Active Cells Research Articles

Determining conditions for inactivation of multispecies biofilm cells by peracetic acid applying response surface methodology

Biofilms can form on various surfaces and are typically associated with multiple microbial species. Particularly in the food industry, biofilms are often a source of contamination, necessitating mitigation strategies. In this study, Response Surface Methodology (RSM) was utilized to optimize conditions for the inactivation of multispecies biofilm cells, with peracetic acid (PAA) (0.05% to 0.5%), treatment time (5 to 30 minutes), and temperature (25 to 60 ºC) as independent variables. The multispecies biofilm consisted of Gram-positive and Gram-negative bacteria previously isolated from biofilms formed in the presence of raw milk. Stainless steel (SS) coupons were immersed in reconstituted whole milk inoculated with Pseudomonas fluorescens, Rahnella inusitata, Staphylococcus aureus, and Micrococcus aloeverae. Sessile cell counts of approximately 108 CFU/cm2 were obtained after 10 days of incubation at 4 ºC. The response surface model exhibited a good fit, with R2 of 0.949 and adjR2 of 0.883. All independent variables had a positive effect on the inactivation of biofilm cells. The maximum inactivation achieved was approximately 7 log (CFU/cm2), with the highest values observed at 0.5% PAA, for 30 min at 60°C. Epifluorescence microscopy revealed that many cells in the biofilm were dead or injured after treatment at the central point (PAA = 0.275%; Time = 17.5 min; Temperature = 42.5 ºC). RSM helps to predict better conditions for maximum biofilm eradication and proves to be a promising approach for monitoring the inactivation of multispecies biofilm cells, which warrants further exploration.

Hippo pathway in cancer cells induces NCAM1+αSMA+ fibroblasts to modulate tumor microenvironment

Cancer cells adeptly manipulate the tumor microenvironment (TME) to evade host antitumor immunity. However, the role of cancer cell-intrinsic signaling in shaping the immunosuppressive TME remains unclear. Here, we found that the Hippo pathway in cancer cells orchestrates the TME by influencing the composition of cancer-associated fibroblasts (CAFs). In a 4T1 mouse breast cancer model, Hippo pathway kinases, large tumor suppressor 1 and 2 (LATS1/2), promoted the formation of neural cell adhesion molecule 1 (NCAM1)+alpha-smooth muscle actin (αSMA)+ CAFs expressing the transforming growth factor-β, which is associated with T cell inactivation and dysfunction. Depletion of LATS1/2 in cancer cells resulted in a less immunosuppressive TME, indicated by the reduced proportions of NCAM1+αSMA+ CAFs and dysfunctional T cells. Notably, similar Hippo pathway-induced NCAM1+αSMA+ CAFs were observed in human breast cancer, highlighting the potential of TME-manipulating strategies to reduce immunosuppression in cancer immunotherapy.

Thermal Inactivation of Cells of Salmonella spp. in Pot Pies Prepared With a Beef, Chicken, or Meat Alternative Filling, With and Without Gravy, During Cooking in a Convection Oven

