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Hawraa Ali-Jerman,1 Zainab Al-Quraishi,1 Ashish Muglikar,1 Yvonne Perrie,1 Rothwelle J Tate,1 Margaret Mullin,2 Gayle McNeill,1 Graeme Mackenzie,1 Christine Dufès1 1Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G4 0RE, UK; 2Cell Analysis Facility, Medical and Veterinary & Life Sciences Shared Research Facilities, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8QQ, UKCorrespondence: Christine Dufès, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0RE, UK, Tel +44 1415483796, Fax +44 1415522562, Email C.Dufes@strath.ac.ukBackground: Gene therapy is a promising therapeutic approach for treating various disorders by introducing modified nucleic acids to correct cellular dysfunctions or introduce new functions. Despite significant advancements in the field, the effective delivery of nucleic acids remains a challenge, due to biological barriers and the immune system’s ability to target and destroy these molecules. Due to their branched structure and ability to condense negatively charged nucleic acids, cationic dendrimers have shown potential in overcoming these challenges. Despite this, standardized scalable production methods are still lacking. This study investigates the use of microfluidics to formulate generation 3-diaminobutyric polypropylenimine (DAB) dendriplexes and compares their characteristics and in vitro gene delivery efficacy to those prepared using conventional manual mixing.Methods: DAB dendriplexes were produced by both microfluidic and manual approaches and characterized. Their cellular uptake and gene expression were evaluated on C6 glioma cancer cells in vitro.Results: Dendriplexes formed using microfluidics at the optimal flow rate and ratio demonstrated enhanced DNA condensation over time, achieving up to 97% condensation at 24 hours. Both preparation methods produced positively charged dendriplexes, indicating stable formulations. However, dendriplexes prepared through hand mixing resulted in smaller particle sizes, significantly higher cellular uptake and gene expression efficacy compared to those prepared by microfluidics. Nonetheless, microfluidic preparation offers the advantage of standardized and scalable production, which is essential for future applications.Conclusion: This study highlights the potential of microfluidic technology to improve precision and scalability in gene delivery, paving the way for future advancements in gene therapy. Our findings suggest that, with further optimization, microfluidic systems could provide superior control over dendriplex formation, expanding their potential use in gene therapy applications. Keywords: polypropylenimine dendrimer, dendriplex, glioma, microfluidics, cellular uptake, transfection

Introduction: This study aimed to systematically identify the environmental factors that impacted people with disability during the COVID-19 pandemic. Methods: A scoping literature review was conducted using LitCOVID (January 1–July 31, 2020). Sixty-six articles met the inclusion criteria that (1) discussed disability and/or health conditions related to functioning and (2) considered environmental factors. A qualitative content analysis was conducted using codes from the WHO International Classification of Functioning, Disability and Health. Results: A total of 212 International Classification of Functioning, Disability and Health codes were used in the coding process. The most frequent codes referred to health services policies and public health guidelines. These policies, although generally considered facilitators for minimizing infection, were frequently identified as barriers to the health, participation, and human rights of people with disability. The lack of disability-specific population data was identified as a key barrier to planning and decision making. Conclusions: The social determinants of health for people with disability were not adequately considered in the acute phase of infection prevention at the population level. Integrating the International Classification of Functioning, Disability and Health in emergency management provides a tool to evaluate functioning and address barriers for those in need.

Copper (Cu) is typically adequate at 0.5 μM (0.03 ppm) in hydroponics and at 2 μM (0.125 ppm) in soilless media, but elevated levels can be used to inhibit pathogenic fungal growth. We studied the effect of elevated Cu on the growth of lettuce and tomato in peat-based media and deep-flow hydroponics. Lettuce growth in hydroponics was not hindered until a concentration greater than 4 μM (0.25 ppm) Cu was used, which is eight times greater than the adequate level. Tomato was more tolerant of elevated Cu, with no growth suppression up to 8 μM (0.5 ppm) in hydroponics. Organic matter tightly binds Cu, and bioavailability is thus determined by organic components in soilless media. We confirmed an adsorption capacity of 19 mg Cu per g of peat, which explains why there was no inhibition of lettuce or tomato growth up to 1000 μM (64 ppm) Cu in peat-based media. When chelated with ethylenediaminetetraacetic acid, Cu binding to organic matter was reduced and growth was decreased in lettuce but not tomato. Both species tolerated a 100-fold greater concentration of Cu in peat-based media than in deep-flow hydroponics. Elevated Cu in solution increased concentrations 20 times greater in root tissue than in leaves. These solution and tissue concentrations are greater than identified toxicity thresholds of pathogenic fungal and fungal-like organisms, and could thus be used to suppress root-borne fungal and fungal-like diseases.

