Hemolytic Research Articles

Perioperative selection of blood and components for substitution: Immunohaematological assessment of immunization risk and methods for selection of optimally compatible blood

The aim of this study is to assess the risk for immune and hematologic adverse events in perioperative transfusion of blood components. Our second goal is to propose methods for the selection of an optimally compatible donor in order to limit the negative impact of complications following blood component transfusion on the hospital outcome of patients admitted to clinics of general, gastrointestinal, thoracic, and vascular surgery. The largest group of patients included in this study were patients with gastrointestinal disorders (614 patients; 31%) and patients with diseases of the organs of the blood and lymphatic system (589 patients; 30%). This was followed by patients with diseases of the skin and subcutaneous tissue (368 patients; 18%) and patients with benign and malignant neoplasms (365 patients; 18%). The remaining 65 patients (3%) were diagnosed with diseases of the endocrine system, trauma patients, and patients with genitourinary disorders. The methods used for optimal selection of compatible blood components in this study are enzyme tests, Coombs tests, and tests in agglutinating medium. The study shows that clinically significant antibodies, which could provoke a post-transfusion hemolytic reaction, were detected in 4.3% of the screened patients. In the majority of patients, the specificity of antierythrocyte antibodies cannot be established. For those patients where the specificity of anti-erythrocyte antibodies could be established, the type of antibody and the antigen system to which it belonged were as follows (in descending order): 1. Anti-erythrocyte antibodies belonging to the Rh system: 37% (n = 17) of all screened patients (1.6% of all patients), including anti-E type (5 patients), anti-D (8 patients), anti-C-1, anti-C (2 patients), and anti-Cw (1 patient). 2. In 3 (6% or anti-erythrocyte antibodies belonging to other erythrocyte antigen systems—Kidd-1 patients, Lewis-2 patients—6% of the screened patients (0.3% of all patients). 3. In 57% (n = 26) of those screened (2.4% of all patients), alloantibodies without any known specificity were identified. The largest number of identified antibodies were directed against the D antigen belonging to the Rh system. Most of the anti-D alloantibodies found in Rhesus D (-) negative patients with no history of prior transfusion of D (+) positive red blood cell (RBC) concentrate possibly have resulted from prior Rh isoimmunization during pregnancy, provided that anti-D antibodies persist for a long time after birth—in some of the studied patients, 15–20 or more years. A greater number of alloimmunizations were found in women—28 (61%), compared to men—18 (39%), due to previous sensitization with different blood group antigens during pregnancy. This study shows that timely diagnosis is essential for the selection of appropriate RBC concentrate, for the avoidance of adverse reactions, and for the improvement of hematological parameters after transfusion of blood components. Graphical abstract:

PLGA-PEG Nanoparticles Loaded with Cdc42 Inhibitor for Colorectal Cancer Targeted Therapy.

Background/Objectives: An inhibitor of small Rho GTPase Cdc42, CASIN, has been shown to reduce cancer cell proliferation, migration, and invasion, yet it has several limitations, including rapid drug elimination and low bioavailability, which prevents its systemic administration. In this study, we designed and characterized a nanoparticle-based delivery system for CASIN encapsulated within poly(lactide-co-glycolide)-block-poly(ethylene glycol)-carboxylic acid endcap nanoparticles (PLGA-PEG-COOH NPs) for targeted inhibition of Cdc42 activity in colon cancer. Methods: We applied DLS, TEM, and UV-vis spectroscopy methods to characterize the size, polydispersity index, zeta potential, encapsulation efficiency, loading capacity, and in vitro drug release of the synthesized nanoparticles. The CCK-8 cell viability test was used to study colorectal cancer cell growth in vitro. Results: We showed that CASIN-PLGA-PEG-COOH NPs were smooth, spherical, and had a particle size of 86 ± 1 nm, with an encapsulation efficiency of 66 ± 5% and a drug-loading capacity of 5 ± 1%. CASIN was gradually released from NPs, reaching its peak after 24 h, and could effectively inhibit the proliferation of HT-29 (IC50 = 19.55 µM), SW620 (IC50 = 9.33 µM), and HCT116 (IC50 = 10.45 µM) cells in concentrations ranging between 0.025-0.375 mg/mL. CASIN-PLGA-PEG-COOH NPs demonstrated low hemolytic activity with a hemolytic ratio of less than 1% for all tested concentrations. Conclusion: CASIN-PLGA-PEG-COOH NPs have high encapsulation efficiency, sustained drug release, good hemocompatibility, and antitumor activity in vitro. Our results suggest that PLGA-PEG-COOH nanoparticles loaded with CASIN show potential as a targeted treatment for colorectal cancer and could be recommended for further in vivo evaluation.

