Molecular pharmacological analysis of the human MrgD receptor : New insights into signaling

Introduction: MrgD is a member of the MAS-related GPCRs family. It has been proposed to play roles in nociception and cardiovascular control. MrgD has been shown to couple to Gq, Gi, and Gs proteins. The amino acid β-alanine and the peptide alamandine act as agonists. Antipsychotics like chlorpromazine and related structures act as low affinity antagonists. In the present study, the human MrgD receptor signaling was further characterized using potential agonists and antagonists and a possible coupling to the β-arrestin pathway was addressed. Methods: G protein-dependent functional assays including Fluo-4 calcium, Nuclear Factor of Activated T cells (NFAT)-reporter gene, and AlphaScreen cAMP assays were used to study the interaction in cloned Chinese hamster ovary (CHO) Flp-In cells stably expressing a variant of the human MrgD receptor. Receptor coupling to the β-arrestin pathway was studied using the PathHunter β-arrestin recruitment GPCR assay (Pro-link tagged human MrgD and enzyme-acceptor-tagged β-arrestin-2 co-expressed in CHO-K1 cells). Results: In CHO cells expressing the human MrgD receptor, β-alanine (300 µM, 1 mM, and 3 mM) caused strong and concentration-dependent Fluo-4 fluorescence increases which were not observed in non-transfected CHO cells. Interestingly, alamandine (10 µM) induced small, and short-lasting fluorescence increases which were similar both in receptor expressing and in control cells. As expected, and previously reported, β-alanine (300 µM) significantly inhibited forskolin-induced cAMP accumulation only in receptor expressing cells. In contrast, alamandine (0.0001 nM to 10 nM) increased the cAMP accumulation with a biphasic concentration-response curve in both cell lines. In CHO-K1 cells stably co-expressing the human MrgD and β-arrestin-2, β-alanine (30 µM to 1 mM) significantly recruited β-arrestin-2 while alamandine (0.00001 nM to 100 µM) stimulated only a weak response. DL-3-aminoisobutyric acid (30 µM to 3 mM), DL-3-aminobutyric acid (100 µM to 3 mM), and GABA (300 µM to 3 mM) activated the β-arrestin pathway with different potencies. Surprisingly, incubation either with chlorpromazine (10 µM) or thioridazine (10 µM) or rimcazole (20 µM) failed to inhibit β-alanine-induced recruitment of β-arrestin-2. Conclusion: The present work provides first evidence on the human MrgD receptor coupling to the β-arrestin pathway. In addition, the data may indicate the biased coupling of the receptor to signaling transduction pathways

Metagenomic profiling of microbial metal interaction in Red Sea deep-anoxic brine pools

Extreme environmental conditions induce evolution of microbiomes and shape microbial abundance. Different geochemical studies reported high metal abundance in Red Sea deep-anoxic brine pools, especially in Atlantis II Deep, which has the highest metals content. Brine pools show wide diversity of biologically essential and non-essential metals. Several metals known for their toxicity to biological life were detected in these pools. Yet, previous microbiome analyses of the pools demonstrated vast microbiological diversity. In this study, we compare metal-resistant prokaryotic microbiomes in different metal-rich brine water samples from Atlantis II lower convective layer (ATII-LCL), Atlantis II upper convective layer (ATII-UCL), Discovery Deep (DD) and Kebrit Deep (KD). Moreover, we investigate genome evolution of microbial communities in response to such excessive metal abundance. Using 16S rRNA pyrotags and shot-gun 454-pyrosequencing, we perform a comparative analyses of (i)-taxonomic assignment of operational taxonomic units to major bacterial and archaeal groups and (ii)-metal resistant protein-coding genes, of the microbial communities and metagenomes. The ATII-LCL, ATII-UCL, DD and KD brine pools metagenomes protein-coding genes involved in microbial-metal interaction and resistances were assessed for abundance and diversity. We report specific microbial richness of these three brine pools. Functional analyses of the metagenomes revealed different metal resistance mechanisms and different modes of mutual interaction between dissolved metals/sediments and microbial communities. This was supported by the strong correlation between specific high metal/s concentration in selected brine water, where; metal resistance, enrichment of metals metabolism and transport were revealed. As expected, ATII-LCL showed the highest relative abundance of genes involved in microbial-metal interaction. Additionally, we report significant abundance of peroxidases-encoding genes, mainly in ATII-LCL, and we hypothesize that generation of hydrogen peroxide (H2O2) occurs through interaction of pyrite deposits. Moreover, we suggest that genus Paenibacillus, which is highly abundant in ATII-LCL has a role in increasing concentration of dissolved iron in brine water. DD and KD showed relatively lower enrichment of genes involved in microbial-metal interaction. However, geochemistry of these environments together with unique microbial abundance give an inference about mechanisms of microbial metal interaction and metabolism taking place there. Eventually, we successfully identified free living metal-resistant prokaryotic communities, showed how prokaryotes tolerate and induce changes to the surrounding environment, and highlighted how geochemical conditions affected microbial abundance and induced evolution of microbiomes in brine pools