Silicon (Si) is a beneficial element that is usually ample in mineral soil solution, but it is minimally bioavailable from soilless substrates. Several Si additives are commercially available, but the rate of dissolution of Si is not well-characterized. The ideal additive would steadily release bioavailable Si over the crop lifecycle. We report the long-term (120 days) dissolution of Si from soilless substrates and substrate additives. Studies involving gently agitated containers with deionized water indicated that perlite, sphagnum peat, vermiculite, and coconut coir released less than 0.03 mmol Si per liter of substrate per day. Rice hulls and wollastonite (CaSiO3) had 7- to 130-times faster rates of dissolution in this system; therefore, they were further studied in peat-based media. Dissolution of Si from the addition of 1 g wollastonite per liter of peat peaked at day 10 at 2.1 mmol Si per liter of media per leaching event (15% by volume); then, it gradually decreased over 120 days. The peak dissolution of Si amended with 12% rice hulls was similar, but it gradually increased over time. The concentrations of nine heavy metals in plant tissue were compared with untreated control plants to determine wollastonite and steel slag. The concentration of some elements statistically increased, but all concentrations were well below the legal concentration limits of these elements for human consumption in the United States. These results indicate that both wollastonite and rice hulls steadily release Si for up to 4 months; therefore, they are good sources of Si for container-grown crops in soilless media.

Jitkasem Meewan,1 Sukrut Somani,1 Jamal Almowalad,1 Partha Laskar,1 Margaret Mullin,2 Graeme MacKenzie,1 Swapnil Khadke,1 Yvonne Perrie,1 Christine Dufès1 1Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G4 0RE, UK; 2Glasgow Imaging Facility, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8QQ, UKCorrespondence: Christine Dufès, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G4 0RE, UK, Tel +44 1415483796, Fax +44 1415522562, Email C.Dufes@strath.ac.ukBackground: The manufacture of nanoparticles using manual methods is hampered by its challenging scale-up and poor reproducibility. To overcome this issue, the production of zein nanoparticles entrapping a lipophilic drug model, coumarin-6, by using a microfluidic system was assessed in this study. The influence of PEG density and chain length on zein nanoparticle characteristics, as well as their uptake efficacy in melanoma cancer cells, was also evaluated.Methods: Zein nanoparticles were prepared by both manual and microfluidic approaches to allow comparison between the two processes. PEGylated zein nanoparticles with various PEG densities and chain lengths were produced by nanoprecipitation and characterized. Their cellular uptake was evaluated on B16F10 melanoma cancer cells in vitro.Results: Zein nanoparticles have successfully been produced by both manual and microfluidic approaches. Parameters such as total flow rate and flow rate ratio of the aqueous and organic phases in microfluidic process, as well as the method preparation and aqueous to organic phase volume ratio during nanoprecipitation, have been shown to strongly influence the characteristics of the resulting nanoparticles. Continuous microfluidics led to the production of nanoparticles with low yield and drug entrapment, unlike nanoprecipitation, which resulted in zein nanoparticles with an appropriate size and an optimal drug entrapment efficiency of 64%. The surface modification of the nanoparticles produced by nanoprecipitation, with lower PEG density and shorter PEG chain length made mPEG5K-zein (0.5:1) the most favorable formulation in our study, resulting in enhanced stability and higher coumarin-6 uptake by melanoma cancer cells.Conclusion: mPEG5K-zein (0.5:1) nanoparticles prepared by nanoprecipitation were the most promising formulation in our study, exhibiting increased stability and enhancing coumarin-6 uptake by melanoma cancer cells.Graphical Abstract: Keywords: zein nanoparticles, microfluidics, nanoprecipitation, poly(ethylene glycol), cellular uptake

