Applied Biosciences

Candida species, normally part of the healthy human flora, can cause severe opportunistic infections when their population increases. This risk is even greater in immunocompromised individuals. Women using intrauterine contraceptive devices (IUDs) are at higher risk for IUD-associated vulvovaginal candidiasis (VVC) because the device provides a surface for biofilm formation. This biofilm formation allows the normal flora to become opportunistic pathogens, leading to symptoms of VVC such as hemorrhage, pelvic pain, inflammation, itching and discharge. VVC is often linked to IUD use, requiring the prompt removal of these devices for effective treatment. This study evaluated the activity of the arginine-rich peptide “protamine” against Candida albicans, Candida tropicalis and Candida krusei isolated from IUD users who had signs of VVC. The antimicrobial activity was measured using the agar disk diffusion and microbroth dilution methods to determine the minimum inhibitory concentration (MIC). The MIC values of protamine against C. albicans, C. tropicalis and C. krusei are 32 μg mL⁻¹, 64 μg mL⁻¹ and 256 μg mL⁻¹, respectively. The determined MIC of protamine was used for a biofilm inhibition assay by crystal violet staining. Protamine inhibited the biofilm formation of the VVC isolates, and its mechanisms were studied through scanning electron microscopy (SEM) and a reactive oxygen species (ROS) assay. The disruption of cell membranes and the induction of oxidative stress appear to be key mechanisms underlying its anti-candidal effects. The results from an in vitro assay support the potential use of protamine as an antibiofilm agent to coat IUDs in the future for protective purposes. Full article

Cannabidiol (CBD), a non-psychoactive compound derived from Cannabis sativa, is believed to have anxiety-reducing and antidepressant effects. However, existing data are inconsistent, likely due to variations in experimental designs, dosages, and stress models. This study sought to assess the impact of CBD on anxiety and depression-like behaviors in Wistar rats exposed to acute cold stress, as well as its impact on pro- and anti-inflammatory cytokines. Male rats were treated with CBD (2.5, 5, or 10 mg/kg) or vehicle for 14 days and subjected to behavioral tests, including the elevated plus maze, social interaction, and forced swim tests. Serum levels of cytokines (IL-6, TNF-α, IL-1β, and IL-10) were analyzed post-experiment using ELISA. Results demonstrated a dose-dependent anxiolytic effect of CBD, with significant improvements in social interaction and reductions in anxiety-like behaviors at 5 and 10 mg/kg. All doses of CBD decreased immobility in the forced swim test, suggesting antidepressant effects. Furthermore, CBD selectively lowered IL-6 levels, a key cytokine in acute stress and depression pathogenesis. These findings indicate that CBD has anxiety-reducing and antidepressant properties, partially mediated by modulation of inflammatory processes, particularly IL-6. Full article

Cancer pain is a highly prevalent problem and one of the most distressing symptoms in cancer patients. The management of cancer pain is one of the most significant challenges in the care of these patients. Cancer pain must be treated quickly and effectively as it affects the quality of life and reduces the patient’s life expectancy. Major opioids are recognized by the World Health Organization (WHO) as first-line treatment for moderate to severe cancer pain, but their use is often hampered by individual variations, comorbidities, and complications associated with cancer. Since the simultaneous use of two major opioids has become frequent, a narrative review was conducted, whose main objectives were to evaluate whether the combination of two major opioids improves pain and quality of life in cancer patients, considering their pharmacodynamic and pharmacokinetic properties and evaluate the impact of this combination on the frequency and intensity of side effects. The search for information was carried out in evidence-based medicine databases, namely PubMed/MEDLINE, Cochrane Library, Database of Abstracts of Reviews of Effects, National Guideline Clearinghouse, NHS Evidence and Index das Revistas Médicas Portuguesas using the MeSH terms “opioids” and “quality of life”. Articles and documents published between 1 January 2010 and 1 June 2023, in English, Portuguese and Spanish, were considered, including original articles, meta-analyses, systematic reviews and clinical guidelines. A total of 342 articles were retrieved and of these, only 13 were selected for full reading. The combination of opioids is based on the principle that different opioids act through different mechanisms, which can reduce dose-related adverse effects. Simultaneous use of two major opioids may allow for more modest increases in the equivalent dose of the second opioid, providing better pain control and reduced side effects such as nausea, vomiting, and constipation. More studies on the combination of opioids are needed to improve cancer pain treatment. The lack of personalized therapies limits the effectiveness of opioids, and variability in treatment responses requires individualized approaches. Personalized medicine, based on pharmacogenomics, is one of the most promising strategies to optimize pain relief and reduce adverse effects. Full article

