Three-dimensional Au-MnO2 nanostructure as an agent of synergistic cancer therapy: chemo-/photodynamic and photothermal approaches.

The design of multimodal cancer therapy was focused on reaching an efficient process and minimizing harmful effects on patients. In the present study, the Au-MnO2 nanostructures have been successfully constructed and produced as novel multipurpose photosensitive agents simultaneously for photodynamic therapy (PDT), photothermal therapy (PTT), and chemodynamic therapy (CDT). The prepared AuNPs were conjugated with MnO2 NPs by its participation in the thermal decomposition process of KMnO4 confirmed by X-ray diffraction (XRD), transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy (FT-IR). The 16.5 nm Au-MnO2 nanostructure exhibited an absorbance at 438 nm, which is beneficial for application in light induction therapy due to the NIR band, as well as its properties of generating reactive oxygen species (ROS) associated with the 808 nm laser light for PDT. The photothermal transduction efficiency was calculated and compared with that of the non-irradiated nanostructure, in which it was found that the 808 nm laser induced a high efficiency of 83%, 41.5%, and 37.5% for PDT, PTT, and CDT, respectively. The results of DPBF and TMB assays showed that the efficiency of PDT and PTT was higher than that of CDT. The nanostructure also confirmed the time-dependent peroxidase properties at different H2O2, TMB, and H2TMB concentrations, promising good potency in applying nanomedicine in clinical cancer therapy.

Potential usage of boron modified carbon nanodots as a marker candidate for coronavirus disease (COVID-19) antibody detection.

The global outbreak of COVID-19 in December 2019 has highlighted rapid and accurate diagnostic tools for effective intervention. While the RT-PCR test offers 86 % sensitivity, uncertainties often require supplementary screening. This research investigates how carbon dots (CDs) can be utilized as markers for COVID-19 antibodies, taking advantage of their biocompatibility and low toxicity. CDs were synthesized using citric acid (CA) and APBA with boronic acid, enabling the detection of COVID-19 IgG antibodies with increased absorbance and fluorescence. Comprehensive analyses confirmed the successful synthesis of APBA-CDs, prompting further exploration of their impact on SARS-CoV-2 RNA. Increased absorbance levels were observed in categories K1, K2, and K3, attributed to the introduction of CDs into plasma, indicating effective binding of APBA-CDs to COVID-19 antibodies. In addition, the fluorescence tests consistently showed heightened levels across all categories, emphasizing the effective binding of APBA-CDs with COVID-19 antibodies, particularly in positive plasma samples. As a part of our analysis, we conducted a PCA test to validate the data, which revealed that APBA-CDs are specific to IgG+ antibodies. The results showed a sensitivity rate of 74 % and a specificity rate of 53 %, while, when tested for IgM antibodies, the sensitivity and specificity rates were 63 % and 27 %, respectively. These findings highlight the potential of APBA-CDs as a sensitive and specific marker for COVID-19 antibody detection, offering potential for diagnostic tool development.

Changes in Combined Lifestyle Risks and the Transition of Activities of Daily Living in the Elderly Population of Taiwan: Evidence from the Taiwan Longitudinal Study on Aging.

Functional ability decline occurs with age. This study aims to investigate the associations between the lifestyle factors-exercising, food consumption, and smoking-and the functional ability of the activities of daily living (ADL) by gender. The data were obtained from the Taiwan longitudinal study on aging, a national cohort study. The cross-sectional results demonstrated that the frequency of exercising was negatively associated with ADL in both men and women. Dairy products were positively associated with ADL in men. The longitudinal results illustrated that current and consistent exercising were negatively associated with changes in ADL scores over 4- and 8-year periods in both men and women. Seafood consumption was negatively associated with changes in the subsequent 4-year ADL scores. Past smoking was positively associated with changes in subsequent 4-year ADL scores in men, while current smoking was positively associated with changes in subsequent 8-year ADL scores in women. Therefore, consistent exercising, food consumption, and smoking were associated with ADL functional ability in elderly people, and the associations differed by gender. Elders exercising consistently had good ADL performance and maintained their ADL ability better, especially women. Seafood consumption decreased the risk of ADL decline, while smoking increased the risk of ADL decline.

