Half Maximal Inhibitory Concentration Research Articles

Cascade-Activatable Nanoprodrug System Augments Sonochemotherapy of Bladder Cancer.

Sonochemotherapy (SCT) has emerged as a powerful modality for cancer treatment by triggering excessive production of reactive oxygen species (ROS) and controlled release of chemotherapeutic agents under ultrasound. However, achieving spatiotemporally controlled release of chemotherapeutic agents during ROS generation is still an enormous challenge. In this work, we developed a cascade-activated nanoprodrug (CAN) system that utilizes a reversible covalent Schiff base mixed with a hypoxia-activatable camptothecin (CPT) prodrug. Briefly, the designed fluorinated CAN system is self-assembled into nanoparticles under aqueous conditions, which could penetrate deep tumors to offer sufficient oxygen for ultrasound-triggered ROS production. Consequently, the nanoparticles substantially exacerbated the hypoxia of the tumor microenvironment (TME) by elevating oxygen consumption. The aggravated hypoxia in turn served as a positive amplifier to boost the tumor-specific CPT release of Azo-CPT prodrug, which made up for the insufficient treatment efficacy of sonodynamic therapy (SDT). On this basis, we observed a substantial reduction, approximately 3.5-fold, in the half-maximal inhibitory concentration (IC50) of the CAN system compared to that of free CPT in bladder cancer cell lines (T24). Furthermore, the CAN system demonstrated potent antitumor efficacy with reduced side effects, resulting in regression and eradication of T24 tumors in various mouse models. In summary, the CAN system can be easily extended by incorporating different chemotherapeutic agents, showing great potential to revolutionize the clinical management paradigm of bladder cancer.

Exploration of (R)-[11C]YH168 as a PET tracer for imaging monoacylglycerol lipase in the brain: from mice to non-human primates.

The monoacylglycerol lipase (MAGL) plays a pivotal role in modulating the endocannabinoid system and is considered an attractive therapeutic target for diseases in both the central nervous system and periphery. The current study aimed to develop and evaluate a suitable carbon-11 labeled tracer for imaging MAGL in preclinical studies. (R)-YH168 was synthesized via a multi-step pathway and its half-maximal inhibitory concentration (IC50) values were measured using an enzymatic assay. Radiosynthesis of (R)-[11C]YH168 was accomplished by 11C-methylation via Suzuki cross-coupling of a pinacol boron precursor. In vitro autoradiography was performed using brain tissues from MAGL knockout and the corresponding wild-type mice. The metabolic stability of (R)-[11C]YH168 in mouse brain and plasma was assessed 5min after injection. Dynamic PET scans were conducted on anesthetized mice and rhesus monkey. For studies in non-human primates, arterial blood samples were analyzed to obtain the input function for kinetic modeling. Blocking studies with the irreversible MAGL inhibitor PF-06795071 were performed to assess the binding specificity of (R)-[11C]YH168. (R)-[11C]YH168 was synthesized via Suzuki coupling of the phenyl boronic ester with [11C]CH3I in the presence of palladium catalyst. In vitro autoradiography revealed a heterogeneous distribution pattern of (R)-[11C]YH168 with higher binding to MAGL-rich brain regions in wild-type mouse brain slices compared to that of MAGL knockout mice. Dynamic PET imaging in wild-type and MAGL knockout mice confirmed its high specificity and selectivity in mouse brains. In the rhesus monkey, (R)-[11C]YH168 displayed good brain permeability. High levels of radioactivity uptake were seen in the cingulate cortex, frontal cortex, cerebellum, occipital cortex, and hippocampus, consistent with MAGL expression. The one-tissue compartment model was appropriate for fitting the regional time-activity curves and provided reliable volume of distribution values across all brain regions. Pretreatment with PF-06795071 (0.1mg/kg) resulted in almost complete blockade (> 95%) of radioactivity uptake, demonstrating binding specificity of (R)-[11C]YH168 to MAGL in the non-human primate brain. The regional non-displaceable binding potential follows the rank order of cingulate cortex ~ frontal cortex ~ insula > putamen > temporal cortex > caudate ~ occipital cortex ~ thalamus > nucleus accumbens ~ hippocampus ~ cerebellum ~ globus pallidus > substantia nigra > amygdala. (R)-[11C]YH168 is a promising PET probe for imaging and quantifying MAGL in the brains of mice and non-human primates. This 11C-labeled tracer holds great potential for translation into human subjects and offers the possibility of performing multiple PET scans on the same subject within a single day.