Drug discovery is inherently a multi-criteria optimization problem. In the first instance, it involves a tremendously large chemical space, where each compound can be characterized by multiple molecular and biological properties. Modern computational approaches try to efficiently explore the chemical space in search of molecules with the desired combination of properties. For example, Pareto optimizers identify a so-called “Pareto front”, a set of non-dominated solutions. From a qualitative perspective, all solutions on the front are potentially equally desirable, each expressing a trade-off between the goals. However, often there is a need to weight the objectives differently, depending on their perceived importance. To address this, we recently implemented a new Multi-Criteria Decision Analysis (MCDA) method as part of the AI-powered Drug Design (AIDD^TM) technology initiative. This allows the user to weight various objective functions differently, which, in turn, efficiently directs the generative chemistry process toward the desired areas in chemical space. Full article

Immune thrombocytopenia, formerly idiopathic thrombocytopenia purpura (ITP), is an autoimmune disease characterized by the depletion of platelets below 100,000/µL when other causes of thrombocytopenia have been ruled out. It is associated with several infectious pathologies, disease states, and as a known side effect and complication of several drugs and chemotherapies. Standard treatment calls for glucocorticoid-mediated immunosuppression, intravenous immunoglobin transfusion, platelet stimulation, platelet transfusion, and splenectomy in instances of chronic and severe disease. While standard treatments are often effective, some cases prove resistant, and more commonly, some patients are unable to tolerate standard treatment protocols or opt out of surgical intervention. In addition, second-line therapies can be unfeasibly expensive and are associated with side effects themselves. Therefore, for a subset of patients afflicted by immune thrombocytopenia, the exploration of alternative treatment methods is needed in order to ease their burden of disease. Emerging evidence suggests that plant-derived extracts, traditionally used in regions such as Asia and Africa to manage acute thrombocytopenia, hold promise as alternative or adjunctive therapies for the mentioned subset of patients. These natural compounds may provide a cost-effective and less invasive option, potentially bridging gaps in current treatment regimens. We propose these extracts may play a role in fulfilling this deficiency in current treatment protocols. With this review, we aim to characterize and compile evidence that various organic extracts and compounds may be utilized to improve outcomes in these patients. By highlighting their clinical relevance and potential for integration into ITP treatment protocols, this manuscript underscores the importance of expanding the alternative therapies for ITP to improve patient outcomes and reduce treatment burdens. Full article

This article describes the basics of chemical thermodynamics and its application to the study of plant biomass and its main components, cellulose, hemicelluloses, lignin, etc. The energy potential of various biomass types, as well as biomass-based solid, liquid, and gaseous biofuels, is determined. A method of additive contributions of combustion enthalpies of main components is proposed to calculate the combustion enthalpy of biomass samples. It is also established that the potential of thermal energy of the initial biomass is higher than the energy potential of secondary biofuels released from this biomass. The thermodynamic functions of plant biopolymers are calculated. Moreover, the thermodynamic stability of various crystalline allomorphs of cellulose and amorphous cellulose is studied. The melting enthalpies of crystallites with different types of crystalline structures are estimated. A thermochemical method for determining the degree of crystallinity of cellulose is proposed. The most important biomass components are cellulose and other polysaccharides. The thermodynamics of the enzymatic hydrolysis of polysaccharides and their conversion into glucose are described. In addition, the thermodynamic analysis of the conversion process of glucose into bioethanol is performed. Considerable attention is also paid to the thermochemistry of cellulose alkalization, etherification, and esterification. Full article