Cooking parameters elaborated in the U.S. Department of Agriculture’s Food Safety and Inspection Service Cooking Guideline for Meat and Poultry Products (Appendix A) were evaluated for inactivation of Salmonella spp. in pot pies. To prepare dough for pot pies, flour, butter, sugar, salt, and water were mixed, portioned into balls (65 or 85 g each), flattened (ca. 13 or 15 cm diameter, ca. 0.5 cm thick), and hand-pressed into pans (ca. 19.4 cm diameter). Next, a 100-g portion of beef, chicken, or a meat alternative, with or without added gravy (55 g of protein and 45 g of gravy), was inoculated with a cocktail of Salmonella spp. (ca. 6.5 or 7.9 log CFU/g of filling) and distributed onto the pie crust. After covering with a 65-g sheet of dough, pies were heat sealed in nylon polyethylene bags and stored at −20 °C for up to 72 h. Frozen pot pies containing a beef or meat alternative filling were cooked in a convection oven to an internal temperature of 57.8 °C (136°F) instantaneous, 62.8 °C (145°F) and held for 4 min, 67.2 °C (153°F) and held for 34 sec, or 71.1 °C (160°F) instantaneous; whereas chicken pot pies were cooked to an internal temperature of 57.8 °C (136°F) instantaneous, 62.8 °C (145°F) and held for 13 min, 67.2 °C (153°F) and held for 96 sec, or 73.9 °C (165°F) instantaneous. Cells of Salmonella spp. were recovered from uncooked or cooked pot pies by stomaching each pie in peptone water and enumerating pathogen levels via direct plating. Cooking delivered Salmonella spp. reductions of ca. 3.6 to ≥6.3 log CFU/g of pot pie. With few exceptions, when pathogen levels decreased to below detection by direct plating (0.5 log CFU/g of pot pie), cells of Salmonella were not recovered by enrichment. In addition, there were minimal differences in the aw and moisture content of the protein filling before and after cooking, suggesting that enclosing the meat, poultry, or meat alternative filling within a dough wrapping maintains moisture in the filling during the cooking process.

Abstract B062: Low dose dual epigenetic therapy utilizing hypomethylating agents and HDAC inhibitors prolong survival in an orthotopic PDAC model and alter the tumor microenvironment

Abstract Background: We have previously studied the effect of hypomethylating agent Decitabine (5- aza-dC; DAC) on pancreatic cancer using the KPC model (KrasG12D/+;Trp53R172H/+;Pdx1- Cre) and observed increased CD4+/CD8+ TILs, greater PD1 expression and slowed tumor growth with increased tumor necrosis. Subsequently we showed a significant survival benefit with DAC+αPD1 compared to either single agent. However, we identified a specific myeloid cell population initially identified by Chi3l3 expression that may be responsible for treatment resistance or escape. Based on prior observations of synergy specifically affecting tumor infiltrating myeloid and lymphoid cells, we focused on identifying a dual epigenetic strategy (a combination of DAC plus histone deacetylase inhibitors (HDACi)) to enhance immunological synergy and test their treatment efficacy with αPD1. Methods: We used a combination primary KPC-derived cell lines, bone marrow (BMDM) and peritoneum derived (PM) myeloid cells and T cells in co-culture systems to assess cytotoxic synergy (Bliss/Lowe consensus) and immune effects (cytokine release, myeloid cell polarization and T cell activation) to evaluate combinations of DAC+HDACi. The KPC orthotopic model relying on US monitoring of tumor size for standardized initiation of therapy was used for survival analysis and profiling in treatment combinations of DAC+HDACi+αPD1. Results: We showed that hypomethylation therapy exerts a primarily paracrine effect on myeloid cells and this polarization results in a decrease in DAC–induced T cell activation. We therefore selected HDAC inhibitors that in combination with DAC alter this paracrine effect. We selected specific HDAC inhibitors based on induction of class of HDAC expression following hypomethylation therapy, highest Bliss synergy scores at low doses and reversal of cytokine release, M2 polarization and T cell inactivation. Three HDAC inhibitors were chosen for in vivo testing, Domatinostat, Pracinostat and Entinostat. Using the treatment paradigm of ultra-low dose DAC+ HDACi (mainly to minimize toxicity) combined with αPD1 in the KPC orthotopic model we show that the triple combination (DAC + Entinostat + αPD1) is well-tolerated and results in a significant survival benefit compared to single and dual agent combinations. In addition, we also demonstrate a reduction in the Chi3l3 expressing myeloid cell population. Conclusions: Our results show that the combination of hypomethylating agent plus HDAC inhibitor has several beneficial effects on the tumor immune response in PDAC. HDAC inhibitors specifically reduce the suppressive myeloid cell population we previously identified after DAC therapy. The combination of dual low dose epigenetic therapy using a hypomethylating agent plus HDAC inhibitor followed by αPD1 results in significant survival benefit in the orthotopic KPC model. Citation Format: Stephen Muzyka, Arturo Orlacchio, Amanda Ackermann, Yasmine Chinda, Yoona Shim, Igor Dolgalev, Tamas Gonda. Low dose dual epigenetic therapy utilizing hypomethylating agents and HDAC inhibitors prolong survival in an orthotopic PDAC model and alter the tumor microenvironment [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B062.

