Significant Activity Research Articles

Functional characterisation of a new halotolerant seawater active glycoside hydrolase family 6 cellobiohydrolase from a salt marsh

Realising a fully circular bioeconomy requires the valorisation of lignocellulosic biomass. Cellulose is the most attractive component of lignocellulose but depolymerisation is inefficient, expensive and resource intensive requiring substantial volumes of potable water. Seawater is an attractive prospective replacement, however seawater tolerant enzymes are required for the development of seawater-based biorefineries. Here, we report a halophilic cellobiohydrolase SMECel6A, identified and isolated from a salt marsh meta-exo-proteome dataset with high sequence divergence to previously characterised cellobiohydrolases. SMECel6A contains a glycoside hydrolase family 6 (GH6) domain and a carbohydrate binding module family 2 (CBM2) domain. Characterisation of recombinant SMECel6A revealed SMECel6A to be active upon crystalline and amorphous cellulose. Mono- and oligosaccharide product profiles revealed cellobiose as the major hydrolysis product confirming SMECel6A as a cellobiohydrolase. We show SMECel6A to be halophilic with optimal activity achieved in 0.5X seawater displaying 80.6 ± 6.93% activity in 1 × seawater. Structural predictions revealed similarity to a characterised halophilic cellobiohydrolase despite sharing only 57% sequence identity. Sequential thermocycling revealed SMECel6A had the ability to partially reversibly denature exclusively in seawater retaining significant activity. Our study confirms that salt marsh ecosystems harbour enzymes with attractive traits with biotechnological potential for implementation in ionic solution based bioprocessing systems.

Process optimization and identification of antioxidant peptides from enzymatic hydrolysate of bovine bone extract, a potential source in cultured meat

Bone protein is a significant secondary product of the meat industry, comprising a substantial quantity of protein. These proteins could be broken down through enzymatic hydrolysis to generate antioxidant peptides. This study aimed to produce antioxidant peptides from bovine bone extract by enzymatic hydrolysis utilizing Flavourzyme and Protamex by optimizing enzyme amounts and time using the Box–Behnken design. The final optimized conditions obtained through the model were as follows: The amount of Flavourzyme was 1,100 U, the amount of Protamex was 2,814 U, and the time was 3.77 (h). Bovine bone extract hydrolysate (BBEH) was purified stepwise using ultrafiltration membranes with molecular cutoffs of 5, 3, and 1 kDa. To assess the antioxidant capacity of the fractions, several methods were used, including radical scavenging activity “1,1-diphenyl-2-picrylhydrazyl (DPPH),” “2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS),” metal chelating activity (MCA), reducing power (RP), and thiobarbituric acid assay (TBA). The results indicated that the ultrafiltration fraction with a molecular weight of less than 1 kDa showed significant antioxidant activity, with 48, 42, and 50% inhibition rates for DPPH, ABTS, and metal chelating, respectively. Using size exclusion chromatography, the fraction with a molecular weight less than 1 kDa was further separated into five sub-fractions: Frac-I, Frac-II, Frac-III, Frac-IV, and Frac-V. Sub-Frac-III, which exhibits significant DPPH radical scavenging activity (55%) and a reducing power of 0.8 at 700 nm, was separated into six sub-sub-fractions using reversed-phase HPLC (RP-HPLC) based on molecular weight and hydrophobicity. The sub-sub-fraction with the highest value for DPPH radical scavenging activity was sub-Fra-III-6, which exhibited approximately 69.45% activity. The sub-Fra-III-6 was analyzed using LC–MS/MS, which identified two specific peptides: Ala-Pro-Phe with a mass of 333.12 Da and Asp-His-Val with a mass of 369.14 Da. These two peptides are likely the primary peptides that might have a crucial role in antioxidant capacity. It can be concluded that BBEH is a valuable source of natural antioxidants and has the potential to serve as a viable resource in the cultured meat industry.