Cultivated yam (Dioscorea rotundata) is a staple tuber crop in West Africa which is mainly vegetatively propagated. Although the majority of yam cultivars flower, the control of their sexual reproduction remains largely unknown despite its importance for plant-breeding programs. While described as a dioecious species, yam has several monoecious (mix) cultivars that are often subject to spontaneous sex changes. This study aims to evaluate the impact of phytohormones on sex expression and flower development in cultivated yams D. rotundata. Exogenous applications of 1 mM 6-benzylaminopurine (BAP) and 1 mM methyl jasmonate (MeJa) were performed on seedlings of female (Gnidou, Anago), male (Flou) and mix (Katala and Laboko) cultivars. The mix cultivars produced monoecious, male and female plants while the female and male cultivars had rather stable sex. Our results showed that MeJa exhibited a significant masculinising effect in mix cultivars and induced inflorescence and flower malformations in female cultivars (56% in Gnidou and 37% in Anago). Most malformations were inflorescence branching and sterile flowers (non-differentiated ovary) with extra cycles of sepals. Moreover, MeJa reduced flowering time in the cultivars of all sexes and increased the number of inflorescences per plant as well as the number of flowers per plant. Our results showed that BAP reduced the flowering time, synchronized flowering in female plants and increased the number of inflorescences per plant in monoecious plants. However, our results did not allow for strong conclusions regarding the effect of BAP on sex expression due to the high proportion of female flowering in both the control and BAP-sprayed plants. Nevertheless, we did not observe any masculinising effect for BAP. Further research that would highlight hormone and homeotic gene interactions in flowering could be of key interest in understanding the hormonal control of sex in cultivated yams D. rotundata. Full article

Microplastics (MPs), defined as plastic particles of less than 5 mm, pose a significant global environmental threat, particularly in aquatic ecosystems, due to their persistence and potential harmful effects on wildlife and human health. They can absorb persistent organic pollutants (POPs), like polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), raising concerns about their impact on biota. To elucidate this impact, the present study employed attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) to analyse the characteristics of MPs sourced from commercial cosmetics. We investigated the toxicity of MPs on Caenorhabditis elegans and two entomopathogenic nematode species, Steinernema feltiae (the enviroCORE strain SB12(1)) and Steinernema carpocapsae (a commercial strain from e-NEMA) in laboratory bioassays. Nematodes were exposed to various concentrations of MPs and other pollutants, including atrazine, 1,3-dichloropropene, naphthalene, and fluorene, in controlled settings over 72 to 96 h. Additionally, high-throughput 18S rDNA sequencing was used to analyse nematode biodiversity in sediments from the River Barrow (RB) in SE Ireland. Our findings revealed that MPs increased nematode mortality and adversely affected community structure, as indicated by nematode maturity and sigma maturity indices, suggesting a potential disruption of the ecological balance in river sediments. This highlighted the ecological risks posed by MP pollution and emphasised the urgent need for further research into the health of benthic ecosystems in Ireland, particularly in relation to how MPs may influence nematode community dynamics and biodiversity. Full article

Rivers are crucial hydrological cycle components, supporting ecosystems and human activities. Managing and protecting river water quality is essential. Diatoms, microscopic algae, are widespread and sensitive to changes in water quality, making them effective bioindicators. This study focused on the Kuils River in the Western Cape, South Africa, where diatom sampling was conducted at four sites. Their communities are affected by different physicochemical parameters, such as changes in pH, salinisation, eutrophication, and organic enrichment. A total of 98 diatom species were identified. The Omnidia software was used to calculate the Generic Diatom Index, Specific Pollution Index, and Trophic Diatom Index. Historical physicochemical data such as pH, chemical oxygen demand, and phosphorus were obtained from the Department of Water and Sanitation between 2019 and 2021 and were used to provide a reference for comparing diatoms. The results showed clear signs of pollution, as shifts in diatom species composition were observed. Pollution-tolerant species such as the Nitzschia palea (Kützing) W Smith 1856, Navicula viridula (Kützing) Ehrenberg 1838, Eunotia bilunaris (Ehrenberg) Schaarschmidt 1880, and Ulnaria ulna (Nitzsch) Compère 2001 dominated, while less pollution-tolerant species like Gomphonema parvulum f.saprophilum Lange-Bertalot and E. Reichardt 1993 and Stephanocylus meneghinianus (Kutzing) Kulikovskiy, Genkal, and Kociolek 2022 were less abundant. Both diatom indices and physicochemical data indicated poor water quality in the Kuils River. The study concludes that diatoms are a valuable tool for biomonitoring river water quality and recommends their use alongside traditional physicochemical methods for future assessments of river systems. Full article