Four butyrolactones and diverse bioactive secondary metabolites from terrestrial Aspergillus flavipes MM2: isolation and structure determination

The chemical constituents and biological activities of the terrestrial Aspergillus flavipes MM2 isolated from Egyptian rice hulls are reported. Seven bioactive compounds were obtained, of which one sterol: ergosterol (1), four butyrolactones: butyrolactone I (2), aspulvinone H (3), butyrolactone-V (6) and 4,4'-diydroxypulvinone (7), along with 6-methylsalicylic acid (4) and the cyclopentenone analogue; terrien (5). Structures of the isolated compounds were deduced by intensive studies of their 1D & 2D NMR, MS data and comparison with related structures. The strain extract and the isolated compounds (1-7) were biologically studied against number of microbial strains, and brine shrimp for cytotoxicity. In this article, the taxonomical characterization of A. flavipes MM2 along with its upscale fermentation, isolation and structural assignment of the obtained bioactive metabolites, and evaluate their antimicrobial and cytotoxic activities were described

Ecological Agro-ecosystem Sustainable Development in Relationship to Other Sectors in the Economic System, and Human Ecological Footprint and Imprint☆

Abstract Sustainable agriculture is the major economic sector (i.e. about 30% of Global economy) with the industrial and trading system in the world's economy. It is important to understand why the sustainable development is very important to the point of view of improving of human life and reducing the poverty. Additionally, we need to sustain our natural resources to be replenished and continue support our human population growth that is continued to increase in alarming rate rather than development, which is in a slow rate that does not meet the demands. This paper is to discuss the importance of global agro-ecosystems, to support humans' needs for feeding and continue their lives in a healthy and sustainable life and to function within the society. In addition, the paper will show the availability of the agriculture natural resources in terms of global ecological biological capacities in hectares and the trends in using these resources in terms of an ecological footprint in hectares. Additionally, we study the term of ecological human imprint in relation to the agro-ecosystem as suggested by Shakir Hanna et al., 2014 . Further the paper will address the impacts of agro-ecosystem on global economy and, further discuss the impacts of human technological advances on agro-ecosystems ecologically, economically, and social importance. Our results show that the global population will be 10.50 billion people in 2050 (i.e. 1.1% the current population growth). The available global cropped land is 2.36 billion global hectares in 2008.The question is the Earth able to provide food and other agricultural products to support the healthy living of all human beings in year 2050 at the current growth rate? The paper is discussing these concerns

Three Presentations of Takayasu’s Arteritis in Hispanic Patients

Takayasu’s arteritis (TA) is a medium and large vessel vasculitis, defined as a nonspecific aortitis that usually involves the aorta and its branches Kobayashi and Numano (2002). Its etiology remains unclear, and its complications are diverse and severe, including stenosis of the thoracic and abdominal aorta, aortic valve damage and regurgitation, and stenosis of the branches of the aorta. Carotid stenosis, coronary artery aneurysms, and renal artery stenosis resulting in renovascular hypertension are also reported sequellae of TA Kobayashi and Numano (2002). The disease was first described in Japan, but has also been diagnosed in India and Mexico Johnston (2002). Its incidence in the United States has been quoted as 2.6 patients per 1,000,000 people/year Johnston (2002). In Japan, its incidence is 3.6 patients per 1,000,000 patients/year and prevalence is 7.85 patients per 100,000 per year Morita et al. (1996). The natural history of this disease, which is commonly present in Asian populations, has only recently been studied in Hispanic patients despite the notable incidence and prevalence of TA in Mexican, South American, and Indian populations (Johnston 2002, Gamarra et al. 2010 ). We present three cases of Hispanic patients who presented with TA at Olive-View-UCLA Medical Center (OVMC). We review their clinical and radiographic presentations. Finally, we review the literature to compare the clinical features of our three patients with data regarding the presentation of TA in more traditional Asian populations

Hypolipidemic effect of vegetable and cereal dietary mixtures from Egyptian sources

