Toxins from harmful algal blooms: How copper and iron render chalkophore a predictor of microcystin production.

Research on harmful algal blooms has focused on macronutrients, yet recent research increasingly indicates that understanding micronutrient roles is also important in the development of effective environmental management interventions. Here, we report results on metallophore production from mesocosms amended with copper and iron (enzymatic co-factors in photosynthetic electron transport) to probe questions of how cyanobacteria navigate the divide between copper nutrition, copper toxicity, and issues with iron bioavailability. These experiments utilized Microcystis, Chlorella and Desmodesmus spp., in mono- and mixed-cultures in lake water from a large, hypereutrophic lake (Taihu, China). To initiate experiments, copper and iron amendments were added to mesocosms containing algae that had been acclimated to achieve a state of copper and iron limitation. Mesocosms were analyzed over time for a range of analytes including algal growth parameters, algal assemblage progression, copper/iron concentrations and biomolecule production of chalkophore, siderophore and total microcystins. Community Trajectory Analysis and other multivariate methods were used for analysis resulting in our findings: 1) Microcystis spp. manage copper/iron requirements though a dynamically phased behavior of chalkophore/siderophore production according to their copper and iron limitation status (chalkophore correlates with Cu concentration, R2 = 0.99, and siderophore correlates with the sum of Cu and Fe concentrations, R2 = 0.98). 2) A strong correlation was observed between the production of chalkophore and the cyanobacterial toxin microcystin (R2 = 0.76)-Chalkophore is a predictor of microcystin production. 3) Based on our results and literature, we posit that Microcystis spp. produces microcystin in response to copper/iron availability to manage photosystem productivity and effect an energy-saving status. Results from this work underscore the importance of micronutrients in influencing harmful algal bloom progression and represents a major advance in understanding the ecological function for the cyanobacterial toxin microcystin as a hallmark of micronutrient limitation stress.

Intravenous Polymyxin B as Adjunctive Therapy to High-Dose Tigecycline for the Treatment of Nosocomial Pneumonia Due to Carbapenem-Resistant Acinetobacter baumannii and Klebsiella pneumoniae: A Propensity Score-Matched Cohort Study.

Although the combination of polymyxin and tigecycline is widely used in treating carbapenem-resistant bacterial infections, the benefit of this combination is still uncertain. To assess whether adding polymyxin B to the high-dose tigecycline regimen would result in better clinical outcomes than the high-dose tigecycline therapy in patients with pneumonia caused by carbapenem-resistant Klebsiella pneumoniae and Acinetobacter baumannii, we conducted a propensity score-matched cohort study in a single center between July 2019 and December 2021. Of the 162 eligible patients, 102 were included in the 1:1 matched cohort. The overall 14-day mortality in the matched cohort was 24.5%. Compared with high-dose tigecycline, the combination therapy was not associated with better clinical outcomes, and showed similar 14-day mortality (OR, 0.72, 95% CI 0.27-1.83, p = 0.486), clinical cure (OR, 1.09, 95% CI 0.48-2.54, p = 0.823), microbiological cure (OR, 0.96, 95% CI 0.39-2.53, p = 0.928) and rate of nephrotoxicity (OR 0.85, 95% CI 0.36-1.99, p = 0.712). Subgroup analyses also did not demonstrate any statistical differences. Based on these results, it is reasonable to recommend against adding polymyxin B to the high-dose tigecycline regimen in treating pneumonia caused by carbapenem-resistant K. pneumoniae and A. baumannii.

We need to plan streamlined environmental impact assessment for the future X-Press Pearl disasters.

The X-Press Pearl disaster illustrates the urgent needs for streamlined environmental impact assessment to inform decision making. The environmental contamination caused by the disaster is complex, and the biological impact of different environmental stressors, and at different biological scales, needs to be determined. Traditional methods for analyzing complex environmental stressors are often inefficient and do not reflect the biological impact of pollution. The combination of chemical stressors and biological impacts is the key to environmental impact assessment based on integrated monitoring. Whole-cell bioreporters are tools for rapid, efficient and quantitative detection of the bioavailability, stressor effects, and toxicity of pollutants, i.e., spanning a wide range of applications. Here we propose the view that using whole-cell bioreporter technology to streamline short-term environmental impact assessment for maritime disasters such as the X-Press Pearl is more fit-for-purpose/practical than other approaches in use.

The Mycobacterium tuberculosis prolyl dipeptidyl peptidase cleaves the N-terminal peptide of the immunoprotein CXCL-10.

Dipeptidyl peptidases constitute a class of non-classical serine proteases that regulate an array of biological functions, making them pharmacologically attractive enzymes. With this work, we identified and characterized a dipeptidyl peptidase from Mycobacterium tuberculosis (MtDPP) displaying a strong preference for proline residues at the P1 substrate position and an unexpectedly high thermal stability. MtDPP was also characterized with alanine replacements of residues of its active site that yielded, for the most part, loss of catalysis. We show that MtDPP catalytic activity is inhibited by well-known human DPP4 inhibitors. Using MALDI-TOF mass spectrometry we also describe that in vitro, MtDPP mediates the truncation of the C-X-C motif chemokine ligand 10, indicating a plausible role in immune modulation for this mycobacterial enzyme.

Pragmatic Comparison of Piperacillin/Tazobactam versus Carbapenems in Treating Patients with Nosocomial Pneumonia Caused by Extended-Spectrum ß-Lactamase-Producing Klebsiella pneumoniae.

