Abstract: The therapeutic potential of prodigiosin as a hydrophobic anticancer agent can be enhanced by various approaches. One of which is the loading of PG into extracellular vesicles. Both, drug distribution and stability in aqueous media are important for targeting and accumulation, which, in turn, is important to achieve an effective drug concentration. Extracellular vesicles (EVs) are nano-sized, cell-derived vesicles with lipid bilayer membrane. EVs can be utilized as drug carriers for both water-soluble and non-water-soluble therapeutic agents. We hypothesized that EVs could effectively address the current challenges of prodigiosin delivery. Several different techniques have been developed for fabricating extracellular vesicles. These include EVs induction with cytochalasin B, prodigiosin and cultivation in serum depleted media (serum-free cultivation). Treatment with cytochalasin B has been shown to be effective for the generation of vesicles. Prodigiosin encapsulated extracellular vesicles (PG-EVs) compared to unmodified naïve ones demonstrated slightly higher charge and hydrodynamic dimensions. Which probably contributes to better stability. Overall, PG –EVs complexes might be more suitable for medical and clinical applications compared to pure forms of PG due to their cell membrane compatibility

Abstract: Endophytic and marine-derived fungi represent prolific and structurally innovative sources of steroidal natural products. In recent years, extensive chemical investigations of diverse fungal taxa—including Aspergillus, Penicillium, and numerous other genera—have revealed an extraordinary variety of steroids and steroid-like metabolites featuring unprecedented carbon skeletons, unusual ring rearrangements, heterocyclic fusions, and hybrid architectures. These metabolites encompass ergostane-, lanostane-, pregnane-, and abeo-type derivatives; secosteroids; polyoxygenated and polycyclic frameworks; meroterpenoid–steroid hybrids; and rare Diels–Alder adducts. Many of these compounds exhibit significant biological activities, including cytotoxic, anti-inflammatory, antimicrobial, antiviral, immunosuppressive, antioxidant, enzyme-inhibitory, larvicidal, neuroprotective, and herbicidal effects. Several metabolites, such as cordycepsterols, citristerones, rubensteroid A, anicequol, and various hybrid steroidal structures, demonstrate potent inhibition of key molecular targets (e.g., COX-2, NF-κB, PTP1B, AChE, NO production) and show promise as leads for anticancer, anti-infective, anti-neuroinflammatory, and metabolic disease therapeutics. Collectively, the rapidly expanding diversity of fungal steroids underscores the remarkable biosynthetic capabilities of fungi and highlights their continued potential as reservoirs of structurally novel and biologically valuable natural products. This review summarizes recent discoveries and structural classes of fungal-derived steroids, emphasizing their chemical diversity, biosynthetic features, and bioactivity profiles.