Distraction osteogenesis (DO) is a process which uses the bone’s natural healing tendencies to repair and lengthen pathologic, missing, or malformed bone. The mechanism of DO mimics the pathway that the body uses in any other fracture repair however the location of the fracture is carefully controlled by a surgical osteotomy. Postoperatively, the bone is allowed to begin its natural healing process, with the lengthener applying constant tension and thus re-initiating the process of healing along the length of the distraction gap. Current clinical indications for DO include limb length discrepancy, congenital bone length deformity, large bone defects, and extremity reconstruction due to hypoplasia or limb salvage procedures. The risks of DO include soft tissue complications, relapse or improper correction, cost or resource-related challenges, and psychosocial stigmas surrounding long treatment durations and the necessity of wearing the distraction lengthening hardware. Future directions for DO include supplements to the bone regeneration process (such as growth factors and/or mechanical stimulation) or improvements to the distractor device itself (changes in material and/or the structure of the device itself). This review aims to offer a comprehensive summary of the indications, underlying biological mechanisms, and practical considerations when implementing the use of distraction osteogenesis in clinical practice. Full article

Integrated crop and livestock systems (ICLSs) have been considered an important management-based decision to improve soil health by carbon sequestration. A two-year study (2019–2021) at CPBES in Newton, MS, was conducted to evaluate the effect of an ICLS on soil microbial diversity in the southeastern region of the USA, representing agroclimatic conditions that are warm and humid. Amplicons targeting bacterial 16S rRNA genes and fungal ITS2 regions were sequenced. Taxonomic assignment and characterization of microbial diversity were performed using QIIME2^®. Soil fungal diversity pattern showed significant difference (alpha diversity, p = 0.031 in 2020 and beta diversity, p = 0.037 in 2021). In contrast, no significant differences were observed in bacterial diversity. However, there were several beneficial bacterial phyla, such as Proteobacteria and Actinobacteria, and fungal phyla such as Ascomycota, which were dominant in both years and did not show significant differences due to cover crop treatments. Canonical Correspondence Analysis (CCA) and Mantel test showed significant influence on fungal diversity due to carbon (rm = 0.2581, p = 0.022), nitrogen (rm = 0.2921, p = 0.0165), and electrical conductivity (rm = 0.1836, p = 0.0583) in 2021, and on bacterial diversity due to EE-GRSP (rm = 0.22, p = 0.02) in 2020. However, the results showed that there were no significant differences between the cover crop treatments that were consistent over a two-year study period. However, the mix of different cover crops such as oats (Avena sativa L.), crimson clover (Trifolium incarnatum L.), and tillage radish (Raphanus sativus L.) demonstrated higher positive correlation and lower negative correlation with different bacterial and fungal phyla. Long term study of ICLS is suggested to understand the shift in microbiome that would help in understanding the role of cover crops and grazing in improving crop production sustainably. Full article

This study explores the interaction between Vemurafenib (VEM), a potent BRAF inhibitor, and calf thymus DNA (ctDNA) using a comprehensive array of biophysical and computational techniques. The primary objective is to understand the potential off-target effects of VEM on DNA, given its established role in melanoma therapy targeting the BRAF V600E mutation. The investigation employed methods such as ultraviolet–visible absorption spectroscopy, steady-state fluorescence, circular dichroism, isothermal titration calorimetry, and advanced molecular dynamics simulations. The results indicate that VEM interacts with DNA primarily through a minor groove-binding mechanism, causing minimal structural disruption to the DNA double helix. Viscosity measurements and melting temperature analyses further confirmed this non-intercalative mode of binding. Calorimetry data revealed an exothermic, thermodynamically favorable interaction between VEM and ctDNA, driven by both enthalpic and entropic factors. Finally, computer simulations identified the most probable binding site and mode of VEM within the minor groove of the nucleic acid, providing a molecular basis for the experimental findings. Full article

