Publications by authors named "John Jackson"

283 Publications

A standardized dataset for conservation prioritization of songbirds to support CITES.

Data Brief 2021 Jun 7;36:107093. Epub 2021 May 7.

Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark.

In this article we present a standardized dataset on 6659 songbirds (Passeriformes) highlighting information relevant to species conservation prioritization with a main focus to support the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Data were collected from both scientific and grey literature as well as several online databases. The data are structured into six knowledge categories: Conventions and Treaties, Human Use, Extinction Risk, Management Opportunities, Biological Information, and Intrinsic Values. The Conventions and Treaties category includes the listings for two international conventions, CITES and the Convention on the Conservation of Migratory Species of Wild Animals (CMS), as well as EU listings for the EU Wildlife Trade Regulations and the EU Birds Directive. The Human Use category contains information on both regulated trade collected from the CITES Trade Database and the United States' Law Enforcement Management Information System (LEMIS), and highly aggregated data on seizures which we obtained from TRAFFIC, the United Nations Office on Drugs and Crime (UNODC) and two data sources on traditional medicine. We also present, for the first time, the complete Songbirds in Trade Database (SiTDB), a trade database curated by taxon expert S. Bruslund based on expert knowledge, literature review, market surveys and sale announcements. Data on the types of human use, including traditional medicine are also provided. The knowledge area on Extinction Risk contains data on the species' IUCN Red List status, the Alliance for Zero Extinction Trigger Species status, site and population at the site, the species' IUCN Climate Change Vulnerability Assessment, and the listing of priority species at the Asian Songbird Crisis Summit. In the Management Opportunities category, we gathered data on ex-situ management from Species360 zoo holdings as well as species management plans from the European and North American Zoo Associations (EAZA and AZA, respectively). Biological Information includes data on body mass, clutch size, diet, availability of data from the IUCN Red List on habitat systems, extent of occurrence, generation length, migration pattern, distribution, and biological data from the Demographic Species Knowledge Index, number of occurrences recorded by the Global Biodiversity Information Facility (GBIF) as well as genomic data from the Bird 10 000K Genomes (B10K) project, Vertebrate Genome Project (VGP) and GenBank. Information on invasive species is also part of this knowledge area. The Intrinsic Value category refers to two measures of the species' intrinsic value, namely Ecological and Evolutionary Distinctiveness. In order to make these knowledge areas comparable, we standardized data following the taxonomy of the Handbook of the Birds of the World and Birdlife (Version 4, 2019). The data enable a broad spectrum of analyses and will be useful to scientists for further research and to policymakers, zoos and other conservation stakeholders for future prioritization decisions.
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http://dx.doi.org/10.1016/j.dib.2021.107093DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8144656PMC
June 2021

Regenerative Medicine Approaches in Bioengineering Female Reproductive Tissues.

Reprod Sci 2021 Jun 20;28(6):1573-1595. Epub 2021 Apr 20.

Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC, 27101, USA.

Diseases, disorders, and dysfunctions of the female reproductive tract tissues can result in either infertility and/or hormonal imbalance. Current treatment options are limited and often do not result in tissue function restoration, requiring alternative therapeutic approaches. Regenerative medicine offers potential new therapies through the bioengineering of female reproductive tissues. This review focuses on some of the current technologies that could address the restoration of functional female reproductive tissues, including the use of stem cells, biomaterial scaffolds, bio-printing, and bio-fabrication of tissues or organoids. The use of these approaches could also be used to address issues in infertility. Strategies such as cell-based hormone replacement therapy could provide a more natural means of restoring normal ovarian physiology. Engineering of reproductive tissues and organs could serve as a powerful tool for correcting developmental anomalies. Organ-on-a-chip technologies could be used to perform drug screening for personalized medicine approaches and scientific investigations of the complex physiological interactions between the female reproductive tissues and other organ systems. While some of these technologies have already been developed, others have not been translated for clinical application. The continuous evolution of biomaterials and techniques, advances in bioprinting, along with emerging ideas for new approaches, shows a promising future for treating female reproductive tract-related disorders and dysfunctions.
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http://dx.doi.org/10.1007/s43032-021-00548-9DOI Listing
June 2021

Analysis of the demographics and characteristics of the Australian pharmacist workforce 2013-2018: decreasing supply points to the need for a workforce strategy.

Int J Pharm Pract 2021 Mar;29(2):178-185

Centre for Medicine Use and Safety, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.

Objectives: An adequate workforce is necessary for health care delivery. The last official analysis of the Australian pharmacist workforce was in 2014 and the results of recent studies are contradictory. The objective of this work was to determine current demographic details and recent trends of the pharmacy workforce and assess the impact of changes in student numbers and migration policy.

Methods: Longitudinal and descriptive analysis was undertaken of National Health Workforce Datasets and registrant data available from the Australian Health Practitioner Regulation Agency and the Pharmacy Board of Australia from 2013 to 2018.

Key Findings: There was an increase in females and a trend towards hospital practice but no change in the geographic distribution of pharmacists over the period. However, the pharmacist workforce grew more slowly than comparable health professions and while the youngest pharmacist cohort (20-34 years) remains the largest, the next oldest cohort increased at a greater rate. The youngest cohort reported a decrease in intention to remain working in pharmacy.

Conclusions: A fall in student numbers and changes to immigration policy have contributed to a low growth rate and ageing of the pharmacist workforce compared with other professions. Whether these factors along with the intentions of young pharmacists will result in a shortage is dependent on developments in demand for pharmacists and a workforce strategy is required to monitor these developments.
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http://dx.doi.org/10.1093/ijpp/riaa022DOI Listing
March 2021

Comparative assessment of competency frameworks for pharmacists in the Western Pacific region including the potential for a regional framework.

Int J Pharm Pract 2021 Mar;29(2):192-195

Department of Pharmacy, National University of Singapore, Singapore.

Objectives: To assess the potential for a regional competency framework for pharmacists in the Western Pacific using the Global Competency Framework (GbCF) as a reference.

Methods: Mixed-methods approach used a self-administered survey and semi-structured interviews of 13 countries to evaluate the perceived benefits, existence and content of competency frameworks.