Insight into the Course of the Ferrier Rearrangement Used to Obtain Untypical Diosgenyl Saponins.

The Ferrier rearrangement was utilized to obtain 2,3-unsaturated diosgenyl glycosides. This reaction proceeded with high stereoselectivity, yielding mostly saponins with an α configuration (hexoses) or predominantly with a β configuration (pentoses). The diversity of the glycals used and the glycosides obtained enabled a deep discussion of the Ferrier rearrangement mechanism. The mechanism was supported by DFT calculations concerning the intermediate ions. It was concluded that the vinylogous anomeric effect may influence the reactivity of the glycals. Two possible Ferrier rearrangement intermediates, dioxolenium and allyloxycarbenium ions, were hypothesized to exist in thermodynamic equilibrium that shifted toward the former. The allyloxycarbenium ion participates in the final rearrangement step and determines the reaction regioselectivity. Furthermore, the conformational stability of the 2,3-unsaturated pyranose ring determines the stereoselectivity of the reaction. Factors influencing this stability, as well as the NMR data enabling recognition of the 0H5 and 5H0 conformations, were identified. Chemoselective hydrogenation of 2,3-unsaturated diosgenyl glycosides provided a series of 2,3-dideoxy analogues. The anticancer, hemolytic, and antibacterial activities of the synthesized saponins are presented alongside a discussion of the structure-activity relationships.

Peptide hemolytic activity analysis using visual data mining of similarity-based complex networks

Peptides are promising drug development frameworks that have been hindered by intrinsic undesired properties including hemolytic activity. We aim to get a better insight into the chemical space of hemolytic peptides using a novel approach based on network science and data mining. Metadata networks (METNs) were useful to characterize and find general patterns associated with hemolytic peptides, whereas Half-Space Proximal Networks (HSPNs), represented the hemolytic peptide space. The best candidate HSPNs were used to extract various subsets of hemolytic peptides (scaffolds) considering network centrality and peptide similarity. These scaffolds have been proved to be useful in developing robust similarity-based model classifiers. Finally, using an alignment-free approach, we reported 47 putative hemolytic motifs, which can be used as toxic signatures when developing novel peptide-based drugs. We provided evidence that the number of hemolytic motifs in a sequence might be related to the likelihood of being hemolytic.

The roles of cell wall inhibition responsive protein CwrA in the pathogenicity of staphylococcus aureus

ABSTRACT The ability to form robust biofilms and secrete a diverse array of virulence factors are key pathogenic determinants of Staphylococcus aureus, causing a wide range of infectious diseases. Here, we characterized cwrA as a VraR-regulated gene encoding a cell wall inhibition-responsive protein (CwrA) using electrophoretic mobility shift assays. We constructed cwrA deletion mutants in the genetic background of methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) strains. Phenotypic analyses indicated that deletion of cwrA led to impaired biofilm formation, which was correlated with polysaccharide intercellular adhesin (PIA). Besides, the results of real-time quantitative PCR (RT-qPCR) and β-galactosidase activity assay revealed that CwrA promoted biofilm formation by influence the ica operon activity in S. aureus. Furthermore, cwrA deletion mutants released less extracellular DNA (eDNA) in the biofilm because of their reduced autolytic activity compared to the wild-type (WT) strains. We also found that cwrA deletion mutant more virulence than the parental strain because of its enhanced haemolytic activity. Mechanistically, this phenotypic alteration is related to activation of the SaeRS two-component system, which positively regulates the transcriptional levels of genes encoding membrane-damaging toxins. Overall, our results suggest that CwrA plays an important role in modulating biofilm formation and haemolytic activity in S. aureus.

