stereotypical threat displays and progressed to overt attacks. Postfight plasma levels of the androgen 11-ketotestosterone were elevated as compared to controls, regardless of the type of stimuli. Cortisol also increased in response to the video images, as previously described for live fights in this species. These results show that the interactive component of a fight and its resolution are not needed to trigger an endocrine response to aggression in this species. The study also demonstrates for the first time in a fish a robust endocrine response to video stimuli and supports the use of this technique for researching aggressive behavior in B. splendens.

Cannabinoids: Emerging sleep modulator.

Sleep is an essential biological phase of our daily life cycle and is necessary for maintaining homeostasis, alertness, metabolism, cognition, and other key functions across the animal kingdom. Dysfunctional sleep leads to deleterious effects on health, mood, and cognition, including memory deficits and an increased risk of diabetes, stroke, and neurological disorders. Sleep is regulated by several brain neuronal circuits, neuromodulators, and neurotransmitters, where cannabinoids have been increasingly found to play a part in its modulation. Cannabinoids, a group of lipid metabolites, are regulatory molecules that bind mainly to cannabinoid receptors (CB1 and CB2). Much evidence supports the role of cannabinoid receptors in the modulation of sleep, where their alteration exhibits sleep-promoting effects, including an increase in non-rapid-eye movement sleep and a reduction in sleep latency. However, the pharmacological alteration of CB1 receptors is associated with adverse psychotropic effects, which are not exhibited in CB2 receptor alteration. Hence, selective alteration of CB2 receptors is also of clinical importance, where it could potentially be used in treating sleep disorders. Thus, it is crucial to understand the neurobiological basis of cannabinoids in sleep physiology. In this review article, the alteration of the endocannabinoid system by various cannabinoids and their respective effects on the sleep-wake cycle are discussed based on recent findings. The mechanisms of the cannabinoid receptors on sleep and wakefulness are also explored for their clinical implications and potential therapeutic use on sleep disorders.

Medication Use Questionnaire for Older Adults in Malaysia: Translation, Cultural Adaptation, and Reliability.

OBJECTIVES: This study aimed to translate the Medication Use Questionnaire into a Malay version, adapt it to Malaysia's culture, and verify its reliability among Malaysia's older adults. METHODS: Methodological approaches were used to translate, validate, and modify the questionnaire. The subjects were older adults aged ≥ 60 years in primary care settings in Penang, Malaysia. Two forward translations (English to Malay) were developed, reviewed, and back translated to English. The reconciliation phase was conducted to compare the translated and original questionnaires. Five older adults were then interviewed for the cognitive debriefing of the reconciled questionnaire to assess the linguistic and cultural equivalence. Two experts assessed content validity, and the translated questionnaire was proofread and finalized. After that, pilot test was done to examine the internal consistency among 20 older adults. RESULTS: Translation of the questionnaire was done with no major disagreements. The main issues identified in cognitive debriefing and content validity were terms, number of questions, and phrases used in the questionnaire. Most participants reported that the questionnaires were not difficult to complete during the cognitive debriefing phase. The issues were then judged and revised accordingly. Further pilot testing on 20 older adults demonstrated good internal consistency reliability, Cronbach α (0.902). CONCLUSIONS: This study findings suggest promising data supporting the use of translated version of the Medication Use Questionnaire that can be used to identify medication use problems among older adults in Malaysia.

Helping older adults with their medication use problems: A qualitative study on perspectives and challenges of primary health care providers.