Key Findings: Variations in structure, components and emphasis of the four frameworks that do exist indicate significant tailoring to local requirements. Based on these four and the GbCF, 32 competencies allocated into four themes has been proposed.

Conclusion: Varying national requirements mitigate against a single regional competency framework.
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http://dx.doi.org/10.1093/ijpp/riaa005DOI Listing
March 2021

Optimized culture system to maximize ovarian cell growth and functionality in vitro.

Cell Tissue Res 2021 Feb 13. Epub 2021 Feb 13.

Wake Forest Institute for Regernative Medicine, Wake Forest School of Medicine, Medical Center Boulvard, Winston Salem, NC, 27157, USA.

Ovaries are the primary physiological source of female sex hormones, which play a crucial role in maintaining ovarian cycle, determining secondary sexual characteristics and preparing the endometrium for implantation. In vitro follicle engineering has been used to investigate follicle development, including ovarian hormone production and gamete maturation. To engineer functional follicles, culture and expansion of the primary ovarian cells are essential. However, the phenotypic and functional characteristics of primary ovarian cells are often lost during culture. The objective of this study is to develop an optimized culture system for maintaining ovarian cell growth and functionality. Granulosa cells (GCs) and theca cells (TCs) were isolated from female rats. The addition of follicle-stimulating hormone (FSH) or luteinizing hormone (LH) to the basal culture media significantly enhanced the secretion of estradiol from GCs and androstenedione from TCs. Serum concentrations of 5% and 10% had a similar role in promoting ovarian cell expansion and secretion of estradiol and androstenedione hormones from both types of cells. Growth differentiation factor 9 (GDF9), bone morphogenic protein 15 (BMP15), BMP7 and basic fibroblast growth factor (bFGF) enhanced GC proliferation and estradiol production, respectively. Among them, the effect of bFGF was most significant. bFGF also enhanced TC proliferation. When GCs and TCs were cultured in 5% serum, gonadotropin and bFGF-containing medium, they proliferated exponentially throughout the culture period of up to 40 days while maintaining their functional characteristics. Taken together, these results indicate that our medium formula is optimal for maximizing proliferation of functionally differentiated ovarian cells.
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http://dx.doi.org/10.1007/s00441-021-03415-wDOI Listing
February 2021

Engineering Functional Rat Ovarian Spheroids Using Granulosa and Theca Cells.

Reprod Sci 2021 Jun 28;28(6):1697-1708. Epub 2021 Jan 28.

Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC, 27101, USA.

Although menopausal hormone therapy (MHT) is the most effective approach to managing the loss of ovarian activity, serious side effects have been reported. Cell-based therapy is a promising alternative for MHT. This study constructed engineered ovarian cell spheroids and investigated their endocrine function. Theca and granulosa cells were isolated from ovaries of 10-week-old rats. Two types of engineered ovarian cell spheroids were fabricated through forced aggregation in microwells, multilayered spheroids with centralized granulosa aggregates surrounded by an outer layer of theca cells and mixed ovarian spheroids lacking spatial rearrangement. The ovarian cell spheroids were encapsulated into a collagen gel. Non-aggregated ovarian cells served as controls. The endocrine function of the engineered ovarian spheroids was assessed over 30 days. The structure of the spheroids was well maintained during culture. The secretion of 17β-estradiol from both types of engineered ovarian cell spheroids was higher than in the control group and increased continuously in a time-dependent manner. Secretion of 17β-estradiol in the multi-layered ovarian cell spheroids was higher than in the non-layered constructs. Increased secretion of progesterone was detected in the multi-layered ovarian cell spheroids at day 5 of culture and was sustained during the culture period. The initial secretion level of progesterone in the non-layered ovarian cell spheroids was similar to those from the controls and increased significantly from days 21 to 30. An in vitro rat model of engineered ovarian cell spheroids was developed that was capable of secreting sex steroid hormones, indicating that the hormone secreting function of ovaries can be recapitulated ex vivo and potentially adapted for MHT.
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http://dx.doi.org/10.1007/s43032-020-00445-7DOI Listing
June 2021

The Design, Characterization and Antibacterial Activity of Heat and Silver Crosslinked Poly(Vinyl Alcohol) Hydrogel Forming Dressings Containing Silver Nanoparticles.

Nanomaterials (Basel) 2021 Jan 4;11(1). Epub 2021 Jan 4.

UBC Faculty of Pharmaceutical Sciences, 2045 Wesbrook Mall, UBC, Vancouver, BC V6T1Z3, Canada.

The prompt treatment of burn wounds is essential but can be challenging in remote parts of Africa, where burns from open fires are a constant hazard for children and suitable medical care may be far away. Consequently, there is an unmet need for an economical burn wound dressing with a sustained antimicrobial activity that might be manufactured locally at low cost. This study describes and characterizes the novel preparation of a silver nitrate-loaded/poly(vinyl alcohol) (PVA) film. Using controlled heating cycles, films may be crosslinked with in situ silver nanoparticle production using only a low heat oven and little technical expertise. Our research demonstrated that heat-curing of PVA/silver nitrate films converted the silver to nanoparticles. These films swelled in water to form a robust, wound-compatible hydrogel which exhibited controlled release of the antibacterial silver nanoparticles. An optimal formulation was obtained using 5% (/) silver nitrate in PVA membrane films that had been heated at 140 °C for 90 min. Physical and chemical characterization of such films was complemented by in vitro studies that confirmed the effective antibacterial activity of the released silver nanoparticles against both gram positive and negative bacteria. Overall, these findings provide economical and simple methods to manufacture stable, hydrogel forming wound dressings that release antibiotic silver over prolonged periods suitable for emergency use in remote locations.
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http://dx.doi.org/10.3390/nano11010096DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7824382PMC
January 2021

The Development of Solvent Cast Films or Electrospun Nanofiber Membranes Made from Blended Poly Vinyl Alcohol Materials with Different Degrees of Hydrolyzation for Optimal Hydrogel Dissolution and Sustained Release of Anti-Infective Silver Salts.

Nanomaterials (Basel) 2021 Jan 3;11(1). Epub 2021 Jan 3.

