Unlocking the Therapeutic Potential of BCL-2 Associated Protein Family: Exploring BCL-2 Inhibitors in Cancer Therapy.

Apoptosis, programmed cell death pathway, is a vital physiological mechanism that ensures cellular homeostasis and overall cellular well-being. In the context of cancer, where evasion of apoptosis is a hallmark, the overexpression of anti-apoptotic proteins like Bcl2, Bcl-xL and Mcl-1 has been documented. Consequently, these proteins have emerged as promising targets for therapeutic interventions. The BCL-2 protein family is central to apoptosis and plays a significant importance in determining cellular fate serving as a critical determinant in this biological process. This review offers a comprehensive exploration of the BCL-2 protein family, emphasizing its dual nature. Specifically, certain members of this family promote cell survival (known as anti-apoptotic proteins), while others are involved in facilitating cell death (referred to as pro-apoptotic and BH3-only proteins). The potential of directly targeting these proteins is examined, particularly due to their involvement in conferring resistance to traditional cancer therapies. The effectiveness of such targeting strategies is also discussed, considering the tumor's propensity for anti-apoptotic pathways. Furthermore, the review highlights emerging research on combination therapies, where BCL-2 inhibitors are used synergistically with other treatments to enhance therapeutic outcomes. By understanding and manipulating the BCL-2 family and its associated pathways, we open doors to innovative and more effective cancer treatments, offering hope for resistant and aggressive cases.

PLGA-Gold Nanocomposite: Preparation and Biomedical Applications.

A composite system consisting of both organic and inorganic nanoparticles is an approach to prepare a new material exhibiting "the best of both worlds". In this review, we highlight the recent advances in the preparation and applications of poly(lactic-co-glycolic acid)-gold nanoparticles (PLGA-GNP). With its current clinically use, PLGA-based nanocarriers have promising pharmaceutical applications and can "extract and utilize" the fascinating optical and photothermal properties of encapsulated GNP. The resulting "golden polymeric nanocarrier" can be tracked, analyzed, and visualized using the encapsulated gold nanoprobes which facilitate a better understanding of the hosting nanocarrier's pharmacokinetics and biological fate. In addition, the "golden polymeric nanocarrier" can reveal superior nanotherapeutics that combine both the photothermal effect of the encapsulated gold nanoparticles and co-loaded chemotherapeutics. To help stimulate more research on the development of nanomaterials with hybrid and exceptional properties, functionalities, and applications, this review provides recent examples with a focus on the available chemistries and the rationale behind encapsulating GNP into PLGA nanocarriers that has the potential to be translated into innovative, clinically applicable nanomedicine.

Prediction of qualitative antibiofilm activity of antibiotics using supervised machine learning techniques.

Although biofilm-specific antibiotic susceptibility assays are available, they are time-consuming and resource-intensive, and hence they are not usually performed in clinical settings. Herein, we introduce a machine learning-based predictive modeling approach that uses routinely available and easily accessible data to qualitatively predict in vitro antibiofilm activity of antibiotics with relatively high accuracy. Three optimized models based on logistic regression, decision tree, and random forest algorithms were successfully developed in this study using data manually collected from published literature. In these models, independent variables that serve as significant predictors of antibiofilm activity are minimum inhibitory concentration, bacterial Gram type, biofilm formation method, in addition to antibiotic's mechanism of action, molecular weight, and pKa. The cross-validation method showed that the optimized models exhibit prediction accuracy of 67% ± 6.1% for the logistic regression model, 73% ± 5.8% for the decision tree model, and 74% ± 5% for the random forest model. However, the one-way ANOVA test revealed that the difference in prediction accuracy between the 3 models is not statistically significant, and hence they can be considered to have comparable performance. The presented modeling approach can serve as an alternative to the resource-intensive biofilm assays to rapidly and properly manage biofilm-associated infections, especially in resource-limited clinical settings.

Aqueous core microcapsules as potential long-acting release systems for hydrophilic drugs.