Apodanthera glaziovii (Cucurbitaceae) Shows Strong Anti-Inflammatory Activity in Murine Models of Acute Inflammation.

Background/objectives:Apodanthera glaziovii is an endemic species from the semi-arid Brazilian, which has limited toxicological and pharmacological studies. This species belongs to a well-studied family known for its bioactive compounds used in treating inflammatory. This study aimed to identify secondary metabolites in the stems from A. glaziovii, evaluate toxicity, and investigate the anti-inflammatory potential of the stem hydroalcoholic extract (SHE-Ag). Methods: qualitative and quantitative assays were employed to identify secondary metabolites, along with chromatographic analyses and 1H and 13C NMR. Toxicity was assessed through in vitro hemolytic toxicity, in vivo genotoxicity, and oral acute toxicity tests before the pharmacological assays were conducted. Results: phytochemical screening, HPLC and NMR analyses suggested the presence of saponins of the norcucurbitacin class. The SHE-Ag exhibited no hemolytic activity and no mutagenic potential. However, in vivo toxicity at a dose of 2000 mg/kg revealed hematological and biochemical alterations, while the 500 mg/kg dose was safe. In the anti-inflammatory assays, SHE-Ag at 100 mg/kg reduced paw edema by 55.8%, and leukocyte and neutrophil migration by 62% and 68% in the peritonitis model, respectively; inflammatory cell migration by 70% in the air pouch model, outperforming indomethacin, which showed a 54% reduction. Conclusions: these findings indicate that SHE-Ag is rich in saponins, confirmed through HPLC and 1H and 13C NMR analyses. The SHE-Ag also demonstrated low toxicity. The inflammation models used showed a reduction in inflammation, pro-inflammatory cells, and edema, highlighting the significant anti-inflammatory activity of hydroethanolic extract A. glaziovii stems.

Individual variation in Macrovipera lebetinus obtusa (blunt-nosed viper) venom from a limited area in Southeastern Anatolia (Türkiye)

Intraspecific variation in snake venom composition has been the subject of many studies, revealing the differences in venom proteomes according to age, diet, sex, and geographic region. Previous research on M. lebetinus venom mainly focused on protein purification, proteomic characterization, and biological activities. Although venom proteomes of its subspecies from different geographical areas were compared, it is a widespread species and there is limited knowledge regarding the individual venom variation of M. lebetinus in the same area. In this study, individual variation in M. lebetinus obtusa venom was investigated in terms of protein profiles, protease, phospholipase A2, ʟ-amino acid oxidase, hyaluronidase and phosphodiesterase, and hemolytic activities based on nine individuals collected in the province of Şanlıurfa (southeastern Türkiye). HPLC analysis indicated more striking differences compared to SDS-PAGE. Phosphodiesterase activity varied most between the individuals, followed by hyaluronidase and phospholipase A2. Hemolytic activity did not show significant alteration based on the blood agar method. The results of this initial study revealed that M. lebetinus venom shows variation between the individuals inhabiting the same habitat, which should be taken into account in the development of antivenoms and the clinical assessment of the bite cases.

Synthesis and characterization of new antimicrobial peptides derived from Temporin F.

Antimicrobial peptides (AMPs) are a promising source of new compounds against resistant bacteria. Temporins are a class of AMPs found on the amphibian Rana temporaria and show activity against Gram-positive and Gram-negative bacteria. There are few studies on how these antimicrobials have been used, but new Temporin-F derivatives were engineered with Lys-substitutions to assess the impact of the net charge on antimicrobial activity and toxicity. We demonstrated through some assays that it is possible to increase the antibacterial activity while maintaining a reduced peptide hemolytic activity with specific substitutions. Our lead synthetic peptide, G6K-Temporin F, has shown higher antimicrobial activity against Gram-negative and Gram-positive bacteria in vitro (MIC range 2 to 32 μmol L-1), with low hemolytic activity maintained, resulting in an increase in the therapeutic window (TW), of 12.5. Also, it showed more resistant to enzymatic degradation. On the other hand, more significant increases in net charges, such as in P3K-G11K-Temporin F, result in a severe increase in toxicity with lower gains in antimicrobial activity (TW of 0.65). In conclusion, we demonstrated that a moderate increase in net charge can lead to a more active analog and G6K-Temporin F is revealed to be promising as a candidate for new AMP therapeutics.