Abstract A045: Disrupting local immunosuppression through combination myCAF/myeloid targeting in pancreatic cancer

Abstract Immunotherapy has revolutionized clinical care for many cancers, yet the approach has not been fully realized in immunologically cold tumors like Pancreatic Ductal Adenocarcinoma (PDAC). One striking hallmark of PDAC is local immunosuppression (LIS) which leads to cytotoxic T cell inactivation and exclusion. There is great need for an increased understanding of the cellular crosstalk within tumors and the identification of stromal and immune populations involved in shaping the tumor microenvironment (TME). Oncogenic KRAS activation in tumor cells promotes the invasion and proliferation of tumor-supporting stromal cells, while excluding cancer-targeted cytotoxic T cells. LIS is mediated by multiple subtypes of cancer-associated fibroblasts (CAFs) and myeloid cells resident within the tumor parenchyma. Multiple prior attempts to reverse LIS in PDAC by targeting individual stromal cell populations have been unsuccessful, alluding to the complexity of stromal crosstalk within the TME. The stromal diversity of PDAC complicates investigating paracrine cascades involving multiple cell types. To decipher diverse drug effects on altering the TME, we employ in vivo studies in mouse models recapitulating the human disease, as well as a novel tumor explant model that enables the short-term culture of intact human or murine PDAC. Importantly, PDAC explants maintain their histopathological architecture and cellular diversity over time. This medium-throughput platform allows for testing of multiple drugs and mechanistic hypotheses in the native PDAC TME. We show in preliminary data that Smoothened inhibition (SMOi) decreases proliferation and activity of myCAFs, but provokes the expansion of CD11b-positive myeloid cells in vivo. Thus, we hypothesize that LIS in PDAC is maintained by a delicate balance between myCAFs and myeloid cells, preventing effective T cell invasion. Single cell RNA-seq data comparing ctrl vs. SMOi-treated murine PDAC elucidates stromal subpopulations involved in the LIS phenotype and guides the identification of myeloid subtypes emerging after SMOi. Strikingly, we demonstrated that simultaneous SMOi and targeting myeloid cells via anti-Gr1 or CCR1 inhibition (CCR1i) significantly elevates cytotoxic T cell numbers up to 20x within the TME. We are currently investigating whether the activity of these T cells may be further potentiated through combination with immunomodulatory agents. By testing various treatment combination in the same TME, we will identify the best synergistic effects for future immunotherapy approaches in human PDAC. In summary, we are elucidating the complex mechanism behind LIS in PDAC by employing our novel explant culture system alongside in vivo studies. We aim to develop a translatable regimen to neutralize LIS, reactivating the cytotoxic T cells in the tumor periphery to invade, proliferate, and attack cancer cells. Citation Format: Marie C Hasselluhn, Lukas Vlahos, Dafydd Thomas, Quin T Waterbury, Alvaro Curiel Garcia, Amanda R Decker-Farrell, Tanner C Dalton, Stephen A Sastra, Carmine F Palermo, Timothy C Wang, Andrea Califano, Kenneth P Olive. Disrupting local immunosuppression through combination myCAF/myeloid targeting in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A045.

Cold induces brain region-selective cell activity-dependent lipid metabolism.

It has been well documented that cold is an enhancer of lipid metabolism in peripheral tissues, yet its effect on central nervous system lipid dynamics is underexplored. It is well recognized that cold acclimations enhance adipocyte functions, including white adipose tissue (WAT) lipid lipolysis and beiging, and brown adipose tissue (BAT) thermogenesis in mammals. However, it remains unclear whether and how lipid metabolism in the brain is also under the control of cold acclimations. Here, we show that cold exposure predominantly increases the expressions of the lipid lipolysis genes and proteins within the paraventricular nucleus of the hypothalamus (PVH). Mechanistically, we find that cold activates cells within the PVH and pharmacological inactivation of cells blunts cold-induced effects on lipid peroxidation, accumulation of lipid droplets (LDs), and lipolysis in the PVH. Together, these findings suggest that PVH lipid metabolism is cold sensitive and integral to cold-induced broader regulatory responses.