Hyperlipidemia is a predominant risk factor for atherosclerosis and associated cardiovascular diseases (CVD). The international guidelines issued by the World Health Organization recommend a reduction in dietary saturated fat and cholesterol intake as a means to prevent hypercholesterolemia and CVD; however, only limited data are available on the benefits of vegetable consumption on CVD risk factors. The aim of this study was to prepare two powder mixtures containing vegetables and cereals and to evaluate their effect in hyperlipidemic rats. The first mixture was prepared from whole wheat, cabbage, parsley and pepper, while the second mixture was prepared from whole wheat, red beet root, parsley and pepper. Whole wheat was used as a source of dietary fiber, while cabbage and beetroot were used as sources of glucosinolates (GLS) and betalains respectively as well as dietary fiber. The chemical compositions of these mixtures were determined. The safety of these mixtures was also evaluated by examining liver and kidney functions. The chemical compositions of the powder mixtures revealed that mixtures (1) and (2) contain 19.1% and 13.3% protein, 2.1% and 2.5 % fat, 69.6% and 77.5% carbohydrates, 1.8% and 1.2% crude fibers, 7.4% and 5.5% ash and 18.3% and 16.8% dietary fibers respectively. Vitamin E was 7.4 and 4.5 mg/100g in mixtures (1) and (2) respectively. β-carotene was 830 and 786μg/100g in mixtures (1) and (2) respectively. Total phenolic compounds were 1910 and 1710 mg as gallic acid equivalents/100g in mixtures (1) and (2) respectively. The results of the animal experiment showed a non-significant reduction in final body weight and body weight gain in rats fed the control diet containing mixture (1) or (2) when compared with different groups. Rats fed the control diet containing mixture (1) or (2) showed a significant reduction in plasma total lipids, T-Ch, LDL-Ch, TG and the ratio of T-Ch /HDLCh in different degrees, while HDL-Ch increased significantly. The studied mixtures showed a hypolipidemic effect, which may be due to the presence of dietary fibers, plant protein, and phenolic compunds.<br><br>La hiperlipidemia es un factor de riesgo predominante en la ateroesclerosis y enfermedades cardiovasculares asociadas (CVD). Las directrices Internacionales dictadas por la Organización Mundial de la Salud recomiendan una reducción en la ingesta de grasas saturadas y colesterol como medio para prevenir la hipercolesterolemia y las CVD; sin embargo, solamente hay un número limitado de datos disponibles sobre el beneficio del consumo de verduras en los factores de riesgo de las CVD. El objetivo de este estudio fue preparar dos mezclas en polvo que contenían verduras y cereales. El efecto hypolipidémico de estas dos mezclas fue evaluado en ratas hiperlipidémicas. La primera mezcla fue preparada con trigo, col, perejil y pimiento mientras que la segunda mezcla fue preparada con trigo, remolacha, perejil y pimiento. El trigo fue usado como fuente de fibra, mientras que la col y la remolacha como fuente de glucosinolatos (GLS) y betalinas, respectivamente y fibra también. La composición química de estas mezclas fue determinada. La seguridad de estas muestras fue también evaluada a través de las funciones del hígado y del riñón. La composición química de las mezclas en polvo indica que la mezcla (1) y (2) contienen un 19.1% y un 13.3% de proteina, un 2.1% y un 2.5 % de grasa, un 69.6% y un 77.5% de azúcares, un 1.8% y un 1.2% de fibra cruda, un 7.4% y un 5.5% de cenizas y un 18.3% y 16.8% fibra, respectivamente. El contenido de vitamin E fue de 7.4 y 4.5 mg/100g de mezcla (1) y (2) respectivamente. El contenido de β-carotene fue de 830 y 786μg/100g de mezcla (1) y (2) respectivamente. Los compuestos fenólicos totales fueron 1910 y 1710 mg como equivalentes de ácido gálico /100g de mezcla (1) y (2) respectivamente. Los resultados de los experimentos con animales mostraron una reducción no significativa en el peso final y en la ganancia de peso en ratas alimentadas con dietas control conteniendo la mezcla (1) y (2) cuando se compara con diferentes grupos. Las ratas alimentadas con dietas control conteniendo mezcla (1) y (2) mostraron una reducción significativa en los lípidos totales del plasma, T-Ch, LDL-Ch, TG y la relación de T-Ch /HDL-Ch con diferentes grados, mientras que HDL-Ch aumento significativamente. Las mezclas estudiadas mostraron un efecto hipolipidémico, que puede ser debido a la presencia de fibra, proteínas de plantas y compuestos fenólicos

Mapping 123 million neonatal, infant and child deaths between 2000 and 2017

Since 2000, many countries have achieved considerable success in improving child survival, but localized progress remains unclear. To inform efforts towards United Nations Sustainable Development Goal 3.2—to end preventable child deaths by 2030—we need consistently estimated data at the subnational level regarding child mortality rates and trends. Here we quantified, for the period 2000–2017, the subnational variation in mortality rates and number of deaths of neonates, infants and children under 5 years of age within 99 low- and middle-income countries using a geostatistical survival model. We estimated that 32% of children under 5 in these countries lived in districts that had attained rates of 25 or fewer child deaths per 1,000 live births by 2017, and that 58% of child deaths between 2000 and 2017 in these countries could have been averted in the absence of geographical inequality. This study enables the identification of high-mortality clusters, patterns of progress and geographical inequalities to inform appropriate investments and implementations that will help to improve the health of all populations

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p<0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p<0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised