Natural variation reveals that intracellular distribution of ELF3 protein is associated with function in the circadian clock

Natural selection of variants within the Arabidopsis thaliana circadian clock can be attributed to adaptation to varying environments. To define a basis for such variation, we examined clock speed in a reporter-modified Bay-0 x Shakdara recombinant inbred line and localized heritable variation. Extensive variation led us to identify EARLY FLOWERING3 (ELF3) as a major quantitative trait locus (QTL). The causal nucleotide polymorphism caused a short-period phenotype under light and severely dampened rhythm generation in darkness, and entrainment alterations resulted. We found that ELF3-Sha protein failed to properly localize to the nucleus, and its ability to accumulate in darkness was compromised. Evidence was provided that the ELF3-Sha allele originated in Central Asia. Collectively we showed that ELF3 protein plays a vital role in defining its light-repressor action in the circadian clock and that its functional abilities are largely dependent on its cellular localization

Glucose-6 Phosphate Dehydrogenase Deficiency in terms of hemolysis indicators and management

Background: Glucose-6 Phosphate Dehydrogenase (G6PD) Deficiency is one of the commonest inherited enzyme abnormalities in humans, caused by many mutations that reduce the stability of the enzyme and its level as red cells progress in age. Objectives: To determine the useful hematologic indicators of hemolysis, observe an early detection of G6PD enzyme deficiency (if any), and the available therapeutic measures. Patients and Methods: 123 patients with G6PD deficiency and hemolysis after exposure to fava beans whom visited AL-Elwiya Pediatric Teaching Hospital from the 1st of February 2016 till 31st of May 2016 were entered this study retrospectively. Hemolysis laboratory indicators were observed. Management supportive measures were put in consideration also. Results: We found that 10-20% levels of hematocrit and normochromic normocytic anemia were the most frequent on presentation, while a range of 15.1-20% of reticulocyte counts was the most common with lower rates in females group. Hyperbilirubinemia was seen with nil patients had abnormal renal function tests. About three quarters (76.4%) of the total number of involved cases had glucose-6-phosphate dehydrogenase (G6PD) deficiency. Only 4 patients required no blood transfusion, 102 patients (82.9%) needed transfusion once, and the rest 17 (13.8%) had more than one blood transfusion. Most of cases (91.1%) recovered within the first 3 days. However; all cases were recovered by the fourth day of admission. Conclusion: Hemoglobin and blood morphology with hyperbilirubinemia were useful hematologic indicators of hemolytic process, while blood transfusion was the most used therapeutic measure, and recovery was expected within 2-3 days

FORMULATION AND CHARACTERIZATION OF FLUCONAZOLE LOADED OLIVE OIL NANOEMULSIONS

Present study was carried out to develop and evaluate olive oil based nano-emulsion for transdermal delivery of fluconazole, a bistriazole based antifungal agent with poor water solubility and lipophilicity. Olive oil, a natural non-irritating, non-toxic proposed permeation enhancer, is known to have some antifungal activity as well. Screening of common emulsifiers like Tweens (Tween 20, tween 60, tween 80), Spans (span 60, span 80), brij 35, puronic 127, and poloxamer 188 were done based on solubility of fluconazole in these surfactants followed by their efficiency to emulsify olive oil in water. Co-emulsifiers such as glycols (polyethylene glycol 200, polyethylene glycol 400, propylene glycol), and short chain alcohols (ethanol, propanol, butanol and octanol) were also screened similarly. Tween 80 and butanol were selected as emulsifier and co-emulsifier respectively to formulate nano-emulsion by aqueous titration method. However, separation was observed after 24 hours. Therefore, span 80 was added as an auxiliary emulsifier to improve emulsification efficiency. Finally, a blend of tween 80, span 80 and butanol was optimized as emulsifier (56 % wt/wt) to emulsify 9 % wt/wt of olive oil in 33 % wt/wt water. Pseudo-ternary phase diagram was employed to identify and optimize the components. Optimized formulation based on phase separation and thermokinetic stability was characterized for globule size, size distribution, zeta potential, viscosity, refractive index and pH. Globule size analysis by zetasizer nano ZS was further confirmed by transmission electron microscopy. Permeation flux of fluconazole from optimized formulation through artificial skin was approximately three fold higher than the control. In conclusion, developed olive oil based nano-emulsion of fluconazole demonstrated promising solubility, permeability and stability. Keywords: Fluconazole, olive oil, nano-emulsion, transdermal permeatio

Characterization of seed oils from different varieties of watermelon [<i>Citrullus lanatus</i> (Thunb.)] from Pakistan

This paper reports the physicochemical characteristics of the seed oils from different varieties of watermelon (<i>Citrullus lanatus</i>) cultivated in Pakistan, namely Sugar Baby, Q-F-12, D-W-H-21 and Red Circle-1885. The oil and crude protein contents from watermelon seeds, within the range of 28.25 to 35.65% and 20.50 to 35.00%, respectively, varied significantly (<i>p</i> < 0.05) among the varieties tested. The levels of moisture, ash, and crude fiber in the seeds tested were found to be 2.16-3.24%, 1.95-3.42% and 4.29-6.60%, respectively. The physicohemical characteristics of the extracted oils were: free fatty acid contents (1.17-2.10% as oleic acid), iodine value (97.10-116.32 g of I2 100g^-1 of oil), saponification index (190.20-205.57 mg KOH g^-1 of oil), unsaponifiable matter (0.54-0.82%) and color (1.12-4.30 R + 12.20-33.40 Y). The oils revealed a reasonable oxidative parameter range as depicted by the determinations of specific extinction at 232 and 270 nm (2.90-4.40 and 2.05- 3.09, respectively), <i>p</i>-anisidine value (5.60-7.70) and peroxide value (2.90-5.06 meqO₂ kg^-1 of oil). Linoleic acid was the major fatty acid found in all the seed oils with contributions of 45.30-51.80% of the total fatty acids (FA). Other fatty acids detected were known to be oleic acid (20.2- 23.5%), palmitic acid (15.1-16.9%) and stearic acid (11.5- 14.4%). The contents of α- and δ-tocopherol in the oils accounted for 120.6-195.6 and 9.1-58.3 mg kg^-1, respectively. The physicochemical attributes of the watermelon seed oils showed a wider variation among the varieties tested. The results of the present study indicate that the seeds of the tested watermelon varieties from Pakistan are a potential source of high-linoleic oil and thus can be explored for commercial use and value addition.<br><br>Se presentan las características físico-químicas de aceites de diferentes variedades de semillas de sandías (Citrullus lanatus) cultivadas en Pakistán: Sugar Baby, QF-12, DWH-21 y Círculo rojo-1885. El aceite y el contenido de proteína cruda de las semillas de sandía están dentro de los rangos: 28,25-35,65% y 20,50-35,00%, respectivamente y varian significativamente (p < 0,05) entre las variedades ensayadas. Los niveles de humedad, fibra cruda y cenizas en las semillas se encontró entre 2.16-3.24%, 1.95-3.42% y 4.29-6.60%, respectivamente. Las características fisico-químicas estudiadas de los aceites extraídos fueron: contenido de ácidos grasos libres (1.17-2.10% de ácido oleico), índice de yodo (97,10-116,32 g de I2 100 g^-1 de aceite), índice de saponificación (190,20-205,57 mg de KOH g^-1 de aceite), insaponificable (0,54-0.82%) y color (1.12-4.30 de I + 12.20- 33.40 y). Los aceites presentaron unos rangos de los parámetros de oxidación razonables, como se muestra en las determinaciones de la extinción específica a 232 y 270 nm (2.90-4.40 y 2.05-3.09 respectivamente), valores de <i>p</i>-anisidina (5.60-7.70) e índice de peróxidos (2,90-5,06 meqO₂ kg^-1 de aceite). El ácido linoleico es el principal ácido graso que se encuentra en todos los aceites de las semillas, con una contribución del 45.30-51.80% del total de ácidos grasos. Otros ácidos grasos determinados fueron oleico, 20,2-23,5%, palmítico, 15,1-16,9%) y esteárico, 11,5-14,4%. El contenido de α- y δ-tocoferol en los aceites fué de 120,6-195,6 y 9,1- 58,3 mg kg^-1, respectivamente. Los atributos físico-químicos de los aceites de semillas de sandía variaron significativamente entre las variedades ensayadas. Los resultados del presente estudio indican que las semillas de las variedades de sandía ensayadas procedentes de Pakistán son una fuente potencial aceites con alto contenido en ácido linoleico y, por lo tanto, se puede explorar para usos comerciales y productos con valor añadido

Adventitial transplantation of blood outgrowth endothelial cells in porcine haemodialysis grafts alleviates hypoxia and decreases neointimal proliferation through a matrix metalloproteinase-9-mediated pathway—a pilot study

Purpose. We hypothesized that adventitial transplantation of blood outgrowth endothelial cells (BOEC) to the vein-to-graft anastomosis of polytetrafluoroethylene grafts will reduce neointimal hyperplasia by reducing hypoxia inducible factor-1α (HIF-1α), by increasing angiogenesis in a porcine model of chronic renal insufficiency with haemodialysis polytetrafluoroethylene grafts. Because matrix metalloproteinases (MMPs) have been shown to be involved with angiogenesis, the expression of MMPs and their inhibitors was determined

Targeted gene delivery in tumor xenografts by the combination of ultrasound-targeted microbubble destruction and polyethylenimine to inhibit survivin gene expression and induce apoptosis

<p>Abstract</p> <p>Background</p> <p>Noninvasive and tissue-specific technologies of gene transfection would be valuable in clinical gene therapy. This present study was designed to determine whether it could enhance gene transfection <it>in vivo </it>by the combination of ultrasound-targeted microbubble destruction (UTMD) with polyethylenimine (PEI) in tumor xenografts, and illuminate the effects of gene silencing and apoptosis induction with short hairpin RNA (shRNA) interference therapy targeting human survivin by this novel technique.</p> <p>Methods</p> <p>Two different expression vectors (pCMV-LUC and pSIREN) were incubated with PEI to prepare cationic complexes (PEI/DNA) and confirmed by the gel retardation assay. Human cervical carcinoma (Hela) tumors were planted subcutaneously in both flanks of nude mice. Tumor-bearing mice were administered by tail vein with PBS, plasmid, plasmid and SonoVue microbubble, PEI/DNA and SonoVue microbubble. One tumor was exposed to ultrasound irradiation, while the other served as control. The feasibility of targeted delivery and tissue specificity facilitated by UTMD and PEI were investigated. Moreover, immunohistochemistry analyses about gene silencing and apoptosis induction were detected.</p> <p>Results</p> <p>Electrophoresis experiment revealed that PEI could condense DNA efficiently. The application of UTMD significantly increases the tissue transfection. Both expression vectors showed that gene expressions were present in all sections of tumors that received ultrasound exposure but not in control tumors. More importantly, the increases in transgene expression were related to UTMD with the presence of PEI significantly. Silencing of the survivin gene could induce apoptosis effectively by downregulating survivin and bcl-2 expression, also cause up-regulation of bax and caspase-3 expression.</p> <p>Conclusions</p> <p>This noninvasive, novel combination of UTMD with PEI could enhance targeted gene delivery and gene expression in tumor xenografts at intravenous administration effectively without causing any apparently adverse effect, and might be a promising candidate for gene therapy. Silencing of survivin gene expression with shRNA could be facilitated by this non-viral technique, and lead to significant cell apoptosis.</p

THG113.31, a specific PGF2alpha receptor antagonist, induces human myometrial relaxation and BKCa channel activation

BACKGROUND: PGF2alpha exerts a significant contractile effect on myometrium and is central to human labour. THG113.31, a specific non-competitive PGF2alpha receptor (FP) antagonist, exerts an inhibitory effect on myometrial contractility. The BKCa channel is ubiquitously encountered in human uterine tissue and plays a significant role in modulating myometrial cell membrane potential and excitability. The objective of this study was to investigate potential BKCa channel involvement in the response of human myometrium to THG113.31. METHODS: Single and whole-cell electrophysiological BKCa channel recordings from freshly dispersed myocytes, were investigated in the presence and absence of THG113.31. Functional studies investigated the effects of THG113.31 on isolated spontaneous myometrial contractions, in the presence and absence of the BKCa channel blocker, iberiotoxin. RESULTS: Single channel recordings identified the BKCa channel as a target of THG113.31. THG113.31 significantly increased the open state probability of these channels [control 0.023+/-0.006; 10 microM THG113.31 0.087+/-0.012 (P = 0.009); and 50 microM THG113.31 0.1356+/-0.018 (P = 0.001)]. In addition, THG113.31 increased whole-cell BKCa currents over a range of membrane potentials, and this effect was reversed by 100 nanoM IbTX. Isometric tension studies demonstrated that THG113.31 exerted a significant concentration-dependent relaxant effect on human myometrial tissue and pre-incubation of strips with IbTX abolished this effect on spontaneously occurring contractions. CONCLUSION: These data suggests that activation of the BKCa channel may contribute, at least partially, to the uterorelaxant effect of THG113.31

Anemia prevalence in women of reproductive age in low- and middle-income countries between 2000 and 2018

Anemia is a globally widespread condition in women and is associated with reduced economic productivity and increased mortality worldwide. Here we map annual 2000–2018 geospatial estimates of anemia prevalence in women of reproductive age (15–49 years) across 82 low- and middle-income countries (LMICs), stratify anemia by severity and aggregate results to policy-relevant administrative and national levels. Additionally, we provide subnational disparity analyses to provide a comprehensive overview of anemia prevalence inequalities within these countries and predict progress toward the World Health Organization’s Global Nutrition Target (WHO GNT) to reduce anemia by half by 2030. Our results demonstrate widespread moderate improvements in overall anemia prevalence but identify only three LMICs with a high probability of achieving the WHO GNT by 2030 at a national scale, and no LMIC is expected to achieve the target in all their subnational administrative units. Our maps show where large within-country disparities occur, as well as areas likely to fall short of the WHO GNT, offering precision public health tools so that adequate resource allocation and subsequent interventions can be targeted to the most vulnerable populations