BACKGROUND: Primary care providers help older adults with medication use problems in Malaysia and globally. They help older adults with medication management, appropriate use, and administration; however, their perspectives and challenges regarding medication use problems in older adults have not been adequately explored. METHODS: The study used a qualitative methodology comprising 30 in-depth interviews among general practitioners and pharmacists in Penang, Malaysia, in public and private primary care settings. Participants were recruited based on purposive sampling. Interviews were transcribed verbatim, and data were coded based on the principles of thematic analysis in NVivo. OBJECTIVE: This study aims to understand primary care providers' perspectives and challenges regarding medication use problems experienced by older adults. RESULTS: Six themes emerged from the study. Theme one highlighted the pharmaceutical care needs of older adults with sensory impairments and accessibility issues. The second and third themes explored medicines management support and potentially inappropriate medication use. Theme four supported collaborative practice, prescribing, and deprescribing among primary health care providers. Theme five discussed health service delivery aligned to older adults' health care needs. The final theme emphasised social and welfare support. CONCLUSION: This study identified various challenges professional primary care providers face in providing aligned healthcare services for older adults and proposed recommendations for further strengthening healthcare quality. Inputs from the primary healthcare system frontier are essential to reduce the challenges and uplift the quality of ageing populations' healthcare in Malaysia.

Medication use problems and factors affecting older adults in primary healthcare.

BACKGROUND: As the largest demographic group utilizing primary healthcare facilities, older adults often face the challenge of managing multiple chronic illnesses, leading to numerous medications. OBJECTIVE: The present study aims to assess medication use problems among older adults and explore the factors affecting them in primary healthcare settings. METHODS: A mixed-method study was conducted to establish a baseline understanding of the perspectives and challenges faced by older adults, with regards to medication use. Translated Medication Use Questionnaire (MedUseQ), a patient-centered tool, was distributed to older adults above 60 in primary healthcare settings to assess their frequency of problems related to medication use. Fifteen semi-structured interviews were conducted to explore this topic in depth. Descriptive and inferential analyses were conducted with quantitative data. Interviews were transcribed verbatim, and thematic analysis was conducted. Quantitative and qualitative findings data were triangulated. RESULTS: The study involved 393 participants. The most common problems with regards to medication use were polypharmacy (55.4%), administration difficulties (48.4%), limited awareness about adverse drug events (47.3%), issues with adherence (46.5%), and accessibility to primary healthcare (42.7%). Approximately 55% were satisfied with the older adult-centered medication use services by pharmacist and doctors. The qualitative findings showed that major factors affecting medication use were forgetfulness, language barriers, lack of awareness, transportation problems, long waiting times, and multiple visits to healthcare facilities. CONCLUSION: The findings of this study reveal that a significant proportion, around 50%, of the older adult population face challenges with medication use in Penang. These difficulties mainly stem from medication administration, adherence, accessibility, polypharmacy, and inadequate medication knowledge. The qualitative analysis further highlighted several factors that contribute to such medication-related problems. Given the rapidly aging Association of Southeast Asian Nations (ASEAN) population, it is essential to devise effective solutions and strategies to tackle medication use-related issues among older adults.

Medication use problems among older adults at a primary care: A narrative of literature review.

Appropriate medication use is one of the most significant challenges among the older population. Although medication use problems are well documented at the secondary and tertiary health care level, the evidence at the primary care level of OECD region is limited. A narrative review of existing literature was conducted through a nonsystematic search for original articles through electronic search databases, Ovid Medline, Google Scholar from 2001 to 2021, and a combination of citation references. Medication use problems are prevalent in older adults at the primary care level. The main issues of medication use identified were as follows; nonadherence, adverse drug events, accessibility, polypharmacy, inappropriate medications, belief about medications, lack of knowledge and awareness, and lack of deprescribing. In addition, the current review has identified the possibilities of the problems: many medications, forgetfulness, lack of deprescribing, lack of communication, poor understanding, and limited awareness of inappropriate medications. This review found that various medication use problems subclusters were identified to impact the health care need among older adults. Therefore, effective interventions targeting these issues need to be developed to reduce medication use problems among older adults at a primary care level.

Nobiletin, a polymethoxylated flavonoid from citrus, shows anti-angiogenic activity in a zebrafish in vivo model and HUVEC in vitro model.