Simultaneous quantification of TB-500 and its metabolites in in-vitro experiments and rats by UHPLC-Q-Exactive orbitrap MS/MS and their screening by wound healing activities in-vitro

BackgroundTB-500 (Ac-LKKTETQ), derived from the active site of thymosin β4 (Tβ4), has various biological functions in its unacetylated form, LKKTETQ. These functions include actin binding, dermal wound healing, angiogenesis, and skin repair. The biological effects of TB-500, however, have not been documented. And the analysis of TB-500 and its metabolites have been neither simultaneously quantified nor structurally identified using synthesized authentic standards. MethodsThis study was aimed to investigating simultaneous analytical methods of TB-500 and its metabolites in in-vitro and urine samples by using UHPLC-Q-Exactive orbitrap MS, and to comparing the biological activity of its metabolites with the parent TB-500. The metabolism of TB-500 was investigated in human serum, various in-vitro enzyme systems, and urine samples from rats treated with TB-500, and their biological activities measured by cytotoxicity and wound healing experiments were also evaluated in fibroblasts. ResultsThe simultaneous analytical method for TB-500 and its metabolites was developed and validated. The study found that Ac-LK was the primary metabolite with the highest concentration in rats at 0–6 h intervals. Also, the metabolite Ac-LKK was a long-term metabolite of TB-500 detected up to 72 hr. No cytotoxicity of the parent and its metabolites was found. Ac-LKKTE only showed a significant wound healing activity compared to the control. ConclusionThe study provides a valuable tool for quantifying TB-500 and its metabolites, contributing to the understanding of metabolism and potential therapeutic applications. Our results also suggest that the previously reported wound-healing activity of TB-500 in literature may be due to its metabolite Ac-LKKTE rather than the parent form.

Insight into the Catalytic Performance of a Zinc-Pillared Curcumin/Bentonite Composite for Enhanced Oxidation of Ibuprofen Residuals into Environmental Products: The Pathway and Toxicity

Environmental organo bentonite synthesis using curcumin-derived chemicals is used as catalyst support for zinc with a Zn-pillaring structure (Zn@CU/BEN). The obtained composite was assessed as an affordable, highly effective, and multifunctional photocatalyst for enhanced oxidation of ibuprofen (IBU) residuals in water supplies. The Zn@CU/BEN composite (0.4 g/L) displayed significant catalytic activities, resulting in the complete oxidation of IBU (25 mg/L) after 80 min. Then, the complete mineralization based on the full elimination of TOC content was recognized after 160 min, with significant indications about the formed intermediates. The identified intermediates, together with the identification of hydroxyl radicals as the essential oxidizing agent, declared an oxidation pathway of IBU over Zn@CU/BEN that involved three steps: hydroxylation, decarboxylation/demethylation, and ring-opening processes. The toxic properties of raw pollutants as well as the oxidizing product at different durations were assessed based on the cell viability results of kidney (HEK293T) and liver (HepG2) cell lines. The partially oxidized sample in the initial duration displayed a higher toxicity impact than the raw IBU. However, the treated sample after 160 min reflected high biosafety and non-toxic properties (cell viability > 97%). The synergetic impact of bentonite and bentonite organo-modified by curcumin extract reflects enhancements in the adsorption as well as the oxidation performance of pillared zinc as a catalyst.

Enhancing Chitosan Fibers: A Dual Approach with Tripolyphosphate and Ursolic Acid.

Chitosan, a well-established biomaterial known for its biocompatibility, biodegradability, and bioactivity, has been the focus of extensive research in recent years. This study explores the enhancement of chitosan fibers' properties through wet impregnation with either ursolic acid (UA) or cross-linking with tripolyphosphate (TPP). In the first experiment, chitosan fibers were treated with UA, for varying immersion set points (1, 2, 4, 6, and 8 h). FTIR, SEM, and UV-Vis spectroscopy analyses demonstrated a chemical reaction between chitosan and UA, with stability reached after 2 h of immersion. Antibacterial testing revealed that chitosan fibers impregnated with UA exhibited significant antibacterial activity against Gram-positive bacteria, notably Staphylococcus aureus. The second experiment involved modifying chitosan fibers' surfaces with a 1% w/v TPP solution for the same periods of time (1, 2, 4, 6, and 8 h). Subsequently, the investigation involved FTIR, SEM, and dynamometry analyses, which revealed successful cross-linking between chitosan and TPP ions, resulting in improved tensile strength after 2 h of immersion. This dual-approach study highlights the potential of chitosan fibers for diverse applications, from wound-healing dressings to antibacterial materials against Gram-positive bacteria.

