In the field of biomaterials for prosthetic reconstructive surgery, there is the lack of advanced innovative methods to investigate the potentialities of smart biomaterials before in vivo tests. Despite the complex osteointegration process being difficult to recreate in vitro, this study proposes an advanced in vitro tissue culture model of osteointegration using human bone. Cubic samples of trabecular bone were harvested, as waste material, from hip arthroplasty; inner cylindrical defects were created and assigned to the following groups: (1) empty defects (CTRneg); (2) defects implanted with a cytotoxic copper pin (CTRpos); (3) defects implanted with standard titanium pins (Ti). Tissues were dynamically cultured in mini rotating bioreactors and assessed weekly for viability and sterility. After 8 weeks, immunoenzymatic, microtomographic, histological, and histomorphometric analyses were performed. The model was able to simulate the effects of implantation of the materials, showing a drop in viability in CTR+, while Ti appears to have a trophic effect on bone. MicroCT and a histological analysis supported the results, with signs of matrix and bone deposition at the Ti implant site. Data suggest the reliability of the tested model in recreating the osteointegration process in vitro with the aim of reducing and refining in vivo preclinical models. Full article

Tibetan pig is a unique pig breed native to the Qinghai–Tibet Plateau. To investigate viral communities associated with porcine respiratory disease complex (PRDC), 167 respiratory samples were collected from Tibetan pigs in the Ganzi Tibetan autonomous prefecture of Sichuan province. Following library construction and Illunima Novaseq sequencing, 18 distinct viruses belonging to 15 viral taxonomic families were identified in Tibetan pigs with PRDC. Among the 18 detected viruses, 3 viruses were associated with PRDC, including porcine circovirus type 2 (PCV-2), Torque teno sus virus (TTSuV), and porcine cytomegalovirus (PCMV). The genomic sequences of two PCV-2 strains, three TTSuV strains, and one novel Porprismacovirus strain were assembled by SOAPdenovo software (v2). Sequence alignment and phylogenetic analysis showed that both PCV-2 strains belonged to PCV-2d, three TTSuVs were classified to TTSuV2a and TTSuV2b genotypes, and the Porprismacovirus strain PPMV-SCgz-2022 showed a close genetic relationship with a virus of human origin. Recombination analysis indicated that PPMV-SCgz-2022 may have originated from recombination events between Human 16,806 × 66-213 strain and Porcine 17,668 × 82-593 strain. Furthermore, the high proportion of single infection or co-infection of PCV2/TTSuV2 provides insight into PRDC infection in Tibetan pigs. This is the first report of the viral communities in PRDC-affected Tibetan pigs in this region, and the results provides reference for the prevention and control of respiratory diseases in these animals. Full article

Cracks in tunnel lining surfaces directly threaten structural integrity; therefore, regular inspection of cracks is essential. Lightweight convolutional neural networks (LCNNs) have recently offered a promising alternative to conventional manual inspection. However, the effectiveness of LCNNs is still adversely affected by the lack of sufficient crack images, which limits the potential detection performance. In this paper, transfer learning was used to optimize deep convolutional generative adversarial networks (DCGANs) for crack image synthesis to significantly improve the accuracy of LCNNs. In addition, an improved LCNN model named ShuffleNetV2-1.0-SE was proposed, incorporating the squeeze–excitation (SE) attention mechanism into ShuffleNetV2-1.0 and realizing highly accurate classification results while maintaining lightness. The results show that the DCGAN-based data enhancement method can significantly improve the classification accuracy of ShuffleNetV2-1.0-SE for tunnel lining cracks. ShuffleNetV2-1.0-SE achieves an accuracy of 98.14% on the enhanced dataset, which is superior to multiple advanced LCNN models. Full article