Traditional Chinese medicinal herbs are a rich source of compounds with reported anti-inflammatory and anti-carcinogenic effects. Growing evidence shows the codependence of chronic inflammation and angiogenesis, and the potential benefits of targeting angiogenesis in the treatment of chronic inflammation and targeting inflammation in the treatment of diseases with impaired angiogenesis. We hypothesized that the anti-inflammatory activity of the natural compounds may owe at least some of its efficacy to their anti-angiogenic activity and hence we investigated the anti-angiogenic activity of these compounds in vivo in zebrafish embryos and in vitro in human umbilical vein endothelial cells (HUVECs). Nobiletin, a polymethoxylated flavonoid from citrus fruits, showed anti-angiogenic activity in both assays. Nobiletin inhibited the formation of intersegmental vessels (ISVs) in live transgenic zebrafish embryos expressing green fluorescent protein (GFP) in the vasculature. Cell cycle analysis of dissociated zebrafish embryo cells showed that nobiletin induced G0/G1 phase accumulation in a dose-dependent manner in GFP-positive endothelial cells. Nobiletin also dose-dependently induced VEGF-A mRNA expression. In HUVECs, nobiletin inhibited endothelial cell proliferation and, to a greater extent, tube formation in a dose-dependent manner. As in the in vivo study, nobiletin induced G0/G1 cell cycle arrest in HUVECs. However, this arrest was not accompanied by an increase in apoptosis, indicating a cytostatic effect of nobiletin. This study, for the first time, identifies nobiletin as having potent anti-angiogenic activity and suggests that nobiletin has a great potential for future research and development as a cytostatic anti-proliferative agent.

Inhibition of TNF-α-mediated endothelial cell-monocyte cell adhesion and adhesion molecules expression by the resveratrol derivative, trans-3,5,4'-trimethoxystilbene.

Resveratrol (RSV) has been shown to have anti-inflammatory activity and to have a protective role against atherosclerosis. Here it is shown, for the first time, that its derivative trans-3,5,4'-trimethoxystilbene (TMS) may be a more potent anti-inflammatory agent than resveratrol. A comparative analysis of the inhibitory activities of related stilbenes, resveratrol, TMS and polydatin (PD), on monocyte adhesion to TNF-α-activated endothelial cells showed TMS to be the most effective, with PD being the least effective. RSV and its analogues inhibited, albeit differentially, the protein and mRNA expression levels of inducible cell adhesion molecules, ICAM-1 and VCAM-1, in cultured endothelial cells. The mechanism of the inhibitory effects of these stilbenes on endothelial cell-monocyte cell adhesion can be attributed mainly to inhibition of NF-κB pathway activation. The results demonstrate that all three investigated stilbene compounds, especially TMS, exhibit a potent inhibitory effect on inflammation-induced cell-cell adhesion, expression of adhesion molecules and activation of the NF-κB pathway.

Clinical and Humanistic Outcomes of Community Pharmacy-Based Healthcare Interventions Regarding Medication Use in Older Adults: A Systematic Review and Meta-Analysis.

This review and meta-analysis aimed to determine the clinical and humanistic outcomes of community pharmacy-based interventions on medication-related problems of older adults at the primary care level. We identified randomized controlled trials (RCTs) examining the impact of various community pharmacy-based interventions from five electronic databases (namely, MEDLINE (Ovid), EMBASE (Ovid), CINAHL, APA PSYInfo, and Scopus) from January 2010 to December 2020. Consequently, we assessed these interventions' clinical and humanistic outcomes on older adults and compared them with non-intervention. We included 13 RCTs in the current review and completed a meta-analysis with six of them. The included studies had a total of 6173 older adults. Quantitative analysis showed that patient education was significantly associated with an increase in the discontinuation of sedative-hypnotics use (risk ratio 1.28; 95% CI (1.20, 1.36) I2 = 0%, p < 0.00001). Moreover, the qualitative analysis showed that medication reviews and education with follow-ups could improve various clinical outcomes, including reducing adverse drug events, reducing uncontrolled health outcomes, and improving appropriate medication use among the elderly population. However, medication review could not significantly reduce the number of older adults who fall (risk ratio 1.25; 95% CI (0.78, 1.99) I2 = 0%, p = 0.36) and require hospitalization (risk ratio 0.72; 95% CI (0.47, 1.12) I2 = 45%, p = 0.15). This study showed that community pharmacy-based interventions could help discontinue inappropriate prescription medications among older adults and could improve several clinical and humanistic outcomes. However, more effective community pharmacy-based interventions should be implemented, and more research is needed to provide further evidence for clinical and humanistic outcomes of such interventions on older adults.