Targeted delivery of letrozole-loaded Mg-doped cobalt ferrite nanoparticles for breast cancer treatment

Breast cancer is the most common cancer in women globally. Letrozole is an aromatase inhibitor used as an anticancer drug. Conventional Letrozole therapies for breast cancer often cause adverse effects when administered orally or parenterally. Efforts have been made to develop an advanced drug delivery system for breast cancer treatment, aiming to improve drug release accuracy and therapeutic efficacy while minimizing side effects. This study focuses on magnesium-doped cobalt ferrite nanoparticles (Mg-doped CFNPs) as potential drug delivery carrier for breast cancer treatment. Mg-doped CFNPs with the chemical formula Co1-xMgxFe2O4 (where x = 0.0, 0.05, 0.10, 0.15, 0.20 and 0.25) were synthesized with different concentrations of 5 %, 10 %, 15 %, 20 %, and 25 % by the sol gel method with a fine size of 30–70 nm and their characteristics such as surface morphology, structural properties, chemical composition, and charge were investigated using scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and zeta potential analysis respectively. After synthesis, the Mg-doped CFNPs were coated with Poly ethylene glycol (PEG) and encapsulated with Letrozole, which exhibited <10 % hemolytic activity across all concentrations, as well as minimal cytotoxic effects on non-cancerous cells (HEK-293) but exhibited significant toxicity towards breast cancer cell lines (MCF-7, MDA-MB-231).The best results were obtained at a concentration of 25 % in both the hemolytic assay and the MTT assay, which indicated that increasing the amount of magnesium in cobalt decreased the toxicity and increased the drug release efficiency of nanoparticles.

Branched oncolytic peptides target HSPGs, inhibit metastasis, and trigger the release of molecular determinants of immunogenic cell death in pancreatic cancer.

Immunogenic cell death (ICD) can be exploited to treat non-immunoreactive tumors that do not respond to current standard and innovative therapies. Not all chemotherapeutics trigger ICD, among those that do exert this effect, there are anthracyclines, irinotecan, some platinum derivatives and oncolytic peptides. We studied two new branched oncolytic peptides, BOP7 and BOP9 that proved to elicit the release of damage-associated molecular patterns DAMPS, mediators of ICD, in pancreatic cancer cells. The two BOPs selectively bound and killed tumor cells, particularly PANC-1 and Mia PaCa-2, but not cells of non-tumor origin such as RAW 264.7, CHO-K1 and pgsA-745. The cancer selectivity of the two BOPs may be attributed to their repeated cationic sequences, which enable multivalent binding to heparan sulfate glycosaminoglycans (HSPGs), bearing multiple anionic sulfation patterns on cancer cells. This interaction of BOPs with HSPGs not only fosters an anti-metastatic effect in vitro, as demonstrated by reduced adhesion and migration of PANC-1 cancer cells, but also shows promising tumor-specific cytotoxicity and low hemolytic activity. Remarkably, the cytotoxicity induced by BOPs triggers the release of DAMPs, particularly HMGB1, IFN-β and ATP, by dying cells, persisting longer than the cytotoxicity of conventional chemotherapeutic agents such as irinotecan and daunorubicin. An in vivo assay in nude mice showed an encouraging 20% inhibition of tumor grafting and growth in a pancreatic cancer model by BOP9.