Evaluation of asphalt film thickness and heavy metal leaching of oxidizing slag used as an aggregate material in dense-graded asphalt concrete.

Electric-arc-furnace (EAF) steelmaking uses scrap iron and steel as raw materials. Scrap iron and steel originate from complex sources and may contain heavy metal components which can leach into the environment over time due to wear-and-tear. A by-product of the EAF steelmaking process is oxidizing slag, and approximately 1.2 million metric tons is produced every year in Taiwan alone. This study investigated substitution of natural aggregates with oxidizing slag in dense-graded asphalt concrete. We evaluated the water resistance and asphalt film thickness of the oxidizing slag substituted asphalt concrete and further explored the performance of oxidizing slag as paving material. We determined the dissolved and total amounts of heavy metals in the oxidizing slag, comparing these results with current regulatory controls to assess the environmental compatibility of the oxidizing slag. We found that due to the complicated sources of oxidizing slag, the basic properties should be analyzed on a batch-to-batch basis. Furthermore, we recommend trial mixing before upscaling the production of oxidizing slag substituted dense-graded asphalt concrete to confirm the mixing time required to achieve uniformity. The results also show that in comparison to natural aggregates used in asphalt concrete, oxidizing slag exhibits superior performance in terms of increased asphalt film thickness and improved water resistance. Furthermore, oxidizing slag as an aggregate material was associated with decreased heavy metal leaching and reduced environmental pollution. The results of the toxicity characteristic leaching procedure (TCLP) met regulatory requirements. However, the microwave-assisted aqua-regia digestion procedure showed heavy metal concentrations exceeding the monitoring standards for food crops. Considering environmental compatibility, it is recommended that controlling the total amount of heavy metals in oxidizing slag should be included in regulatory requirements. Furthermore, we should prohibit the use of materials such as oxidizing slag and other steel furnace slag in the roadways adjacent to edible crop farmlands.

Mass Spectrometry-Based Analyses of Carbon Nanodots: Structural Elucidation.

Carbon nanodots (CNDs) are nanomaterials with ubiquitous applications in health for diagnosis and treatments. The key to enhancing the applications of carbon nanodots in various fields lies on how deep its structure is understood. Here, we review the mass spectroscopy (MS) techniques employed for carbon nanodot analysis. We aimed to revive the use of MS to support the structural elucidation of carbon nanodots. General techniques used in nanomaterials characterization include laser desorption/ionization (LDI), matrix-assisted LDI (MALDI), inductively coupled plasma (ICP), and electrospray ionization (ESI) MS. For CNDs characterization, LDI-MS, MALDI-MS, and ESI-MS were employed. The techniques required further instrumentations of time-of-flight (TOF), for MALDI, and TOF, quadrupole (Q), and tandem (MS/MS) for ESI. LDI-MS could be applied to prove the surface and core structural composition of carbon nanodots. Meanwhile, MALDI-MS was used to elucidate the surface structures of CNDs. Finally, ESI-MS could provide significant insight into the carbon nanodots' structural composition and bonding patterns. In summary, MS could be combined with other techniques to unambiguously elucidate the structure of carbon nanodots.

Chief complaints and computed tomography results in the emergency department: a three-year retrospective cohort study.

BACKGROUND: Computed tomography (CT) is frequently performed in the patients who admitted to the emergency department (ED), discharged but returned to ED within 72 h. It is unknown whether the main complaints of patients assist physicians to use CT effectively. This study aimed to find the association between chief complaints and the CT results. METHODS: This three-year retrospective cohort study was conducted in the ED of a tertiary medical center. Adult patients who returned to the ED after the index visit were included from 2019 to 2021. Demographics, pre-existing diseases, chief complaints, and CT region were recorded by independent ED physicians. A logistic regression model with an odds ratio (OR) and 95% confidence interval (CI) was used to determine the relationship between chief complaints and positive CT results. RESULTS: In total, 7,699 patients revisited ED after the index visit; 1,202 (15.6%) received CT. The top chief complaints in patients who received CT were abdominal pain, dizziness, and muscle weakness. Patients with abdominal pain or gastrointestinal symptoms had a significantly higher rate of positive abdominopelvic CT than those without it (OR 2.83, 95% CI 1.98-4.05, p < 0.001), while the central nervous system and cardiopulmonary chief complaints were not associated (or negatively associated) with new positive CT findings. CONCLUSION: Chief complaints of patients on revisit to the ED are associated with different yields of new findings when CT scans of the chest, abdomen and head are performed. Physicians should consider these differential likelihoods of new positive findings based on these data.