Quantitative Assessment of Bony Fish Tropomyosin Using a Monoclonal Antibody Competitive ELISA.

Bony fish is one of the big-nine allergenic foods in the US. This study established a monoclonal antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (ELISA) for the detection of bony fish tropomyosin (TM), a fish allergen. Immunoassay (Western blot and ELISA) was performed to characterize anti-TM mAb8F5 (target configuration, immunoaffinity, and species selectivity). Atlantic cod (Gadus morhua) tropomyosin (CTM) and mAb8F5 were used for assay development. The immunosignal was captured by two light sources, i.e., colorimetry (cELISA) and luminescence (lELISA). During assay validation, cELISA had a wider working range (1-250 ng/μL) than lELISA (0.5-62.5 ng/μL). However, lELISA showed better sensitivity because of its lower half-maximal inhibitory concentration (IC50, 8.8 ng/μL) than cELISA (37.1 ng/μL). The optimal cELISA was applied to measure CTM-equivalents in 21 bony fish samples (0.03-4.2 mg/g) and laboratory-spiked samples (recovery rates of CTM-spiked shrimp samples ranged from 104.7 to 120.8%). lELISA was used to quantify 5.25 μg CTM residues on four food contact surfaces (3.4-6.9 μg from CTM residues and 3.2-4.8 μg from Atlantic cod were detected). This immunoassay has the potential to surveil undeclared allergenic bony fish residues and validate the hygiene of food contact surfaces.

Molecular mechanisms of action of curaxin CBL0137 on breast cancer cells in vitro

Introduction. Breast cancer (BC) represents a group of malignant neoplasms with various molecular profiles, which are characterized by aberrations in the mechanisms of epigenetic transcription regulation. One of these disruptions associated with a worse prognosis is the overexpression of the BET protein family, responsible for the interaction of transcription factors with histone-rich acetylated regions. Previously, for the multitargeted epigenetically active agent curaxin CBL0137 (CBL), we have shown the ability to inhibit the expression of BRD2, BRD3, BRD4 proteins in HeLa TI cells and BRD3, BRD4 proteins in BC cells.Aim. To analyze the molecular mechanisms of CBL0137 action on BC cells in vitro, including: 1) assessment of cytotoxicity, 2) analysis of the effect on the cell cycle, 3) assessment of the ability to trigger apoptosis, 4) cause DNA damage, and 5) analyze the effect on the expression of genes involved in proliferation, apoptosis and repair processes.Materials and methods. The cytotoxicity of CBL on BC cells (MCF7, MDA-MB-231, SKBR3) was assessed using the MTT assay. The effect of the agent on the cell cycle and activation of apoptosis was analyzed by flow cytometry. Analysis of DNA damage under the action of CBL was performed using the comet assay. Changes in the expression of genes associated with proliferation, repair and apoptosis were evaluated using real-time polymerase chain reaction.Results. The half maximal inhibitory concentration (IC50) on BC cells under the action of CBL0137 were 1 μM at 72-hour exposure, 14–25 μM at 24-hour. Curaxin CBL (0.5 and 1 μM) caused G2/M cell cycle arrest, and also triggered apoptosis in all cell lines. Under the action of 1 μM CBL, an increase in the degree of DNA damage in MCF7 and SKBR3 cells was recorded, and under the action of both concentrations in MDA-MB-231 cells. The gene expression profile involved in proliferation also corresponded to the arrest in the G2/M phase of the cell cycle. Also, under the action of CBL, the activation of p53-dependent repair genes occurred. An increase in the expression of pro-apoptotic and a decrease in anti-apoptotic genes was shown.Conclusion. Curaxin CBL0137 showed differential effects on molecular processes in BC cells. We have identified a cytostatic effect of the compound, confirmed at various molecular levels. The data obtained indicate the ability of CBL0137 to inhibit tumor processes in BC cells, which makes it a potentially interesting agent for combination therapy.