Discovery of the Active Compounds of the Ethyl Acetate Extract Site of Ardisia japonica (Thunb.) Blume for the Treatment of Acute Lung Injury.

The objective of this study was to identify and evaluate the pharmacodynamic constituents of Ardisiae Japonicae Herba (AJH) for the treatment of acute lung injury (ALI). To fully analyze the chemical contents of various extraction solvents (petroleum ether site (PE), ethyl acetate site (EA), n-butanol site (NB), and water site (WS)) of AJH, the UPLC-Orbitrap Fusion-MS technique was employed. Subsequently, the anti-inflammatory properties of the four extracted components of AJH were assessed using the lipopolysaccharide (LPS)-induced MH-S cellular inflammation model. The parts that exhibited anti-inflammatory activity were identified. Additionally, a technique was developed to measure the levels of specific chemical constituents in the anti-inflammatory components of AJH. The correlation between the "anti-inflammatory activity" and the constituents was analyzed, enabling the identification of a group of pharmacodynamic components with anti-inflammatory properties. ALI model rats were created using the tracheal drip LPS technique. The pharmacodynamic indices were evaluated for the anti-inflammatory active portions of AJH. The research revealed that the PE, EA, NB, and WS extracts of AJH included 215, 289, 128, and 69 unique chemical components, respectively. Additionally, 528 chemical components were discovered after removing duplicate values from the data. The EA exhibited significant anti-inflammatory activity in the cellular assay. A further analysis was conducted to determine the correlation between anti-inflammatory activity and components. Seventeen components, such as caryophyllene oxide, bergenin, and gallic acid, were identified as potential pharmacodynamic components with anti-inflammatory activity. The pharmacodynamic findings demonstrated that the intermediate and high doses of the EA extract from AJH exhibited a more pronounced effect in enhancing lung function, blood counts, and lung histology in a way that depended on the dosage. To summarize, when considering the findings from the previous study on the chemical properties of AJH, it was determined that the EA contained a group of 13 constituents that primarily contributed to its pharmacodynamic effects against ALI. The constituents include bergenin, quercetin, epigallocatechingallate, and others.

A Review of Current and Pipeline Drugs for Treatment of Melanoma.

Malignant melanoma is the most aggressive form of skin cancer. Standard treatment options include surgery, radiation therapy, systemic chemotherapy, targeted therapy, and immunotherapy. Combining these modalities often yields better responses. Surgery is suitable for localized cases, sometimes involving lymph node dissection and biopsy, to assess the spread of the disease. Radiation therapy may be sometimes used as a standalone treatment or following surgical excision. Systemic chemotherapy, while having low response rates, is utilized as part of combination treatments or when other methods fail. The development of resistance to systemic chemotherapies and associated side effects have prompted further research and clinical trials for novel approaches. In the case of advanced-stage melanoma, a comprehensive approach may be necessary, incorporating targeted therapies and immunotherapies that demonstrate significant antitumor activity. Targeted therapies, including inhibitors targeting BRAF, MEK, c-KIT, and NRAS, are designed to block the specific molecules responsible for tumor growth. These therapies show promise, particularly in patients with corresponding mutations. Combination therapy, including BRAF and MEK inhibitors, has been evidenced to improve progression-free survival; however, concerns about resistance and cutaneous toxicities highlight the need for close monitoring. Immunotherapies, leveraging tumor-infiltrating lymphocytes and CAR T cells, enhance immune responses. Lifileucel, an FDA-approved tumor-infiltrating lymphocyte therapy, has demonstrated improved response rates in advanced-stage melanoma. Ongoing trials continue to explore the efficacy of CAR T-cell therapy for advanced melanoma. Checkpoint inhibitors targeting CTLA-4 and PD-1 have enhanced outcomes. Emerging IL-2 therapies boost dendritic cells, enhancing anticancer immunity. Oncolytic virus therapy, approved for advanced melanoma, augments treatment efficacy in combination approaches. While immunotherapy has significantly advanced melanoma treatment, its success varies, prompting research into new drugs and factors influencing outcomes. This review provides insights into current melanoma treatments and recent therapeutic advances.