Resveratrol derivative, trans-3,5,4'-trimethoxystilbene, exerts antiangiogenic and vascular-disrupting effects in zebrafish through the downregulation of VEGFR2 and cell-cycle modulation.

Angiogenesis plays an important role in the development of neoplastic diseases such as cancer. Resveratrol and its derivatives exert antiangiogenic effects, but the mechanisms of their actions remain unclear. The aim of this study was to evaluate the antiangiogenic activity of resveratrol and its derivative trans-3,5,4'-trimethoxystilbene in vitro using human umbilical vein endothelial cells (HUVECs) and in vivo using transgenic zebrafish, and to clarify their mechanisms of action in zebrafish by gene expression analysis of the vascular endothelial growth factor (VEGF) receptor (VEGFR2/KDR) and cell-cycle analysis. trans-3,5,4'-Trimethoxystilbene showed significantly more potent antiangiogenic activity than that of resveratrol in both assays. In zebrafish, trans-3,5,4'-trimethoxystilbene caused intersegmental vessel regression and downregulated VEGFR2 mRNA expression. Trans-3,5,4'-trimethoxystilbene also induced G2/M cell-cycle arrest, most specifically in endothelial cells of zebrafish embryos. We propose that the antiangiogenic and vascular-targeting activities of trans-3,5,4'-trimethoxystilbene result from the downregulation of VEGFR2 expression and cell-cycle arrest at G2/M phase.

Online Multi-Domain Geriatric Health Screening in Urban Community Dwelling Older Malaysians: A Pilot Study.

Introduction: With a rapidly aging population, the Malaysian health care system needs to develop solutions to address the lack of resources that are required for the assessment of the older person. The complex nature of geriatric syndromes coupled with the occurrence of multiple comorbid illnesses with aging, make geriatric assessment a resource intensive process. Digital health solutions could play an important role in supporting existing health care systems, especially in low and middle income countries, with limited speciality services in geriatrics. Objective: This is a pilot study aimed at screening for geriatric syndromes through self-administered online surveys in urban community dwelling older Malaysians and assessing the pattern of geriatric syndromes in relation to the frailty status of the study participants. Methods: This is a cross-sectional pilot study conducted between July-September 2020. Community dwelling adults aged 60 years and over were invited to take part in an online survey. Information on sociodemographic variables, comorbidities, and the self-reported results of geriatric syndromes (frailty, sarcopenia, anorexia of aging, urinary incontinence, falls, and cognitive impairment), were collected through the survey. Results: Data was collected for 162 participants over a period of 2 months. The mean (SD) age of the respondents was 66.42 (5.25) years with 64.9% females. Majority of the respondents were of Chinese ethnic origin (67.9%) and had tertiary level of education (75.9%). The average time taken by participants to complete the survey was 16.86 min. Urinary incontinence was the highest reported geriatric syndrome (55.1%) followed by falls (37.6%), anorexia of aging (32.8%), cognitive impairment (27.8%), and sarcopenia (8.3%). Frailty was detected in 4.5% of the study population. Loss of weight in the previous year was the highest reported component of the frailty assessment tool. The presence of sarcopenia, anorexia of aging, poor/fair self-rated health, urinary incontinence, and multimorbidity were significantly higher in older adults who were frail or prefrail. Conclusion: Screening for geriatric syndromes through online surveys is a feasible approach to identify older adults in the community who are likely to benefit from geriatric assessment. However, the demographic profile of the older population that are accessible through such digital platforms is limited.

Factors Associated With Falls Among Urban-Dwellers Aged 55 Years and Over in the Malaysian Elders Longitudinal Research (MELoR) Study.