Measuring universal health coverage based on an index of effective coverage of health services in 204 countries and territories, 1990–2019 : a systematic analysis for the Global Burden of Disease Study 2019

Background: Achieving universal health coverage (UHC) involves all people receiving the health services they need, of high quality, without experiencing financial hardship. Making progress towards UHC is a policy priority for both countries and global institutions, as highlighted by the agenda of the UN Sustainable Development Goals (SDGs) and WHO's Thirteenth General Programme of Work (GPW13). Measuring effective coverage at the health-system level is important for understanding whether health services are aligned with countries' health profiles and are of sufficient quality to produce health gains for populations of all ages. Methods: Based on the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, we assessed UHC effective coverage for 204 countries and territories from 1990 to 2019. Drawing from a measurement framework developed through WHO's GPW13 consultation, we mapped 23 effective coverage indicators to a matrix representing health service types (eg, promotion, prevention, and treatment) and five population-age groups spanning from reproductive and newborn to older adults (>= 65 years). Effective coverage indicators were based on intervention coverage or outcome-based measures such as mortality-to-incidence ratios to approximate access to quality care; outcome-based measures were transformed to values on a scale of 0-100 based on the 2.5th and 97.5th percentile of location-year values. We constructed the UHC effective coverage index by weighting each effective coverage indicator relative to its associated potential health gains, as measured by disability-adjusted life-years for each location-year and population-age group. For three tests of validity (content, known-groups, and convergent), UHC effective coverage index performance was generally better than that of other UHC service coverage indices from WHO (ie, the current metric for SDG indicator 3.8.1 on UHC service coverage), the World Bank, and GBD 2017. We quantified frontiers of UHC effective coverage performance on the basis of pooled health spending per capita, representing UHC effective coverage index levels achieved in 2019 relative to country-level government health spending, prepaid private expenditures, and development assistance for health. To assess current trajectories towards the GPW13 UHC billion target-1 billion more people benefiting from UHC by 2023-we estimated additional population equivalents with UHC effective coverage from 2018 to 2023. Findings: Globally, performance on the UHC effective coverage index improved from 45.8 (95% uncertainty interval 44.2-47.5) in 1990 to 60.3 (58.7-61.9) in 2019, yet country-level UHC effective coverage in 2019 still spanned from 95 or higher in Japan and Iceland to lower than 25 in Somalia and the Central African Republic. Since 2010, sub-Saharan Africa showed accelerated gains on the UHC effective coverage index (at an average increase of 2.6% [1.9-3.3] per year up to 2019); by contrast, most other GBD super-regions had slowed rates of progress in 2010-2019 relative to 1990-2010. Many countries showed lagging performance on effective coverage indicators for non-communicable diseases relative to those for communicable diseases and maternal and child health, despite non-communicable diseases accounting for a greater proportion of potential health gains in 2019, suggesting that many health systems are not keeping pace with the rising non-communicable disease burden and associated population health needs. In 2019, the UHC effective coverage index was associated with pooled health spending per capita (r=0.79), although countries across the development spectrum had much lower UHC effective coverage than is potentially achievable relative to their health spending. Under maximum efficiency of translating health spending into UHC effective coverage performance, countries would need to reach $1398 pooled health spending per capita (US$ adjusted for purchasing power parity) in order to achieve 80 on the UHC effective coverage index. From 2018 to 2023, an estimated 388.9 million (358.6-421.3) more population equivalents would have UHC effective coverage, falling well short of the GPW13 target of 1 billion more people benefiting from UHC during this time. Current projections point to an estimated 3.1 billion (3.0-3.2) population equivalents still lacking UHC effective coverage in 2023, with nearly a third (968.1 million [903.5-1040.3]) residing in south Asia. Interpretation: The present study demonstrates the utility of measuring effective coverage and its role in supporting improved health outcomes for all people-the ultimate goal of UHC and its achievement. Global ambitions to accelerate progress on UHC service coverage are increasingly unlikely unless concerted action on non-communicable diseases occurs and countries can better translate health spending into improved performance. Focusing on effective coverage and accounting for the world's evolving health needs lays the groundwork for better understanding how close-or how far-all populations are in benefiting from UHC