Precursor-Directed Biosynthesis of Antialgal Fluorinated Bacillamide Derivatives in Bacillus atrophaeus.

The bacillamides are a class of indole alkaloids produced by the Bacillus genus that possess significant antialgal activity. Incorporation of fluorine into the bacillamides was carried out using a precursor-directed biosynthesis approach, with 4-, 5-, and 6-fluorotryptophan added to growing cultures of Bacillus atrophaeus IMG-11. This yielded the corresponding fluorinated analogues of bacillamides A and C, in addition to new derivatives of the related metabolite N-acetyltryptamine, thus demonstrating a degree of plasticity in the bacillamide biosynthetic pathway. The bacillamide derivatives were tested for activity against bloom-forming algae, which revealed that fluorination could improve the antialgal activity of these compounds in a site-specific manner, with fluorination at the 6-position consistently resulting in improved activity.

Conamonin A and dihydrochalcones from the whole plants of Conamomum rubidum (Lamxay & N.S.Lý) Škorničk. & A.D. Poulsen showing anti-inflammatory and cytotoxic activities

A new compound, conamonin A (1), was isolated from the whole plants of Conamomum rubidum with eight known dihydrochalcones (2-9). Their structures were elucidated by a combination of spectroscopic methods as well as by comparison with previously reported data. The absolute configuration of 1 was assigned by TDDFT-ECD method. Compounds 1 and 8 showed inhibitory activity against LPS-induced NO production in the RAW 264.7 cells, with IC50 values of 58.29 ± 2.88 and 81.77 ± 5.99 μM, respectively. Compounds 3/4 and 5/6 exhibited inhibitory effects, with IC50 values of 28.76 ± 1.16 and 29.89 ± 1.79 μg/mL, respectively. Compounds 2, 7-9 exhibited significant cytotoxic activity against human lung carcinoma (the SK-LU-1 cell line) with IC50 values ranging from 9.87 to 17.99 µM. This study offers valuable insights into the chemical constituents and biological activities of Conamomum rubidum, highlighting its potential as a source for discovering new anti-inflammatory and cytotoxic agents.

Regiodivergent Synthesis and Biological Activities of Indoloquinoline Based Compounds.

Cryptolepine, neocryptolepine, and isocryptolepine have remained popular synthetic targets ever since their isolation from the aqueous extracts of the West African climbing shrub Cryptolepis sanguinolenta. These natural alkaloids were found to contain significant antimalarial, antiproliferative and antimicrobial activities, making them ideal starting points for the development of novel drug candidates. As natural product synthesis is often plagued with step-heavy procedures and poor atom economy, the discovery of synthetic protocols addressing these concerns are sorely needed. In our laboratories, we have devoted our efforts into the development of regiodivergent synthesis whereby two of the indoloquinoline natural products, namely neocryptolepine and 11H-indolo[3,2-c]quinolines, could be assembled in only a few steps from a common and readily available starting material. Our synthetic endeavors to meet these goals include a cascade palladium-catalyzed Suzuki-Miyuara cross-coupling and intramolecular C-N bond formation and a photochemical nitrene insertion strategy. Furthermore, our methods also allowed for the construction of several diversely functionalized natural product derivatives which were subjected to biological evaluations.