Integrating eugenol with intensive care in leukemia patients: exploration of pro-apoptotic potential against HL-60, human leukemia cell line

Objective: The objective of this in-vitro study was to explore the pro-apoptotic potential of eugenol (4-allyl-2-methoxyphenol) on Human Leukemia-60 (HL-60) cell line as a potent phytochemical in intensive care setting to leukemia patients. Methodology: After formal approval by all of the respective ethical committees, this study was simultaneously conducted at all respective institutional departments. The study included culturing HL-60 cell line and its treatment with serial concentrations of eugenol for calculation of subsequent IC50 values (half-maximal inhibitory concentration) via MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, expression analysis of gene, comparative analysis of relative gene fold (Intrinsic Biomarkers Caspase-3, Caspase-9) as apoptosis mediator markers followed by RT-qPCR. Cellular apoptotic morphology was confirmed via Hoechst 333258 staining. Results: For HL-60 cell line, the IC50 of eugenol (14.1 uM) showed high gene expression of pro-apoptotic biomarkers (Caspase-3 and Caspase-9). Hoechst 333258 staining showed prominent apoptotic bodies leading to nuclear fragmentations. Conclusion: Eugenol proved to possess robust pro-apoptotic potential leading to diagnostic efficacy against leukemia HL-60 cell line. Further studies would help in identifying key mechanisms by which eugenol exhibits anti-cancer potential against HL-60 cell lines. Abbreviations: HL-60 - Human Leukemia-60; IC50 - Half-Maximal Inhibitory Concentration Keywords: Eugenol, Pro-apoptotic Effects, Intensive Care, Anti-Cancer Mechanisms, HL-60, Natural Chemotherapeutics Citation: Khaliq HMH, Bughio R, Nangdev P, Aziz O, Javed W. Integrating eugenol with intensive care in leukemia patients: exploration of pro-apoptotic potential against HL-60, human leukemia cell line. Anaesth. pain intensive care 2024;28(5):871−875; DOI: 10.35975/apic.v28i5.2491 Received: June 18, 2024; Reviewed: August 03, 2024; Accepted: August 11, 2024

Design, synthesis, X-ray crystal structure, and antifungal evaluation of new acetohydrazide derivatives containing a 4-thioquinazoline moiety.

To find efficient agricultural fungicides, 29 new 4-thioquinazoline-containing acetohydrazide derivatives were prepared and tested for their fungicidal properties. All of the target compounds were characterized by 1H and 13C nuclear magnetic resonance and high-resolution mass spectrometry techniques, and the molecular structure of compound A2 was verified by single-crystal X-ray diffraction measurement. The experimental results revealed that many compounds from this series had impressive inhibition efficacies in vitro against the tested fungi. For example, compound A25 was identified as the best fungicidal agent against Rhizoctonia solani with an EC50 (half-maximal effective concentration) value of 0.66 μg mL-1, superior to those of the commercial fungicides chlorothalonil, carbendazim and boscalid. Additionally, this compound displayed favorable protection and curative activities in vivo against rice sheath blight caused by R. solani. Antifungal mechanistic studies on compound A25 indicated that this compound exerted its strong anti-R. solani effects probably through an effective inhibition of fungal succinate dehydrogenase activity [half-maximal inhibitory concentration (IC50) = 4.88 μm] and the impairment of cell membrane integrity, based on the results from enzymatic bioassays, molecular docking studies, and scanning and transmission electron microscopy observations. Acetohydrazide derivatives containing the 4-thioquinazoline moiety had the potential to be employed as lead compounds for developing more efficient agricultural fungicides in the near future. © 2024 Society of Chemical Industry.

Efficient Selection of the 2,4-Dichlorophenoxyacetic Acid Nanobody Gene from the Phage Library Constructed with Sorted Specific Cells and Expression in Plants to Confer Herbicide Resistance.

Immunomodulation in biotechnological processes requires an adequate level of specific, high-affinity recombinant antibodies expressed in the affected cells. Here, we report a new strategy to obtain a sensitive nanobody against the 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide. Unlike the previous methods that used total peripheral blood lymphocytes, specific peripheral lymphocytes stained with a fluorescein isothiocyanate-labeled 2,4-D coating antigen were isolated via fluorescence-activated cell sorting and used for phage display library construction. This strategy significantly reduced interference of anticarrier protein nanobody phages to small-molecule nanobody development and required only two cycles of panning to obtain the desired positive clones. Nb4-11, one of the most sensitive phage clones, showed good sensitivity and specificity against 2,4-D. Compared with previously reported 2,4-D nanobodies, the sequence of Nb4-11 exhibited completely different complementarity-determining regions (CDRs). The half-maximum inhibition concentration of the Nb4-11-based ic-ELISA was 29.3 ± 1.9 ng/mL. The Nb4-11 gene was subsequently transferred into Arabidopsis thaliana to confer herbicide resistance, and homozygous transgenic T3 lines were obtained. Stable expression of the 2,4-D nanobody in the model plant was confirmed via PCR, ic-ELISA, and Western blot analysis. In the dose-response bioassay, the transgenic T3 lines were resistant to 2 g of 2,4-D ai/ha. This work offers a new way to sort specific peripheral lymphocytes, efficiently develop nanobodies against small molecules, and create a novel mechanism for herbicide resistance based on the expression of nanobodies in plants.