BATF is a major driver of NK cell epigenetic reprogramming and dysfunction in AML.

Myelodysplastic syndrome and acute myeloid leukemia (AML) belong to a continuous disease spectrum of myeloid malignancies with poor prognosis in the relapsed/refractory setting necessitating novel therapies. Natural killer (NK) cells from patients with myeloid malignancies display global dysfunction with impaired killing capacity, altered metabolism, and an exhausted phenotype at the single-cell transcriptomic and proteomic levels. In this study, we identified that this dysfunction was mediated through a cross-talk between NK cells and myeloid blasts necessitating cell-cell contact. NK cell dysfunction could be prevented by targeting the αvβ-integrin/TGF-β/SMAD pathway but, once established, was persistent because of profound epigenetic reprogramming. We identified BATF as a core transcription factor and the main mediator of this NK cell dysfunction in AML. Mechanistically, we found that BATF was directly regulated and induced by SMAD2/3 and, in turn, bound to key genes related to NK cell exhaustion, such as HAVCR2, LAG3, TIGIT, and CTLA4. BATF deletion enhanced NK cell function against AML in vitro and in vivo. Collectively, our findings reveal a previously unidentified mechanism of NK immune evasion in AML manifested by epigenetic rewiring and inactivation of NK cells by myeloid blasts. This work highlights the importance of using healthy allogeneic NK cells as an adoptive cell therapy to treat patients with myeloid malignancies combined with strategies aimed at preventing the dysfunction by targeting the TGF-β pathway or BATF.

Dielectric Resonators for Microwave Hyperthermia

AbstractHyperthermia, utilizing various types of energy sources such as microwaves, holds great promise for cancer treatment. The intense interaction between microwaves and substances during hyperthermia can produce heat efficiently, leading to the inactivation of cancer cells. Mie‐type resonance is widely studied in the regulation of electromagnetic waves, offering a new perspective for microwave hyperthermia. In this study, a Mie‐type dielectric resonator is reported for microwave hyperthermia using high‐permittivity and temperature‐stable ceramic. The proposed dielectric particles have the characteristics of compact size, low cost, and excellent biocompatibility. The resonant frequency of the system can be finely tuned through material design and arrangement, thus expanding its potential application scenarios. The dielectric particles can be rapidly heated under microwave irradiation, thereby enabling effective hyperthermia to eliminate cancer cells. This work demonstrates the potential of a safe and tunable Mie‐type dielectric resonator in microwave hyperthermia, which is expected to promote further research on Mie‐type resonance in the biomedical field.

Evaluation of the removal of cyanobacteria and cyanotoxins with a composite hydrogel based on chemically modified gelatin and PVA-containing graphene oxide nanoparticles

After being applied to pharmaceutics removal from water, the newly synthesized composite hydrogel based on chemically modified gelatin and PVA-containing graphene nanoparticles (CHGP-GOn), was now assessed for another application. The hydrogels were able to interact with Microcystis aeruginosa cells, resulting in the formation of small cell colonies and cell lysis related to exposure time. In the removal tests, the best general removal efficiency of cyanobacterial cells was achieved with the highest adsorbent mass at natural pH, achieving values of removal of 90 % for cells, 75 % chlorophyll-a, 63 % and 43 % for turbidity and visible colour removals, respectively. From the kinetic study, the results showed that cell inactivation has achieved removal equilibrium in 19 h, with a qe of 106 × 106 cells/g of CHGP-GOn. Furthermore, with the results from the other parameters, the tests presented a removal equilibrium of just 14 h, there was a removal of 4670 µg/L of Chlorophyll-a and 6450 µg/L/g of MC-LR microcystin per gram of adsorbent. The experimental data best fitted to the Elovich model, indicating possible removal by chemisorption. Analysis of cellular integrity and morphology showed that within just 12 h, few cyanobacterial cells showed membrane disruption and release of intracellular toxins, with an increase in the toxicity medium related to extended exposure time, and presentation of morphological and superficial cellular damage within 24 h. In addition, other characteristic measurements presented an exceptional mechanical strength and resistance that was slightly reduced after swelling. However, showing good development during adsorption tests under agitation, without any detachment of material and an absence of GO leaching. Therefore, it can be concluded that the synthesized hydrogel should be applied in removing M. aeruginosa cells and their toxins from water treatment, serving as an excellent alternative to traditional adsorbents.