Diversifying the benzenesulfonamide scaffold for potential V-ATPase inhibitors: synthesis and insecticidal activity evaluation.

Celangulin V is a natural β-dihydroagarofuran derivative isolated from Celastrus angulatus which shows insecticidal activity in many agricultural pests. Using celangulin V as a molecular probe, we find out a new pesticide target: subunit H of V-ATPase. To explore the potential application of this novel target, lead sulfonamides have been found through virtual screening. Combined with the previous work, 46 sulfonamide derivatives are designed and synthesized. All target compounds are first screened for their insecticidal activities against Mythimna separata. The results of bioassay reveal that most of the designed compounds exhibit significant insecticidal activities against third-instar larvae of M. separata under the concentration of 10mg/mL, and compound 8.4 shows the highest activity with LC50 value of 1.72mg/mL, 15-fold smaller than that of celangulin V (25.89mg/mL). Molecular docking results further indicated that compound 8.4 might serve as a potential inhibitor of the subunit H of V-ATPase. This study provides a potential sulfonamide candidate compound for the M. separata control.

Association of peripheral B cells and delirium: combined single-cell sequencing and Mendelian randomization analysis.

Delirium seriously affects the prognosis of patients and greatly reduces the ability to work and live. Peripheral inflammatory events may contribute to the development of delirium, the mechanism of which is still unclear. There is a lack of effective diagnostic and treatments for delirium in clinical practice. The study aims to investigate alterations in peripheral immune cell subsets under inflammatory stress and to explore causal associations with delirium. Single-cell transcriptional sequencing data of human peripheral blood mononuclear cells (PBMC) before and after lipopolysaccharide (LPS) intervention were processed by the Seurat package in R software. PBMC subsets and cellular markers were defined after downscaling and clustering by the Harmony algorithm to identify characteristic subsets in the context of inflammatory stress. Subsequently, a two-sample Mendelian randomization (MR) study was used to explore the causal associations of these inflammation-related PBMC subsets and their molecular phenotypes with delirium. Based on publicly available genetic data, the study incorporated 70 PBMC-associated immune traits, including 8 types of circulating immune cells, 33 B cell subsets and molecular phenotypes, 13 T cell subsets, and 16 B cell-associated cytokines. The results were also validated for robustness, heterogeneity, and horizontal pleiotropy. Under LPS-induced inflammatory stress, B cells, T cells, monocytes, and dendritic cells in human PBMC showed significant activation and quantitative changes. Of these, only lymphocyte and B cell counts were causally associated with delirium risk. This risk link is also seen in the TNF pathway. Further studies of B cells and their subsets revealed that this association may be related to unswitched memory B cells and CD27 expressed on memory B cells. Annotation of the screened SNPs revealed significant polymorphisms in CD27 and CD40 annotated by rs25680 and rs9883798, respectively. The functions of the key annotated genes may be related to the regulation of immune responses, cell differentiation, proliferation, and intercellular interactions. The present study revealed the potential possibility that B cell, memory B cell subset, and TNF-related molecules may be involved in the development of delirium due to peripheral inflammation, which can provide clues for further investigation of delirium prevention and treatment strategies.