In vitro antidiabetic evaluation of Bombax buonopozense methanol leaf extract

Diabetes mellitus (DM) is a major worldwide health burden that requires research into cost-effective treatment alternatives. It has been claimed that herbal medicines, have improved glucose metabolism in diabetics. It has been showed that several plant extracts are useful in preserving glucose homeostasis. Therefore, the present study aimed to evaluate the phytochemical and antidiabetic activities of Bombax buonopozense methanol leaf extracts. Standard methods were used for the identification of alkaloids, tannins, flavonoids, saponins, phenolics, cardiacglycosides and steroids. The enzyme inhibitory activities of the extracts of B. buonopozense was evaluated on α-amylase and α-glucosidase. The crude leaf extract of B. buonopozense exhibited a more effective inhibition on α-glucosidase with IC50 (Half maximal inhibitory concentration) values 888.20±35.06 μg/mL when compared to control (ascorbic acid) with values 1076±2.77 μg/mL. Fractions of ethyl acetate showed lower inhibitory property of alpha glucosidase with IC50 value 18.44±2.63 μg/mL compared to the control (ascorbic acid with IC50 value 16325±1318 μg/mL). This study showed the scientific basis for the traditional therapeutic usage of B. buonopozense by proven its antidiabetic activity in vitro. This is the first time that B. buonopozense’s antidiabetic efficacy and possible mechanisms have been documented.

Mimicking Retinoblastoma Treatment With Repeated Topotecan or Melphalan Develops Cross-Resistance to Classic Agents But Not to Repurposed Drugs.

Refractory or recurrent retinoblastoma results from acquired chemoresistance and the management of these eyes often requires surgical removal. Our objective was to develop retinoblastoma models resistant to chemotherapy by exposing cancer cells to repeated chemotherapy mimicking the clinical scenario. These newly resistant cells were used to evaluate potential novel therapies. Chemoresistant cells were obtained by exposing two primary retinoblastoma cell cultures to three weekly doses of melphalan or topotecan. The sensitivity of these resistant cells to each chemotherapy was evaluated, and cross-resistance to topotecan, melphalan, and carboplatin was assessed. Genomic alterations and differential expression of efflux/influx transporters between chemoresistant and parental cells were analyzed. Subsequently, sensitivity of both resistant and parental cells to the repurposed agents digoxin, methylene blue, and gemcitabine was assessed. Four chemoresistant models were successfully established, showing significantly higher half-maximal inhibitory concentration (IC50) values for melphalan and topotecan compared to their corresponding parental cells (P < 0.05). Cross-resistance between melphalan and topotecan was demonstrated, with a 3-fold increase in the IC50. Chemoresistant cells also showed reduced sensitivity to carboplatin (P < 0.05) compared to parental cells, whereas sensitivity to the evaluated repurposed agents remained unchanged. Genomic analysis revealed no selective alterations in the resistant cells, although differential expression of influx/efflux transporters was observed across all chemoresistant models. In vitro simulation of patient treatment was useful to establish chemoresistant retinoblastomas and to identify strategies to overcome resistance to topotecan or melphalan through drug repurposed. Our results warrant further investigation to support the clinical translation.

Small molecules against viruses causing hemorrhagic fevers in Russia

This review is devoted to small molecules (compounds with molecular weights of up to 800–900 Da) that show activity against viruses causing hemorrhagic fevers. Despite the wide occurrence of hemorrhagic fevers, effective medications for their prevention and treatment are virtually absent. Currently, the search for a new drug to fight these viruses is still a relevant task, since the synthetic nucleoside ribavirin used in clinical practice is not sufficiently effective. The review presents compounds that have antiviral action against the causative agents or alleviate symptoms of three acute infectious diseases occurring in Russia: Omsk hemorrhagic fever, Crimean-Congo hemorrhagic fever, and hemorrhagic fever with renal syndrome. The structures and activity parameters (half maximal inhibitory concentration) of the compounds are given and hypothesized mechanisms of their action are considered. The data published during the whole period of studies of these diseases are summarized. &lt;br&gt;The bibliography includes 160 references.