The aim of this in vitro study was to investigate the effect of different toothpaste ingredients on biofilm volume and vitality in an established non-contact biofilm removal model. A multi-species biofilm comprising Porphyromonas gingivalis, Streptococcus sanguinis, and Fusobacterium nucleatum was grown on protein-coated titanium disks. Six disks per group were exposed to 4 seconds non-contact brushing using a sonic toothbrush. Four groups assessed slurries containing different ingredients, i.e., dexpanthenol (DP), peppermint oil (PO), cocamidopropyl betaine (CB), and sodium hydroxide (NaOH), one positive control group with the slurry of a toothpaste (POS), and a negative control group with physiological saline (NEG). Biofilm volume and vitality were measured using live-dead staining and confocal laser scanning microscopy. Statistical analysis comprised descriptive statistics and inter-group differences. In the test groups, lowest vitality and volume were found for CB (50.2 ± 11.9%) and PO (3.6 × 10⁵ ± 1.8 × 10⁵ µm³), respectively. Significant differences regarding biofilm vitality were found comparing CB and PO (p = 0.033), CB and NEG (p = 0.014), NaOH and NEG (p = 0.033), and POS and NEG (p = 0.037). However, no significant inter-group differences for biofilm volume were observed. These findings suggest that CB as a toothpaste ingredient had a considerable impact on biofilm vitality even in a non-contact brushing setting, while no considerable impact on biofilm volume was found. Full article

Fabry disease (FD) is an X-linked lysosomal storage disorder, characterised by the cellular accumulation of globotriaosylceramide due to impaired alpha-galactosidase A enzyme activity. FD may manifest with multisystem pathology, including reduced bone mineral density (BMD). Registry data suggest that the introduction of Fabry-specific therapies (enzyme replacement therapy or chaperone therapy) has led to significant improvements in overall patient outcomes; however, there are limited data on the impact on bone density. The aim of this study was to describe the effect of Fabry-specific therapies on longitudinal changes in bone mineral density (BMD) in FD. We performed a retrospective observational study analysing bone densitometry (DXA) in patients with genetically confirmed FD. Patients were grouped based on the use of Fabry-specific therapies. The between-group longitudinal change in BMD Z-score was analysed using linear mixed effects models. A total of 88 FD patients were analysed (50 untreated; 38 treated). The mean age at first DXA was 38.5 years in the untreated group (84% female) and 43.7 years in the treated group (34% female). There was no significant longitudinal between-group difference in the BMD Z-score at the lumbar spine. However, the Z-score per year at the total hip (β = −0.105, p < 0.001) and femoral neck (β = −0.081, p = 0.001) was significantly lower over time in the treated than the untreated group. This may reflect those receiving therapy having a more severe underlying disease. Nevertheless, this suggests that Fabry-specific therapies do not reverse all disease mechanisms and that the additional management of BMD may be required in this patient population. Full article

Anthropometric assessments are commonly used to diagnose sarcopenia in older adults. However, the ongoing exploration of novel approaches aims to improve the early detection of sarcopenia. This study investigated the association between the height-changing score (HCS) and the risk of sarcopenia defined by anthropometric measurements in 340 older adults (mean age: 66.2 years). The HCS derived from the difference in height and demi-span equivalent height (DEH) was used as an indicator of declining height in the older adults. Multivariate logistic regression analysis revealed a significant association between the HCS and the risk of sarcopenia in both male and female older adults (OR = 1.146, 95% CI [1.021, 1.286], p = 0.021). In addition, income, BMI, and nutritional status were significantly associated with the risk of sarcopenia (OR = −1.933, 95% CI [0.271, 0.986], p = 0.045; OR = −2.099, 95% CI [0.386, 0.587], p < 0.001; OR = −1.443, 95% CI [0.555, 0.866], p = 0.001, respectively). The white blood cell count, lymphocyte count, and HDL cholesterol were blood biomarkers significantly correlated with calf circumference. It can be suggested that the HCS acts as an indicator and screening tool for sarcopenia risk in older adults, highlighting the potential impact of decreased height on muscle mass loss. Encouraging nutritional support can help mitigate the risk of sarcopenia. Full article

The aim of this study was to determine the effects of sex and hunting location on carcass characteristics and meat quality of wild-living mallard (Anas platyrhynchos) from Croatia. Twenty-eight mallards (14 ♂; 14 ♀) were hunted at two hunting locations (HL I = 8 ♂, 8 ♀; HL II = 6 ♂, 6 ♀) in the Croatian lowlands. The carcasses were eviscerated, dressed, and dissected, and the individual internal organs and carcass parts were weighed. The breast muscle (m. pectoralis major) was sampled and used to determine color, pH, drip loss, cooking loss, shear force, and proximate chemical and fatty acid composition. Sex and HL had a significant effect on the majority of carcass characteristics analyzed, but they had no effect on the physical meat parameters. The protein and ash content of mallard meat was significantly higher in HL II (23.16 %vs. 22.67%; 1.45% vs. 1.36%, respectively) and the moisture content in females (72.40% vs. 71.59%). HL had a significant effect on SFA (II 33.96% vs. I 29.91%), PUFA n-3 (II 3.55% vs. I 2.69%), PUFA/SFA and n-6/n-3 ratios, and all lipid indices. Females had a significantly higher C22:6n-3 content, a higher PI index and a lower n-6/n-3 ratio. The data presented in this study contribute to a better understanding of game-bird meat quality originating from different regions. Full article

This study’s aim is to examine the effect of a combined balance and multidirectional plyometric training intervention on postural balance ability and lower limb explosive performance in U-13 male and female soccer athletes. Twenty pre-adolescent (age: 12.6 ± 1.6 years) soccer athletes followed a 6-week training intervention combining balance exercises, dynamic stabilization tasks and multidirectional plyometric exercises at a frequency of twice/week for 20–25 min, based on a progressive increase in exercise difficulty from phase A (week 1–3) to phase B (week 4–6). Pre- and post-training measurements were carried out to assess the following: (a) static balance performance in single (left, right)-legged and two-legged quiet stance trials with eyes open and eyes closed (two trials per stance and vision condition of 30 s duration) and (b) lower limb explosive performance in countermovement and squat jumps without arm swing (three trials/jump). The vertical GRF was recorded by a customized force plate (Wii, 1.000 Hz, Biovision) and offline, CoP and explosive performance parameters were calculated. The overall results showed that the static balance ability of athletes remained unaffected, while restricting their vision deteriorated their postural control. The lower limb explosive performance showed a trend for improvement; however, inter-individual variations in athletes’ responses might have obscured any effect. Full article

Accurately predicting the phase behavior and physical properties of carbon dioxide (CO₂) in pure water/NaCl mixtures is crucial for the design and implementation of carbon capture, utilization, and storage (CCUS) technology. However, the prediction task is complicated by CO₂ liquefaction, CO₂ hydrate formation, multicomponent and multiphase coexistence, etc. In this study, an improved method that combines Benedict–Webb–Rubin–Starling equation of state (BWRS EOS) + hydrate thermodynamic theories was proposed to predict CO₂ solubility and phase equilibrium conditions for a mixed system across various temperature and pressure conditions. By modifying the interaction coefficients in BWRS EOS and the Van der Waals–Platteeuw model, this new method is applicable to complex systems containing two liquid phases and a CO₂ hydrate phase, and its high prediction accuracy was verified through a comparative evaluation with a large number of reported experimental data. Furthermore, based on the calculation results, the characteristics of CO₂ solubility and the variation of phase equilibrium conditions of the mixture system were discussed. These findings highlight the influence of hydrates and NaCl on CO₂ solubility characteristics and clearly demonstrate the hindrance of NaCl to the formation of CO₂ hydrates. This study provides valuable insights and fundamental data for designing and implementing CCUS technology that contribute to addressing global climate change and environmental challenges. Full article

The studies carried out address an important problem for the concrete and asphalt industry, referring to the wear phenomena that affect the elements (flights) of rotary dryers used to dry mineral aggregates. In this article, the authors propose a lifetime estimation method for rotary dryer flights. In order to benefit from greater credibility, the proposed method was applied based on the experimental results obtained, by the authors, in the laboratory, on a stand that reproduces a portion of the real equipment. Starting from these results, the authors identified the mathematical functions used to model the failure rates (wear) of the flights by referring to characteristic quantities for wear evaluation: mass loss of material; reduction in the thickness of the part; surface affected by wear. The experimental data—the input data for the proposed algorithm—correspond to six steels used in industrial applications. The wear phenomenon was modeled considering both the situation of uniform wear and the situation of differentiated wear on the flight surface. Therefore, the steel type with the highest service life values could be identified, namely, steel grades E and G. The method proposed in this paper can be extended to any category of material or part provided that the failure rate function is identified beforehand, possibly based on a laboratory experiment. Full article

Vitamin C (VC), also known as ascorbic acid, plays a crucial role as a water-soluble nutrient within the human body, contributing to a variety of metabolic processes. Research findings suggest that increased doses of VC demonstrate potential anti-tumor capabilities. This review delves into the mechanisms of VC absorption and its implications for cancer management. Building upon these foundational insights, we explore modern delivery systems for VC, evaluating its use in diverse cancer treatment methods. These include starvation therapy, chemodynamic therapy (CDT), photothermal/photodynamic therapy (PTT/PDT), electrothermal therapy, immunotherapy, cellular reprogramming, chemotherapy, radiotherapy, and various combination therapies. Full article