COVID-19 and Advice Speech Act: A Syntactic-Pragmatic Study

Advising is a significant activity in a number of organizational health settings, including face-to-face counseling and the dissemination of information via pamphlets, posters, television, and Internet. Health-related advice includes anything from medicine to stopping smoking, getting immunized, and modifying one's diet. A strange sickness initially manifested itself in Wuhan, China, in late 2019. This disease spread rapidly over the world and was eventually named coronavirus disease (COVID-19). Due to the disease's global expansion, the World Health Organization (WHO) has declared it a pandemic, which indicates that it is spreading among individuals in a large number of nations, resulting in mortality. In early May, the WHO announced that more than 200 thousand individuals had died. The situation is worse, as the number of reported cases continues to rise daily. The purpose of this study is to shed light on the language of acts and advising actions, as well as the right method of advising. The advising act verbs and COVID-19 can be investigated syntactically and pragmatically. There is an effective function for advice speeches in relation to COVID-19 announcements and advice. Additionally, there is a strong correlation between the number of mistakes and accuracy in the eight questions across the eight groups. The study's findings indicate that Iraqi students do not follow health recommendations supplied by foreign health organizations. Concerning the COVID-19 pandemic, WHO is one of the leaders capable of effectively managing its concerns and taking proactive measures to contain the disease by issuing directives addressing the viruses as contagious agents. By including these characteristics of syntactic-pragmatic advising acts, these sorts of actions may be comprehended and executed by everyone in any scenario when speaking. Iraqi EFL learners had an unsatisfactory performance evaluation result as a result of inadequate teaching of PV in connection with public advice during the COVID-19 epidemic.

Anti-parkinsonian, anti-inflammatory, anti-microbial, analgesic, anti-hyperglycemic and anticancer activities of poly-fused ring pyrimidine derivatives

Purpose: To investigate the anti-inflammatory, anti-cancer, anti-parkinsonian, anti-microbial, analgesic, and anti-hyperglycemic properties of a variety of poly-fused pyrimidine derivatives.
 Methods: A novel series of fused pyrimidine derivatives 1-6 were synthesized by reacting amino pyrazole derivatives with active methylene derivatives to give the corresponding compounds 1-6. Anti-parkinsonian activity was investigated with benzotropene as a reference drug, anti-inflammatory effect in mice was evaluated using carrageenan in paw edema with indomethacin as reference drug, anti-microbial activity was assessed using nutritional agar with ciprofloxacin and ketoconazole as reference drugs, analgesic activity was evaluated using valdecoxib as reference drug, anti-hyperglycemic activity was investigated using alloxan and sucrose models (SLM) with pioglitazone as reference drug and anticancer activity was investigated using the MTT micro-cultured tetrazolium assay method with doxorubicin as reference drug.
 Results: Pyrimidine derivatives 1-6 possess significant active anti-parkinsonian, anti-inflammatory, anti-microbial, analgesic, anti-hyperglycemic and anticancer activities in comparison to the reference drugs used for each model. Compared to Benzotropene®, compounds 1 and 3 showed a significantly stronger anti-parkinsonian activity (p < 0.03). Compound 3 showed a significantly stronger anti-inflammatory effect than Indomethacin® (p < 0.05). Compounds 5 and 6 exhibited significant anti- microbial activity compared to ciprofloxacin® (p < 0.05). Compounds 4 and 6 exhibited significantly improved analgesic activity as compared to Valdecoxib® (p < 0.01). Compounds 1 and 3 exhibited significantly higher anti-hyperglycemic effects in SLM model when compared to pioglitazone® (p < 0.02). Compound 5 demonstrated the highest activity against human colon cancer cell line (HT-29) and human prostate cancer cell line (DU145), and also, significantly improved level of efficacy against human lung cancer cell line (A549) compared to Doxorubicin® (p < 0.02). 
 Conclusion: Using the six pyrimidine derivatives 1 - 6 as a lead molecule, a novel class of clinically beneficial anti-cancer, anti-inflammatory, anti-microbial, analgesic, and anti-hyperglycemic drugs may be produced.

Cold maceration extraction of wild fruit Terminalia bellirica (Gaertn.) Roxb.: exploring its bioactives for biomedical applications