Osteoarthritis is a joint disease that causes pain, stiffness, and reduced joint function because the protective cushioning inside the joints, called cartilage, gradually wears away. This condition is caused by various factors and complex processes in the joint’s environment, involving different types of cells producing factors that can either maintain the joint health or contribute to osteoarthritis. This study aimed to understand the factors influencing both healthy and diseased joints in DDD strategies for the in vitro preconditioning of MSCs. An electronic search in the PubMed, Scopus, and Web of Science databases was carried out using the terms (cartilage OR chondr*) AND (repair OR regeneration OR healing) AND (niche OR microenvironment)) AND (“growth factor” OR GF OR cytokine). Researchers used various methods, including macroscopic examinations, histology, immunohistochemistry, and microCT. Molecules associated with joint inflammation were identified, like macrophage markers, MMP-13, TNF, apoptotic markers, and interleukins. Chondrogenesis-related factors such as aggrecan GAG, collagen type II, and TGF beta family were also identified. This study suggests that balancing certain molecules and ensuring the survival of joint chondrocytes could be crucial in improving the condition of osteoarthritic joints, emphasizing the importance of chondrocyte survival and activity. Future preconditioning methods for MSC- and EV-based therapies can find suitable strategies in the described microenvironments to explore co-culture systems and soluble or extracellular matrix factors. Full article

Metallo-β-lactamases (MBLs) are members of the structurally conserved but functionally diverse MBL-fold superfamily of metallohydrolases. MBLs are a major concern for global health care as they efficiently inactivate β-lactam antibiotics, including the “last-resort” carbapenems, and no clinically suitable inhibitors are currently available. Increasingly, promiscuous β-lactamase activity is also observed in other members of the superfamily, including from viruses, which represents an underexplored reservoir for future pathways to antibiotic resistance. Here, two such MBL-fold enzymes from Bacillus phages, the cyclic mononucleotide-degrading proteins Apyc_Goe3 and Apyc_Grass, are shown to degrade β-lactam substrates efficiently in vitro. In particular, Apyc_Grass displays a distinct preference for carbapenem substrates with a catalytic efficiency that is within one order of magnitude of the clinically relevant MBL NDM-1. Mutagenesis experiments also demonstrate that the loss of a metal-bridging aspartate residue reduces nuclease activity up to 35-fold but improves carbapenemase activity. In addition, we hypothesise that the oligomeric state significantly influences β-lactamase activity by modifying access to the active site pocket. Together, these observations hint at a possible new avenue of resistance via the spread of phage-borne MBL-fold enzymes with β-lactamase activity. Full article

Papaya is a globally important crop, with production primarily based on hermaphrodite plants. Papaya has three sex types—male, female, and hermaphrodite—determined by flower morphology, but this is only distinguishable at the flowering stage. In this study, a recombinase polymerase amplification (RPA) assay was developed and optimized to identify the three sexes of papaya. Recombinant uvsX, uvsY, gp32, and Bsu DNA polymerase were used to study the effects of temperature, reaction time, and sensitivity conditions for RPA reaction efficiency. The optimal conditions were found to be 41 °C and a 30 min reaction time, allowing the detection of the target sex from specific DNA markers, even when using crude extract. This study shows that RPA could be used for sex determination in papaya, and the findings could contribute to developing a point-of-need strategy due to their sensitivity and specificity. Full article

The olive tree is currently cultivated worldwide, with higher incidence in Mediterranean Basin. Its climate is highly favorable to the synthesis of phenolic compounds, stored in olive leaves; their consumption has been linked to a lower incidence of cancer and cardiovascular disorders for which the research interest upon this feature has increased in last decade. This study aimed (i) to review evidence about the importance of olive leaf extract (OLE) on human health and the physiological effect of its major compounds; (ii) to update the state of the art of studies conducted on the health and technological usage of olive leaf extract; (iii) to report potential uses of OLE in pharmaceuticals, food production, and cosmetics; and (iv) to prospect the future of clinical applications of OLE from diverse cultivars, especially in metabolic inflammatory conditions such as polycystic ovary syndrome (PCOS). Overall, cultivars richer in TPC, including TFC, αT, omega-3 and omega-9, present a main research target for supplementation alone or in conjunction with vitaminic compounds, due to their nutraceutical value in metabolic disorders, chronic inflammatory diseases, and anti-aging treatments, whereas cultivars with less water content might be useful as substrates for food preservation. With regard to future prospects, it would be of great interest to clarify the specific mechanisms underlying the beneficial effects of OLE on neuro-immune and cardiovascular health to design safer and healthier nature-based medicine for a wide array of costly and highly prevalent chronic diseases, such as inflammatory and metabolic-related syndromes, namely, PCOS. Full article