PAFerroptosis Combined with Metabolic Disturbance of Mito by p52-ZER6 for Enhanced Cancer Immunotherapy induced by Nano-Bacilliform-Enzyme.

The confused gene expressions and molecular mechanisms for mitochondrial dysfunction of traditional nanoenzymes is a challenge for tumor therapy. Herein, a nano-bacilliform-enzyme obtains the ability to inhibit p52-ZER6 signal pathway, regulate the genes related to mitochondrial metabolism, and possess the GOx/CAT/POD-like property. NBE acquires catalytic activity from the electronic energy transition. The tannin of NBE as a mitochondrial (Mito)-targeting guide overloads MitoROS, and then metabolic disorder and lipid peroxidation of Mito membrane occurs, thus leading to a novel death pathway called PAFerroptosis (pyroptosis, apoptosis, and Ferroptosis). Simultaneously, in order to refrain from mitophagy, hydroxychloroquine is mixed with NBE to form a combo with strength pyroptosis. As a result, NBE/combo improves the PAFerroptosis obviously by activation of CD8+T cells and inactivation of MDSC cells, up-regulating expression of caspase-3 signal pathway, intercepting DHODH pathway to arrive excellent antitumor effect (93%). Therefore, this study establishes a rational nanoenzyme for mitochondrial dysfunction without mitophagy for effective antitumor therapy.

Feasibility assessment and underlying mechanisms of metabisulfite pretreatment for enhanced volatile fatty acids production from anaerobic sludge fermentation

Employing chemical pretreatment for waste activated sludge (WAS) fermentation is crucial to achieving sustainable sludge management. This study investigated the feasibility of metabisulfite (MS) pretreatment for enhancing volatile fatty acids (VFAs) production from WAS. The results show that after 24-h MS pretreatment, the content of soluble organic matter and loosely bound extracellular polymeric substances (LB-EPS), especially proteins, increased significantly. During the fermentation, MS pretreatment under alkaline conditions was more efficient, with VFA peaking on the fifth day, showing a 140 % increase compared to the alkaline control group. Correlation analysis suggests that the dosage of MS, rather than pH, is closely related to the levels of soluble protein, polysaccharides, LB-EPS, and subsequential VFAs production, while alkaline conditions facilitate the dissolution of total organic carbon. Furthermore, sulfite radicals (SO3•−) are attributed to cell inactivation and lysis, while alkaline conditions initially reduce the size of the flocs, further promoting MS for attacking flocs, thereby improving the performance of fermentation. The study also found that MS pretreatment reduced microbial community diversity, enriched hydrolytic and fermentation bacteria (Actinobacteriota and Firmicutes), and suppressed methanogens (Methanobacteriaceae and Methanosaetaceae), making it a safe, viable, and cost-effective chemical agent for sustainable sludge management.