Falls are major issues affecting the older population with potentially serious complications, including fractures, head injury, institutionalization, fear of falling and depression. While risk factors for falls have been established across Western Europe and North America, geographical differences in falls risk have not been well researched. We aim to examine the clinical and physical risk factors for falls in a middle-income South East Asian country. Cross-sectional data from the Malaysian Elders Longitudinal Research (MELoR) study involving 1,362 community dwelling individuals aged 55 years and above was utilized. Information on sociodemographic and medical history was obtained by computer-assisted questionnaires completed during home visits and hospital-based detailed health checks. Univariate and multivariate analyses compared non-fallers and fallers in the previous 12 months. Urinary incontinence, hearing impairment, depression, arthritis and cognitive impairment were risk factors for falls in the past 12 months after adjustment for age in our study population. Awareness about the risk factors in a population helps the design of fall prevention strategies that target specific or multiple risk factors.

Cross-sectional analysis of ethnic differences in fall prevalence in urban dwellers aged 55 years and over in the Malaysian Elders Longitudinal Research study.

OBJECTIVES: Falls represent major health issues within the older population. In low/middle-income Asian countries, falls in older adults remain an area which has yet to be studied in detail. Using data from the Malaysian Elders Longitudinal Research (MELoR), we have estimated the prevalence of falls among older persons in an urban population, and performed ethnic comparisons in the prevalence of falls. DESIGN: Cross-sectional analysis was carried out using the first wave data from MELoR which is a longitudinal study. SETTING: Urban community dwellers in a middle-income South East Asian country. PARTICIPANTS: 1565 participants aged ≥55 years were selected by simple random sampling from the electoral rolls of three parliamentary constituencies. OUTCOME MEASURES: Consenting participants from the MELoR study were asked the question 'Have you fallen down in the past 12 months?' during their computer-assisted home-based interviews. Logistic regression analyses were conducted to compare the prevalence of falls among various ethnic groups. RESULTS: The overall estimated prevalence of falls for individuals aged 55 years and over adjusted to the population of Kuala Lumpur was 18.9%. The estimated prevalence of falls for the three ethnic populations of Malays, Chinese and Indian aged 55 years and over was 16.2%, 19.4% and 23.8%, respectively. Following adjustment for ethnic discrepancies in age, gender, marital status and education attainment, the Indian ethnicity remained an independent predictor of falls in our population (relative risk=1.45, 95% CI 1.08 to 1.85). CONCLUSION: The prevalence of falls in this study is comparable to other previous Asian studies, but appears lower than Western studies. The predisposition of the Indian ethnic group to falls has not been previously reported. Further studies may be needed to elucidate the causes for the ethnic differences in fall prevalence.

RGG-boxes of the EWS oncoprotein repress a range of transcriptional activation domains.

The Ewings Sarcoma Oncoprotein (EWS) interacts with several components of the mammalian transcriptional and pre-mRNA splicing machinery and is also found in the cytoplasm and even on the cell surface. The apparently diverse cellular functions of EWS are, however, not well characterized. EWS harbours a potent N-terminal transcriptional activation domain (the EAD) that is revealed in the context of oncogenic EWS-fusion proteins (EFPs) and a C-terminal RNA-binding domain (RBD) that recruits pre-mRNA splicing factors and may couple transcription and splicing. In contrast to EFPs, the presumed transcriptional role of normal EWS remains enigmatic. Here, we report that multiple RGG-boxes within the RBD are necessary and sufficient for cis-repression of the EAD and that RGG-boxes can also repress in-trans, within dimeric partners. Lys can functionally substitute for Arg, indicating that the basic nature of the Arg side chain is the critical determinant of RGG-box-mediated repression. In addition to the EAD, RGG-boxes can repress a broad range of activation domains (including those of VP16, E1a and CREB), but repression can be alleviated by the simultaneous presence of more than one activation domain. We therefore propose that a key function of RGG boxes within native EWS is to restrict promiscuous activation by the EAD while still allowing EWS to enter functional transcription complexes and participate in other transactions involving pre-mRNAs.

In vitro and in vivo structure and activity relationship analysis of polymethoxylated flavonoids: identifying sinensetin as a novel antiangiogenesis agent.