Abstract: Background/Objectives: The delayed effects of subchronic and chronic organophosphate poisoning may manifest years after exposure, often masked by age-related diseases. The aim of this retrospective cohort study was to identify the biochemical "trace" that could remain in the patients decades after the poisoning. To achieve this goal, we determined a wide range of biochemical parameters, along with the spectrum of esterified and non-esterified fatty acids (EFA and NEFA, respectively), in the blood plasma of a cohort of elderly patients diagnosed with occupational pathology (OP) due to subacute or (sub)chronic exposure to organophosphates in the 1980s. Methods: Elderly patients with and without a history of exposure to organophosphates were retrospectively divided into two groups: controls (n=59, aged 73±4, men 29% and women 71%) and having the OP (n=84, aged 74±4, men 29% and women 71%). The period of neurological examination and blood sampling for subsequent analysis was approximately one and a half years - from mid-2022 to the end of 2023. Determination of the content of biomarkers of metabolic syndrome, NEFA and EFA in blood plasma was performed by HPLC-MS/MS and GC-MS. Results: The medical history of the examined elderly individuals with OP and the aged control group includes common age-related diseases. However, patients with OP more often have hepatitis (33 vs 7%, p<0.001) and other gastrointestinal diseases (54 vs 32%, p<0.05), total with diagnosis of polyneuropathy (87 vs 66%, p<0.01) and polyneuropathy of the upper and lower extremities (29 vs 12%, p<0.05), impaired distal sensitivity (83 vs 56%, p<0.001), increased body mass index (30.0±4.6 vs 28.4±4.1, p<0.05), and also they have an increased drug load (5 vs 4, p<0.001). Patients with OP more often have balance problems (84 vs 69%, p<0.05) and difficulties in performing daily activities (75 vs 49%, p<0.01). No differences were found between the groups when assessing cognitive function using the MMSE (mini mental state examination), SAGE (self-administered gerocognitive exam), and “Clock” tests. Of the 34 basic biochemical parameters determined on the biochemical analyzer, statistically significant changes were found for only 11, with their median values not going beyond the reference range (clinical norm), and the differences being due to significant individual deviations. Analysis of some metabolic biomarkers with HPLC-MS/MS revealed in the OP group a 1.4-fold decrease in the concentration of 3-hydroxybutyrate (p<0.05, AUC=0.62) and 1.6-fold decrease in the concentration of 2-hydroxybutyrate (p<0.0001, AUC=0.72). In the OP group, a 26% decrease in acetyl-L-carnitine concentration was found (p<0.001, AUC=0.69). An additional study of the esterase profile of patients revealed a decrease in the activity of butyrylcholinesterase (BChE) by 14% (p<0.05, AUC=0.62) though increase in the esterase activity of albumin by 29% (p<0.05, AUC=0.62) in the OP group. Correlation analysis revealed the most significant relationships between albumin esterase activity and arachidonic acid concentration in the OP group (0.64, p<0.0001). A study of a wide range of fatty acids in patients with OP revealed reciprocal relationships between EFA and NEFA. A decrease in concentration was shown for esters of margaric (p<0.01, AUC=0.65), stearic (p<0.01, AUC=0.65), eicosadienoic (p<0.01, AUC=0.63), eicosatrienoic (p<0.01, AUC=0.66), arachidonic (p<0.001, AUC=0.67), eicosapentaenoic (p<0.05, AUC=0.60), and docosahexaenoic (p<0.0001, AUC=0.74) fatty acids. An increase in the concentration was shown for free (non-esterified) fatty acids: heptadecenoic (p<0.0001, AUC=0.72), eicosapentaenoic (p<0.05, AUC=0.62), eicosatrienoic (p<0.001, AUC=0.68), docosahexaenoic (p<0.01, AUC=0.66), γ-linolenic (p<0.01, AUC=0.66), myristic (p<0.0001, AUC=0.71), eicosenoic (p<0.01, AUC=0.65), arachidonic (p<0.01, AUC=0.66), eicosadienoic (p<0.001, AUC=0.67), oleic (p<0.01, AUC=0.66), linoleic (p<0.01, AUC=0.65), palmitic (p<0.001, AUC=0.68), linoelaidic (p<0.01, AUC=0.65), stearic (p<0.01, AUC=0.64), palmitoleic (p<0.05, AUC=0.61), pentadecanoic (p<0.05, AUC=0.61), and margaric (p<0.01, AUC=0.64). Decrease in the ratios of omega-3 to other unsaturated fatty acids were observed only for the esterified forms: by 29% for n3/n6 (p<0.0001, AUC=0.70), by 33% for n3/(n6+n9) (p<0.0001, AUC=0.72), and by 26% for n3/(all fatty acids) (p<0.0001, AUC=0.72). Finally, we have found out how the most reliable indicators correlate with each other. The esterified docosahexaenoic acid – lactate dehydrogenase (LDH) (0.67, p <0.0001) and LDH – total antioxidant status (TAS) (-0.61, p <0.0001) correlations are stronger in the OP group compared to the correlation in the overall sample and lose their strength and statistical significance in the control group. In the OP group, esterified docosahexaenoic fatty acid, LDH activity, and TAS concentration have a moderate correlation with calcium concentration (-0.66, p <0.0001; -0.67, p <0.0001; 0.63, p <0.0001, respectively). In the control group, this correlation is significantly weakened. Esterified docosahexaenoic acid in the OP group also demonstrated strong positive correlations with a row of other esterified fatty acids, though these correlations were not significantly weakened in the control group. Conclusions: The data obtained allow us to consider an increased level of NEFA as one of the main cytotoxic factors for the vascular endothelium, which can determine the specificity of age-related diseases. Modification of albumin properties by arachidonic acid decreased bioavailability of docosahexaenoic acid could be molecular links that cause specific manifestations of OP-induced pathology at late stages after exposure. Epigenetic changes, the contribution of dietary patterns and intestinal microbiota to the spectrum of EFA and NEFA require additional research.

Abstract: Calcium signaling dysfunction is a central contributor to neuronal hyperexcitability and seizure propagation in epilepsy, yet the intracellular mechanisms underlying the actions of valproic acid (VPA) remain incompletely understood. In this study, we investigated whether VPA modulates calcium homeostasis at the level of the endoplasmic reticulum (ER) and how this action influences cytosolic calcium dynamics associated with epileptiform activity. ER calcium levels were directly measured using ER-targeted aequorin in HeLa and PC12 cells, while cytosolic Ca²⁺ signals were monitored by fura-2 fluorescence imaging in bovine chromaffin cells exposed to veratridine, a model of sustained sodium channel activation and calcium oscillations. VPA induced a concentration-dependent release of Ca²⁺ from the ER, with an IC₅₀ of approximately 17 µM. This effect was preserved in permeabilized cells and exhibited activation kinetics comparable to those elicited by inositol 1,4,5-trisphosphate (InsP₃). Pharmacological inhibition of InsP₃ receptors (InsP₃Rs), but not ryanodine receptors or SERCA, abolished VPA-induced ER Ca²⁺ release, supporting a selective InsP₃R-mediated mechanism. Functionally, VPA suppressed the repetitive cytosolic Ca²⁺ oscillations induced by veratridine, while simultaneously producing a sustained elevation of cytosolic Ca²⁺ originating from ER stores and facilitating depolarization-evoked catecholamine secretion. Together, these results support the conclusion that VPA acts as a novel functional agonist of InsP₃Rs and identify ER Ca²⁺ mobilization as a previously unrecognized intracellular mechanism contributing to its modulatory effects on calcium signaling and excitability in epilepsy.

Abstract: In recent years, the global use of dietary supplements has surged, largely due to their appeal as tools for improving well-being and preventing disease. While these products may offer clear advantages—such as addressing micronutrient shortages and enhancing physical or mental performance—they also carry significant risks, including toxicity, potential drug interactions, and limited clinical validation. This paper explores the current body of scientific literature on dietary supplements, offering a nuanced analysis of their advantages and drawbacks, particularly in general and military populations. Drawing upon peer-reviewed studies, regulatory documents, and expert guidelines, the review outlines key safety considerations and presents practical recommendations for evidence-based use.

Abstract: Multiple myeloma is a malignant tumor disease that affects plasma cells and is one of the most common tumors of lymphoid origin. In the process of oncogenesis, that is, the formation of a tumor, there is a significant change in the immune balance in the body, which leads to the suppression of the immune response to the tumor. This suppression is one of the reasons why the tumor can progress and cause serious health problems for the patient. One of the key factors that affect immune balance is innate lymphoid cells (ILCs). ILCs play an important role in regulating the immune response and can both promote and hinder the development of tumor processes, depending on their functional state and interaction with other cells of the immune system. It has been shown that the number of immature CD5+ILC2s cells in the peripheral blood of patients with multiple myeloma is comparable to that of healthy individuals. However, autologous hematopoietic stem cell transplantation in MM patients leads to an increase in the relative number of CD5+ILC2s.

Abstract: Tuberculosis (TB) persists as a devastating global health crisis, exacting a disproportionate burden on low-middle-income countries (LMIC), within which the burgeoning pediatric TB population is especially vulnerable to severe TB manifestations. Rifampicin, a cornerstone of the first-line TB regimen, is indispensable; yet its therapeutic potential is substantially constrained by intrinsic poor aqueous solubility, limited permeability, physicochemical instabilities, and deleterious drug-drug interactions with drugs such as isoniazid. Although current research focuses on formulating rifampicin into fixed-dose combinations – some reporting improved outcomes – the variable and often suboptimal rifampicin bioavailability in such formulations remains a critical concern. Consequently, the development of a stable, independent rifampicin oral formulation should be prioritized to ensure effective TB treatment. There are currently no commercially available rifampicin-only liquid preparations, despite the urgent need for palatable, flexible, age-appropriate formulations. This paucity constitutes a profound and persistent gap in pediatric TB, compromising accurate weight-based dosing, treatment adherence, and therapeutic outcomes while contributing to rifampicin resistance. This paper highlights the key characteristics and formulation challenges of rifampicin, with particular emphasis on pediatric TB. It further discusses liquid self-emulsifying drug delivery systems (SEDDSs) as a novel approach to enhance oral rifampicin delivery. These feasible lipid-based delivery systems offer notable promise, as they are capable of maintaining rifampicin in a solubilized and stable state throughout the gastrointestinal tract, which in turn will likely improve bioavailability. The possible development of a stable liquid rifampicin SEDDS represents a paradigm shift in addressing the longstanding neglect of pediatric TB treatment.

Abstract: Erythropoietin-producing hepatocellular receptor A2 (EphA2) has emerged as a key mediator that promotes tumor malignant progression. EphA2 overexpression and its non-canonical signaling lead to oncogenic transformation, metabolic reprogramming, resistance to treatments, and metastasis. Therefore, strategies targeting EphA2 have been evaluated in clinical trials. However, the clinical effects were not sufficient. An anti-EphA2 monoclonal antibody (mAb), Ea2Mab-7 (mouse IgG1, κ), demonstrated high affinity and specificity among Eph receptors. In this study, we produced recombinant class-switched Ea2Mab-7 variants, including Ea2Mab-7-mG2a (mouse IgG2a) and Ea2Mab-7-hG1 (human IgG1). Both Ea2Mab-7-mG2a and Ea2Mab-7-hG1 recognized human triple-negative breast cancer MDA-MB-231, pancreatic cancer MIA PaCa-2, and colorectal cancer HCT-15 in flow cytometry. Furthermore, both Ea2Mab-7-mG2a and Ea2Mab-7-hG1 exerted significant antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity against these tumors. In mouse xenograft models of breast, pancreatic, and colorectal cancers, both mAbs demonstrated antitumor activity. These results indicate the potential of Ea2Mab-7 variants for the treatment of EphA2-positive cancers.

Abstract:

This study extracted polysaccharides from Adansonia Suarezensis fruit pulp using various methods: hot water (ASP-HW), acid (ASP-AC), alkaline (ASP-AL), ultrasound-assisted hot water (ASP-HWU), ultrasound-assisted acid (ASP-ACU), and ultrasound-assisted alkaline (ASP-ALU). A comparison was conducted on the yield, chemical composition, structural properties, and biological activities. The findings indicated that the extraction solvent significantly influenced various essential properties of the ASPs, such as yield, monosaccharide composition, molecular weight, particle size, and thermal stability. The polysaccharides consisted of galacturonic acid, galactose, xylose, and arabinose.ASP-AL and ASP-ALU exhibited smaller molecular weight and particle size, with molecular weights of 19,813 Da and 19,600 Da, and particle sizes of 146.67 ± 22.46 nm and 140.97 ± 20.38 nm, respectively. The ASPs displayed characteristic polysaccharide structures, with ASP-AC, ASP-ACU, ASP-AL, and ASP-ALU possessing a triple-helix conformation. Bioactivity assays demonstrated that ASP-AL and ASP-ALU had enhanced DPPH radical scavenging activity (IC₅₀ 116.67 ± 0.58 μg/mL and 113.67 ± 2.31 μg/mL, respectively), ABTS radical scavenging activity (IC₅₀ 79.67 ± 0.58 μg/mL and 79.33 ± 1.15 μg/mL, respectively), and α-glucosidase inhibitory activity (IC₅₀ 0.146 ± 0.01 mg/mL and 0.206 ± 0.01 mg/mL, respectively).These findings indicate that ASPs possess significant bioactivity, supporting their potential use in pharmaceuticals and functional foods.

Abstract: Background: Erectile dysfunction (ED) affects approximately 52% of men aged 40–70 years in population-based cohorts and is strongly associated with vascular and metabolic disease, particularly diabetes mellitus.[1] In Saudi Arabia, diabetes prevalence in adults is estimated at over 20%, magnifying the burden of vasculogenic ED.[2] Phosphodiesterase-5 inhibitors (PDE5i) improve erectile function but have limitations in patients with endothelial dysfunction, neurogenic compromise, or contraindications to nitrates.[3]Hypothesis: Helospectins I and II are 37–38–amino-acid peptides isolated from Heloderma (Gila monster and Mexican beaded lizard) venom that belong to the glucagon/VIP superfamily and exhibit potent vasodilatory, smooth muscle–relaxant, and hypotensive effects.[4,5] Building on the successful translation of the venom peptide exendin-4 into the antidiabetic drug exenatide,[6,7] we hypothesize that synthetic helospectin analogues could serve as selective agonists at VPAC1/VPAC2 receptors in genital vascular smooth muscle, promoting corpus cavernosum and clitoral vasodilation independent of endothelial nitric oxide (NO), and thereby treating ED and clitoral engorgement (female sexual arousal disorder, FSAD).[8–10]Methods (theoretical proposal): We outline (i) a biochemical and physiological rationale based on helospectin/VIP homology, distribution of VIP-family peptides in male and female genital tissues, and venom-derived pharmacology; (ii) a conceptual mechanism of action centred on cAMP/PKA-mediated relaxation of cavernosal and clitoral smooth muscle; and (iii) a Phase I/IIa randomized, double-blind, placebo-controlled crossover trial of intranasal synthetic helospectin in men with ED and women with FSAD.Expected outcomes: We anticipate that helospectin analogues could augment genital blood flow and erection/engorgement responses in a manner that is at least partly independent of endothelial NO, potentially benefiting patients with diabetic and neurogenic ED. Safety concerns include systemic hypotension, off-target vasodilation, and theoretical effects on exocrine pancreas and gut. This preprint is offered as a hypothesis-generating framework to encourage collaborative medicinal chemistry and preclinical work, not as evidence for clinical use.

Abstract:

Background: Human Immunodeficiency Virus type 1 (HIV-1) remains a major global health challenge. Despite advances in antiretroviral therapy, new prevention strategies are needed, particularly topical microbicides capable of blocking the earliest steps of viral entry. HIV-1 attachment relies on interactions with heparan sulfate proteoglycans on host cell surfaces; therefore, sulfated heparan-mimetic polymers have been explored as antiviral agents. In this context, sulfated chitosan microparticles are designed to mimic natural glycosaminoglycan receptors, acting as biomimetic decoys that prevent viral attachment and entry. Methods: Low molecular weight sulfated chitosan (LMW Ch-S) microparticles were synthesized and characterized (FTIR, SEM) following US Patent No. 11,246,839 B2 (Bucarey et al., 2022). Their antiviral activity was evaluated by incubating the microparticles with high-viral-load HIV-1–positive plasma to enable viral binding and removal by pull-down. The performance of the synthesized Ch-S microparticles was compared with established heparinoid controls, including soluble heparin and heparin microparticles. Results: The Ch-S microparticles exhibited stronger virus-binding and neutralizing capacity than all heparinoid comparators, achieving the highest overall reduction in viral load. Subsequent evaluation across multiple tested concentrations confirmed a consistent antiviral effect, indicating that the synthesized Ch-S microparticles maintain robust virus–particle interactions throughout the concentration range examined. Conclusions: These findings demonstrate that LMW Ch-S microparticles possess potent antiviral properties and outperform classical heparinoid materials, supporting their potential application as topical microbicides targeting early HIV-1 entry mechanism.

Abstract:

Background/Objectives. Pulmonary arterial hypertension (PAH) is a rare but severe disease which leads to right ventricular (RV) maladaptation, failure and death. Currently approved drugs have limited impact on disease progression. A multitarget strategy consisted of activation of adenosine A₂B receptor and inhibition of phosphodiesterase-4 (PDE4) in combination with human mesenchymal stromal cells (hMSCs) was tested in PAH animal model. The main objective was to determine whether the combination improves pulmonary hemodynamics, vascular remodeling, and RV function, given the limited disease-modifying effects of approved vasodilators. Methods. Vascular reactivity was assessed in isolated rat pulmonary artery rings exposed to the dual target compound named LASSBio-1860 alone or in the presence of either A₂A (ZM-241385) or A₂B (MRS-1706) antagonist. PAH was induced in male Wistar rats through the administration of monocrothaline (MCT, 60 mg·kg⁻¹). After confirmation of PAH, by the decrease of the ratio of pulmonary artery acceleration time and ejection time ratio (PAAT/TET), animals were randomized divided to receive vehicle, hMSC (single i.v. dose, 1×10⁵ cells), LASSBio-1860 (62 mg·kg⁻¹·day⁻¹, p.o., 14 days), or the combination. Outcomes included PAAT/TET and RV cardiac output (RV-CO) using echocardiography, RV systolic pressure (RVSP) by direct puncture, Fulton index and RV wall thickness, lung histology (perivascular cell counts and wall thickness), and RV protein expression (TGF-β, CaMKII) by Western blot. Results. LASSBio-1860 produced endothelium-independent vasorelaxation of rat pulmonary arteries, consistent with A₂B agonism and PDE4 inhibition responses. In MCT-induced PAH, the combination of LASSBio-1860 with hMSCs promoted: 1. Recovery of PAAT/TET and RV-CO; 2. Decrease of RVSP; 3. Reduction of RV hypertrophy, vascular inflammation and remodeling; 4. Downregulation of ventricular TGF-β and CaMKII. Conclusions. Combination of LASSBio-1860 with hMSC improved RV function, attenuated pulmonary hypertension, RV and vascular remodeling; reduced inflammatory/proliferative signaling in MCT induced-PAH, supporting a promising multitarget therapeutic strategy for PAH.

Abstract: The aging of the population and the increasing prevalence of multimorbidity contribute to the widespread use of polypharmacotherapy, which in turn elevates the risk of adverse drug reactions and clinically significant drug–drug interactions. One of the key yet frequently underestimated issues in clinical practice is the prescribing cascade, which occurs when an adverse drug reaction is misinterpreted as a new medical condition, leading to the initiation of an additional medication. This phenomenon is particularly relevant in the elderly population, in whom altered pharmacokinetics and pharmacodynamics, together with reduced organ reserve, increase susceptibility to adverse drug events, including nephrotoxicity. The objective of this paper is to review current evidence on the mechanisms and clinical consequences of the prescribing cascade-with particular emphasis on renal function deterioration—as well as strategies for its prevention in the geriatric population. Analysis of the literature indicates that prescribing cascades remain insufficiently recognized in clinical practice, despite the availability of pharmacotherapy assessment tools such as the Beers Criteria and the STOPP/START criteria. Documented prescribing cascades have been shown to contribute to deterioration in health status and quality of life, an increased frequency of hospitalizations, and a greater burden on healthcare systems. Particularly concerning are cascades involving cardiovascular, neurological, and analgesic medications, which may induce or exacerbate renal injury, ultimately leading to chronic kidney disease and organ failure. Prescribing cascades represent a significant yet frequently underestimated threat to the efficacy and safety of pharmacotherapy in older adults. Their consequences may extend beyond reduced quality of life and increased treatment costs to include serious complications such as the development of renal failure. Enhancing clinicians’ awareness, conducting systematic medication reviews, and employing validated assessment tools are essential for the identification and prevention of prescribing cascades, thereby reducing the risk of renal injury and improving clinical outcomes.

Abstract:

Lactococcus lactis, a safe food-grade lactic acid bacterium, has attracted increasing attention as a live biotherapeutic platform for mucosal vaccine development. Its genetic simplicity, absence of endotoxins, and availability of well-characterized inducible systems have enabled controlled expression and delivery of heterologous antigens and therapeutic molecules. This review highlights recent advances in the use of genetically modified L. lactis for mucosal immunization, focusing on expression technologies, routes of administration, and immune mechanisms relevant to protection or tolerance. Preclinical studies demonstrate its capacity to induce both mucosal and systemic immune responses against diverse pathogens, underscoring its potential as a safe and versatile vaccine chassis. Remaining challenges include regulatory harmonization, biosafety concerns, and the need for standardized manufacturing and evaluation frameworks. Together, these developments position L. lactis as a promising candidate for next-generation mucosal vaccines and live biotherapeutic products.

Abstract: Pharmacy-based point-of-care (POC) services have evolved from pilot initiatives to an essential component of decentralized healthcare delivery. These services—ranging from rapid infectious-disease screening to chronic-disease monitoring—improve access, re-duce diagnostic delays and empower pharmacists as front-line healthcare providers. The present paper is the first comprehensive review of the evolution of pharmacy POC services worldwide, combined with the analysis of the regulatory and educational frameworks supporting implementation, technological drivers such as biosensors, mo-bile health and artificial intelligence and in-depth socioeconomic considerations. Bene-fits for patients, pharmacies and healthcare systems are contrasted with challenges in-cluding variable reimbursement, uneven regulatory oversight and workforce preparedness.

Abstract: Background/objectives: The COVID-19 pandemic prompted an unprecedented global response, including the rapid development and deployment of vaccines using novel technologies such as mRNA and adenoviral vectors. Vaccination has proven highly effective in reducing severe COVID-19 outcomes. This study investigated whether receiving COVID-19 vaccines could reduce the risk of developing long COVID symptoms. Methods: This retrospective observational study analysed electronic health records from 627 adults diagnosed with COVID-19 in London to assess the impact of COVID vaccines on the risk of long COVID. We evaluated both the number of vaccine doses received and whether participants received heterologous booster vaccines or the homologous regimens. The statistical methods employed multinomial propensity score modelling and inverse probability of treatment weighting to minimise confounding variables and strengthen causal inference. Results: The study included 627 participants, with an average age of 59. The sample consisted of 55% women, and 63% individuals of them received a COVID-19 vaccine. Older age and higher BMI were both consistently linked to a greater risk of developing long COVID symptoms. While one vaccine dose did not significantly change the odds of developing long COVID (odds ratio, 0.907; p = 0.60), receiving two or more doses (odds ratios ranging from 2.135 to 2.319, all p < 0.001) significantly increased the risk. Participants vaccinated after contracting COVID-19 (OR:5.93, p< 0.001) had nearly six times higher odds of developing long COVID compared to those vaccinated beforehand. Neither heterologous booster vaccines nor homologous vaccine strategies were found to reduce the odds of long COVID significantly. However, a 41% reduction in the odds of long COVID was observed with heterologous booster vaccines (OR = 0.59, 95% CI: 0.34–1.03, p = 0.07). Conclusion: Two or more vaccine doses correlated with increased odds of long COVID, particularly after infection. The heterologous booster vaccine approach showed a possible protective effect. These findings highlight the complex relationship and emphasise the need for further research on future vaccine policies.

Abstract:

Background/Objectives: Burn injuries pose a significant challenge due to tissue damage and impaired healing. Cell-based therapies offer promise by delivering therapeu-tic cells to the wound site. However, effective cell delivery remains a critical hurdle. This study investigates the potential of non-cross-linked hyaluronic acid (HA) as a simple and versatile carrier for delivering autologous keratinocytes and fibroblasts to treat early burn wounds. Methods: Primary keratinocytes and fibroblasts were isolated from uninjured adult skin. In addition, fibroblasts and adipose stem cells (ASC) from polydactyly and progenitor fibroblasts were used. Non-cross-linked HA Redensity 1^® (RD1) solutions of varying concentrations were prepared and applied on various in vitro models. Cell viabil-ity, proliferation, migration and collagen stimulation were assessed using standard as-says. Additionally, cells were suspended in Redensity 1 and applied a the in vitro de-epidemalized dermis (DED) wound model to examine cell delivery and tissue refor-mation. Results: Preliminary data demonstrated the feasibility of using non-cross-linked HA RD1 gel as a cell carrier. RD1 gel enhanced cell viability, retention, migration and col-lagen deposition. Histological analysis revealed improved cell adhesion and migration. Conclusions: This study provides valuable insight into the potential of non-cross-linked HA RD1 as a simple and effective delivery vehicle for cell therapies in early burn care. Successful translation of this approach could significantly improve clinical outcomes for burn patients.