Faculty of Pharmaceutical Sciences, UBC, 2405 Wesbrook Mall, Vancouver, BC V6T 1Z3, Canada.

Introduction: We previously described the manufacture and characterization of hydrogel forming, thin film, anti-infective wound dressings made from Poly Vinyl Alcohol (PVA) and silver nanoparticles, crosslinked by heat. However, these films were designed to be inexpensive for simple manufacture locally in Africa. In this new study, we have further developed PVA dressings by manufacturing films or electrospun membranes, made from blends of PVA with different degrees of hydrolyzation, that contain silver salts and degrade in a controlled manner to release silver in a sustained manner over 12 days.

Methods: Films were solvent cast as films or electrospun into nanofibre membranes using blends of 99 and 88% hydrolyzed PVA, containing 1% silver sulphadiazine, carbonate, sulphate, or acetate salts. Dissolution was measured as weight loss in water and silver release was measured using inductively coupled plasma (ICP) analysis.

Results: Cast films generally stayed intact at PVA 99: PVA 88% ratios greater than 40:60 whereas electrospun membranes needed ratios greater than 10:90. Films (40:60 blend ratio) and membranes (10:90) all released silver salts in a sustained fashion but incompletely and to different extents. Electrospun membranes gave more linear release patterns in the 2-12 day period and all salts released well.

Conclusion: Blended PVA cast films offer improved control over hydrogel dissolution and silver release without the need for high temperature crosslinking. Blended PVA electrospun membranes further improve membrane dissolution control and silver release profiles. These blended PVA films and membranes offer improved inexpensive systems for the manufacture of long lasting anti-infective hydrogel wound dressings.
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http://dx.doi.org/10.3390/nano11010084DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7824092PMC
January 2021

Meaningful Causal Decompositions in Health Equity Research: Definition, Identification, and Estimation Through a Weighting Framework.

Authors:
John W Jackson

Epidemiology 2021 03;32(2):282-290

From the Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD.

Causal decomposition analyses can help build the evidence base for interventions that address health disparities (inequities). They ask how disparities in outcomes may change under hypothetical intervention. Through study design and assumptions, they can rule out alternate explanations such as confounding, selection bias, and measurement error, thereby identifying potential targets for intervention. Unfortunately, the literature on causal decomposition analysis and related methods have largely ignored equity concerns that actual interventionists would respect, limiting their relevance and practical value. This article addresses these concerns by explicitly considering what covariates the outcome disparity and hypothetical intervention adjust for (so-called allowable covariates) and the equity value judgments these choices convey, drawing from the bioethics, biostatistics, epidemiology, and health services research literatures. From this discussion, we generalize decomposition estimands and formulae to incorporate allowable covariate sets (and thereby reflect equity choices) while still allowing for adjustment of non-allowable covariates needed to satisfy causal assumptions. For these general formulae, we provide weighting-based estimators based on adaptations of ratio-of-mediator-probability and inverse-odds-ratio weighting. We discuss when these estimators reduce to already used estimators under certain equity value judgments, and a novel adaptation under other judgments.
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http://dx.doi.org/10.1097/EDE.0000000000001319DOI Listing
March 2021

Extramammary Borderline Phyllodes Tumor Presenting as an Umbilical Mass.

Int J Surg Pathol 2020 Dec 21:1066896920981632. Epub 2020 Dec 21.

Royal Devon and Exeter Hospital, Exeter, UK.

Phyllodes tumors (PTs) represent a spectrum of rare, fibroepithelial neoplasms of the breast, which can be subcategorized as benign, borderline, or malignant based on their histological appearance. Accessory breast tissue may present anywhere along the embryological mammary ridge, and at distant locations as aberrant breast tissue. We present the case of a 56-year-old lady with an umbilical mass, thought to represent a strangulated hernia. Sections showed a fibroepithelial tumor with leaf-like ducts, conspicuous mitotic activity (up to 8 per 10 high-power fields), and focal infiltration into fat. Immunohistochemical studies showed diffuse positivity of epithelial cells for estrogen receptor, mammaglobin, GCDFP-15, and CK7. These findings were consistent with a borderline PT. This is the first case report of PT presenting as an umbilical mass, and the first extramammary borderline PT described.
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http://dx.doi.org/10.1177/1066896920981632DOI Listing
December 2020

Oxygen limitation fails to explain upper chronic thermal limits and the temperature size rule in mayflies.

J Exp Biol 2021 01 12;224(Pt 1). Epub 2021 Jan 12.

Stroud Water Research Center, Avondale, PA 19311, USA.

An inability to adequately meet tissue oxygen demands has been proposed as an important factor setting upper thermal limits in ectothermic invertebrates (especially aquatic species) as well as explaining the observed decline in adult size with increased rearing temperature during the immature stages (a phenomenon known as the temperature size rule, or TSR). We tested this by rearing three aquatic insects (the mayflies and two species of the complex) through their entire larval life under a range of temperature and oxygen concentrations. Hyperoxia did not extend upper thermal limits, nor did it prevent the loss of size or fertility experienced near upper chronic thermal limits. At moderate temperatures, the TSR pattern was observed under conditions of hyperoxia, normoxia and hypoxia, suggesting little or no influence of oxygen on this trend. However, for a given rearing temperature, adults were smaller and less fecund under hypoxia as a result of a lowering of growth rates. These mayflies greatly increased the size of their gills in response to lower dissolved oxygen concentrations but not under oxygen-saturated conditions over a temperature range yielding the classic TSR response. Using ommatidium diameter as a proxy for cell size, we found the classic TSR pattern observed under moderate temperature conditions was due primarily to a change in the number of cells rather than cell size. We conclude overall that a failure to meet tissue oxygen demands is not a viable hypothesis for explaining either the chronic thermal limit or TSR pattern in these species.
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http://dx.doi.org/10.1242/jeb.233338DOI Listing
January 2021

A polymeric paste-drug formulation for local treatment of upper tract urothelial carcinoma.

Urol Oncol 2021 03 26;39(3):194.e1-194.e7. Epub 2020 Nov 26.