We have previously optimized the internal phase separation process to give rise to aqueous core microcapsules with polymeric shells composed of poly(lactide-co-glycolide) (PLGA) or poly(lactide) (PLA). In this study, the ability of these microcapsules to act as controlled release platforms of the model hydrophilic drug phenobarbital sodium was tested. Furthermore, the effect of the initial amounts of drug and water added to the system during microcapsule synthesis was investigated. Finally, the effect of varying polymer properties such as end functionalities, molecular weights, and lactide to glycolide ratios, on the characteristics of the produced microcapsules was studied. This was done by utilizing seven different grades of the polyester polymers. It was demonstrated that, within certain limits, drug loading is nearly proportional to the initial amounts of drug and water. Furthermore, drug encapsulation studies demonstrated that ester termination and increases in polymeric molecular weight result in lower drug loading and encapsulation efficiency. Moreover, drug release studies demonstrated that ester termination, increases in molecular weight, and increases in the lactide to glycolide ratio all result in slower drug release; this grants the ability to tailor the drug release duration from a few days to several weeks. In conclusion, such minor variations in polymer characteristics and formulation composition can result in dramatic changes in the properties of the produced microcapsules. These changes can be fine-tuned to obtain desirable long-acting microcapsules capable of encapsulating a variety of hydrophilic drugs which can be used in a wide range of applications.

Synergistic antibacterial activity of silver nanoparticles and hydrogen peroxide.

The increasing challenge of antibiotic resistance requires not only the discovery of new antibiotics, but also the development of new alternative approaches. Herein, the synergistic antibacterial activity of silver nanoparticles and hydrogen peroxide combination is reported. Unlike the bacteriostatic or slightly bactericidal activity achieved by using each agent alone, using these two agents in combination, even at relatively low concentrations, resulted in complete eradication of both the Gram negative Escherichia coli and the Gram positive Staphylococcus aureus in short treatment times indicating a clear synergistic effect between them. Modifying the surface chemistry of silver nanoparticles and the accompanied change in their surface charge enabled a further enhancement of such synergistic effect implying the importance of this aspect. Mechanistically, a Fenton-like reaction between silver nanoparticles and hydrogen peroxide is discussed and hypothesized to be the basis of the observed synergy. Achieving such a significant antibacterial activity at low concentrations reduces the potential toxicity of these agents and hence enables their utilization as an alternative antibacterial approach in wider range of applications.

Facile Hydrophobication of Glutathione-Protected Gold Nanoclusters and Encapsulation into Poly(lactide-co-glycolide) Nanocarriers.

We report a simple surface functionalization of glutathione-capped gold nanoclusters by hydrophobic ion pairing with alkylamine followed by a complete phase transfer to various organic solvents with maintained colloidal stability and photoluminescence properties. The described surface hydrophobication enables efficient encapsulation of gold nanoclusters into PLGA nanocarriers allowing their visualization inside cultured cells using confocal fluorescent microscopy. The simplicity and efficiency of the described protocols should extend the biomedical applications of these metallic nanoclusters as a fluorescent platform to label hydrophobic polymeric nanocarriers beyond conventional organic dyes.

Self-evaluation and professional status as predictors of burnout among nurses in Jordan.

The aim of this study is to evaluate the contribution of self-evaluation, professional status and several demographic factors in predicting burnout among nurses in Jordan. This study was performed on a stratified convenience sample of 350 nurses from 6 hospitals. Modified versions of burnout inventory, self-evaluation scale, and professional status scale were developed, validated and used in this study. Burnout, self-evaluation, and professional status are all found to be at moderate levels. Furthermore, self-evaluation and professional status are found to be important predictors of burnout. On the other hand, although type of hospital and educational level are found to be associated with the level of burnout, overall none of the studied demographic factors was found to be a significant predictor of burnout among nurses in Jordan.

Tunable sustained release drug delivery system based on mononuclear aqueous core-polymer shell microcapsules.