Construction of 2-azidacetic acid functionalized high-crystallinity fluorescent covalent organic framework: Applications in mitoxantrone and Fe3+ sensing and adsorption

Due to the dual functions of fluorescence detection and adsorption, fluorescent covalent organic frameworks (COFs) have attracted significant attention. However, common fluorescent COFs often exhibit unsatisfactory fluorescence properties and selectivity, coupled with poor solution dispersibility, which limit their effectiveness in detection and adsorption applications. In response, a novel post-modified fluorescent COF (named AZC-COF) was synthesized by connecting a fluorescent COF (COF-TB) with 2-azidacetic acid through a copper-catalyzed aide-alkyne cycloaddition (CuAAC) reaction. AZC-COF demonstrated excellent solution dispersibility and robust green fluorescence, boasting an absolute fluorescence quantum yield (QY) of 7.58%, which was 13.5 times higher than that of COF-TB. Furthermore, leveraging the active carboxylic acid and triazole sites, AZC-COF exhibited remarkable binding abilities for mitoxantrone (MIX) and Fe3+, enabling sensitive detection and efficient adsorption of them. In contrast, due to the absence of these functional sites, COF-TB showed poor detection and enrichment capabilities for MIX and Fe3+. The impressive detection and adsorption efficiencies of MIX and Fe3+ in environmental water, aquatic organism (fish) and plasma samples underscore the potential of AZC-COF as a detection-adsorption platform. Additionally, AZC-COF demonstrated low toxicity and hemolytic activity, alongside promising potential for cell imaging and detection of MIX and Fe3+, highlighting its considerable application prospect in biological systems.

Intraspecific cooperation allows the survival of Staphylococcus aureus staff: a novel strategy for disease relapse

BackgroundThe contribution of interspecies interactions between coinfecting pathogens to chronic refractory infection by affecting pathogenicity is well established. However, little is known about the impact of intraspecific interactions on infection relapse, despite the cross-talk of different strains within one species is more common in clinical infection. We reported a case of chronic refractory pulmonary infection relapse, caused by two methicillin-sensitive S. aureus (MSSA) strains (SA01 and SA02) and revealed a novel strategy for relapse via intraspecific cooperation.MethodsThe hemolytic ability, growth curve, biofilm formation, virulence genes and response of G. mellonella larvae to S. aureus infection were analysed to confirm this hypothesis.ResultsSA02 hemolytic activity was inhibited by SA01, along with the expression of hemolysin genes and the virulence factor Hla. Additionally, SA01 significantly enhanced the biofilm formation of SA02. AIP-RNAIII may be a possible pathway for this interaction. Compared with mono-infection, a worse outcome (decreased larval survival and increased microbial burden) of the two MSSA strains coinfected with G. mellonella confirmed that intraspecific interactions indeed enhanced bacterial survival in vivo.ConclusionThe intraspecific interaction of S. aureus could lead to chronic refractory infection via pathogenicity changes.

Synergistic Effect of Silver Nanoparticles with Antibiotics for Eradication of Pathogenic Biofilms.

The increase in nosocomial multidrug resistance and biofilm-forming bacterial infections led to the search for new alternative antimicrobial strategies other than traditional antibiotics. Silver nanoparticles (AgNP) could be a viable treatment due to their wide range of functions, rapid lethality, and minimal resistance potential. The primary aim of this study is to prepare silver nanoparticles and explore their antibacterial activity against biofilms. AgNPs with specific physicochemical properties such as size, shape, and surface chemistry were prepared using a chemical reduction technique, and then characterized by DLS, SEM, and FTIR. The activity of AgNPs was tested alone and in combination with some antibiotics against MDR Gram-negative and Gram-positive planktonic bacterial cells and their biofilms. Finally, mammalian cell cytotoxicity and hemolytic activity were tested using VERO and human erythrocytes. The findings of this study illustrate the success of the chemical reduction method in preparing AgNPs. Results showed that AgNPs have MIC values against planktonic organisms ranging from 0.0625 to 0.125 mg/mL, with the greatest potency against gram-negative bacteria. It also effectively destroyed biofilm-forming cells, with minimal biofilm eradication concentrations (MBEC) ranging from 0.125 to 0.25 mg/ml. AgNPs also had lower toxicity profiles for the MTT test when compared to hemolysis to erythrocytes. Synergistic effect was found between AgNPs and certain antibiotics, where the MIC was dramatically reduced, down to less than 0.00195 mg/ml in some cases. The present findings encourage the development of alternative therapies with high efficacy and low toxicity.