Vancouver Prostate Centre, Department of Urologic Sciences, University of British Columbia, Vancouver, BC, Canada. Electronic address:

Background: Intravesical instillation of chemo- or immunotherapy is commonly used in bladder cancer. Upper tract urothelial carcinoma (UTUC) shares similar pathological features, but current formulations are not suitable for direct instillation to the upper urinary tract.

Objective: To evaluate in vivo applicability, characteristics and toxicity of ST-UC, a mucoadhesive polymeric paste formulation of gemcitabine, for upper urinary tract instillation.

Material And Methods: Three pigs received 10 ml of ST-UC (100 mg/ml gemcitabine) retrogradely into 1 renal pelvis for pharmacokinetic studies. Four days later, a second injection into the contralateral renal pelvis was followed by serial euthanasia of the pigs and nephroureterectomy after 1, 3, and 6 hours. Adverse effects were monitored. Urine, serum, and tissue gemcitabine concentrations were measured, along with histologic examination of the upper urinary tract.

Results: Retrograde instillation of ST-UC was well tolerated with mild, completely receding hydronephrosis. Urine gemcitabine concentrations were highest in the first 3-hour collection interval. Hundred percent of gemcitabine was recovered in the urine within 24 hours. Serum peak concentrations (c) of gemcitabine were low at 5.5 µg/ml compared to the 10 to 30 µg/ml levels observed after a single intravenous dose of 1,000 mg/m gemcitabine. The formulation was still traceable after one hour and gemcitabine tissue concentrations are supportive of this extended drug exposure. No major histopathological changes were observed. The main limitation of this study is the lack of antitumor activity data.

Conclusion: This preclinical evaluation of ST-UC demonstrated feasible instillation in the renal pelvis, no significant safety concerns, and sustained release of gemcitabine.
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http://dx.doi.org/10.1016/j.urolonc.2020.07.028DOI Listing
March 2021

Author Correction: Multicellular 3D Neurovascular Unit Model for Assessing Hypoxia and Neuroinflammation Induced Blood-Brain Barrier Dysfunction.

Sci Rep 2020 Nov 18;10(1):20384. Epub 2020 Nov 18.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, 27101, USA.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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http://dx.doi.org/10.1038/s41598-020-77348-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7673116PMC
November 2020

Meta-analysis of Total Effect Decomposition in the Presence of Multiple Mediators: The Example of Schizophrenia Treatment.

Epidemiology 2021 01;32(1):120-130

Department of Biostatistics, Columbia Mailman School of Public Health, New York, New York.

Background: Causal mediation analysis addresses mechanistic questions by decomposing and quantifying effects operating through different pathways. Because most individual studies are underpowered to detect mediating effects, we outlined a parametric approach to meta-analyzing causal mediation and interaction analyses with multiple mediators, compared it with a bootstrap-based alternative, and discussed its limitations.

Methods: We employed fixed- and random-effects multivariate meta-analyses to integrate evidence on treatment-mediators and mediators-outcome associations across trials. We estimated path-specific effects as functions of meta-analyzed regression coefficients; we obtained standard errors using the delta method. We evaluated the performance of this approach in simulations and applied it to assess the mediating roles of positive symptoms of schizophrenia and weight gain in the treatment effect of paliperidone ER on negative symptoms across four efficacy trials.

Results: Both simulations and the application showed that the meta-analytic approaches increased statistical power. In the application, we observed substantial mediating effects of positive symptoms (proportions mediated from fixed-effects meta-analysis: (Equation is included in full-text article.)). Weight gain may have beneficial mediating effects; however, such benefit may disappear at high doses when metabolic side effects were excessive.

Conclusions: Meta-analyzing causal mediation analysis combines evidence from multiple sources and improves power. Targeting positive symptoms may be an effective way to reduce negative symptoms that are challenging to treat. Future work should focus on extending the existing methods to allow for more flexible modeling of mediation.
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http://dx.doi.org/10.1097/EDE.0000000000001269DOI Listing
January 2021

Transcriptomic and life history responses of the mayfly Neocloeon triangulifer to chronic diel thermal challenge.

Sci Rep 2020 11 5;10(1):19119. Epub 2020 Nov 5.

Graduate Toxicology Program, Department of Biological Sciences, North Carolina State University, 850 Main Campus Drive, Rm1112, Raleigh, NC, 27695, USA.

To better understand the effects of transient thermal stress in an aquatic insect, we first identified static temperatures associated with fitness deficits, and then reared larvae from egg hatch to adulthood under diurnally variable regimens including daily forays into deleterious temperatures. We sampled mature larvae at the coolest and warmest portions of their respective regimens for RNA-seq analysis. Few transcripts (28) were differentially expressed when larvae oscillated between favorable temperatures, while 614 transcripts were differentially expressed when experiencing daily transient thermal stress. Transcripts associated with N-glycan processing were downregulated while those associated with lipid catabolism and chitin turnover were significantly upregulated in heat stressed larvae. An across-regimen comparison of differentially expressed transcripts among organisms sampled at comparable temperatures demonstrated that the effects of daily thermal stress persisted even when larvae were sampled at a more optimal temperature (806 differentially expressed transcripts). The chronically stressed population had reduced expression of transcripts related to ATP synthesis, mitochondrial electron chain functions, gluconeogenesis and glycolytic processes while transcripts associated with cell adhesion, synaptic vesicle transport, regulation of membrane potential and lipid biosynthesis increased. Comparisons of constant vs. variable temperatures revealed that the negative consequences of time spent at stressful temperatures were not offset by more time spent at optimal temperatures.
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http://dx.doi.org/10.1038/s41598-020-75064-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7644658PMC
November 2020

Standardized data to support conservation prioritization for sharks and batoids (Elasmobranchii).

Data Brief 2020 Dec 24;33:106337. Epub 2020 Sep 24.

Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark.