Effects of Platycodon grandiflorus on doxorubicin resistance and epithelial-mesenchymal transition of breast cancer cells via the p38 mitogen-activated protein kinase pathway.

With the increase of chemotherapy frequency for breast cancer, the drug resistance rate of patients is rising, accompanied by cell invasion and metastasis, thus causing mortality. We aimed to explore the mechanism by which Platycodon grandiflorus affects breast cancer cells in terms of the doxorubicin (Dox) resistance and epithelial-mesenchymal transition (EMT) via the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. MCF-7/R cell lines with resistance to Dox were established. After 24h of culture with DMEM (blank group), they were divided into Platycodon grandiflorus, Platycodon grandiflorus + Ophiopogon japonicus, Platycodon grandiflorus + Curcumae Rhizoma, Platycodon grandiflorus + Curcumae Rhizoma + U46619 groups. Flow cytometry, colony formation assay, as well as Transwell assay were performed to examine the cells for apoptosis, proliferation, and invasion, respectively. Western blotting was performed to measure the phosphorylated (p)-p38 MAPK-to-p38 MAPK ratio together with N-cadherin, vimentin, β-catenin, and E-cadherin protein expressions. Compared with the blank group, the half maximal inhibitory concentration (IC50), number of cell colonies, number of invading cells and expressions of proteins related to EMT (i.e. N-cadherin, vimentin, and β-catenin) significantly reduced, but increases in apoptosis rate, p-p38 MAPK/p38 MAPK ratio and E-cadherin protein expression were observed in different groups (P < 0.05). Compared with the Platycodon grandiflorus + Curcumae Rhizoma group, the Platycodon grandiflorus + Curcumae Rhizoma + U46619 group had significantly decreased IC50, cell colony count, invading cell count and β-catenin, N-cadherin, and vimentin expressions, in addition to elevated E-cadherin protein expression, apoptosis rate, and p-p38 MAPK/p38 MAPK ratio (P < 0.05). Platycodon grandiflorus can reverse the resistance of breast cancer cells to Dox and regulate their biological activities by activating the p38 MAPK signaling pathway.

Assessment of Phytochemical Compounds, Enzyme Production, and Antifungal, Antioxidant, and Anti-mutagenic Activities in Endophytic Fungi Derived from Mangrove Trees

Mangrove forest is a unique wetland ecosystem that is highly productive and provides an environment for a variety of microorganisms. Endophytic fungi derived from mangrove plants provide the plants with protection from adverse environmental conditions, while also allowing the fungi to produce valuable bioactive compounds. The present study sampled 11 mangrove trees and isolated, screened, and identified the potent endophytic fungi and their bioactive substances showing anti-pathogenic, anti-mutagenic and antioxidant activities, while the endophytes were investigated for their enzymatic potential. In total, 47 endophytic fungi were isolated from the leaves (36) and stems (11) of the host plants and all isolates were tested for antagonistic activities against selected plant pathogens. Based on the results, isolates BgS-04 and BcL-05 had the highest anti-pathogenic activities against Curvularia sp., Fusarium sp., and Colletotrichum sp. Therefore, the ethyl acetate crude extracts from these two fungi were further investigated for their antioxidant and anti-mutagenic activities and their phenolic compound contents, based on phytochemical analysis. Based on the results, the crude extracts of BgS-04 and BcL-05 contained 5.24 and 4.8 mg gallic acid equivalent/g of total phenolic compounds, respectively, and had antioxidant activity (half maximal inhibitory concentration) levels of 7.4 and 4.26 mg/mL, respectively. The preliminary qualitative phytochemical analysis of the fungal crude extracts identified tannins and coumarins. The anti-mutagenic activity levels of BgS-04 and BcL-05 against the mutagenic compounds, Trp-P-1 and DMBA, were determined using the Ames test, which revealed that the crude extracts of BgS-04 and BcL-05 had moderate-to-high antimutagenic potential against TA98 and TA100.All 47 endophytic isolates were assessed for their potential role in producing extracellular enzyme; they were capable of producing protease (53%), pectinase (28%), amylase (26%) and cellulase (19%) but none of them produced lipase. Among the isolates, RmL-01 derived from the leaves of Rhizophora mucronata had the significantly highest amylase production. Maximum amylase production (141.2 U/mL) was observed at 30 °C, pH 7.0 and 120 hours of incubation time. Molecular identification of the isolates BgS-04, BcL-05 and RmL-01 using nuclear ribosomal DNA internal transcribed spacer sequences revealed that they were Pestalotiopsis parva, Collectotrichum perseae, and Aspergillus oryzae, respectively, with high bootstrap support. It was concluded that the distinct groups of mangrove endophytes were potential sources of novel and valuable bio-based compounds with impressive anti-plant pathogen, anti-mutagenic, and antioxidant activities and capable of producing multi-industrial enzyme cocktails that might be important and useful for biotechnological applications.