Efficacy of gaseous ozone and UVC radiation against Candida auris biofilms on polystyrene surfaces

The growth and attachment of highly resistant biofilms on/to polymer-based surfaces, including reusable medical devices is a significant health concern in infection control. The increasing prevalence of the multidrug-resistant yeast, Candida auris, has recently garnered immense interest as a contributor to healthcare-associated infections. In this study, we investigate the potential of disinfection processes (ozone, UVC, and ozone + UVC) to inactivate C. auris biofilms (NCPF 8971), grown on polystyrene surfaces. A bespoke decontamination chamber is used to expose the substrate for 20–60 mins, corresponding to ozone doses between 1000 and 3000 ppm.min and UVC doses between 2864 and 11,592 mJ/cm2, respectively. The performance of these 3 treatment methods against the biofilms and vegetative/planktonic cells of the organism is comparatively evaluated. While complete inactivation (> 8 log10 reduction, CFU/mL) of the vegetative cells is observed within 40 mins by all methods, the biofilms proved considerably difficult to inactivate. Only 3.3 log10 reduction was achieved using ozone for 40 min, whereas 7.2 log10 reduction was obtained using UVC for 40 min. A hybrid application of ozone and UVC improved the decontamination efficacy compared to their independent applications (i.e., ozone only and UVC only). Scanning electron microscopy results yielded new insights into the inactivation mechanisms; thus providing the needed foundation for newer developments in oxidation-based processes for biofilm mitigation and control.

A hurdle strategy based on the combination of non-thermal treatments to control diarrheagenic E. coli in cheese

This study aimed to assess the efficacy of a multi-hurdle process combining mild High Hydrostatic Pressure (HHP) treatments and Thyme Oil (TO) edible films as a non-thermal method to combat pathogenic E. coli (aEPEC and STEC) in raw cow's-milk cheese stored at 7 °C and packaged under modified atmosphere. Changes in headspace atmosphere of cheese packs and treatment effects on Lactic Acid Bacteria (LAB) counts and diarrheagenic E. coli strains (aEPEC and STEC) were evaluated over a 28 d storage period. The results demonstrated that the combined treatment exhibited the most significant antimicrobial effect against both strains compared to individual treatments, achieving reductions of 4.30 and 4.80 log cfu/g after 28 d of storage for aEPEC and STEC, respectively. Notably, the synergistic effect of the combination treatment resulted in the complete inactivation of intact cells for STEC and nearly completed inactivation for aEPEC by the end of the storage period. These findings suggest that the combination of HHP with selected hurdles could effectively enhance microbial inactivation capacity, offering promising alternatives for improving cheese safety without affecting the starter microbiota.

ALK5/VEGFR2 dual inhibitor TU2218 alone or in combination with immune checkpoint inhibitors enhances immune-mediated antitumor effects

Transforming growth factor β (TGFβ) is present in blood of patients who do not respond to anti-programmed cell death (ligand) 1 [PD-(L)1] treatment, and through synergy with vascular endothelial growth factor (VEGF), it helps to create an environment that promotes tumor immune evasion and immune tolerance. Therefore, simultaneous inhibition of TGFβ/VEGF is more effective than targeting TGFβ alone. In this study, the dual inhibitory mechanism of TU2218 was identified through in vitro analysis mimicking the tumor microenvironment, and its antitumor effects were analyzed using mouse syngeneic tumor models. TU2218 directly restored the activity of damaged cytotoxic T lymphocytes (CTLs) and natural killer cells inhibited by TGFβ and suppressed the activity and viability of regulatory T cells. The inactivation of endothelial cells induced by VEGF stimulation was completely ameliorated by TU2218, an effect not observed with vactosertib, which inhibits only TGFβ signaling. The combination of TU2218 and anti-PD1 therapy had a significantly greater antitumor effect than either drug alone in the poorly immunogenic B16F10 syngeneic tumor model. The mechanism of tumor reduction was confirmed by flow cytometry, which showed upregulated VCAM-1 expression in vascular cells and increased influx of CD8 + CTLs into the tumor. As another strategy, combination of anti-CTLA4 therapy and TU2218 resulted in high complete regression (CR) rates in CT26 and WEHI-164 tumor models. In particular, immunological memory generated by the combination of anti-CTLA4 and TU2218 in the CT26 model prevented the development of tumors after additional tumor cell transplantation, suggesting that the TU2218-based combination has therapeutic potential in immunotherapy.

Theranostic dye entrapped in an optimized blended-polymer matrix for effective photodynamic inactivation of diseased cells.