We collated and synthesized information on 1,226 Elasmobranch species (i.e., sharks, rays, and skates) globally from a wide range of sources. We obtained curated and standardized data from online databases, legal documents, press releases, and websites. All data were standardized according to the taxonomic nomenclature described in the Catalogue of Life. We grouped data into five categories: 1) biological information, 2) conservation status, 3) management opportunities, 4) use, and 5) inclusion in international conventions and treaties. For species biological information, we included migration, habitat, species characteristics such as length & body weight, their threat to humans, life-history trait data availability from FishBase, whether the species was listed on the Global Register of Migratory Species, the presence of occurrence data from the Global Biodiversity Information Facility (GBIF), information on genomics from GenBank, and species evolutionary distinctiveness scores. For conservation status, we recorded threat status from the International Union for Conservation of Nature Red List of Threatened Species™ and inclusion in the Alliance for Zero Extinction (AZE). For management opportunities, we identified species under human care in zoos and aquariums in the Species360 network, species under management in studbooks from the European Association of Zoos and Aquaria (EAZA), the American Association for Zoos and Aquariums (AZA), and the Zoo and Aquarium Association Australasia (ZAA), as well as data on recovery, management, and action plans at the class, family, and species levels. For use, we collated species-level data on international trade levels from the CITES (the Convention on International Trade in Endangered Species of Wild Fauna and Flora) Trade Database, as used in aquaculture, as bait, and as gamefish, recording the purpose of the trade according to the IUCN Red List and the global catches reported to the FAO (Food and Agriculture Organization of the United Nations). Finally, we collated information from seven international conventions and treaties: CITES, UNCLOS (the United Nations Convention for the Law of the Sea), CMS (the Convention on the Conservation of Migratory Species of Wild Animals), Shark MoU (the Memorandum of Understanding on the Conservation of Migratory Sharks), BERN (the Convention on the Conservation of European Wildlife and Natural Habitats), OSPAR (Protecting and conserving the North-East Atlantic and its resources), and the Barcelona Convention for the Protection of the Marine Environment and the Coastal Region of the Mediterranean. Our data are comparable across databases and will assist further research on in-situ and ex-situ population management for sharks and batoids. Our data can be of use to international policy makers, aquarium curators, management authorities, conservation practitioners, and scientists interested in prioritizing Elasmobranchs for conservation.
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http://dx.doi.org/10.1016/j.dib.2020.106337DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7554021PMC
December 2020

Induction immunosuppression and the risk of incident malignancies among older and younger kidney transplant recipients: A prospective cohort study.

Clin Transplant 2020 12 20;34(12):e14121. Epub 2020 Nov 20.

Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.

Background: Older (≥65) KT recipients differ from their younger counterparts in their immune response to immunosuppression (IS) and may have a different risk of malignancy after receiving induction.

Methods: We identified 66 700 adult KT recipients treated with anti-thymocyte globulin (ATG) (n = 40 443) or interleukin-2 receptor antagonist (IL-2RA) (n = 26 327) induction (1/1/1999-12/31/2014) using USRDS/Medicare data. We estimated the risk of first-diagnosed post-KT malignancy associated with induction (ATG vs. IL-2RA) using Cox proportional hazard models. We then tested whether these risks differed between older and younger recipients (Wald test for interaction). Models incorporated inverse probability of treatment weights to adjust for confounders.

Results: The 3-year cumulative incidences of any diagnosed malignancy were 11.5%. ATG was associated with a higher malignancy risk (HR = 1.12, 95%CI:1.06-1.18). This association differed (p  = 0.04) between younger (HR = 1.12, 95%CI:1.06-1.18) and older recipients (HR = 1.03, 95%CI:0.96-1.09). ATG was also associated with higher risk of skin (HR = 1.18, 95%CI:1.08-1.29), lung (HR = 1.24, 95%CI:1.05-1.47), and ovary malignancies (HR = 1.94, 95%CI:1.08-3.48). However, only the association of ATG with post-KT skin malignancy differed (p  = 0.01) between younger (HR = 1.18; 95%CI:1.08-1.29) and older (HR = 1.01; 95%CI:0.93-1.09) recipients.

Conclusions: Compared with IL-2RA induction, ATG was associated with elevated post-KT malignancy risk but only among younger recipients. Transplant centers may need to tailor induction IS for younger recipients to mitigate malignancy risk.
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http://dx.doi.org/10.1111/ctr.14121DOI Listing
December 2020

Part 1. Evaluation of Epigallocatechin Gallate or Tannic Acid Formulations of Hydrophobic Drugs for Enhanced Dermal and Bladder Uptake or for Local Anesthesia Effects.

J Pharm Sci 2021 02 8;110(2):796-806. Epub 2020 Oct 8.

Faculty of Pharmaceutical Sciences, University of British Columbia, 2045 Wesbrook Mall, Vancouver, BC, Canada.

Epigallocatechin gallate (EGCG) and tannic acid (TA) are known to increase the aqueous solubility and cellular uptake of the hydrophobic drugs docetaxel, paclitaxel, amphotericin B, and curcumin. In this study the practical application of gallate-based solubilization phenomena for the uptake of these drugs into dermal and bladder tissue and of lidocaine for wound healing application was studied. The penetration of all these drugs into pig skin or docetaxel into pig bladder using EGCG or TA formulations was measured. Overall, EGCG and TA particulate or propylene glycol paste formulations of drugs allowed for greatly increased levels of drug uptake into skin as compared to control formulations. EGCG/propylene glycol pastes allowed for rapid lidocaine uptake into skin. EGCG and TA formulations of docetaxel allowed for approximately 10 fold increases in bladder tissue uptake of docetaxel over tween based solutions. Morphologically, both EGCG and TA caused a mild, dose dependent exfoliation of the bladder wall. Both EGCG and TA formed injectable viscous pastes with propylene glycol which solidified in water and degraded and released lidocaine over 2-35 days. These data support the use of EGCG and TA based formulations of certain drugs for improved dermal, bladder and wound applications.
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http://dx.doi.org/10.1016/j.xphs.2020.10.001DOI Listing
February 2021

Paper 2. Epigallocatechin Gallate and Tannic Acid Based Formulations of Finasteride for Dermal Administration and Chemoembolization.

J Pharm Sci 2021 02 6;110(2):807-814. Epub 2020 Oct 6.