Biocompatibility, antimicrobial efficacy, and therapeutic potential of cobalt carbonate nanoparticles in wound healing, sex hormones, and metabolic regulation in diabetic albino mice

Nanotechnology enables the manipulation of materials at the nanoscale, offering innovative solutions in various fields. Nanoparticles, with their small size and unique properties, have significant applications in the biomedical filed. The current study was designed to assess the biological applications of self-synthesized cobalt carbonate (CoCO3) nanoparticles. The crystalline structure and chemical composition of the CoCO3-NPs were confirmed by SEM, XRD, and FTIR techniques. We observed the 16.58 nm size of novelly synthesized CoCO3 NPS. The scanning electron microscope study confirmed a uniform cubic spinel structure. The biocompatibility and antimicrobial activity were checked in an invitro setup. We exposed albino mice to these synthesized NPs to study wound healing and metabolic effects. The results of biocompatibility analysis indicated hemolytic activity in a dose-dependent way, which showed no cytotoxic effect except at a higher concentration. Furthermore, the results showed enhanced wound healing processes in CoCO3-NP-treated albino mice as compared to the control group. CoCO3-NPs have considerable effect on the thyroid hormone and insulin levels in albino mice. The levels of T3, T4, and insulin were increased in a dose-dependent manner. Interactions between CoCO3-NPs and thyroxine and insulin were confirmed through molecular docking. We confirmed the antimicrobial efficiency of the nanoparticles using MIC values and zones of inhibition against Staphylococcus haemolyticus and Staphylococcus aureus. Despite their concentration-dependent biocompatibility concerns, the results are promising, as CoCO3-NPs hold potential for use in medical practice, particularly in advanced wound management and microbe inhibition.

High-temperature emulsification coupled with low-temperature gelation for fabrication of agarose microsphere implants with well-controlled size for skin tissue enhancement.

Soft tissue deficiencies profoundly impact the daily lives, and mental well-being of patients. Microspheres facilitate collagen synthesis by establishing a conducive environment for fibroblast growth. Existing synthetic polymer microspheres are typically prepared through emulsification and cross-linking at normal temperatures. However, residues of cross-linking agents can adversely affect biocompatibility, thereby limiting their biomedical applications. Agarose has garnered significant attention owing to its biodegradability and excellent biocompatibility. We have for the first time developed high-temperature emulsification coupled with low-temperature gelation for fabrication of agarose microsphere implants with well-controlled size for skin tissue enhancement. The agarose microspheres exhibited favorable sphericity and dispersion, possessing a uniform particle size with an average diameter of 37.24 μm. Furthermore, the microspheres demonstrated commendable injectability and biodegradability. Additionally, the implants displayed remarkable biocompatibility, effectively promoting the proliferation of human foreskin fibroblast-1 (HFF-1) cells. The microspheres exhibited no systemic toxicity and induced no hemolytic or thermogenic reactions. In photoaged mice skin models, the agarose microspheres augmented dermal density, and enhanced skin elasticity. The microspheres showed the capacity to stimulate the regeneration of collagen fibers. The agarose microspheres offer a novel avenue for soft tissue filling and hold significance in the field of tissue engineering and skin regeneration.

Characterization of Photobacterium damselae subsp. damselae isolated from diseased Asian seabass (Lates calcarifer) and the preliminary development of a formalin-killed cell vaccine.

Asian seabass (Lates calcarifer) is an economically important fish species that is widely cultivated in Thailand. However, aquaculture of Asian seabass is limited by infectious diseases. One of the most serious diseases is photobacteriosis, caused by Photobacterium damselae. Vaccination is recognized as an efficient disease prevention and pathogen control method for strengthening the aquaculture industry. To promote vaccine development, the characterization of pathogenic bacteria and their pathogenesis is required. In this study, isolates of P. damselae were obtained from commercial aquaculture farms in Thailand during 2019-2021. Analyses of 16S rRNA and the urease subunit alpha genes identified the isolates as P. damselae subsp. damselae (Phdd). Antibiotic susceptibility analyses showed that all Phdd isolates were resistant to amoxicillin (10 μg). Haemolysis and phospholipase activities were used to categorize P. damselae into three groups based on their biological activities. The pathogenicity of four candidates (SK136, PD001, PD002 and T11L) was tested in Asian seabass. Isolate SK136 showed the highest virulence, with a lethal dose (LD50) of 1.47 × 105 CFU/fish, whereas isolate PD001 did not show any virulence. Genotypic characterization, based on multi-locus sequence typing analysis, demonstrated that all candidates were novel strains with new sequence types (64, 65, 66 and 67). Preliminary vaccination using formalin-killed cells (FKCs) protected Asian seabass from artificial challenges. Taken together, these results provide fundamental knowledge for vaccine development against Phdd infection in Asian seabass.