Despite the wide range of treatment options available for cancer therapy, including chemotherapy, radiation therapy, and surgical procedures, each of these treatments has a different side-effect profile and leaves the patient with no option but to choose. Due to their insensitivity and nonspecificity, conventional treatments damage normal cells together with cancer cells. In recent years, a significant amount of attention has been focused on photodynamic therapy (PDT) as a treatment for cancer and drug-resistant microbes. An activated photosensitizer is used as a part of the procedure along with oxygen molecules and a specific wavelength of light belonging to the visible or NIR spectral zone. A light-sensitive laser dye, rhodamine 6G (R6G), was used in the present study as a photosensitizer, taking a challenge to improve the aqueous solubility and ROS quantum yield using optimum concentration (160mg/ml) of chitosan-alginate (Cs-Alg) blended polymeric nanoformulations. As evidenced by steady-state spectrophotometric and fluorometric measurements, ROS quantum yield increases three-fold over aqueous solution along with solubility gaining that was validated by PDT experiment using human epithelial carcinoma (KB) cell line. Phantom optical imaging was taken using the IVIS imaging system to establish the formulations as a fluorescence-based optical contrast agent, and zebrafish embryos were used to establish their safe in vivo use. The release profile of R6G was fitted using kinetic models, which followed the Non-Fickian kinetic profile. In conclusion, we recommend the formulations as a potential theranostic agent that will aid in PDT-based therapy in conjunction with optical imaging-based diagnosis.

Oxidation of Microcystis aeruginosa and Microcystins with Peracetic Acid.

Peracetic acid (PAA) shows potential for use in drinking water treatment as an alternative to prechlorination, such as for mussel control and disinfection by-product precursor destruction, though its impact as a preoxidant during cyanobacterial blooms remains underexplored. Here, Microcystis aeruginosa inactivation and microcystin-LR and -RR release and degradation using PAA were explored. The toxin degradation rates were found to be higher in alkaline conditions than in neutral and acidic conditions. However, all rates were significantly smaller than comparable rates when using free chlorine. The inactivation of M. aeruginosa cells using PAA was faster at acidic pH, showing immediate cell damage and subsequent cell death after 15-60 min of exposure to 10 mg/L PAA. In neutral and alkaline conditions, cell death occurred after a longer lag phase (3-6 h). During cell inactivation, microcystin-LR was released slowly, with <35% of the initial intracellular toxins measured in solution after 12 h of exposure to 10 mg/L PAA. Overall, PAA appears impractically slow for M. aeruginosa cell inactivation or microcystin-LR and -RR destruction in drinking water treatment, but this slow reactivity may also allow it to continue to be applied as a preoxidant for other purposes during cyanobacterial blooms without the risk of toxin release.

Single-cell RNA sequencing reveals reduced intercellular adhesion molecule crosstalk between activated hepatic stellate cells and neutrophils alleviating liver fibrosis in hepatitis B virus transgenic mice post menstrual blood-derived mesenchymal stem cell transplantation.

Liver fibrosis can cause hepatitis B virus (HBV)-associated hepatocellular carcinoma. Menstrual blood-derived mesenchymal stem cells (MenSCs) can ameliorate liver fibrosis through paracrine. Single-cell RNA sequencing (scRNA-seq) may be used to explore the roadmap of activated hepatic stellate cell (aHSC) inactivation to target liver fibrosis. This study established HBV transgenic (HBV-Tg) mouse model of carbon tetrachloride (CCl4)-induced liver fibrosis and demonstrated that MenSCs migrated to the injured liver to improve serological indices and reduce fibrotic accumulation. RNA-bulk analysis revealed that MenSCs mediated extracellular matrix accumulation and cell adhesion. Liver parenchymal cells and nonparenchymal cells were identified by scRNA-seq in the control, CCl4, and MenSC groups, revealing the heterogeneity of fibroblasts/HSCs. A CellChat analysis revealed that diminished intercellular adhesion molecule (ICAM) signaling is vital for MenSC therapy. Specifically, Icam1 in aHSCs acted on Itgal/Itgb2 and Itgam/Itgb2 in neutrophils, causing decreased adhesion. The expression of Itgal, Itgam, and Itgb2 was higher in CCl4 group than in the control group and decreased after MenSC therapy in neutrophil clusters. The Lcn2, Pglyrp1, Wfdc21, and Mmp8 had high expression and may be potential targets in neutrophils. This study highlights interacting cells, corresponding molecules, and underlying targets for MenSCs in treating HBV-associated liver fibrosis.