Poly(d,l-lactide-co-glycolide) (PLGA) and poly(d,l-lactide) (PLA) polymers were used successfully in the preparation of polymer shell microcapsules with mononuclear aqueous cores by the internal phase separation method. These microcapsules were prepared with varying amounts of polymer and water and loaded with fluorescein sodium as a model water soluble drug. Evaluation of drug loading and encapsulation efficiency reveals an optimum polymer to water ratio of around 1:3. Prepared PLGA and PLA microcapsules exhibit sustained drug release over 7 and 49 days, respectively. Drug release from both microcapsule types follow zero order kinetics over the first 90% release. Further tuning of release rate is found possible by preparing microcapsules with mixtures of PLGA and PLA polymers at varying ratios. These results suggest that aqueous core-PLGA and PLA microcapsules would be promising platforms for a wide range of sustained drug delivery systems for many hydrophilic drugs.

Evaluation of paromomycin sulphate permeation using ex vivo human skin model.

Ex vivo evaluation of drug release and skin permeation from topical formulations of antileishmanial drug paromomycin sulphate was carried out using intact full thickness human skin. Potency-based microbiological assay was used for the analysis of paromomycin concentrations. A total percentage drug recovery of 86 ± 26% was obtained. Incubation periods of 1 and 3 h resulted in percentage drug permeation into deep skin layers ranging from 1.3 ± 0.04% to 5.3 ± 2.0% with paraffin-based ointment and from 1.6 ± 0.8% to 3.9 ± 1% with microemulsion-based emulgel. Although a small percentage, this is still significantly higher than those previously reported using animal skin models.

Controlling the internal morphology of aqueous core-PLGA shell microcapsules: promoting the internal phase separation via alcohol addition.

The ability to control the internal core architecture of polymeric microcapsules has a direct impact on their applications. However, this task, especially to produce microcapsules with a high percentage of mononuclear aqueous cores, proved to be challenging. In this work, and in continuation to our previous studies, we report a facile protocol to prepare poly(D,L-lactide-co-glycolide) (PLGA) microcapsules with unprecedented percentage (almost 100%) of mononuclear aqueous cores by the internal phase separation method via adding alcohols. Different types of alcohols (methanol, ethanol, propanol, isopropanol, butanol, and octanol) were incorporated into the internal phase solution and then emulsified into mineral oil. In situ monitoring of emulsion droplets was performed by phase contrast microscopy at different time points and the percentage of mononuclear droplets was measured. While alcohol-free formulation ended up with only around 51% of mononuclear microcapsules, incorporating alcohols resulted in the formation of more than 90% of mononuclear microcapsules. Octanol, in particular, exhibited an outstanding performance as its incorporation led to an immediate (at 0 h) formation of almost entirely mononuclear microcapsules. Final microcapsules exhibited spherical shape with mean particle size in the range of 1-2 µm as depicted by scanning electron microscopy and dynamic light scattering analysis.

Cellular uptake of nanoparticles: journey inside the cell.

Nanoscale materials are increasingly found in consumer goods, electronics, and pharmaceuticals. While these particles interact with the body in myriad ways, their beneficial and/or deleterious effects ultimately arise from interactions at the cellular and subcellular level. Nanoparticles (NPs) can modulate cell fate, induce or prevent mutations, initiate cell-cell communication, and modulate cell structure in a manner dictated largely by phenomena at the nano-bio interface. Recent advances in chemical synthesis have yielded new nanoscale materials with precisely defined biochemical features, and emerging analytical techniques have shed light on nuanced and context-dependent nano-bio interactions within cells. In this review, we provide an objective and comprehensive account of our current understanding of the cellular uptake of NPs and the underlying parameters controlling the nano-cellular interactions, along with the available analytical techniques to follow and track these processes.

Preparation of Aqueous Core-Poly(d,l-Lactide-co-Glycolide) Shell Microcapsules With Mononuclear Cores by Internal Phase Separation: Optimization of Formulation Parameters.

Previously, our group employed the internal phase separation method to produce aqueous core-PLGA [poly(d,l-lactide-co-glycolide)] shell microcapsules with polynuclear core morphologies. This report describes the preparation of the more desired and challenging architecture with mononuclear cores. Optimization of formulation parameters including (1) varying the composition of the internal phase and (2) incorporating selected organic solvents (dichloromethane, chloroform, methanol, and acetonitrile) into the internal phase was systematically evaluated. Varying the composition of the internal phase (i.e., PLGA and water levels) failed to produce mononuclear microcapsules. However, incorporating methanol or acetonitrile into the internal phase produced microcapsules with mononuclear cores as confirmed by phase-contrast microscopy, transmission electron microscopy, and scanning electron microscopy. Stability of the prepared emulsions (internal phase of PLGA, acetone, acetonitrile, and water) was optimized by evaluating different types of surfactants with varying concentrations. Among them, lecithin in the range of 0.5%-5% wt/wt provided the best emulsion stability. Interestingly, increasing lecithin concentrations led to the production of microcapsules with smaller sizes (from 2.4 ± 1.6 to 1.1 ± 0.8 µm) and higher percentage of mononuclear cores. The resulting aqueous core-PLGA shell microcapsules are expected to have interesting applications in drug delivery systems with controlled release for hydrophilic drugs and proteins.

Eradication and phenotypic tolerance of Burkholderia cenocepacia biofilms exposed to atmospheric pressure non-thermal plasma.

Chronic lung infection with bacteria from the Burkholderia cepacia complex (BCC), and in particular B. cenocepacia, is associated with significant morbidity and mortality in patients with cystic fibrosis (CF). B. cenocepacia can spread from person to person and exhibits intrinsic broad-spectrum antibiotic resistance. Recently, atmospheric pressure non-thermal plasmas (APNTPs) have gained increasing attention as a novel approach to the prevention and treatment of a variety of hospital-acquired infections. In this study, we evaluated an in-house-designed kHz-driven plasma source for the treatment of biofilms of a number of clinical CF B. cenocepacia isolates. The results demonstrated that APNTP is an effective and efficient tool for the eradication of B. cenocepacia biofilms but that efficacy is highly variable across different isolates. Determination of phenotypic differences between isolates in an attempt to understand variability in plasma tolerance revealed that isolates which are highly tolerant to APNTP typically produce biofilms of greater biomass than their more sensitive counterparts. This indicates a potential role for biofilm matrix components in biofilm tolerance to APNTP exposure. Furthermore, significant isolate-dependent differences in catalase activity in planktonic bacteria positively correlated with phenotypic resistance to APNTP by isolates grown in biofilms.

Potential cellular targets and antibacterial efficacy of atmospheric pressure non-thermal plasma.

Atmospheric pressure non-thermal plasma (APNTP) has been gaining increasing interest as a new alternative antibacterial approach. Although this approach has demonstrated promising antibacterial activity, its exact mechanism of action remains unclear. Mechanistic elucidation of the antimicrobial activity will facilitate development and rational optimisation of this approach for potential medical applications. In this study, the antibacterial efficacy of an in-house-built APNTP jet was evaluated alongside an investigation of the interactions between APNTP and major cellular components in order to identify the potential cellular targets involved in plasma-mediated bacterial destruction mechanisms. The investigated plasma jet exhibited excellent, rapid antibacterial activity against a selected panel of clinically significant bacterial species including Bacillus cereus, meticillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and Pseudomonas aeruginosa, all of which were completely inactivated within 2 min of plasma exposure. Plasma-mediated damaging effects were observed, to varying degrees, on all of the investigated cellular components including DNA, a model protein enzyme, and lipid membrane integrity and permeability. The antibacterial efficacy of APNTP appears to involve a multiple-target mechanism, which potentially reduces the likelihood of emergence of microbial resistance towards this promising antimicrobial approach. However, cellular membrane damage and resulting permeability perturbation was found to be the most likely rate-determining step in this mechanism.

Eradication of Pseudomonas aeruginosa biofilms by atmospheric pressure non-thermal plasma.

Bacteria exist, in most environments, as complex, organised communities of sessile cells embedded within a matrix of self-produced, hydrated extracellular polymeric substances known as biofilms. Bacterial biofilms represent a ubiquitous and predominant cause of both chronic infections and infections associated with the use of indwelling medical devices such as catheters and prostheses. Such infections typically exhibit significantly enhanced tolerance to antimicrobial, biocidal and immunological challenge. This renders them difficult, sometimes impossible, to treat using conventional chemotherapeutic agents. Effective alternative approaches for prevention and eradication of biofilm associated chronic and device-associated infections are therefore urgently required. Atmospheric pressure non-thermal plasmas are gaining increasing attention as a potential approach for the eradication and control of bacterial infection and contamination. To date, however, the majority of studies have been conducted with reference to planktonic bacteria and rather less attention has been directed towards bacteria in the biofilm mode of growth. In this study, the activity of a kilohertz-driven atmospheric pressure non-thermal plasma jet, operated in a helium oxygen mixture, against Pseudomonas aeruginosa in vitro biofilms was evaluated. Pseudomonas aeruginosa biofilms exhibit marked susceptibility to exposure of the plasma jet effluent, following even relatively short (≈ 10's s) exposure times. Manipulation of plasma operating conditions, for example, plasma operating frequency, had a significant effect on the bacterial inactivation rate. Survival curves exhibit a rapid decline in the number of surviving cells in the first 60 seconds followed by slower rate of cell number reduction. Excellent anti-biofilm activity of the plasma jet was also demonstrated by both confocal scanning laser microscopy and metabolism of the tetrazolium salt, XTT, a measure of bactericidal activity.

Synergistic phage-antibiotic combinations for the control of Escherichia coli biofilms in vitro.

The potential application of phage therapy for the control of bacterial biofilms has received increasing attention as resistance to conventional antibiotic agents continues to increase. The present study identifies antimicrobial synergy between bacteriophage T4 and a conventional antibiotic, cefotaxime, via standard plaque assay and, importantly, in the in vitro eradication of biofilms of the T4 host strain Escherichia coli 11303. Phage-antibiotic synergy (PAS) is defined as the phenomenon whereby sub-lethal concentrations of certain antibiotics can substantially stimulate the host bacteria's production of virulent phage. Increasing sub-lethal concentrations of cefotaxime resulted in an observed increase in T4 plaque size and T4 concentration. The application of PAS to the T4 one-step growth curve also resulted in an increased burst size and reduced latent period. Combinations of T4 bacteriophage and cefotaxime significantly enhanced the eradication of bacterial biofilms when compared to treatment with cefotaxime alone. The addition of medium (10(4) PFU mL(-1)) and high (10(7) PFU mL(-1)) phage titres reduced the minimum biofilm eradication concentration value of cefotaxime against E. coli ATCC 11303 biofilms from 256 to 128 and 32 µg mL(-1), respectively. Although further investigation is needed to confirm PAS, this study demonstrates, for the first time, that synergy between bacteriophage and conventional antibiotics can significantly improve biofilm control in vitro.

Application of atmospheric pressure nonthermal plasma for the in vitro eradication of bacterial biofilms.

The use of atmospheric pressure nonthermal plasma represents an interesting and novel approach for the decontamination of surfaces colonized with microbial biofilms that exhibit enhanced tolerance to antimicrobial challenge. In this study, the influence of an atmospheric pressure nonthermal plasma jet, operated in a helium and oxygen gas mixture under ambient pressure, was evaluated against biofilms of Bacillus cereus, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Within < 4 min of plasma exposure, complete eradication of the two gram-positive bacterial biofilms was achieved. Although gram-negative biofilms required longer treatment time, their complete eradication was still possible with 10 min of exposure. Whilst this study provides useful proof of concept data on the use of atmospheric pressure plasmas for the eradication of bacterial biofilms in vitro, it also demonstrates the critical need for improved understanding of the mechanisms and kinetics related to such a potentially significant approach.