Fabrication, microstructure and hemostatic activity of Cu-Zn manganite nanoparticles

Copper subbed zinc manganite Zn(1-x)CuxMn2O4 (x = 0.05,0.15,0.25,0.35,0.45,0.55) (BCZMO) NPs were prepared by sol–gel co-precipitation technique at surrounding temperature. BCZMO NPs were analyzed by XRD, SEMfor microstructure characterizations. XRD study confirms the tetragon structure of BCZMO NPs and effect of copper on ZMO NPs. SEM micrographs supports our findings. The BCZMO NPs interfered in blood coagulation and exhibited pro-coagulant property. BCZMO NPs decreased the coagulation time of citrated plasma from control 4.31 min to 1.35 min. In addition, the BCZMO NPs aggregated platelets up to 80 % at a concentration of 600 µg while the agonist epinephrine prompted platelet collection of around 90 % at 10 mM concentration.No kind of hemolytic activity in RBC cells shown by these BCZMO-NPs insisting their non-toxic nature. These BCZMO-NPs exhibited pro-coagulant effect with their interference in blood coagulation cascade and initiation in platelet aggregation. Thus, BCZMO NPs can be a potential candidate in biomedical field as hemostatic agents.

Modified mesoporous silica nanocarriers containing superparamagnetic iron oxide nanoparticle, 5-fluorouracil or oxaliplatin, and metformin as a radiosensitizer, significantly impact colorectal cancer radiation therapy

This study investigates the anticancer effects of SPION-based silica nanocarriers carrying 5-fluorouracil (5-FU) or oxaliplatin (OX), and metformin (MET) on colorectal cancer cells. Nanocarriers were equipped with pH-responsive gold gatekeepers for controlled release, PEGylation for longer circulation, and folic acid (FA) for targeted delivery. The effects were evaluated by investigating cell viability, cellular uptake, flow cytometry, and clonogenic assay in vitro. The efficacy of the system was also tested in vivo on C57BL/6 mice bearing HT-29 tumors, and potential side effects were evaluatedNanocarriers were synthesized with hydrodynamic diameters of 79.8 nm for 5-FU and 85.2 nm for OX; zeta potentials of −21 and –22 mV, respectively, and remained stable after 72 h. Encapsulation efficiencies were 85 % for 5-FU, 80 % for OX, and 83 % for MET, with loading capacities of 44 %, 38 %, and 41 %, respectively. Drug release in acidic buffer was 38.7 % for 5-FU, 32.8 % for OX, and 43.5 % for MET. MTT assay showed increased toxicity due to FA conjugation, while PEGylation reduced the hemolysis activity. Targeted nanocarriers demonstrated superior cellular uptake and tumor localization compared to non-targeted variants. The combination of 5-FU-MET and OX-MET nanocarriers with radiation therapy (RT) demonstrated the greatest effect on their antitumor activity, accompanied by minimal side effects indicating effective tumor targeting in vivo. MRI and CT imaging further supported these findings. This study underscores the synergistic impact of MET alongside RT on the inhibition of cancer cells and tumor growth for both targeted 5-FU and OX nanocarriers reflecting the significant radiosensitizing properties of MET.