Terminalia bellirica (T. bellirica) (Gaertn.) Roxb. is a well-known traditional medicinal plants that show promising treatment because of fewer side effects in humans. In the present study, the total phenol, flavonoid, condensed and hydrolyzable tannins extracted and analyzed from cold macerated (CM) T. bellirica (Gaertn.) Roxb. fruit (TBF) and leaves (TBL) extract with the identification of bioactive compounds using GC-MS/MS technique. The highest amount of bioactive content was found in ethanolic extract than toluene. Current experimental data of TBF extract shows the maximum and significant biological activity like free radical scavenging activity against DPPH and FRAP assays with IC50 values of 51.07 ± 0.52 μg/ml and 63.14 ± 0.59 μg/ml respectively. However, IC50 cytotoxicity values of TBF extract on MCF-7 cells for 24 hrs was found to be 6.34 ± 0.72 μg/ml. Minimum inhibitory concentration (MIC) for infectious pathogens Escherichia coli and Bacillus cereus was >12.5 μg/ml and >100 μg/ml respectively, however, anti-inflammatory activity was demonstrated as an IC50 value of 509.1 ± 1.72 μg/ml. Cold macerated fruit extract revealed threatening inhibitory potential against the α-amylase and α-glucosidase enzymes, with IC50 of 50.98 ± 0.23 μg/ml and 46.70 ± 1.38 μg/ml respectively. Finally, the outcome of this study showed that T. bellirica (Gaertn.) Roxb. fruit extract could be an effective source of bioactives with efficient biomedical properties.

Efficacy and Mechanism of Hypofractionation Radiotherapy Combined with PD-1 Inhibitors in a Model of Head and Neck Melanoma

Head and neck mucosal melanoma is one of the most common types of melanoma in China, but the prognosis is worse than other types, and there is no effective treatment plan to improve patient survival. This study analyzes the efficacy of hypofractionation radiotherapy combined with PD-1 inhibitor in the treatment of head and neck mucosal melanoma, as well as its impact on the tumor immune microenvironment. NPSG mice were used to construct a humanized bilateral lesion tumor model of the humanized immune system. The models were divided into an RT (8 Gy)+anti PD-1 group, an RT (2 GyX4)+anti PD-1 group, an Anti PD-1 group, an RT (8 Gy) group, and a blank group. Differences in efficacy and immune cells in blood, lymph nodes, and tumor tissues were compared between different treatment groups. The treatment effect of RT (8 Gy)+anti PD-1 was better than the other groups with a tumor growth inhibition value (TGI) over 60%. Significant recruitment and activation of CD8+T cells were found in the blood, lymph nodes, and tumor tissues and significantly inhibited the level of PD-1+CD8+T cells in the group of RT (8 Gy)+anti PD-1. This study confirmed the efficacy of hypofractionation radiotherapy combined with PD-1 inhibitors, which can inhibit tumor growth and produce distant effects. The appearance of a distant effect is related to the enhancement in the number and activity of CD8+T cells in the local tumor and peripheral blood and lymph nodes. This study confirms the therapeutic and immune regulatory effect of hypofractionation radiotherapy combined with PD-1 inhibitors.

Anti-Aspergillus activities of olorofim at sub-MIC levels during early-stage growth.

Among antifungal compounds in clinical development for systemic disease, the orotomide olorofim is one of only two that target a completely new mechanism of action. Olorofim is highly potent against pathogenic Aspergillus species including cryptic species that frequently show increased resistance to current agents. In this study, our primary focus was on evaluating in detail the inhibitory activity of voriconazole and olorofim against different pathogenic Aspergillus species employing high-throughput microscopy. Compared to standardized, less-sensitive visual assessment-based methods, microscopy-assisted growth monitoring allowed us to detect sub-MIC drug concentration ranges with significant inhibitory activity at early-stage growth. This revealed that olorofim exerts growth inhibition at concentrations that are several magnitudes below those of voriconazole.

Rutin/Sulfobutylether-β-Cyclodextrin as a Promising Therapeutic Formulation for Ocular Infection.

Ocular pathologies present significant challenges to achieving effective therapeutic results due to various anatomical and physiological barriers. Natural products such as flavonoids, alone or in association with allopathic drugs, present many therapeutic actions including anticancer, anti-inflammatory, and antibacterial action. However, their clinical employment is challenging for scientists due to their low water solubility. In this study, we designed a liquid formulation based on rutin/sulfobutylether-β-cyclodextrin (RTN/SBE-β-CD) inclusion complex for treating ocular infections. The correct stoichiometry and the accurate binding constant were determined by employing SupraFit software (2.5.120) in the UV-vis titration experiment. A deep physical-chemical characterization of the RTN/SBE-β-CD inclusion complex was also performed; it confirmed the predominant formation of a stable complex (Kc, 9660 M-1) in a 1:1 molar ratio, with high water solubility that was 20 times (2.5 mg/mL) higher than the free molecule (0.125 mg/mL), permitting the dissolution of the solid complex within 30 min. NMR studies revealed the involvement of the bicyclic flavonoid moiety in the complexation, which was also confirmed by molecular modeling studies. In vitro, the antibacterial and antibiofilm activity of the formulation was assayed against Staphylococcus aureus and Pseudomonas aeruginosa strains. The results demonstrated a significant activity of the formulation than that of the free molecules.

Phytochemical analysis, antioxidant, antimicrobial, and toxicity studies of Schima wallichii growing in Nepal

ABSTRACT The present study focuses on qualitative and quantitative phytochemical and biological analysis in bark and leaf extracts of Schima wallichii growing in Gulmi, Nepal. TPC and TFC were measured using the Folin-Ciocalteu reagent and the aluminum chloride colorimetric method, respectively. The DPPH assay was used for antioxidant activity, and antimicrobial activity was studied with the agar-well diffusion method. The presence of polyphenols, flavonoids, quinones, coumarins, saponins, terpenoids, and tannins was detected by qualitative phytochemical analysis. Total phenolic content in an aqueous leaf extract and methanol bark extract ranged from 199 ± 5 to 219 ± 8 mg GAE/g. Similarly, the flavonoid content found between 6.0 ± 1.7 in bark to 33 ± 2 mg QE/g in leaf extract. The antioxidant activity of IC50 36 ± 3, 39 ± 1, 52 ± 3, and 73 ± 4 µg/mL for methanolic bark extract, methanolic leaf, aqueous bark, and aqueous leaf extracts, respectively. The bark methanol extract showed the most antibacterial activity with ZOI of 10.0 ± 0.3, 8.0 ± 0.3, 14 ± 0, and 11 ± 1 mm against Klebsiella pneumoniae, Staphylococcus aureus, Bacillus subtilis, and Escherichia coli, respectively. Plant extracts were found to be nontoxic to mildly toxic to the brine shrimp nauplii. The plant is found to be rich in phenolics, and flavonoids showing significant antioxidant and antibacterial activities, and is nontoxic against brine shrimp nauplii. The plant imparts a health-promoting effect that needs to be explored in pharmaceutical research.

Interspecies Brain PBPK Modeling Platform to Predict Passive Transport through the Blood-Brain Barrier and Assess Target Site Disposition.

The high failure rate of central nervous system (CNS) drugs is partly associated with an insufficient understanding of target site exposure. Blood-brain barrier (BBB) permeability evaluation tools are needed to explore drugs' ability to access the CNS. An outstanding aspect of physiologically based pharmacokinetic (PBPK) models is the integration of knowledge on drug-specific and system-specific characteristics, allowing the identification of the relevant factors involved in target site distribution. We aimed to qualify a PBPK platform model to be used as a tool to predict CNS concentrations when significant transporter activity is absent and human data are sparse or unavailable. Data from the literature on the plasma and CNS of rats and humans regarding acetaminophen, oxycodone, lacosamide, ibuprofen, and levetiracetam were collected. Human BBB permeability values were extrapolated from rats using inter-species differences in BBB surface area. The percentage of predicted AUC and Cmax within the 1.25-fold criterion was 85% and 100% for rats and humans, respectively, with an overall GMFE of <1.25 in all cases. This work demonstrated the successful application of the PBPK platform for predicting human CNS concentrations of drugs passively crossing the BBB. Future applications include the selection of promising CNS drug candidates and the evaluation of new posologies for existing drugs.