Abstract: Background: Infections caused by the multidrug-resistant pathogen Mycobacterium abscessus (Mab) are notoriously difficult to treat. The novel β-lactamase inhibitor durlobactam, in combination with β-lactams, shows potent bactericidal activity against Mab, but the potential for acquired resistance remains a clinical concern. Objectives: To identify and characterize mechanisms of acquired resistance to durlobactam in Mab. Methods: In vitro single-step resistance selection was performed by plating wild-type Mab ATCC 19977 on agar containing durlobactam. Resistant mutants were isolated, and their genomes were sequenced. The resistance phenotype was confirmed by constructing a targeted gene deletion mutant and by transcriptional silencing using a CRISPR interference (CRISPRi) system. Minimum inhibitory concentrations (MICs) were determined by both, an agar-based method and broth microdilution. Results: Whole-genome sequencing of durlobactam-resistant mutants identified loss-of-function mutations in ponA1, a gene encoding a class A penicillin-binding protein involved in cell wall synthesis. Targeted deletion of ponA1 (ΔponA1) and CRISPRi-mediated knockdown of ponA1 expression both recapitulated the resistance phenotype, resulting in a significant increase in the durlobactam MIC on solid agar media. Strikingly, broth microdilution MICs remained largely unaffected. Conclusions: Inactivation of the peptidoglycan synthase PonA1 is a novel mechanism of resistance to durlobactam in Mab that is phenotypically expressed only during growth on solid surfaces. This finding identifies a specific genetic pathway for resistance and highlights that standard broth-based susceptibility testing could miss clinically relevant resistance mechanisms.

Abstract: Drug discovery is a complex, multi-parameter optimization process. I argue that a greater emphasis on optimizing binding affinity will accelerate the development of new medicines. Note that “optimizing” is not always synonymous with “maximizing.” Optimizing affinity provides seven distinct benefits: achieving potent tool compounds more quickly; making compounds with increased potency; making more selective compounds; optimizing drug candidates more quickly; encouraging the pursuit of more synthetically challenging compounds; expanding chemical diversity during lead optimization; and minimizing interactions with "avoid-ome” targets that lead to poor ADME and tox properties. While affinity is certainly not the only thing that matters, the value of optimizing drug – receptor interactions is profound and often underappreciated.

Abstract:

Background/Objectives: Male infertility is one of the major problems in Iraqi health and society, caused by several factors, such as acquired, environmental, and genetic factors. Awareness of the crucial role of telomeres and mitochondria in sperm production and fertility has increased in recent years. This study aimed to evaluate the association between mitochondrial DNA (mtDNA) copy number and telomere length in sperm and the degree of infertility in Iraqi males. Methods: Of the 200 study participants, 50 were healthy controls and 150 were infertile. Sperm count, motility, and morphology were assessed by collecting and analyzing semen samples. After DNA extraction, the mitochondrial ND1 gene and the reference nuclear gene GAPDH were analyzed by quantitative PCR (qPCR) to determine the mtDNA copy number. To determine telomere length, another qPCR analysis was used. Results: The mtDNA copy number of infertile men was significantly higher than that of healthy controls with a P-value (0.001). In addition, sperm of the patients group showed a significant reduction in telomere length, (P=0.001). According to the results of the study, male infertility in Iraqi men is associated with a higher number of mtDNA copies and shorter telomere length. DNA damage or a disruption in the mitochondria energy production pathway could be the cause of this association. Conclusions: This study reveals that a higher number of mtDNA copies and shorter telomere lengths are associated with male infertility in Iraqi men. These results highlight the importance of continuing research and exploring new avenues in the field of male infertility.