Isolation and Preliminary Characterization of Probiotic Isolates from Healthy Thai Adult Feces

Probiotics are known for health-promoting microorganisms, for examples, assisting gastrointestinal digestion and short chain fatty acid producers. Several current oral human probiotics were derived from human samples; yet, of various ethnic (genetics) and dietary patterns. As probiotics from similar ethnic (genetics) and dietary pattern might confer certain advantages (e.g., more compatibility with hosts), this study isolated and characterized probiotics from healthy Thai adults. To screen for potential probiotic isolates from healthy Thai adult feces, and preliminary characterize these probiotic isolates by biochemical tests and 16S rRNA gene sequencing, following the established Thailand’s Food and Drug Administration (FDA) and National Center for Genetic Engineering and Biotechnology (BIOTEC) guidelines. Twenty fecal samples with clinically verified healthy Thai males and females aged 22-42 years were cultivated on MRS (or M17) media along the broad probiotic screenings (pH 4.0 and 0.1% oxgall) for bacterial probiotic propagation. All derived colonies of different morphologies were continued colony PCR using universal probiotic primers (specific for genera Bifidobacterium, Enterococcus, Lactobacillus, Pediococcus and Streptococcus thermophilus), and only PCR-positive colonies from any of these probiotic primer pair were further cultivation for isolates. Then, these isolates were screened and characterized general probiotic properties that included acid tolerance (pH 2.0), bile salt tolerance (0.3% bile salt), hydrophobicity, antibiotic (ampicillin, ciprofloxacin, erythromycin and vancomycin) susceptibility, no hemolysis activity, catalase, and potassium hydroxide (KOH). The final passing isolates represented potential probiotic isolates that could perhaps maintain in gastrointestinal tract with safety, and were full-length 16S rRNA gene sequenced to identify the genus and species by BLASTN against non-redundant GenBank database. A total of 295 bacterial isolates were obtained from the cultivated on MRS (or M17) media along the broad probiotic screenings from 20 fecal samples. Then, following the general probiotic property characterizations and the full-length 16S rRNA gene sequence analysis, 7 potential isolates were finally derived: 2 Bifidobacterium animalis subsp. lactis, 1 Lactobacillus (new genus name as Limosilactobacillus) reuteri, 1 L. reuteri or L. vaginalis, and 3 Pediococcus pentosaceus. These isolates exhibited strong acid and bile salt tolerances, and relatively high cell surface hydrophobicity. These isolates were susceptible to ampicillin and erythromycin, similar to a reference probiotic control Lactobacillus casei strain Shirota; and the CU13-450 and CU13-451 were also susceptible to vancomycin, similar to a reference control Bifidobacterium breve. Catalase-negative consistent with these bacteria in intestinal tract as anaerobes or facultative anaerobes, and KOH-negative result inferred the Gram-positive bacteria consistent with the full-length 16S rRNA gene sequence identification (Bifidobacterium, Lactobacillus, and Pediococcus are Gram-positive bacteria). No hemolysis activity was observed for these isolates. These 7 probiotic isolates had general probiotic properties and with confirmed the 16S rRNA gene sequences, that follow the Thailand’s FDA and BIOTEC guidelines. However, additional beneficial function investigations, such as in vitro epithelial cell adhesion and in vivo animal model, and whole genome sequencing, should be performed to better validate their possible human health benefits and safety.

Isolation of Lactobacillus spp. from Healthy Infants' Feces and Study of their Antibiotic Susceptibility

This study aimed to isolate Lactobacillus spp. bacteria from the feces of healthy newborns and investigate their susceptibility to antibiotic resistance. 120 fecal samples were obtained from infants under 12 months of age and susceptibility was identified using biochemical tests and molecular identification methods. A total of 30 positive isolates, chosen randomly from the 90 positive samples, were selected to conduct a safety assessment. the isolates were analyzed for their hemolytic activity and susceptibility to antibiotics. Using 16S rRNA sequencing, a total of 30 positive isolates were determined to be lactobacillus spp. The results of the antibiotic resistance test indicated that the majority of the isolates showed susceptibility to tetracycline, chloramphenicol, gentamicin, clindamycin, and amoxicillin while demonstrating resistance to vancomycin, nalidixic acid, streptomycin, and amikacin. conclusion: Based on these specific traits, the isolates of Lactobacillus spp. demonstrated favorable qualities as probiotics.