A hierarchal model for bacterial cell inactivation in solution by direct and indirect treatment using cold atmospheric plasmas

Cold atmospheric plasma devices have shown promise for a variety of plasma medical applications, including wound healing and bacterial inactivation often performed in liquids. In the latter application, plasma-produced reactive oxygen and nitrogen species (RONS) interact with and damage bacterial cells, though the exact mechanism by which cell damage occurs is unclear. Computational models can help elucidate relationships between plasma-produced RONS and cell killing by enabling direct comparison between dissimilar plasma devices and by examining the effects of changing operating parameters in these devices. In biological applications, computational models of plasma-liquid interactions would be most effective in design and optimization of plasma devices if there is a corresponding prediction of the biological outcome. In this work, we propose a hierarchal model for planktonic bacterial cell inactivation by plasma produced RONS in liquid. A previously developed reaction mechanism for plasma induced modification of cysteine was extended to provide a basis for cell killing by plasma-produced RONS. Results from the model are compared to literature values to provide proof of concept. Differences in time to bacterial inactivation as a function of plasma operating parameters including gas composition and plasma source configuration are discussed. Results indicate that optimizing gas-phase reactive nitrogen species production may be key in the design of plasma devices for disinfection.

Anticancer transmission effects induced by hybrid plasma bubble-activated medium

As an indirect application of cold atmospheric plasma, the use of plasma-activated solutions has recently attracted significant attention for intracavity tumor perfusion therapy. Here, an underwater plasma-bubble reactor with three discharge modes was applied to activate a cell culture medium, with the aim of inhibiting the growth of bladder cancer T24 cells in vitro. The results showed that although the reactive species in cell culture medium generated by the underwater bubble plasma varied between the dielectric barrier discharge (DBD) mode, spark mode, and hybrid mode, the plasma-activated cell culture medium (PAM) corresponding to all three discharge modes effectively decreased the viability of T24 cells. However, after co-culturing the PAM-pretreated T24 cells with normal T24 cells, it was observed that normal T24 cells were not affected by either the DBD or spark mode; however, the hybrid mode PAM-pretreated T24 cells further induced inactivation and apoptosis of normal T24 cells. Further studies suggested that the aqueous reactive species generated by the underwater bubble plasma in hybrid mode not only induced the apoptosis of T24 cells directly but also triggered PAM-pretreated T24 cell-derived exosome-mediated anticancer activity toward cancer cells. These results may provide a strategy for enhancing the anticancer extent and effect of PAM as well as advancing its clinical application.

Scatalysis: Exploring the mechanism of photocatalytic algal removal

Photocatalytic algal removal is an environmentally friendly and low-cost algal bloom control technology. In this work, carbon nitride nanosheets based on morphology control were synthesized, and a ternary Z-scheme photocatalyst was designed through heterojunction engineering for photocatalytic removal of harmful algae and their organic matter, and the removal rate of chlorophyll a was close to 100 % in 300 min. Three-dimensional fluorescence spectrograms and cellular SEM maps revealed the removal of algae and their organic matter, and the mechanism of Ag/AgCl/ZIF-8/SCN photocatalytic algal removal was explored in conjunction with the changes in membrane permeability and various physiological functions. Mechanistic studies showed that excess O2·− played a crucial role in cell inactivation and organic matter degradation. Oxidative stress caused by O2·− promoted the accumulation of H2O2 in cells and accelerates cell death. This study provides new ideas and support for the application of harmful algal bloom control and the mechanism of algal cell inactivation in real water bodies.