One-Pot Synthesis and &lt;i&gt;In Vitro&lt;/i&gt; Studies of Calix[4]-2-methylresorcinarene Derivatives as Antimalarial Agents Against &lt;i&gt;Plasmodium falciparum&lt;/i&gt; Chloroquine-Resistant Strain FCR-3

Malaria is an endemic disease in Indonesia caused by infection from the Plasmodium parasite. Recently, antimalarial resistance significantly contributed to the decline in the cure rate of malaria sufferers. In this work, three calix[4]resorcinarenes have been synthesized from 2-methylresorcinol and different benzaldehyde derivatives, i.e., 4-chlorobenzaldehyde, 4-methoxybenzaldehyde, and 4-dimethylaminobenzaldehyde through the one-pot synthesis procedure. The calix[4]resorcinarenes synthesis was done through a cyclo-condensation reaction by using HCl 37% as the catalyst and ethanol as the solvent in an one-pot reaction. The structures of the synthesized products were confirmed using Fourier transform infrared, proton-nuclear magnetic resonance, and liquid chromatography-mass spectrometry techniques. The antimalarial activity assay was evaluated against the Plasmodium falciparum FCR-3 strain through an in vitro study. Three synthesized compounds, i.e., C-4-chlorophenylcalix[4]-2-methylresorcinarene, C-4-methoxyphenylcalix[4]-2-methylresorcinarene and C-4-dimethylaminophenylcalix[4]-2-methylresorcinarene have been successfully synthesized in up to 97% yield. The C-4-chlorophenylcalix[4]-2-methylresorcinerene exhibited the most potent antimalarial activity with a half-maximal inhibitory concentration (IC50) value of 2.66 µM against P. falciparum FCR-3 while the C-4-methoxyphenylcalix[4]-2-methylresorcinarene and C-4-dimethylaminophenylcalix[4]-2-methylresorcinarene gave the IC50 values of 23.63 and 13.82 µM, respectively. From the results, it could be concluded that the antimalarial activity of calix[4]-2-methylresorcinarenes was influenced by the type of substituent of aromatic rings at the para position.

Bacillus lipopeptides inhibit lipase activity and promote 3T3-L1 preadipocyte differentiation.

Bacillus lipopeptides have been reported to display anti-obesity effects. In the present study, Lipopeptides from Bacillus velezensis FJAT-45028 that consisted of iturin, fengycin and surfactin were reported. The lipopeptides exhibited a strong lipase inhibition activity in a concentration-dependent manner with a half maximal inhibitory concentration of 0.012 mg/mL, and the inhibition mechanism and type were reversible and competitive, respectively. Results of CCK8 assay showed that 3T3-L1 preadipocyte cells were completely viable under treatment of 0.050-0.2 mg/mL lipopeptides for 24 or 48 h. It was found that the lipopeptides could increase lipid droplets in the differentiated 3T3-L1 adipocytes in tested concentration and suppress the expression of peroxisome proliferator-activated receptor gamma (PPARγ). These results indicated the potential anti-obesity mechanism of the tested lipopeptides might be to inhibit lipase activity but not to suppress lipid accumulation in the adipocytes. Moreover, the lipopeptides could elevate glucose utilisation by 14.43%-33.81% in the differentiated 3T3-L1 adipocytes.

Preparation, characterization, and ex vivo evaluation of isoxanthohumol nanosuspension