We performed steered molecular dynamics (SMD) and umbrella sampling simulations of Cl− ion migration through the transmembrane domain of a prototypical E. coli CLC Cl−/H+ antiporter by employing combined quantum-mechanical (QM) and molecular-mechanical (MM) calculations. The SMD simulations revealed interesting conformational changes of the protein. While no large-amplitude motions of the protein were observed during pore opening, the side chain rotation of the protonated external gating residue Glu148 was found to be critical for full access of the channel entrance by Cl−. Moving the anion into the external binding site (Sext) induced small-amplitude shifting of the protein backbone at the N-terminal end of helix F. As Cl− traveled through the pore, rigid-body swinging motions of helix R separated it from helix D. Helix R returned to its original position once Cl− exited the channel. Population analysis based on polarized wavefunction from QM/MM calculations discovered significant (up to 20%) charge loss for Cl− along the ion translocation pathway inside the pore. The delocalized charge was redistributed onto the pore residues, especially the functional groups containing π bonds (e.g., the Tyr445 side chain), while the charges of the H atoms coordinating Cl− changed almost negligibly. Potentials of mean force computed from umbrella sampling at the QM/MM and MM levels both displayed barriers at the same locations near the pore entrance and exit. However, the QM/MM PMF showed higher barriers (~10 kcal/mol) than the MM PMF (~2 kcal/mol). Binding energy calculations indicated that the interactions between Cl− and certain pore residues were overestimated by the semi-empirical PM3 Hamiltonian and underestimated by the CHARMM36 force fields, both of which were employed in the umbrella sampling simulations. In particular, CHARMM36 underestimated binding interactions for the functional groups containing π bonds, missing the stabilizations of the Cl− ion due to electron delocalization. The results suggested that it is important to explore these quantum effects for accurate descriptions of the Cl− transport.

Abstract Background Immigrants to Germany and their children are at particular risk for tuberculosis (TB). Methods 35 Patients (10 male/25 female aged 2 - 59 years (median 33 years) originating mostly from high incidence countries in Asia (19 [54.3%]) in Africa (14 [40.0%] and East Europe (2 [5.7%]), attended at the Tropical Medicine unit were analysed. Results Primary clinical presentation was most frequently lymphadenitis (13 [37.1%]). other organs involved included bones (7 [20.0%]), central nervous system (5 [14.3%]), urogenital organs (3 [8.6%]), lung (3 [8.6%]), mediastinum, (2 [5.7%]) and abdomen (2 [5.7%]). ESR was abnormal in 21/28 (75.0%), CRP in 20/35 (57.1%), and protein electrophoresis in 22/26 (84.6%) cases. The tuberculin skin test was strongly positive in all 15 cases where the test had been performed. Tuberculosis interferon gamma release assay (TB-IGRA) was positive in all 35 cases (100%). PCR for nucleic acids of Mycobacterium (M.) tuberculosis complex was positive in only 7/20 (35.0%) cases. M. tuberculosis was identified in 32/35 (91.4%), M. bovis in 2 (5.7%) cases. 1 case was diagnosed clinically. All patients were negative for HIV. Typical histopathology was seen in the 29 cases, where biopsies had been taken. Chest-X-ray did not reveal specific pulmonary lesions in the majority of cases (22/35 [62.9%]). Diagnosis of TB was mostly delayed (4 to 299 weeks, [median 8]). The most frequent primary suspicion was a malignancy (17/35 [48.6%]) while TB was initially suspected in 5 cases only. Diagnosis of TB is impeded by its multifaceted presentation especially in immigrants.

Research has found that many individuals who believe they have experienced discrimination do not report the incident to a relevant authority figure. Much of this research has focused on gender- and race-based discrimination, with relatively little research examining religion-based discrimination. Using data from a representative sample of U.S. adults, this research note examines the rate of reporting religious discrimination to authorities and the reasons given for non-reporting. Our analysis finds that only 13 percent of individuals perceiving religious discrimination report the experience to a relevant authority figure. The most common reason for non-reporting is a belief that nothing could be done, although the reasons given for non-reporting depend in part on the specific context or type of discrimination experienced. Moreover, relative to Christians, Muslims are significantly more likely to attribute their non-reporting to a belief that nothing could be done and a fear of personal risks. [ABSTRACT FROM AUTHOR]

A heterocycle-to-heterocycle strategy is presented for the preparation of highly fluorescent and solvatochromic dibenzonaphthyridinones (DBNs) via methodology that leads to the formation of a tertiary, spiro-fused carbon center. A linear correlation between the results of photophysical experiments and time dependent density functional theory calculations was observed for the λ(max) of excitation for DBNs with varying electronic character.

Purpose: To identify meaningful educative experiences that contributed to the development of rehabilitation counselors' abilities to provide effective rehabilitation counseling services to lesbian, gay, bisexual, transgender, and queer (LGBTQ) persons living with chronic illness/disabilities (CID). Method: This was a secondary analysis of a larger qualitative data set that examined the educational experiences and practices of 12 certified rehabilitation counselors working with LGBTQ persons. Results: Using a qualitative content analysis methodology, 5 categories were identified: (a) formal didactic education, (b) affirmative supervision and consultation during practica/internship, (c) extracurricular-related training, (d) identification, and (e) family of origin. Conclusion: To help facilitate future counselors' self-perceived abilities to provide effective services to LGBTQ persons living with CID, rehabilitation counselor educators should place more emphasis on LGBTQ-related issues in their curriculum and provide practical (experiential) learning opportunities for their students.