A D-π-A-type ratiometric fluorescent probe to detect polarity changes and inhibition effect during ferroptosis.

To thoroughly understand ferroptosis's biological functions in living cells, it is crucial to investigate the polarity variations that occur during this unique Fe(II)-facilitated oxidative type of cell death. In this work, we report the development of a ratiometric probe (Po-P) to visualize the polarity changes in living cells and the inhibition effect during ferroptosis. The polarity-responsive fluorophore utilized by Po-P has a D-π-A-type structure. Based on theoretical calculations, ICT was proposed as the basis for Po-P's polarity-responsive mechanism. According to cell imaging results, Po-P had a desirable capacity for monitoring polarity fluctuations and erastin-induced ferroptosis. Furthermore, inhibition imaging revealed that dihydrolipoic acid (DHLA) could potentially prevent polarity changes that occur during erastin-induced ferroptosis, just as vitamin E (VE). We anticipate that the probe Po-P could be a valuable tool to quickly monitor polarity fluctuations and inhibition effects during ferroptosis and create new medications for treating disorders related to ferroptosis.

Prediction of high-risk emergency department revisits from a machine-learning algorithm: a proof-of-concept study.

BACKGROUND: High-risk emergency department (ED) revisit is considered an important quality indicator that may reflect an increase in complications and medical burden. However, because of its multidimensional and highly complex nature, this factor has not been comprehensively investigated. This study aimed to predict high-risk ED revisit with a machine-learning (ML) approach. METHODS: This 3-year retrospective cohort study assessed adult patients between January 2019 and December 2021 from National Taiwan University Hospital Hsin-Chu Branch with high-risk ED revisit, defined as hospital or intensive care unit admission after ED return within 72 hours. A total of 150 features were preliminarily screened, and 79 were used in the prediction model. Deep learning, random forest, extreme gradient boosting (XGBoost) and stacked ensemble algorithm were used. The stacked ensemble model combined multiple ML models and performed model stacking as a meta-level algorithm. Confusion matrix, accuracy, sensitivity, specificity and area under the receiver operating characteristic curve (AUROC) were used to evaluate performance. RESULTS: Analysis was performed for 6282 eligible adult patients: 5025 (80.0%) in the training set and 1257 (20.0%) in the testing set. High-risk ED revisit occurred for 971 (19.3%) of training set patients vs 252 (20.1%) in the testing set. Leading predictors of high-risk ED revisit were age, systolic blood pressure and heart rate. The stacked ensemble model showed more favourable prediction performance (AUROC 0.82) than the other models: deep learning (0.69), random forest (0.78) and XGBoost (0.79). Also, the stacked ensemble model achieved favourable accuracy and specificity. CONCLUSION: The stacked ensemble algorithm exhibited better prediction performance in which the predictions were generated from different ML algorithms to optimally maximise the final set of results. Patients with older age and abnormal systolic blood pressure and heart rate at the index ED visit were vulnerable to high-risk ED revisit. Further studies should be conducted to externally validate the model.

Engineering H2O2 Self-Supplying Platform for Xdynamic Therapies via Ru-Cu Peroxide Nanocarrier: Tumor Microenvironment-Mediated Synergistic Therapy.

Of the most common, hypoxia, overexpressed glutathione (GSH), and insufficient H2O2 concentration in the tumor microenvironment (TME) are the main barriers to the advancment of reactive oxygen species (ROS) mediated Xdynamic therapies (X = photo, chemodynamic, chemo). Maximizing Fenton catalytic efficiency is crucial in chemodynamic therapy (CDT), yet endogenous H2O2 levels are not sufficient to attain better anticancer efficacy. Specifically, there is a need to amplify Fenton reactivity within tumors, leveraging the unique attributes of the TME. Herein, for the first time, we design RuxCu1-xO2-Ce6/CPT (RCpCCPT) anticancer nanoagent for TME-mediated synergistic therapy based on heterogeneous Ru-Cu peroxide nanodots (RuxCu1-xO2 NDs) and chlorine e6 (Ce6), loaded with ROS-responsive thioketal (TK) linked-camptothecin (CPT). The Ru-Cu peroxide NDs (RCp NDs, x = 0.50) possess the highest oxygen vacancy (OV) density, which grants them the potential to form massive Lewis's acid sites for peroxide adsorption, while the dispersibility and targetability of the NDs were improved via surface modification using hyaluronic acid (HA). In TME, RCpCCPT degrades, releasing H2O2, Ru2+/3+, and Cu+/2+ ions, which cooperatively facilitate hydroxyl radical (•OH) formation and deactivate antioxidant GSH enzymes through a cocatalytic loop, resulting in excellent tumor therapeutic efficacy. Furthermore, when combined with laser treatment, RCpCCPT produces singlet oxygen (1O2) for PDT, which induces cell apoptosis at tumor sites. Following ROS generation, the TK linkage is disrupted, releasing up to 92% of the CPT within 48 h. In vitro investigations showed that laser-treated RCpCCPT caused 81.5% cell death from PDT/CDT and chemotherapy (CT). RCpCCPT in cancer cells produces red-blue emission in images of cells taking them in, which allows for fluorescence image-guided Xdynamic treatment. The overall results show that RCp NDs and RCpCCPT are more biocompatible and have excellent Xdynamic therapeutic effectiveness in vitro and in vivo.

Gas-Solid Phase Reaction Derived Silver Bismuth Iodide Rudorffite: Structural Insight and Exploring Photocatalytic Potential of CO2 Reduction.

Photocatalytic reduction of CO2 is a promising strategy to mitigate the effects of global warming by converting CO2 into valuable energy-dense products. Silver bismuth iodide (SBI) is an attractive material owing to its tunable bandgap and favorable band-edge positions for efficient CO2 photoreduction. In this study, SBI materials, including AgBi2I7, AgBiI4, Ag2BiI5, and Ag3BiI6 are first synthesized, through gas-solid reaction by controlling the stoichiometric ratio of reactants. The X-ray absorption near edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) results revealed that the distance between Ag-I is proportional to the degree of Ag ions delocalization, which occupies the vacant sites. That greatly retards the charge recombination at vacant sites. In addition, the surface potential via photo-assisted Kelvin probe force measurements of various SBI catalysts shows that Ag3BiI6 exhibits the highest surface potential change due to the rich delocalized Ag ions. This results in effective charge carrier transport and prevention of charge recombination at vacant sites. Taking the above advantages, the averaged CO and CH4 production rates for Ag3BiI6 achieved 0.23 and 0.10 µmol g-1 h-1, respectively. The findings suggest that Ag3BiI6 has a high potential as a novel photocatalyst for CO2 reduction and sheds light on the possibility of solving environmental contamination and sustainable energy crises.

Golgi apparatus-targeting fluorescent probe for the imaging of superoxide anion (O2•-) in living cells during ferroptosis.

Ferroptosis is an emerging iron-dependent oxidative cell death type, and recently has been demonstrated to show close relation with Golgi apparatus (GA). Exploring the fluctuation of superoxide anion (O2•-) level in GA during ferroptosis is of great significance to profoundly study the biological functions of GA in ferroptosis. Here, we present a GA-targeting probe (N-GA) to monitor cellular O2•- during ferroptosis. N-GA employed a triflate group and a tetradecanoic amide unit as the recognition site for O2•- and GA-targeting unit, respectively. After the response of N-GA to O2•-, the triflate unit of N-GA converted into hydroxyl group with strong electron-donating ability, generating bright green fluorescence under UV light. N-GA exhibited excellent sensitivity and selectivity towards O2•-. Fluorescence imaging results showed that N-GA could be applied as a GA-targeting probe to monitor cellular O2•-. The stimulation of cells with PMA and rotenone could result in the massive generation of endogenous O2•- in GA. Erastin-induced ferroptosis can markedly induce the increase of O2•- level in GA. Similar to Fer-1 and DFO, dihydrolipoic acid (DHLA) and rutin were demonstrated to inhibit the enormous production of O2•- in GA of the living cells during ferroptosis.

Redox Molecular Junction Metal-Covalent Organic Frameworks for Light-assisted CO2 Energy Storage.

Visible-light sensitive and bi-functionally favored CO2 reduction (CRR)/evolution (CER) photocathode catalysts that can get rid of the utilization of ultraviolet light and improve sluggish kinetics is demanded to conquer the current technique-barrier of traditional Li-CO2 battery. Here, a kind of redox molecular junction sp2c metal-covalent organic framework (i.e. Cu3-BTDE-COF) has been prepared through the connection between Cu3 and BTDE and can serve as efficient photocathode catalyst in light-assisted Li-CO2 battery. Cu3-BTDE-COF with redox-ability, visible-light-adsorption region, electron-hole separation ability and endows the photocathode with excellent round-trip efficiency (95.2 %) and an ultralow voltage hysteresis (0.18 V), outperforming the Schiff base COFs (i.e. Cu3-BTDA-COF and Cu3-DT-COF) and majority of the reported photocathode catalysts. Combined theoretical calculations with characterizations, Cu3-BTDE-COF with the integration of Cu3 centers, thiazole and cyano groups possess strong CO2 adsorption/activation and Li+ interaction/diffusion ability to boost the CRR/CER kinetics and related battery property.

Development of a turn-on fluorescent probe for the imaging of intracellular hypochlorous acid (HClO) during ferroptosis.

Ferroptosis is a burgeoning iron-dependent cell death form, and has close relation with hypochlorous acid (HClO). Exploring the fluctuation of the HClO level in living cells during ferroptosis could contribute to the profound study of the biological functions of HClO during ferroptosis. Here, we present a turn-on probe (RH-C) for the imaging of intracellular HClO during ferroptosis. The probe RH-C utilized the N,N-dimethylthiocarbamate group as a selective recognition site for HClO, and displayed desirable sensitivity and selectivity to HClO. The probe RH-C could detect the exogenous and endogenous HClO in living cells. Furthermore, RH-C was competent in monitoring the changes of endogenous HClO level during the process of ferroptosis. Biological imaging results suggested that erastin-induced ferroptosis can result in the excessive production of the endogenous HClO, and ferrostatin-1 (Fer-1) and vitamin E (VE) could block the massive accumulation of HClO in living cells.

Strategic Assessment of Boron-Enriched Carbon Dots/Naproxen: Diagnostic, Toxicity, and In Vivo Therapeutic Evaluation.

Cancer is a significant global public health concern, ranking as the leading cause of mortality worldwide. This study thoroughly explores boron-doped carbon dots (B-CDs) through a simple/rapid microwave-assisted approach and their versatile applications in cancer therapy. The result was highly uniform particles with an average diameter of approximately 4 nm. B-CDs exhibited notable properties, including strong fluorescence with a quantum yield of 33%. Colloid stability tests revealed their robustness within a pH range of 6-12, NaCl concentrations up to 0.5 M, and temperatures ranging from 30 to 60 °C. The study also delved into the kinetics of naproxen release from B-CDs as a drug delivery system. The loading efficacy of naproxen exceeded 55.56%. Under varying pH conditions, the release of naproxen from B-CDs conformed to the Peppas-Sahlin model, demonstrating the potential of Naproxen-loaded CDs for cancer drug delivery. In vitro cytotoxicity assessments, conducted using the CCK-8 Assay and flow cytometry, consistently indicated low toxicity with average cell viability exceeding 80%. An in vivo toxicity test on female mice administered 20 mg/kg of B-CDs for 31 days revealed reversible histological changes in the liver and kidneys, while the pancreas remained unaffected. Importantly, B-CDs did not impact the mice's physical behavior, body weight, or survival. In vivo experiments targeting benzo(a)pyrene-induced fibrosarcoma demonstrated the efficacy of B-CDs as naproxen carriers in the treatment of cancer. This in vivo study provides a thorough comprehension of B-CDs synthesis and toxicity and their potential applications in cancer therapy and drug delivery systems.