Faculty of Pharmaceutical Sciences, University of British Columbia, 2045 Wesbrook Mall, Vancouver, BC V6T1Z3, Canada.

Finasteride is used to treat benign prostatic hyperplasia (BPH) and pattern hair loss (androgenetic alopecia or APA). The local administration of formulations with increased solubility and controlled release of finasteride are proposed using gallate-containing compositions within embolic microparticles or paste. Finasteride solubility in either epigallocatechin gallate (EGCG) or tannic acid (TA) solutions was assessed using HPLC. Poly(dl-lactide-co-glycolide) (PLGA) or poly(methylmethacrylate) (PMMA) microspheres (100-400 μm) containing finasteride and EGCG or TA were effectively manufactured. Embolic particles were loaded with finasteride/EGCG/TA. Dermal uptake of TA/EGCG/finasteride topical compositions was measured in pig skin. The solubility of finasteride was dramatically increased using EGCG- or TA-based compositions. Finasteride loaded microspheres released over two months which was increased by EGCG or TA inclusion. Embolic particles soaked up finasteride and EGCG or TA and released the encapsulated drug over two weeks. Dermal uptake of finasteride from EGCG- or TA-based formulations was enhanced between 10 and 50 fold in layers as deep as 500 μm when compared to a generic control formulation. Gallate-based formulations of finasteride increase drug solubility and allow for effective release of the drug from embolic formulations. Paste or powder EGCG- or TA-based formulations of finasteride greatly increase dermal penetration of the drug.
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http://dx.doi.org/10.1016/j.xphs.2020.09.041DOI Listing
February 2021

Structural and Social Determinants of Health Factors Associated with County-Level Variation in Non-Adherence to Antihypertensive Medication Treatment.

Int J Environ Res Public Health 2020 09 14;17(18). Epub 2020 Sep 14.

College of Public Health, Ohio State University, Columbus, OH 43210, USA.

Background: Non-adherence to antihypertensive medication treatment (AHM) is a complex health behavior with determinants that extend beyond the individual patient. The structural and social determinants of health (SDH) that predispose populations to ill health and unhealthy behaviors could be potential barriers to long-term adherence to AHM. However, the role of SDH in AHM non-adherence has been understudied. Therefore, we aimed to define and identify the SDH factors associated with non-adherence to AHM and to quantify the variation in county-level non-adherence to AHM explained by these factors.

Methods: Two cross-sectional datasets, the Centers for Disease Control and Prevention (CDC) Atlas of Heart Disease and Stroke (2014-2016 cycle) and the 2016 County Health Rankings (CHR), were linked to create an analytic dataset. Contextual SDH variables were extracted from the CDC-CHR linked dataset. County-level prevalence of AHM non-adherence, based on Medicare fee-for-service beneficiaries' claims data, was extracted from the CDC Atlas dataset. The CDC measured AHM non-adherence as the proportion of days covered (PDC) with AHM during a 365 day period for Medicare Part D beneficiaries and aggregated these measures at the county level. We applied confirmatory factor analysis (CFA) to identify the constructs of social determinants of AHM non-adherence. AHM non-adherence variation and its social determinants were measured with structural equation models.

Results: Among 3000 counties in the U.S., the weighted mean prevalence of AHM non-adherence (PDC < 80%) in 2015 was 25.0%, with a standard deviation (SD) of 18.8%. AHM non-adherence was directly associated with poverty/food insecurity (β = 0.31, -value < 0.001) and weak social supports (β = 0.27, -value < 0.001), but inversely with healthy built environment (β = -0.10, -value = 0.02). These three constructs explained one-third ( = 30.0%) of the variation in county-level AHM non-adherence.

Conclusion: AHM non-adherence varies by geographical location, one-third of which is explained by contextual SDH factors including poverty/food insecurity, weak social supports and healthy built environments.
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http://dx.doi.org/10.3390/ijerph17186684DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7557537PMC
September 2020

Examining Disparities in Excessive Alcohol Use Among Black and Hispanic Lesbian and Bisexual Women in the United States: An Intersectional Analysis.

J Stud Alcohol Drugs 2020 07;81(4):462-470

Departments of Epidemiology and Health Policy and Management, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland.

Objective: Sexual minority (i.e., lesbian, bisexual) women and racial-ethnic minority groups in the United States are disproportionately harmed by excessive alcohol use. This study examined disparities in excessive alcohol use at the intersection of race-ethnicity and sexual identity for non-Hispanic Black and Hispanic sexual minority women.

Method: Using data from the 2015 National Survey on Drug Use and Health, we compared the age-adjusted prevalence of binge drinking and heavy alcohol use among sexual minority women of color, sexual minority White women, and heterosexual women of color with that of White heterosexual women. The joint disparity is the difference in the prevalence of excessive alcohol use between sexual minority women of color and White heterosexual women. The excess intersectional disparity is the portion of the joint disparity that is due to being both a racial-ethnic minority and a sexual minority woman.

Results: Black and Hispanic sexual minority women reported the highest prevalence of binge drinking (45.4% and 43.4%, respectively), followed by White sexual minority women (35.7%) and White heterosexual women (23%). Black and Hispanic heterosexual women reported the lowest prevalence of binge drinking (20.8% and 20.2%, respectively). The joint disparity in binge drinking between Black sexual minority women and White heterosexual women was 21.2%, and the excess intersectional disparity was 17.7%. The joint disparity in binge drinking between Hispanic sexual minority women and White heterosexual women was 16.8%, and the excess intersectional disparity was 10.8%.

Conclusions: Disparities in excessive alcohol consumption for Black and Hispanic sexual minority women, compared with White heterosexual women, were larger than what would be expected when considering differences by race or sexual identity individually.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7437553PMC
July 2020

Strategies to promote access to medications during the COVID-19 pandemic.

Aust J Gen Pract 2020 08;49(8):530-532

BPharm, GradDipHospPharm, MPH, Director: Project Pharmacist, Centre for Medicine Use and Safety, Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Vic.

Background: During the COVID-19 pandemic, vulnerable and older people with chronic and complex conditions have self-isolated in their homes, potentially limiting opportunities for consultations to have medications prescribed and dispensed.