SCOPE: Polymethoxylated flavonoids are present in citrus fruit in a range of chemical structures and abundance. These compounds have potential for anticarcinogenesis, antitumor, and cardiovascular protective activity, but the effect on angiogenesis has not been well studied. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) in vitro and zebrafish (Danio rerio) in vivo models were used to screen and identify the antiangiogenesis activity of seven polymethoxylated flavonoids; namely, hesperetin, naringin, neohesperidin, nobiletin, scutellarein, scutellarein tetramethylether, and sinensetin. Five, excluding naringin and neohesperidin, showed different degrees of potency of antiangiogenesis activity. Sinensetin, which had the most potent antiangiogenesis activity and the lowest toxicity, inhibited angiogenesis by inducing cell cycle arrest in the G0/G1 phase in HUVEC culture and downregulating the mRNA expressions of angiogenesis genes flt1, kdrl, and hras in zebrafish. CONCLUSION: The in vivo structure-activity relationship (SAR) analysis indicated that a flavonoid with a methoxylated group at the C3' position offers a stronger antiangiogenesis activity, whereas the absence of a methoxylated group at the C8 position offers lower lethal toxicity in addition to enhancing the antiangiogenesis activity. This study provides new insight into how modification of the chemical structure of polymethoxylated flavonoids affects this newly identified antiangiogenesis activity.

Combined in vivo imaging and omics approaches reveal metabolism of icaritin and its glycosides in zebrafish larvae.

Flavonoids isolated from Herba Epimedii such as icaritin, icariin and epimedin C have been suggested as potential bone anabolic compounds. However, the "specific localized effects" of these flavonoids in bone, in vivo, and the metabolism of these flavonoids in zebrafish larvae have never been demonstrated. In this study, we used multiple methods including in vivo imaging, drug metabolites profiling, transcriptomic and proteomic approaches to determine the mechanisms involved in the distribution and metabolism of the flavonoids in zebrafish larvae by measuring the fluorescence emission, in vivo, of icaritin and its glycoside derivatives. The fluorescence emission mechanism of icaritin in vitro was identified by spectrophotometric analysis, and the fluorescent property of icaritin was used as a probe to visualize the metabolism and distribution of icaritin and its glycoside derivatives in zebrafish larvae. Phase I and phase II metabolism of icaritin and its derivatives were identified in zebrafish by mass spectrometry. The combined transcriptomics and proteomics demonstrate a high degree of conservation of phase I and phase II drug metabolic enzymes between zebrafish larvae and mammals. Icaritin and its glycoside derivatives were demonstrated using combined approaches of in vivo imaging, drug metabolites identification, and transcriptomic and proteomic profiling to illustrate phase I and phase II metabolism of the flavonoids and their distribution in bone of zebrafish larvae. This study provides a new methodological model for use of the zebrafish larvae to examine drug metabolism.

Indirubin shows anti-angiogenic activity in an in vivo zebrafish model and an in vitro HUVEC model.

AIM OF THE STUDY: Indirubin is an active ingredient of the traditional Chinese medicine, Dang Gui Long Hui Wan, commonly used for the treatment of chronic myelocytic leukemia (CML) and other inflammatory conditions. These anti-leukemic and anti-inflammatory activities may be mediated by anti-angiogenic action. To investigate the anti-angiogenic activity of indirubin, we tested its inhibitory effect on blood vessel formation in zebrafish embryos and on endothelial cell proliferation in culture. MATERIALS AND METHODS: The anti-angiogenic activity of indirubin was tested using transgenic zebrafish embryos with fluorescent vasculature and human umbilical vein endothelial cells (HUVECs). Apoptosis was analyzed with a terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay. RESULTS: Indirubin dose-dependently inhibited intersegmental vessel formation in zebrafish embryos. It also inhibited HUVEC proliferation by the induction of cellular apoptosis and cell-cycle arrest at the G0/G1 phase. CONCLUSIONS: The anti-angiogenic activity of indirubin may partly contribute to its anti-leukemic and anti-psoriatic properties and may be valuable for the treatment of diseases with excessive angiogenesis. The zebrafish model of angiogenesis was further validated in this study.