Novel molecular classification and prognosis of papillary renal cell carcinoma based on a large-scale CRISPR-Cas9 screening and machine learning.

Papillary renal cell carcinoma (PRCC) is a highly heterogeneous cancer, and PRCC patients with advanced/metastatic subgroup showed obviously shorter survival compared to other kinds of renal cell carcinomas. However, the molecular mechanism and prognostic predictors of PRCC remain unclear and are worth deep studying. The aim of this study is to identify novel molecular classification and construct a reliable prognostic model for PRCC. The expression data were retrieved from TCGA, GEO, GTEx and TARGET databases. CRISPR data was obtained from Depmap database. The key genes were selected by the intersection of CRISPR-Cas9 screening genes, differentially expressed genes, and genes with prognostic capacity in PRCC. The molecular classification was identified based on the key genes. Drug sensitivity, tumor microenvironment, somatic mutation, and survival were compared among the novel classification. A prognostic model utilizing multiple machine learning algorithms based on the key genes was developed and tested by independent external validation set. Our study identified three clusters (C1, C2 and C3) in PRCC based on 41 key genes. C2 had obviously higher expression of the key genes and lower survival than C1 and C3. Significant differences in drug sensitivity, tumor microenvironment, and mutation landscape have been observed among the three clusters. By utilizing 21 combinations of 9 machine learning algorithms, 9 out of 41 genes were chosen to construct a robust prognostic signature, which exhibited good prognostic ability. SERPINH1 was identified as a critical gene for its strong prognostic ability in PRCC by univariate and multiple Cox regression analyses. Quantitative real-time PCR and Western blot demonstrated that SERPINH1 mRNA and protein were highly expressed in PRCC cells compared with normal human renal cells. This study exhibited a new molecular classification and prognostic signature for PRCC, which may provide a potential biomarker and therapy target for PRCC patients.

Immunocompetent mouse models revealed that S100A4+ monocytes/macrophages facilitate long-term Zika virus infection in the testes.

During its global epidemic, Zika virus (ZIKV) attracted widespread attention due to its link with various severe neurological symptoms and potential harm to male fertility. However, the understanding of how ZIKV invades and persists in the male reproductive system is limited due to the lack of immunocompetent small animal models. In this study, immunocompetent murine models were generated by using anti-IFNAR antibody blocked C57BL/6 male mice and human STAT2 (hSTAT2) knock in (KI) male mice. After infection, viral RNA could persist in the testes even after the disappearance of viremia. We also found a population of ZIKV-susceptible S100A4+ monocytes/macrophages that were recruited into testes from peripheral blood and played a crucial role for ZIKV infection in the testis. By using single-cell RNA sequencing, we also proved that S100A4+ monocytes/macrophages had a great impact on the microenvironment of ZIKV-infected testes, thus promoting ZIKV-induced testicular lesions. In conclusion, this study proposed a novel mechanism of long-term ZIKV infection in the male reproductive system.

Tocilizumab Exerts Anti-Tumor Effects on Colorectal Carcinoma Cell Xenografts Corresponding to Expression Levels of Interleukin-6 Receptor.