Objective: The aim of this article is to describe initiatives to ensure ongoing access to medications during the COVID-19 pandemic.

Discussion: Cooperation between wholesalers and purchase limits in pharmacies have helped to ensure supply of essential medications. Therapeutic substitution by pharmacists is permitted for specific products authorised by the Therapeutic Goods Administration. Prescribers are permitted to issue digital image prescriptions, and implementation of electronic prescribing has been fast-tracked. Expanded continued dispensing arrangements introduced during the bushfire crises have been temporarily extended. Pharmacists are permitted to provide medication management reviews via telehealth. A Home Medicines Service has been introduced to facilitate delivery of medications to people who are vulnerable or elderly. Anticipatory prescribing and medication imprest systems are valuable for access to end-of-life medications within residential aged care.
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http://dx.doi.org/10.31128/AJGP-04-20-5390DOI Listing
August 2020

Formation and optimization of three-dimensional organoids generated from urine-derived stem cells for renal function in vitro.

Stem Cell Res Ther 2020 07 22;11(1):309. Epub 2020 Jul 22.

Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston Salem, NC, USA.

Background: Organoids play an important role in basic research, drug screening, and regenerative medicine. Here, we aimed to develop a novel kind of three-dimensional (3D) organoids generated from urine-derived stem cells (USCs) and to explore whether kidney-specific extracellular matrix (kECM) could enable such organoids for renal function in vitro.

Methods: USCs were isolated from human urine samples and cultured with kECM extraction to generate 3D organoids in vitro. Eight densities from 1000 to 8000 cells per organoids were prepared, and both ATP assay and Live/Dead staining were used to determine the optimal USC density in forming organoids and kECM additive concentration. The morphology and histology of as-made organoids were evaluated by hematoxylin and eosin (H.E.) staining, immunofluorescence staining and whole mount staining. Additionally, RT-qPCR was implemented to detect renal-related gene expression. Drug toxicity test was conducted to evaluate the potential application for drug screening. The renal organoids generated from whole adult kidney cells were used as a positive control in multiple assessments.

Results: The optimized cell density to generate ideal USC-derived organoids (USC-organoids) was 5000 cells/well, which was set as applying density in the following experiments. Besides, the optimal concentration of kECM was revealed to be 10%. On this condition, Live/Dead staining showed that USC-organoids were well self-organized without significant cell death. Moreover, H.E. staining showed that compact and viable organoids were generated without obvious necrosis inside organoids, which were very close to renal organoids morphologically. Furthermore, specific proximal tubule marker Aquaporin-1 (AQP1), kidney endocrine product erythropoietin (EPO), kidney glomerular markers Podocin and Synaptopodin were detected positively in USC-organoids with kECM. Nephrotoxicity testing showed that aspirin, penicillin G, and cisplatin could exert drug-induced toxicity on USC-organoids with kECM.

Conclusions: USC-organoids could be developed from USCs via an optimal procedure. Combining culture with kECM, USC-organoid properties including morphology, histology, and specific gene expression were identified to be similar with real renal organoids. Additionally, USC-organoids posed kECM in vitro showed the potential to be a drug screening tool which might take the place of renal organoids to some extent in the future.
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http://dx.doi.org/10.1186/s13287-020-01822-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7374873PMC
July 2020

From Inflammation to Cutaneous Repair: Topical Application of Lupeol Improves Skin Wound Healing in Rats by Modulating the Cytokine Levels, NF-κB, Ki-67, Growth Factor Expression, and Distribution of Collagen Fibers.

Int J Mol Sci 2020 Jul 13;21(14). Epub 2020 Jul 13.

Department of Morphology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu 18618-689, São Paulo, Brazil.

Skin wound healing is a highly complex event that involves different mediators at the cellular and molecular level. Lupeol has been reported to possess different biological activities, such as anti-inflammatory, antioxidant, antidiabetic, and in vitro wound healing properties, which motivated us to proceed with in vivo studies. We aimed to investigate the wound healing effect of lupeol-based cream for 3, 7, and 14 days. Wound excisions were induced on the thoraco-lumbar region of rats and topically treated immediately after injury induction. Macroscopic, histopathological, and immunohistochemical analyses were performed. Cytokine levels were measured by ELISA and gene expression was evaluated by real-time RT-qPCR. Our results showed a strong wound-healing effect of lupeol-based cream after 7 and 14 days. Lupeol treatment caused a reduction in proinflammatory cytokines (TNF-a, IL-1β, and IL-6) and gene and protein NF-κB expression, and positively altered IL-10 levels, showing anti-inflammatory effects in the three treatment periods. Lupeol treatment showed involvement in the proliferative phase by stimulating the formation of new blood vessels, increasing the immunostaining of Ki-67 and gene expression, and immunolabeling of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), and increasing gene expression of transforming growth factor beta-1 (TGF-β1) after seven days of treatment. Lupeol was also involved in the tissue regeneration phase by increasing the synthesis of collagen fibers noted in the three treatment periods analyzed. Our findings suggest that lupeol may serve as a novel therapeutic option to treat cutaneous wounds by regulating mechanisms involved in the inflammatory, proliferative, and tissue-remodeling phases.
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http://dx.doi.org/10.3390/ijms21144952DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7404060PMC
July 2020

Administration of secretome from human placental stem cell-conditioned media improves recovery of erectile function in the pelvic neurovascular injury model.

J Tissue Eng Regen Med 2020 10 31;14(10):1394-1402. Epub 2020 Jul 31.

Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC, USA.