Despite great advances in the treatment of oncological diseases, the development of medical technologies to prevent or reduce complications of therapy, in particular, those associated with surgery and the introduction of antibiotics, remains relevant. The aim of this study is to evaluate the effectiveness of the use of autoprobiotics based on indigenous non-pathogenic strains of Enterococcus faecium and Enterococcus hirae as a personalized functional food product (PFFP) in the complex therapy of colorectal cancer (CRC) in the early postoperative period. A total of 36 patients diagnosed with CRC were enrolled in the study. Study group A comprised 24 CRC patients who received autoprobiotic therapy in the early postoperative period, while the control group C included 12 CRC patients without autoprobiotic therapy. Prior to surgery and between days 14 and 16 post-surgery, comprehensive evaluations were conducted on all patients, encompassing the following: stool and gastroenterological complaints analysis, examination of the gut microbiota (bacteriological study, quantitative polymerase chain reaction, metagenome analysis), and analysis of interleukins in the serum. Results: The use of autoprobiotics led to a decrease in dyspeptic complaints after surgery. It was also associated with the absence of postoperative complications, did not cause any side effects, and led to a decrease in the level of pro-inflammatory cytokines (IL-6 and IL-18) in the blood serum. The use of autoprobiotics led to positive changes in the structure of escherichia and enterococci populations, the elimination of Parvomonas micra and Fusobacterium nucleatum, and a decrease in the quantitative content of Clostridium perfringens and Akkermansia muciniphila. Metagenomic analysis (16S rRNA) revealed an increase in alpha diversity. Conclusion: The introduction of autoprobiotics in the postoperative period is a highly effective and safe approach in the complex treatment of CRC. Future studies will allow the discovery of additional fine mechanisms of autoprobiotic therapy and its impact on the digestive, immune, endocrine, and neural systems. Full article

The aim of this work is to obtain novel and interesting results for mild solutions to a semilinear differential inclusion involving a w-weighted, $\Phi$-Hilfer, fractional derivative of order $\mu \in (1,2)$ with non-instantaneous impulses in Banach spaces with infinite dimensions when the linear term is the infinitesimal generator of a strongly continuous cosine family and the nonlinear term is a multi-valued function. First, we determine the formula of the mild solution function for the considered semilinear differential inclusion. Then, we give sufficient conditions to ensure that the mild solution set is not empty or compact. The desired results are achieved by using the properties of both the w-weighted $\Phi$-Laplace transform, w-weighted $\psi$-convolution and the measure of non-compactness. Since the operator, the w-weighted $\Phi$-Hilfer, includes well-known types of fractional differential operators, our results generalize several recent results in the literature. Moreover, our results are novel because no one has previously studied these types of semilinear differential inclusions. Finally, we give an illustrative example that supports our theoretical results. Full article

Health systems in high-resource countries recognize the importance of making decisions about the services offered to the population based on scientific evidence. Producing this evidence is especially challenging in areas such as newborn care where the frequency of conditions is rare. However, methodological advances in the field of economic evaluation could change how this evidence is used in decision-making. This study aimed to investigate how decision-makers in the Canadian province of Quebec perceive the value of recent advances in economic evaluations for perinatal studies and how these advances might affect the offer of neonatal interventions in the public health care system. A qualitative study was conducted. A total of 10 policymakers were interviewed. A neo-institutional conceptual framework highlighting three dimensions, structure, power, and interpretive schemes, was used for data collection and analyses. Structural factors, interpretative schemes, and power management between the groups concerned concur to ensure that providing services to newborns is not hindered by the difficulty of producing evidence. They also ensure that the decisions regarding which disease to screen for take into consideration the specificity of neonatology, in particular, the social value given to children not captured by available evidence. Full article

The shift towards sustainable transportation is an urgent worldwide issue, leading to the investigation of creative methods to decrease the environmental effects of traditional vehicles. Electric vehicles (EVs) are a promising alternative, but the issue lies in establishing efficient and environmentally friendly charging infrastructure. This review explores the existing research on the subject of photovoltaic-powered electric vehicle charging stations (EVCSs). Our analysis highlights the potential for economic growth and the creation of robust and decentralized energy systems by increasing the number of EVCSs. This review summarizes the current knowledge in this field and highlights the key factors driving efforts to expand the use of PV-powered EVCSs. The findings indicate that MATLAB was predominantly used for theoretical studies, with projects focusing on shading parking lots. The energy usage varied from 0.139 to 0.295 kWh/km, while the cost of energy ranged from USD 0.0032 to 0.5645 per kWh for an on-grid system. The payback period (PBP) values are suitable for this application. The average PBP was demonstrated to range from 1 to 15 years. The findings from this assessment can guide policymakers, researchers, and industry stakeholders in shaping future advancements toward a cleaner and more sustainable transportation system. Full article

The dung beetle optimization (DBO) algorithm, a swarm intelligence-based metaheuristic, is renowned for its robust optimization capability and fast convergence speed. However, it also suffers from low population diversity, susceptibility to local optima solutions, and unsatisfactory convergence speed when facing complex optimization problems. In response, this paper proposes the multi-strategy improved dung beetle optimization algorithm (MDBO). The core improvements include using Latin hypercube sampling for better population initialization and the introduction of a novel differential variation strategy, termed “Mean Differential Variation”, to enhance the algorithm’s ability to evade local optima. Moreover, a strategy combining lens imaging reverse learning and dimension-by-dimension optimization was proposed and applied to the current optimal solution. Through comprehensive performance testing on standard benchmark functions from CEC2017 and CEC2020, MDBO demonstrates superior performance in terms of optimization accuracy, stability, and convergence speed compared with other classical metaheuristic optimization algorithms. Additionally, the efficacy of MDBO in addressing complex real-world engineering problems is validated through three representative engineering application scenarios namely extension/compression spring design problems, reducer design problems, and welded beam design problems. Full article

One of the significant challenges in scaling agile software development is organizing software development teams to ensure effective communication among members while equipping them with the capabilities to deliver business value independently. A formal approach to address this challenge involves modeling it as an optimization problem: given a professional staff, how can they be organized to optimize the number of communication channels, considering both intra-team and inter-team channels? In this article, we propose applying a set of bio-inspired algorithms to solve this problem. We introduce an enhancement that incorporates ensemble learning into the resolution process to achieve nearly optimal results. Ensemble learning integrates multiple machine-learning strategies with diverse characteristics to boost optimizer performance. Furthermore, the studied metaheuristics offer an excellent opportunity to explore their linear convergence, contingent on the exploration and exploitation phases. The results produce more precise definitions for team sizes, aligning with industry standards. Our approach demonstrates superior performance compared to the traditional versions of these algorithms. Full article

According to the Magnus principle, a rotating cylinder experiences a lateral force perpendicular to the incoming flow direction. This phenomenon can be harnessed to boost the lift of an airfoil by positioning a rotating cylinder at the leading edge. In this study, we simulate flapping-wing motion using the sliding mesh technique in a heaving coordinate system to investigate the energy harvesting capabilities of Magnus effect flapping wings (MEFWs) featuring a leading-edge rotating cylinder. Through analysis of the flow field vortex structure and pressure distribution, we explore how control parameters such as gap width, rotational speed ratio, and phase difference of the leading-edge rotating cylinder impact the energy harvesting characteristics of the flapping wing. The results demonstrate that MEFWs effectively mitigate the formation of leading-edge vortices during wing motion. Consequently, this enhances both lift generation and energy harvesting capability. MEFWs with smaller gap widths are less prone to induce the detachment of leading-edge vortices during motion, ensuring a higher peak lift force and an increase in the energy harvesting efficiency. Moreover, higher rotational speed ratios and phase differences, synchronized with wing motion, can prevent leading-edge vortex generation during wing motion. All three control parameters contribute to enhancing the energy harvesting capability of MEFWs within a certain range. At the examined Reynolds number, the optimal parameter values are determined to be = 0.0005, R = 3, and = 0°. Full article

Marine shales from the Niutitang Formation and marine–continental transitional shales from the Longtan Formation are two sets of extremely important hydrocarbon source rocks in South China. In order to quantitatively compare the pore complexity characteristics between marine and marine–continental transitional shales, the shale and kerogen of the Niutitang Formation and the Longtan Formation are taken as our research subjects. Based on organic petrology, geochemistry, and low-temperature gas adsorption analyses, the fractal dimension of their pores is calculated by the Frenkel–Halsey–Hill (FHH) and Sierpinski models, and the influences of total organic carbon (TOC), vitrinite reflectance (R_o), and mineral composition on the pore fractals of the shale and kerogen are discussed. Our results show the following: (1) Marine shale predominantly has wedge-shaped and slit pores, while marine–continental transitional shale has inkpot-shaped and slit pores. (2) Cylindrical pores are common in organic matter of both shale types, with marine shale having a greater gas storage space (CRV) from organic matter pores, while marine–continental transitional shale relies more on inorganic pores, especially interlayer clay mineral pores, for gas storage due to their large specific surface area and high adsorption capacity (CRA). (3) The fractal characteristics of marine and marine–continental transitional shale pores are influenced differently. In marine shale, TOC positively correlates with fractal dimensions, while in marine–continental shale, R_o and clay minerals have a stronger influence. R_o is the primary factor affecting organic matter pore complexity. (4) Our two pore fractal models show that the complexity of the shale in the Longtan Formation surpasses that of the shale in the Niutitang Formation, and type I kerogen has more complex organic matter pores than type III, aiding in evaluating pore connectivity and flow effectiveness in shale reservoirs. Full article

Previous studies have shown that boletes are abundant and diverse in China, especially in tropical and subtropical regions. In the present study, morphological, ecological, host relationship, and a four-locus (28S, tef1, rpb1, and rpb2) molecular phylogenetic analyses were used to study the family Boletaceae in subtropical and tropical China. Four new bluing species are described from three genera, viz. Boletellus verruculosus (Chinese name疣柄条孢牛肝菌), Xerocomellus tenuis (Chinese name细柄红绒盖牛肝菌), Xer. brunneus (Chinese name褐盖红绒盖牛肝菌), and Xerocomus zhangii (Chinese name张氏绒盖牛肝菌). Moreover, the genus Nigroboletus is treated as a synonym of Xerocomellus, and a new combination, namely Xer. roseonigrescens (Chinese name玫瑰红绒盖牛肝菌), is proposed. Full article

Autophagy, a conserved cellular recycling process, plays a crucial role in maintaining homeostasis under stress conditions. It also regulates the development and virulence of numerous filamentous fungi. In this study, we investigated the specific function of ATG8, a reliable autophagic marker, in the opportunistic pathogen Aspergillus flavus. To investigate the role of atg8 in A. flavus, the deletion and complemented mutants of atg8 were generated according to the homologous recombination principle. Deletion of atg8 showed a significant decrease in conidiation, spore germination, and sclerotia formation compared to the WT and atg8^Cstrains. Additionally, aflatoxin production was found severely impaired in the ∆atg8 mutant. The stress assays demonstrated that ATG8 was important for A. flavus response to oxidative stress. The fluorescence microscopy showed increased levels of reactive oxygen species in the ∆atg8 mutant cells, and the transcriptional result also indicated that genes related to the antioxidant system were significantly reduced in the ∆atg8 mutant. We further found that ATG8 participated in regulating the pathogenicity of A. flavus on crop seeds. These results revealed the biological role of ATG8 in A. flavus, which might provide a potential target for the control of A. flavus and AFB1 biosynthesis. Full article

Background: The pathogenic fungus Candida albicans is a leading agent of death in immunocompromised individuals with a growing trend of antifungal resistance. Methods: The purpose is to induce resistance to drugs in a sensitive C. albicans strain followed by whole-genome sequencing to determine mechanisms of resistance. Strains will be assayed for pathogenicity attributes such as ergosterol and chitin content, growth rate, virulence, and biofilm formation. Results: We observed sequential increases in ergosterol and chitin content in fluconazole-resistant isolates by 78% and 44%. Surface thickening prevents the entry of the drug, resulting in resistance. Resistance imposed a fitness trade-off that led to reduced growth rates, biofilm formation, and virulence in our isolates. Sequencing revealed mutations in genes involved in resistance and pathogenicity such as ERG11, CHS3, GSC2, CDR2, CRZ2, and MSH2. We observed an increase in the number of mutations in key genes with a sequential increase in drug-selective pressures as the organism increased its odds of adapting to inhospitable environments. In ALS4, we observed two mutations in the susceptible strain and five mutations in the resistant strain. Conclusion: This is the first study to induce resistance followed by genotypic and phenotypic analysis of isolates to determine mechanisms of drug resistance. Full article