The use of tocilizumab against the interleukin-6 receptor (IL-6R) has been demonstrated as inhibiting the progression of diverse cancers in vitro and in vivo. Nonetheless, evidence regarding the anti-tumor effects of tocilizumab on human colorectal carcinoma (CRC) corresponding to IL-6R expression levels remains scarce. To investigate the influence of IL-6R expression, SW480 and HT-29 cells inoculated subcutaneously into NU/NU mice were used as human CRC xenograft models with anti-IL-6R antibody (tocilizumab) therapy. The IL-6R expression levels, histology of CRC growth/invasiveness, and tumor growth-related signaling pathway were estimated by H&E and immunohistochemical staining. SW480 tumor cells with higher IL-6R expression levels showed better responsiveness in tocilizumab therapy than in the treated HT-29 group. Likewise, therapeutic effects of tocilizumab on the proliferative ability with mitotic index and Ki-67 expressions, invasiveness with MMP-9 proteinase expressions, and ERK 1/2 and STAT3 signaling transduction in the SW480 treatment group were superior to the HT-29 treatment group. In light of our results, IL-6R is the key indicator for the efficacy of tocilizumab treatment in CRC xenografts. From the perspective of precision medicine, tumor response to anti-IL-6R antibody therapy could be predicted on the basis of IL-6R expression levels. In this manner, tocilizumab may serve as a targeted and promising anti-CRC therapy.

Insights into biofilm development on polyhydroxyalkanoate biofilm carrier for anoxic azo dye decolourization of acid orange 7.

Polyhydroxyalkanoates (PHAs) are promising alternatives to non-degradable polymers in various applications. This study explored the use of biologically recovered PHA as a biofilm carrier in a moving bed biofilm reactor for acid orange 7 treatment. The PHA was comprised of 86 ± 1 mol% of 3-hydroxybutyrate and 14 ± 1 mol% of 3-hydroxyhexanoate and was melt-fused at 140 °C into pellets. The net positive surface charge of the PHA biocarrier facilitated attachment of negatively charged activated sludge, promoting biofilm formation. A 236-µm mature biofilm developed after 26 days. The high polysaccharides-to-protein ratio (>1) in the biofilm's extracellular polymeric substances indicated a stable biofilm structure. Four main microbial strains in the biofilm were identified as Leclercia adecarboxylata, Leuconostoc citreum, Bacillus cereus, and Rhodotorula mucilaginosa, all of which exhibited decolourization abilities. In conclusion, PHA holds promise as an effective biocarrier for biofilm development, offering a sustainable alternative in wastewater treatment applications.

Ectopic expression of a bacterial thiamin monophosphate kinase enhances vitamin B1 biosynthesis in plants.

Plants and bacteria have distinct pathways to synthesize the bioactive vitamin B1 thiamin diphosphate (TDP). In plants, thiamin monophosphate (TMP) synthesized in the TDP biosynthetic pathway is first converted to thiamin by a phosphatase, which is then pyrophosphorylated to TDP. In contrast, bacteria use a TMP kinase encoded by ThiL to phosphorylate TMP to TDP directly. The Arabidopsis THIAMIN REQUIRING2 (TH2)-encoded phosphatase is involved in TDP biosynthesis. The chlorotic th2 mutants have high TMP and low thiamin and TDP. Ectopic expression of Escherichia coli ThiL and ThiL-GFP rescued the th2-3 mutant, suggesting that the bacterial TMP kinase could directly convert TMP into TDP in Arabidopsis. These results provide direct evidence that the chlorotic phenotype of th2-3 is caused by TDP rather than thiamin deficiency. Transgenic Arabidopsis harboring engineered ThiL-GFP targeting to the cytosol, chloroplast, mitochondrion, or nucleus accumulated higher TDP than the wild type (WT). Ectopic expression of E. coli ThiL driven by the UBIQUITIN (UBI) promoter or an endosperm-specific GLUTELIN1 (GT1) promoter also enhanced TDP biosynthesis in rice. The pUBI:ThiL transgenic rice accumulated more TDP and total vitamin B1 in the leaves, and the pGT1:ThiL transgenic lines had higher TDP and total vitamin B1 in the seeds than the WT. Total vitamin B1 only increased by approximately 25-30% in the polished and unpolished seeds of the pGT1:ThiL transgenic rice compared to the WT. Nevertheless, these results suggest that genetic engineering of a bacterial vitamin B1 biosynthetic gene downstream of TMP can enhance vitamin B1 production in rice.