Grape pomace is a winery byproduct that is rich in polyphenols with antioxidant capacity. This study investigated the effect of 0, 5, and 10% inclusion of dehydrated grape pomace (DGP) in finishing pig diets on the growth performance, carcass traits, fatty acid profile, fresh meat quality, and fat stability of a local pig genotype. A total of 36 pigs, 18 barrows, and 18 gilts (83.23 ± 6.03 kg and 132.1 ± 5.6 days old) were allotted in a randomized block design considering the initial weight (block) within sex, with six replicates of each sex per treatment. Including DGP in the diets did not affect daily weight gain or the feed-to-gain ratio; however, daily feed intake increased linearly (p < 0.05) and backfat thickness at the last rib, backfat thickness at the first sacral vertebrae, P2 backfat thickness, fat area, and the percentage of lean meat decreased linearly (p < 0.05) in pigs. The inclusion of DGP in pig diets did not affect the antioxidant potential evaluated by thiobarbituric acid-reactive substances in mini hamburgers or the quality characteristics of fresh meat, except for intramuscular fat (EE). The dietary inclusion of DGP linearly increased (p < 0.05) EE, saturated fatty acids, monounsaturated fatty acids, and Σω-3 and reduced the ω-6:ω-3 ratio in a linear way (p < 0.05) in the loin of pigs. We concluded that it is feasible to include up to 10% of DGP in pig diets without affecting growth performance, but carcass quality may be impaired due to increased adiposity. Furthermore, meat quality can be improved by increasing intramuscular fat and ω-3 fatty acid content, but fat stability is not affected when DGP is included at up to 10% of the diet for 49 days prior to slaughter. Full article

The commercial value of kiwifruit is determined mostly by its nutritional composition and antioxidant content. The enrichment of these traits in an era where climate change affects kiwi growth conditions is worth investigating via the application of biostimulants that enhance plant growth. In this work, we evaluated the effects of three commercial biostimulants on the metabolism and growth characteristics of the Actinidia deliciosa cultivar ‘Hayward’: (I) a humic and fulvic-based biostimulant, (II) a glycine–betaine–proline-based biostimulant, and (III) a vegetal amino acid-based biostimulant. A biostimulant-free treatment was used as a control. In the two-year experiment (2022 and 2023), the growth characteristics of kiwi trees were measured, such as stem length, the number of leaves, leaf area, and fresh and dry biomass at the end of each growing season. The leaves’ total chlorophyll, total phenolics, and proline content were detected during the two seasons in order to obtain more insights into plant metabolism. Κiwifruit qualities and antioxidant traits, such as total soluble solids, titratable acidity, firmness, fresh and dry biomass, DPPH, and ascorbic acid (vitamin C), were recorded during the postharvest life for each season. Data analysis illustrated the beneficial impact of some biostimulants on several of the previously mentioned parameters, such as antioxidant kiwifruit traits, especially in the case of glycine–betaine–proline-based acid-based biostimulants. Full article

Stool is a mixture of excrement, microbiota, enzymes, undigested material, and small molecules. Fecal metabolomics has gained interest recently, owing to advances in metabolomics and growing research into both the host’s physiology and the gut microbiome. One challenge with fecal metabolomics is preserving the sample integrity from collection until analysis, as the microbiota and enzymes continue to alter the metabolome following defecation. Currently, flash-freezing or lyophilization are utilized to minimize post-collection metabolome changes; however, this requires complex equipment and immediate processing, precluding the possibility for at-home sampling. Commercial devices containing stabilizing solvents have been developed to facilitate at-home collection, ambient transport, and sample storage. Here, we explore the efficacy of a commercially available stool collection device with a stabilization reagent tailored to fecal metabolomics. Stool samples from six donors were either processed shortly post-collection or stored at room temperature for seven days in the tube, with and without the stabilization reagent. Comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS)-based untargeted metabolomics was utilized for analyzing extracted metabolites. Chemometric analysis was used to evaluate the performance of the device. We found that the device with the stabilization reagent minimized changes in the metabolite profile relative to unstabilized stool left at room temperature for one week. Full article