Human placental stem cells (PSCs) enhance histological and functional recovery in a rodent erectile dysfunction (ED) model. We tested the hypothesis that bioactive factors secreted by PSC (i.e., the secretome) mediate functional recovery and that acellular-conditioned media (CM) from PSC culture (PSC-CM) could be used independently to facilitate functional and histological recovery. To identify factors relative to efficacy of PSC, a comparison of CM from PSC and three additional human stem cell populations was performed. CM from human PSC, amniotic fluid stem cells (AFSCs), adipose-derived stem cells (ADSC), and human umbilical vein endothelial cells (HUVECs) was assayed using a semi-quantitative human cytokine antibody array. Male rats, after surgically created ED by neurovascular injury, were randomly divided into four groups: vehicle control (phosphate-buffered saline [PBS]), PSC, PSC-CM, and serum-free media control (SFM) as control. Functional data on intracorporal and mean arterial pressure were obtained, and histological architecture was examined 6 weeks after single injection. PSCs were found to secrete at least 27 cytokines and growth factors at a significantly higher level than the other three cell types. Either single injection of PSC-CM or PSC significantly improved erectile functional recovery and histological architecture compared with SFM or PBS. Injection of the secretome isolated from human PSC improves erectile functional recovery and histological structure in a rat model of neurovascular injury-induced ED. Further characterization of the unique protein expression within the PSC-CM may help to identify the potential for a novel injectable cell-free therapeutic for applicable patients.
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http://dx.doi.org/10.1002/term.3105DOI Listing
October 2020

Changes in age-structure over four decades were a key determinant of population growth rate in a long-lived mammal.

J Anim Ecol 2020 10 17;89(10):2268-2278. Epub 2020 Jul 17.

Department of Biology, University of Turku, Turku, Finland.

A changing environment directly influences birth and mortality rates, and thus population growth rates. However, population growth rates in the short term are also influenced by population age-structure. Despite its importance, the contribution of age-structure to population growth rates has rarely been explored empirically in wildlife populations with long-term demographic data. Here we assessed how changes in age-structure influenced short-term population dynamics in a semi-captive population of Asian elephants Elephas maximus. We addressed this question using a demographic dataset of female Asian elephants from timber camps in Myanmar spanning 45 years (1970-2014). First, we explored temporal variation in age-structure. Then, using annual matrix population models, we used a retrospective approach to assess the contributions of age-structure and vital rates to short-term population growth rates with respect to the average environment. Age-structure was highly variable over the study period, with large proportions of juveniles in the years 1970 and 1985, and made a substantial contribution to annual population growth rate deviations. High adult birth rates between 1970 and 1980 would have resulted in large positive population growth rates, but these were prevented by a low proportion of reproductive-aged females. We highlight that an understanding of both age-specific vital rates and age-structure is needed to assess short-term population dynamics. Furthermore, this example from a human-managed system suggests that the importance of age-structure may be accentuated in populations experiencing human disturbance where age-structure is unstable, such as those in captivity or for endangered species. Ultimately, changes to the environment drive population dynamics by influencing birth and mortality rates, but understanding demographic structure is crucial for assessing population growth.
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http://dx.doi.org/10.1111/1365-2656.13290DOI Listing
October 2020

Multicellular 3D Neurovascular Unit Model for Assessing Hypoxia and Neuroinflammation Induced Blood-Brain Barrier Dysfunction.

Sci Rep 2020 06 17;10(1):9766. Epub 2020 Jun 17.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, 27101, USA.

The blood-brain barrier (BBB) is a dynamic component of the brain-vascular interface that maintains brain homeostasis and regulates solute permeability into brain tissue. The expression of tight junction proteins between adjacent endothelial cells and the presence of efflux proteins prevents entry of foreign substances into the brain parenchyma. BBB dysfunction, however, is evident in many neurological disorders including ischemic stroke, trauma, and chronic neurodegenerative diseases. Currently, major contributors to BBB dysfunction are not well understood. Here, we employed a multicellular 3D neurovascular unit organoid containing human brain microvascular endothelial cells, pericytes, astrocytes, microglia, oligodendrocytes and neurons to model the effects of hypoxia and neuroinflammation on BBB function. Organoids were cultured in hypoxic chamber with 0.1% O for 24 hours. Organoids cultured under this hypoxic condition showed increased permeability, pro-inflammatory cytokine production, and increased oxidative stress. The anti-inflammatory agents, secoisolariciresinol diglucoside and 2-arachidonoyl glycerol, demonstrated protection by reducing inflammatory cytokine levels in the organoids under hypoxic conditions. Through the assessment of a free radical scavenger and an anti-inflammatory endocannabinoid, we hereby report the utility of the model in drug development for drug candidates that may reduce the effects of ROS and inflammation under disease conditions. This 3D organoid model recapitulates characteristics of BBB dysfunction under hypoxic physiological conditions and when exposed to exogenous neuroinflammatory mediators and hence may have potential in disease modeling and therapeutic development.
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http://dx.doi.org/10.1038/s41598-020-66487-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7299970PMC
June 2020

Decomposing Race and Ethnic Differences in CVD Risk Factors for Mid-life Women.

J Racial Ethn Health Disparities 2021 Feb 27;8(1):174-185. Epub 2020 May 27.

General Preventative Medicine Resident, Preventive Medicine Residency Program, Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street Room WB602, Baltimore, MD, 21205, USA.

Objective: This study decomposes race and ethnic differences in hypertension, waist circumference, obesity and allostatic load between black non-Hispanic (BNH), Mexican American (MA), and white non-Hispanic (WNH) women.

Data: This study uses 10,109 observations from The National Health and Nutrition Examination Survey from years 1999-2014 for BNH, MA women, and WNH between 40 and 75 years old.

Methodology: We used the Oaxaca-Blinder decomposition to explore how demographic, socioeconomic, healthcare access, and health behavior factors are associated with race and ethnic differences in blood pressure, waist circumference, body mass index (BMI), and allostatic load score (ALS).

Results: We found that demographic factors, socioeconomic status, healthcare access, and health behaviors explained from 0 to 50% of the difference in CVD risk factors between BNH and WNH. However, these factors explain from 39 to 100% of the difference in CVD risk factors between MA and WNH. Differences in demographic, socioeconomic, access to care, and health behavior factor variables explained very little of the differences in CVD risk factors between NHB and MA women.

Conclusion: The impact of the determinants on CVD risk factors varies by race and ethnicity. Efforts to address differences in CVD risk factors should promote health equity programs and acknowledge that even race and ethnic groups that have similar demographic, SES, access to care, and health behavior factors can have different outcomes.
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http://dx.doi.org/10.1007/s40615-020-00769-9DOI Listing
February 2021