This study assessed the equilibrium solubility, oil–water distribution coefficient, and dissociation constant of Isoxanthohumol (IXN), and formulated IXN nanoparticles (IXN-Nps) using a micro media grinding method. The research characterized the particle size, polydispersity index, zeta potential, morphology, and structure of the nanoparticles, and evaluated the optimal cryoprotectant. Additionally, the study examined the toxicity and in vitro and in vivo release of IXN on HT-29 cells. IXN is classified as a Biopharmaceutical Classification System (BCS) II class drug with weak acidity. The average particle size of IXN-Nps is 249.500 nm, with a polydispersity index (PDI) of 0.149 and a zeta potential of −25.210 mV. The research identified 5 % mannitol as the optimal cryoprotectant. Compared to IXN, the half-maximal inhibitory concentration of IXN-Nps decreased to one-third, demonstrating a significant inhibitory effect on HT-29 colon cancer cells. The in vitro cumulative release rate of IXN-Nps within 24 h was 3.5 times higher than that of the IXN solution. In vivo pharmacokinetic results revealed that the oral bioavailability of IXN-Nps increased significantly by 2.8 times compared to the IXN solution. The correlation coefficient (r = 0.9227) exceeded the critical value for significance at the 0.01 (r = 0.834) level, indicating a strong correlation between in vivo and in vitro results. Consequently, the nanosuspension overcame the low solubility limitation of IXN and proved to be an effective method for enhancing the oral bioavailability of IXN

Construction and characterization of Laminaria polysaccharide functionalized selenium nanoparticles based on an activity-oriented approach

Laminaria polysaccharides (LP) have been shown to effectively stabilize selenium nanoparticles (SeNPs), forming LP–SeNP complexes with enhanced bioactivity. However, the correlation between their bioactivity and physicochemical properties remains inadequately explored. This study used chemical reduction with LP as stabilizer to investigate how LP-to-selenium mass ratio (LPSMR), reaction time, and temperature influence particle size, selenium content, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of LP–SeNPs. The results showed that enhanced DPPH scavenging correlated with higher selenium content and smaller particle size, primarily modulated by LPSMR and temperature. Under optimal conditions (LPSMR of 1; temperature of 25°C), the resulting LP–SeNPs exhibited uniform morphology with a particle size of 81.41nm and selenium content of 653.91mg/g. This stability was achieved through non-covalent interactions between LP and SeNPs, providing superior light and acid resistance compared to unmodified SeNPs. Notably, LP–SeNPs showed synergistic antioxidant effects, with lower half-maximal inhibitory concentration (IC50) values for scavenging DPPH, hydroxyl, and superoxide anion radicals than LP or SeNPs alone, and enhanced hypoglycemic activity. Cytotoxicity assays confirmed LP–SeNPs had reduced toxicity compared to Na2SeO3 and selenopeptide. These findings provide insights into the structure-activity relationships of LP–SeNPs and support their potential application as antioxidant and hypoglycemic agents.

A multitask interpretable model with graph attention mechanism for activity prediction of low-data PIM inhibitors.

The aberrant expression of proviral integration site for Moloney murine leukemia virus (PIM) kinases is closely related to various tumors and chemotherapy resistance, making them attractive targets for cancer therapy. However, due to the extremely high homology among the three PIM isoforms (PIM1, PIM2, PIM3) and the limited availability of existing bioactivity data, screening and designing selective PIM inhibitors remain a daunting challenge. To address this issue, this study constructed a multitask regression model that can simultaneously predict the half-maximal inhibitory concentration (IC50 values). The model utilizes an attention mechanism to capture effects within local atomic groups and the interactions between different groups of atoms. Through weight sharing, the model enhances the accuracy of predicting PIM3 inhibitors by leveraging the rich and highly correlated data from PIM1 and PIM2 isoforms. Additionally, visualizing the weights of nodes (atoms in the molecule) in the model helps us to intuitively understand the relationship between molecular features and prediction outcomes, thereby enhancing the interpretability of the model. In summary, this work provides new insights and methods for performing activity prediction tasks for multiple similar targets in low-data scenarios.

Unveiling phytochemicals and antioxidant, cytotoxic, anti-thrombotic, anti-inflammatory, and antibacterial activities from the leaves of Dalbergia stipulacea (Roxb.)

