Target Product Research Articles

A loop-mediated isothermal amplification test for yaws: a multi-country diagnostic accuracy evaluation

To meet the WHO target of eradicating yaws by 2030, highly sensitive and specific diagnostic tools are needed. A multiplex Treponema pallidum-Haemophilus ducreyi loop-mediated isothermal amplification (TPHD-LAMP) test holds promise as a near-patient diagnostic tool for yaws and H ducreyi. We conducted a prospective evaluation in Cameroon, Côte d'Ivoire, Ghana, and the Republic of the Congo to determine the diagnostic accuracy of the TPHD-LAMP test, as well as to assess its acceptability, feasibility, and cost. Active case searching within schools and communities was used to locate participants with clinically suspicious laws-like lesions. Individuals with serologically confirmed active yaws provided paired lesion swabs between March, 2021, and April, 2023. For each participant, one swab was tested with the TPHD-LAMP at a local district laboratory and the other with reference quantitative PCR (qPCR) tests conducted at national reference laboratories. The primary outcome was TPHD-LAMP test sensitivity and specificity compared with qPCR. Laboratory technicians were interviewed using a multiple-choice survey to gauge acceptability and feasibility of the TPHD-LAMP test. Costs of each test were calculated. Of 3085 individuals with at least one suspected yaws lesion, 531 (17%) were serologically confirmed. We enrolled 493 participants with seropositive yaws and a further 32 with negative serology. The sensitivity of the TPHD-LAMP test for detecting T pallidum was 63% (95% CI 56-70) and the specificity was 66% (95% CI 61-71). Sensitivity and specificity for T pallidum improved to 73% (63-82; p=0·0065) and 75% (68-80; p=0·0003), respectively, in H ducreyi-negative samples. Interviews highlighted challenges in user-friendliness and practicality of the TPHD-LAMP test. The cost of the test per sample was one third of that of qPCR, although the TPHD-LAMP test entailed higher costs to establish the assay. This was the first multi-country diagnostic evaluation of a molecular test for yaws. The TPHD-LAMP testing, in its current form, falls short of the WHO target product profile criteria for yaws diagnostics. These findings highlight the importance of assessing new diagnostics in real-world conditions to ensure their suitability for programmatic use. The EDCTP2 programme supported by the EU.

Lignin valorization to bioplastics with an aromatic hub metabolite-based autoregulation system

Exploring microorganisms with downstream synthetic advantages in lignin valorization is an effective strategy to increase target product diversity and yield. This study ingeniously engineers the non-lignin-degrading bacterium Ralstonia eutropha H16 (also known as Cupriavidus necator H16) to convert lignin, a typically underutilized by-product of biorefinery, into valuable bioplastic polyhydroxybutyrate (PHB). The aromatic metabolism capacities of R. eutropha H16 for different lignin-derived aromatics (LDAs) are systematically characterized and complemented by integrating robust functional modules including O-demethylation, aromatic aldehyde metabolism and the mitigation of by-product inhibition. A pivotal discovery is the regulatory element PcaQ, which is highly responsive to the aromatic hub metabolite protocatechuic acid during lignin degradation. Based on the computer-aided design of PcaQ, we develop a hub metabolite-based autoregulation (HMA) system. This system can control the functional genes expression in response to heterologous LDAs and enhance metabolism efficiency. Multi-module genome integration and directed evolution further fortify the strain’s stability and lignin conversion capacities, leading to PHB production titer of 2.38 g/L using heterologous LDAs as sole carbon source. This work not only marks a leap in bioplastic production from lignin components but also provides a strategy to redesign the non-LDAs-degrading microbes for efficient lignin valorization.

A comprehensive evaluation of an artificial intelligence based digital pathology to monitor large-scale deworming programs against soil-transmitted helminths: A study protocol.

Manual screening of a Kato-Katz (KK) thick stool smear remains the current standard to monitor the impact of large-scale deworming programs against soil-transmitted helminths (STHs). To improve this diagnostic standard, we recently designed an artificial intelligence based digital pathology system (AI-DP) for digital image capture and analysis of KK thick smears. Preliminary results of its diagnostic performance are encouraging, and a comprehensive evaluation of this technology as a cost-efficient end-to-end diagnostic to inform STH control programs against the target product profiles (TPP) of the World Health Organisation (WHO) is the next step for validation. Here, we describe the study protocol for a comprehensive evaluation of the AI-DP based on its (i) diagnostic performance, (ii) repeatability/reproducibility, (iii) time-to-result, (iv) cost-efficiency to inform large-scale deworming programs, and (v) usability in both laboratory and field settings. For each of these five attributes, we designed separate experiments with sufficient power to verify the non-inferiority of the AI-DP (KK2.0) over the manual screening of the KK stool thick smears (KK1.0). These experiments will be conducted in two STH endemic countries with national deworming programs (Ethiopia and Uganda), focussing on school-age children only. This comprehensive study will provide the necessary data to make an evidence-based decision on whether the technology is indeed performant and a cost-efficient end-to-end diagnostic to inform large-scale deworming programs against STHs. Following the protocolized collection of high-quality data we will seek approval by WHO. Through the dissemination of our methodology and statistics, we hope to support additional developments in AI-DP technologies for other neglected tropical diseases in resource-limited settings. The trial was registered on September 29, 2023 Clinicaltrials.gov (ID: NCT06055530).

Ag–Pt Alloy Nanoparticles Modified Zn‐Based Nanosheets for Highly Selective CO2 Photoreduction to CH4

AbstractMulti‐electron involved photoreduction of CO2 to hydrocarbon fuels is a long‐standing challenge, particularly in manipulating the selectivity of the target products. Here, a novel Ag–Pt bimetallic alloy on Zn‐based supports by photo‐deposition is designed for photocatalytic CO2 reduction to CH4 without any additives. This photocatalyst exhibits ≈ 98.9% selectivity for CH4. Experimental and theoretical calculations demonstrated that the excellent performance of the photocatalyst can be attributed to the localized surface plasmon resonance effect of metals, as well as the synergistic effects of the bimetallic sites. These factors are found to be beneficial not only for capturing solar radiation and facilitating electron migration but also for promoting the adsorption and activation of CO2 and the protonation of *CO. As a result, the photocatalyst achieved high selectivity and activity of photoreduction of CO2 to CH4. This work provides insights into the design of photocatalysts with highly selective target products for CO2 reduction.

Direct Synthesis of Benzothiazoles and Benzoxazoles from Carboxylic Acids Utilizing (o-CF3PhO)3P as a Coupling Reagent.

A general and efficient method for the direct synthesis of benzothiazoles and benzoxazoles from carboxylic acids with 2-aminobenzenethiols or 2-aminophenols using (o-CF3PhO)3P as a simple coupling reagent has been developed. Diverse benzothiazoles and benzoxazoles were synthesized in moderate to excellent yields. And the gram-scale preparation of benzothiazole and benzoxazole also proceeded smoothly under the mild conditions. Moreover, a plausible reaction mechanism was discussed, with (o-CF3PhO)3P and its hydrolysis product (o-CF3PhO)2P(O)H contributing to the formation of the target products as an amide synthesis coupling agent and a cyclization reaction promoter, respectively.

Improving the catalytic performance of carbonyl reductase based on the functional loops engineering.

Vibegron functions as a potent and selective β3-adrenergic receptor agonist, with its chiral precursor (2S,3R)-aminohydroxy ester (1b) being crucial to its synthesis. In this study, loop engineering was applied to the carbonyl reductase (EaSDR6) from Exiguobacterium algae to achieve an asymmetric reduction of the (rac)-aminoketone ester 1a. The variant M5 (A138L/A190V/S193A/Y201F/N204A) was obtained and demonstrated an 868-fold increase in catalytic efficiency (kcat/Km = 260.3 s-1 mM-1) and a desirable stereoselectivity (>99% enantiomeric excess, e.e.; >99% diastereomeric excess, d.e.) for the target product 1b in contrast to the wild-type EaSDR6 (WT). Structural alignment with WT indicated that loops 137-154 and 182-210 potentially play vital roles in facilitating catalysis and substrate binding. Moreover, molecular dynamics (MD) simulations of WT-1a and M5-1a complex illustrated that M5-1a exhibits a more effective nucleophilic attack distance and more readily adopts a pre-reaction state. The interaction analysis unveiled that M5 enhanced hydrophobic interactions with substrate 1a on cavities A and B while diminishing unfavorable hydrophilic interactions on cavity C. Computational analysis of binding free energies indicated that M5 displayed heightened affinity towards substrate 1a compared to the WT, aligning with its decreased Km value. Under organic-aqueous biphasic conditions, the M5 mutant showed >99% conversion within 12 h with 300 g/L substrate 1a (highest substrate loading as reported). This study enhanced the catalytic performance of carbonyl reductase through functional loops engineering and established a robust framework for the large-scale biosynthesis of the vibegron intermediate.

Borane‐Catalyzed Selective Transformation of Levulinic Acid and Hydrazines into Hexahydropyridazines and Tetrahydropyridazin‐3(2H)‐ones Using Hydrosilane

Abstract. Borane‐catalyzed one‐pot tandem cyclization/hydrosilylation of levulinic acid and other γ‐ketoacids with hydrazines for selective access to hexahydropyridazines and tetrahydropyridazin‐3(2H)‐ones has been developed. The combination of B(C6F5)3 and Et3SiH allows for synthesis of various hexahydropyridazines and tetrahydropyridazin‐3(2H)‐ones in 67‐97% yields with tolerance towards various functional groups and easy scale‐up. This reaction can be rendered enantioselective by using a chiral borane catalyst generated in situ from HB(C6F5)2 and a binaphthyl‐based chiral diene and PhMe2SiH, affording the target products in 70‐92% yields with up to >99% ee. The judicious choice of borane catalyst and hydrosilane reductant proves to be critical for catalysis. Mechanistic investigations elucidate the catalytic pathway.

Uracil base PCR implemented for reliable DNA walking

PCR-based DNA walking is of efficacy for capturing unknown flanking genomic sequences. Here, an uracil base PCR (UB-PCR) with satisfying specificity has been devised for DNA walking. Primary UB-PCR replaces thymine base with uracil base, resulting in a primary PCR product composed of U-DNAs. A single-primer (primary nested sequence-specific primer) single-cycle amplification, using the four normal bases (adenine, thymine, cytosine, and guanine) as substrate, is then performed on the primary PCR product. Clearly, only those U-DNAs, ended by the primary nested sequence-specific primer at least at one side, will produce the corresponding normal single strands. Next, the single-cycle product undergoes uracil-DNA glycosylase treatment to destroy the U-DNAs, while the normal single strands are unaffected. Afterward, secondary even tertiary PCR is performed to exclusively enrich the target product. The feasibility of UB-PCR has been checked by obtaining unknown sequences bordering the three selected genetic sites.

A Spin-Labeled Derivative of Gossypol

Gossypol and its derivatives arouse interest due to their broad spectrum of biological activities. Despite its wide potential application, there is no reported example of gossypol derivatives bearing stable radical functional groups. The first gossypol nitroxide hybrid compound was prepared here via formation of a Schiff base. By this approach, synthesis of a gossypol nitroxide conjugate was performed by condensation of gossypol with a 4-amino-TEMPO (4-amino-2,2,6,6-tetramethylpiperidin-1-oxyl) free radical, which afforded the target product in high yield. Its structure was proven by a combination of NMR and EPR spectroscopy, infrared spectroscopy, mass spectrometry, and high-resolution mass spectrometry. In addition, the structure of the gossypol nitroxide was determined by single-crystal X-ray diffraction measurements. In crystals, the paramagnetic Schiff base exists in an enamine–enamine tautomeric form. The tautomer is strongly stabilized by the intra- and intermolecular hydrogen bonds promoted by the resonance of π-electrons in the aromatic system. NMR analyses of the gossypol derivative proved that in solutions, the enamine–enamine tautomeric form prevailed. The gossypol nitroxide at micromolar concentrations suppressed the growth of tumor cells; however, compared to gossypol, the cytotoxicity of the obtained conjugate was substantially lower.

Transition-metal-free decarbonylation-oxidation of 3-arylbenzofuran-2(3H)-ones: access to 2-hydroxybenzophenones.

A transition-metal-free decarbonylation-oxidation protocol for the conversion of 3-arylbenzofuran-2(3H)-ones to 2-hydroxybenzophenones under mild conditions has been developed. NMR studies confirmed the role of in-situ-generated hydroperoxide in the conversion. The protocol was applied to a diverse range of substrates to access the target products in good to excellent yields. A structure-activity study for the 5-substituted-2-hydroxybenzophenones towards UV-protection abilities has been verified by mathematical calculations.

A Novel Biosynthetic Strategy for Ginsenoside Ro: Construction of a Metabolically Engineered Saccharomyces cerevisiae Strain Using a Newly Identified UGAT Gene from Panax ginseng as the Key Enzyme Gene and Optimization of Fermentation Conditions

Ginsenoside Ro, as one of the few oleanane-type ginsenosides, is well known for its unique molecular structure and biological activities. Currently, research on the biosynthesis of ginsenoside Ro is still in its early stages. Therefore, the establishment of a new ginsenoside Ro cell factory is of great significance for the in-depth development and utilization of genes related to ginsenoside Ro synthesis, as well as for the exploration of pathways to obtain ginsenoside Ro. In this study, we cloned endogenous constitutive promoters, terminators, and other genetic elements from S. cerevisiae BY4741. These elements were then sequentially assembled with the uridine diphosphate glucuronic acid transferase gene identified in our previously study (PgUGAT252645) and several other reported key enzyme genes, to construct DNA fragments used for integration into the genome of S. cerevisiae BY4741. By sequentially transferring these DNA fragments into chemically competent cells of engineering strains and conducting screening and target product detection, we successfully constructed an engineered S. cerevisiae strain (BY-Ro) for ginsenoside Ro biosynthesis using S. cerevisiae BY4741 as the host cell. Strain BY-Ro produced 253.32 μg/L of ginsenoside Ro under optimal fermentation conditions. According to subsequent measurements and calculations, this equates to 0.033 mg/g DCW, corresponding to approximately 31% of the ginsenoside Ro content found in plant samples. This study not only included a deeper investigation into the function of PgUGAT252645 but also provides a novel engineering platform for ginsenoside Ro biosynthesis.

Exploring the Impact of Environmental Conditions and Bioreactors on Microalgae Growth and Applications

Microalgae and their bioproducts have diverse applications, including wastewater remediation, CO2 fixation, and the synthesis of nutraceuticals, pharmaceuticals, and biofuels. However, the production of these organisms heavily relies upon environmental conditions, which can significantly impact growth. Furthermore, microalgae cultivation itself can be a source of economic and environmental concerns. Thus, microalgae growth systems have become a critical consideration for both research and industry, to bolster microalgae cultivation and address its accompanying issues. Both open and closed systems, such as raceway ponds and photobioreactors, respectively, are commonly used during the growth process but have their own advantages and drawbacks. However, for microalgae growth, photobioreactors may address most concerns as the system’s design lowers the risk of contamination and provides the ability to control the delivery of desired growth factors. To determine the appropriate system for targeted microalgae cultivation, it is crucial to determine factors such as the scale of cultivation and growth and productivity targets. Additionally, efficient usage of these growth systems and carefully selected incubation factors can aid in addressing some of the economic and environmental issues associated with microalgae production. This review will summarize the current applications of bioreactors in both research and industrial capacities and summarize growth and incubation factors for microalgae.

Recent strategies in target identification of natural products: Exploring applications in chronic inflammation and beyond.

Natural products are a treasure trove for drug discovery, especially in the areas of infection, inflammation and cancer, due to their diverse bioactivities and complex, and varied structures. Chronic inflammation is closely related to many diseases, including complex diseases such as cancer and neurodegeneration. Improving target identification for natural products contributes to elucidating their mechanism of action and clinical progress. It also facilitates the discovery of novel druggable targets and the elimination of undesirable ones, thereby significantly enhancing the productivity of drug discovery and development. Moreover, the rise of polypharmacological strategies, considered promising for the treatment of complex diseases, will further increase the demand for target deconvolution. This review underscores strategies for identifying natural product targets (NPs) in the context of chronic inflammation over the past 5 years. These strategies encompass computational methodologies for early target discovery and the anticipation of compound binding sites, proteomics-driven approaches for target delineation and experimental biology techniques for target validation and comprehensive mechanistic exploration.

Pore-scale modeling of wettability alteration coupled two-phase flow in carbonate porous media

The Middle East marine carbonate reservoirs are the primary targets for global crude oil exploration and production. In contrast to the clastic rock reservoirs, the marine carbonate reservoirs have undergone more complicated deposition, diagenesis and transformation, and the rock surface tends to be oil-wet or mixed-wet. One of the most effective methods for enhancing oil recovery in marine carbonate reservoirs is the use of an ion matched (IM)-surfactant (S) system to alter the wettability of rock surfaces. In this paper, we propose a microscale mathematical model of wettability alteration-coupled fluid flow by considering oil–water two-phase flow, wettability alteration, solute advection- diffusion and surfactant adsorption. The proposed microscale mathematical model is solved using the multi-component lattice Boltzmann model, and its accuracy is further verified. Finally, the effects of displacing fluid, ion-matched water concentration and surfactant concentration on oil mobilization at pore-scale are studied. Results show that, ion-matched water has a positive effect on wettability alteration, and typically shifts the cross-point of oil–water relative permeability curve to the right. The concentration of ion-matched water primarily affects fluid morphology of the wetting-phase and has little effect on phase permeability. After adding surfactant to the ion-matched water, the Euler coefficient of oil-phase gradually increases as the displacement continues, a large amount of remaining oil is gradually dispersed into scattered oil droplets and the oil-phase relative permeability is further improved.

Data Visualization of Bike Store Sales in Europe

This paper presents a comprehensive analysis of bike sales and customer demographics in Europe based on a dataset obtained from Kaggle.com. The dataset covers the period from 2011 to 2016 and includes information on customer age, gender, product categories, order quantities, unit costs, unit prices, profit, and revenue. The analysis utilizes data visualization techniques to identify key trends, including the top-selling products, customer purchasing power across different age groups and customer genders, and product profit margins. The findings suggest opportunities for retailers to target specific customer segments and product categories to maximize profits. Additionally, the report highlights the growing popularity of cycling as a sport and a healthy lifestyle choice, indicating a potential for increased demand in the bike retail market. Overall, the insights provided in this report aim to assist retailer chains in developing effective strategies to capitalize on the opportunities presented by the European bike sales market.

Shell Materials for Chewable Capsules, From Design of Experiments to Optimal Dosage Form Manufacturing

AbstractInnovations in chewable dosage forms provide solutions for swallowing difficulties faced by certain patient populations. Chewable softgel capsules (SGC) offer opportunities to expand the benefits of SGC, excelling in the delivery of lipid‐based formulations by optimizing the mechanical properties of the shell material without compromising industrial‐scale manufacturing. This study employs an original approach, with a target product profile that aims to combine processability and chewability, to develop optimized chewable dosage forms. Through a mixture design of experiments, key factors such as plasticizer content and the addition of acid‐modified thin‐boiling corn starch are explored, focusing on response parameters critical to achieving the desired properties of the capsule shell material. Optimal shell compositions—one starch‐free and another containing 9% w/w starch, with respective gelatin/non‐volatile plasticizer ratios of 0.81 and 0.67 are determined and evaluated by manufacturing chewable SGC at pilot scale using rotary die technology. The successful manufacturing and resulting capsules, which exhibit the intended properties, highlight the efficiency of this approach in striking the right balance between processability, soft texture, and quick disintegration time.

Optimized Economic Production Quantity Inventory Model with Weibull Decay, Dynamic Holding Cost Under Varied Demand Scenario and Demand Series Analysis

This paper presents a sophisticated modification of the economic production quantity (EPQ) model. It incorporates Weibull-distributed progressive deterioration, dynamic holding costs, variable demand, and strong strategies for effectively handling shortages and backlogs. Targeting products that have a short-term expiration but can still be stored for a long time without significant deterioration, the model aims to increase production quantities while lowering overall variable expenses. The extended EPQ model utilizes Maclaurin series approximations to solve ordinary differential equations, effectively capturing the dynamics of inventory under three different demand patterns. The focus of this handle’s situations when the demand for a product changes in a predictable manner over time and considers the likelihood of defective goods during the manufacturing process. This approach provides useful insights for optimizing inventory management and production planning in the manufacturing of goods that are used up or consumed. The usefulness of the model is confirmed in the study through the use of two illustrative instances. Additionally, sensitivity analysis is utilized to suggest potential areas for additional investigation.

Design, Synthesis, and 3D-QASR of Oxazoline Derivatives Containing an S-S Moiety as Potential Acaricide.

To mitigate the detrimental effects of agricultural pest mites on crop yield, an active substructure splicing strategy was employed to modify etoxazole by introducing an S-S moiety. The target products were obtained efficiently via a bilateral disulfurating reagent (DSMO), which was developed by our group. The leaf dip method was used to evaluate the activities of the designed target compounds against the eggs and larvae of the spider mite (Tetranychus cinnabarinus). Most of the target compounds exhibited good efficacy in controlling the larvae and eggs of T. cinnabarinus. Based on these results, a three-dimensional quantitative structure-activity relationship (3D-QSAR) model was established to guide the construction of compound 7l. Notably, compound 7l exhibited a better activity against T. cinnabarinus eggs (LC50 = 0.0035 mg/L) compared to etoxazole (LC50 = 0.2990 mg/L). Greenhouse bioassays indicated that compound 7l exhibits excellent acaricidal activity against egg of T. cinnabarinus, which is better than the etoxazole at 1.0 mg/L. Additionally, some of the compounds showed inhibitory effects against Dickeya zeae (D. zeae), Xanthomonas campestris pv campestris (Xcc), Xanthomonas oryzae pvoryza (Xoo), and Xanthomonas oryzae pvoryzicola (Xoc). Furthermore, compounds 7l not only exhibited relatively potent against Plutella xylostella activities (LC50 = 24.0 mg/L) but also had low toxicity (LC50 > 11.0 μg/bee) to Apis mellifera. In conclusion, the current experimental results suggest that oxazoline derivatives containing an S-S moiety have the potential to serve as lead compounds for the development of novel acaricide agents.

A promise of nose to brain delivery of bevacizumab intranasal sol-gel formulation substantiated in rat C6 glioma model.

Glioblastoma is one of the rapidly spreading cancers, with its potent malignancy often linked to pronounced angiogenesis within tumors. To mitigate this vascularization profile, bevacizumab (Avastin®), a monoclonal antibody, has been utilized for its antiangiogenic activity. However, its effectiveness is hindered by challenges in crossing the blood-brain barrier and the risk of off-target organ toxicity. Delivering drugs directly from the nose to the brain through the olfactory or trigeminal nerves bypassing the blood-brain barrier offers enhanced bioavailability and a more precise targeting strategy. To overcome these challenges, we aimed to develop bevacizumab in situ gel loaded mesoporous silica nanoparticles for intranasal delivery and further examine their pharmacokinetic and pharmacodynamic characteristics. The intranasal gel of mesoporous silica nanoparticles loaded with bevacizumab was optimized and formulated using a factorial and quality by design approach. In the case of bevacizumab mesoporous silica nanoparticles, lower particle size and most negative zeta potential were selected as quality target product profiles which is important for drug loading on the mesoporous silica nanoparticles and also transport of these nanoparticles across the nasal mucosa to the brain. A design space with a multidimensional combination of input variables and process parameters has been demonstrated to assure quality. To optimize the design space and achieve the desired quality standards, the base catalyst and surfactant concentration were chosen as the critical process parameters, while particle size and zeta potential were identified as the critical quality attributes. The novel formulation was assessed for physicochemical parameters such as particle size, zeta potential, entrapment efficiency, appearance, color, consistency, and pH. Additionally, studies on in vitro release, ex vivo permeation, stability, nasal toxicity, organ safety, and bioavailability were conducted. The efficacy study was conducted in an orthotopic murine glioblastoma rat model in which C6 Luc cells were instilled in the striatum of the rat's brain. In vivo, bioluminescence imaging of brain tumors was carried out to observe the tumor regression after treatment with the intranasal and intravenous bevacizumab formulation. Biochemical parameters and histopathology were performed for organ safety studies. The optimized intranasal formulation exhibited an average particle size of 318.8nm and a zeta potential of - 14.7mV for the mesoporous silica nanoparticles. The formulation also demonstrated an entrapment efficiency of 91.34% and a loading capacity of 45.67%. Further pharmacokinetic studies revealed that the optimized intranasal bevacizumab formulation achieved a significantly higher brain concentration Cmax = 147.9ng/ml, indicating improved bioavailability compared to rats administered with intravenous bevacizumab formulation (BEVATAS®), which had a Cmax of 127.2ng/ml. Moreover, this nanoparticle formulation entirely mitigated systemic exposure to bevacizumab. Organ safety evaluation of different biochemical parameters and histopathological analyses revealed that the intranasal bevacizumab-treated group was showing less off-target organ toxicity compared to the group treated with intravenous bevacizumab formulation. Subsequently, the efficacy of this nanosystem was evaluated in an orthotopic glioblastoma rat model, monitoring tumor growth over time through in vivo bioluminescence imaging and assessing anti-angiogenic effects. Twenty-one days post-induction, mesoporous silica nanoparticles loaded with bevacizumab in situ gel exhibited a marked reduction in average bioluminescence radiance (4.39 × 103) from day 7 (1.35 × 107) emphasizing an enhanced anti-angiogenic effect compared to the group treated with intravenous bevacizumab formulation which showed a gradual decrease in average bioluminescence radiance (4.82 × 104) from day 7 (1.42 × 107). These results suggest that the proposed novel formulation of mesoporous silica nanoparticles loaded bevacizumab in situ gel could serve as a promising avenue to enhance glioblastoma treatment efficacy, thereby potentially improving patient quality of life and survival rates significantly. Furthermore, the success of this delivery method could open new avenues for treating other neurological disorders, such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, and stroke. By providing effective brain-targeted therapies, this approach has the potential to revolutionize treatment options and improve outcomes for a broad spectrum of neurological conditions.

Coenzyme A biosynthesis in Bacillus subtilis: discovery of a novel precursor metabolite for salvage and its uptake system.

Vitamins are essential components of the diet of animals and humans. Vitamins are thus important targets for biotechnological production. While efficient fermentation processes have been developed for several vitamins, this is not the case for vitamin B5 (pantothenate), the precursor of coenzyme A. We have elucidated the complete pathway for coenzyme A biosynthesis in the biotechnological workhorse Bacillus subtilis. Moreover, a salvage pathway for coenzyme A synthesis was found in this study. Normally, this pathway depends on pantetheine; however, we observed activity of the salvage pathway on complex medium in mutants lacking the pantothenate biosynthesis pathway even in the absence of supplemented pantetheine. This required rewiring of metabolism by expressing a cystine transporter due to acquisition of mutations affecting the regulation of cysteine metabolism. This shows how the hidden "underground metabolism" can give rise to the rapid formation of novel metabolic pathways.