The study was aimed to conduct phytochemical investigation and assess the in vitro antioxidant, cytotoxic, anti-thrombotic, anti-inflammatory and antibacterial properties of crude methanol extract of Dalbergia stipulacea leaf and its various extractives. All the fractions contain quinines, alkaloids, flavonoids, tannins, resins, glycosides, and flavonoids, while crude methanol, petroleum ether, and ethyl acetate fractions contain saponins and steroids in qualitative phytochemical analysis. In GC-MS/MS technique, a total of 44 phytoconstituents were identified and characterized, where 3-O-Methyl-D-glucose (22.80 %), 8,11,14-Docosatrienoic acid, methyl ester (19.86 %), pentadecanoic acid, methyl ester (14.17 %), 13-docosenamide (6.62 %), nonadecanoic acid, methyl ester (4.62 %), and phytol (3.04 %) were most prominent. In addition, some promising bioactive constituents, such as pantolactone, aromandendrene, D-allose, loliolide, neophytadiene, dibutyl phthalate, kolavenol, and squalene are present in the leaf of D. stipulacea plant species. Petroleum ether soluble fraction (PESF) of the plant had the highest phenolic content (27.34 mg of GAE/gm) and showed the most effective DPPH free radical scavenger with a half maximal inhibitory concentration (IC50) value of 3.87 μg/mL. The PESF also exhibited cytotoxicity with median lethal dose (LC50) value of 2.76 μg/mL compared to the standard medication vincristine sulfate (LC50 = 0.45 μg/mL) in brine shrimp lethality bioassay. Compared to streptokinase (63.34 %), aqueous soluble fraction (AQSF) displayed the highest percentage of clot lysis (38.61 %). The PESF exhibited the most significant suppression of heat-induced hemolysis (32.61 %) and hypotonic solution-induced hemolysis (33.86 %), respectively, revealing promising anti-inflammatory properties. In addition, the methanol extract and its fractions exerted promising antibacterial properties with notable zone of inhibition (6–25 mm) compared to ciprofloxacin (17–37 mm). The current evidence supports the traditional medicinal applications of the plant, particularly its ability to act as an antioxidant, cytotoxic, thrombolytic, anti-inflammatory, and antibacterial agent. However, further phytochemical isolation and in vivo screening is necessary to discover new drugs from the leaf of D. stipulacea plant species based on the current evidence.

In vitro effects of crocin on the possible anticancer properties of Lactococcus lactis against colorectal adenocarcinoma cells.

Probiotics have been suggested to contribute to cancer prevention through various mechanisms. Additionally, recent studies have established a connection between diet, microbiota, and overall health. In this respect, the current study aims to understand the impact of crocin on possible anti-cancer and antibacterial effects of Lactococcus lactis (L. lactis) in colorectal cancer cells and pathogenic bacteria. The study involved collecting cell-free supernatants (CFSs) from untreated bacteria as a control group and bacteria treated with crocin, and then examining their ability to prevent the growth of HCT-116 colon cancer cells. It was demonstrated that L. lactis, when treated with crocin, can effectively combat against various types of pathogenic bacteria and can survive in acidic conditions. Both CFS and cro-CFS exhibited a dose-dependent inhibition of HCT-116 cell growth but crocin-treated bacteria showed more significant effects. The half-maximal inhibitory concentration (IC50) for cell growth inhibition was 97.41 µL/mL in CSF group and 72.07 µL/mL in cro-CFS group. The results of flow cytometry tests confirmed the MTT assay findings, showing that cro-CFS group had a significantly higher rate of apoptosis compared to CFS of control group. The results obtained from qPCR also showed that the Caspase 9 and BAX genes were upregulated, and the BCL-2 expression level was reduced in cells treated with cro-CFS compared to the CFS group. Overall, these findings suggest that crocin may alter the composition of CFS from probiotics that are present in the gut, potentially impacting their ability to combat cancer.

Synthesis and biological evaluation of esculetin derivatives as antidiabetic agents

Esculetin derivatives 4a-4j (including six novel hydroxylated/halogenated derivatives 4a, 4d, 4e, 4f, 4g, 4h) were synthesized through Perkin reaction with 2,4,5-trihydroxybenzaldehyde and appropriate phenylacetic acid as raw materials, and screened for α-glucosidase and α-amylase inhibitory activities. Among these compounds, compound 3-(3-iodophenyl)-6,7-dihydroxy-2H-chromen-2-one (4d) showed promising hypoglycaemic activity with half maximal inhibitory concentration (IC50) values of 0.18±0.03 mM and 1.82±0.18 mM against α-glucosidase and α-amylase, respectively, classifying it as a mixed-type inhibitor. Compound 4d could statically quench the intrinsic fluorescence of enzymes, which bound to enzymes mainly through hydrogen bonding and hydrophobic interactions affecting the microenvironment of proteins. Compound 4d also exhibited antioxidant activity, which demonstrated good scavenging effects for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals. Furthermore, compound 4d still maintained a certain degree of hypoglycemic and antioxidant activities after simulated gastrointestinal digestion. The oral toxicity study showed that compound 4d had no significant effect on the serum biochemical indices and organs of mice, indicating its safety profile. This study suggests that esculetin may be a potential skeleton to the development of hypoglycaemic drugs.