The effectiveness of piperacillin/tazobactam for managing nosocomial pneumonia caused by extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae is unknown. To answer this question, we conducted a retrospective cohort study in two tertiary teaching hospitals of patients admitted between January 2018 and July 2021 with a diagnosis of nosocomial pneumonia caused by ESBL-producing K. pneumoniae receiving either piperacillin/tazobactam or carbapenems within 24 h from the onset of pneumonia for at least 72 h. Clinical outcomes, including 28-day mortality and 14-day clinical and microbiological cure, were analyzed. Of the 136 total patients, 64 received piperacillin/tazobactam and 72 received carbapenems. The overall 28-day mortality was 19.1% (26/136). In the inverse probability of treatment weighted cohort, piperacillin/tazobactam therapy was not associated with worse clinical outcomes, as the 28-day mortality (OR, 0.82, 95% CI, 0.23-2.87, p = 0.748), clinical cure (OR, 0.94, 95% CI, 0.38-2.35, p = 0.894), and microbiological cure (OR, 1.10, 95% CI, 0.53-2.30, p = 0.798) were comparable to those of carbapenems. Subgroup analyses also did not demonstrate any statistical differences. In conclusion, piperacillin/tazobactam could be an effective alternative to carbapenems for treating nosocomial pneumonia due to ESBL-producing K. pneumoniae when the MICs are ≤8 mg/L.

From speciation to toxicity: Using a "Two-in-One" whole-cell bioreporter approach to assess harmful effects of Cd and Pb.

Due to the sheer number of contaminated sites, bioavailability-based measurement and modeling of toxicity is used to triage response; despite advances, both remain relatively cumbersome. Cadmium (Cd) and lead (Pb) are two of the most toxic and globally prevalent pollutants, disproportionately impacting disadvantaged communities. Here we demonstrate the use of high throughput lights-on bioreporter technology to measure both speciation and toxicity. The organism's response is fit-for-purpose to parameterize the Biotic Ligand Model used in risk assessment of aquatic ecotoxicity and setting environmental Water Quality Criteria. Toxicity endpoints for analogous Cd and Pb models reported in literature average 71st and 44th rank-percentile sensitivity of Genus Mean Acute Values for acute toxicity (i.e., insensitive) in comparison to the bioreporter, the unique dual-mode measurement ability of which can predict toxicity endpoints from below the 5th percentile up to the 50th rank-percentile. These results are extensible to other reporters, paving the way to cost-efficient environmental risk assessment of aquatic ecotoxicity for a wide range of priority toxic pollutants.

Whole-cell biosensors for determination of bioavailable pollutants in soils and sediments: Theory and practice.

The bioavailability of pollutants is a key factor affecting environmental risk. Whole-cell bioreporters are a demonstratedly effective tool for the investigation of pollutant bioavailability in water and soil/sediment. Unlike aqueous samples, transmittance of bioreporter optical signal is reduced in direct-contact assays with soil/sediment, which affects the accuracy of bioreporter-detected pollutant bioavailability. No studies have measured the magnitude and variability of soil/sediment effects on signal in direct-contact assays or how associated uncertainties influence results. In this study, we investigate the optical effects of soil/sediment particles in suspensions on bioreporter signal transmittance and quantify how variable these optical effects are from sample-to-sample. We find that neglecting bioreporter signal diminution by soil/sediment, as many studies do, can lead to order-of-magnitude errors in results, underestimating risk. Correction based on methods in ad hoc use (e.g. comparison to signal from non-inducible reporter or use of reference soil/sediment) are also problematic for some types of experiment, and could lead to errors in excess of 30%. Our findings have a sound basis in theory, and we provide recommendations concerning the most suitable type of approach to use for different experimental settings. Generally, if best accuracy is not needed to quantify bioavailability, for samples that have been ground, sieved, and are of reasonably uniform color, it may be possible to use a single or average correction factor, particularly for experiments performed at a single slurry concentration. For investigations studying bioavailability under varying solid-phase:water ratios (e.g., sorption/desorption), detailed compensation measurements are needed for independent variables, including each specific soil/sediment sample, slurry concentration, and in some cases bioreporter signal intensity. Our measurements and calculations indicate that best results are obtained when working in the region of ballistic photon transmittance. Findings herein will be useful in areas that require information on bioavailability, such as ecotoxicology and environmental risk assessment.

Prevalence and factors associated with post-traumatic stress disorder in healthcare workers exposed to COVID-19 in Wuhan, China: a cross-sectional survey.

BACKGROUND: The COVID-19 pandemic has posed significant threats to both the physical and psychological health of healthcare workers working in the front-line combating COVID-19. However, studies regarding the medium to long term impact of COVID-19 on mental health among healthcare workers are limited. Therefore, we conducted this cross-sectional survey to investigate the prevalence, factors and impact of post-traumatic stress disorder (PTSD) in healthcare workers exposed to COVID-19 8 months after the end of the outbreak in Wuhan, China. METHODS: A web-based questionnaire was delivered as a link via the communication application WeChat to those healthcare workers who worked at several COVID-19 units during the outbreak (from December 2019 to April 2020) in Wuhan, China. The questionnaire included questions on social-demographic data, the post-traumatic stress disorder checklist-5 (PCL-5), the family care index questionnaire (Adaptation, Partnership, Growth, Affection and Resolve, APGAR), and the quality-of-life scale (QOL). The prevalence, risk and protective factors, and impact of PTSD on healthcare workers were subsequently analyzed. RESULTS: Among the 659 participants, 90 healthcare workers were still suffering from PTSD 8 months after the end of the outbreak of COVID-19 in Wuhan, in which avoidance and negative impact were the most affected dimensions. Suffering from chronic disease, experiencing social isolation, and job dissatisfaction came up as independent risk factors for PTSD, while obtaining COVID-19 related information at an appropriate frequency, good family function, and working in well-prepared mobile cabin hospitals served as protective factors. The impact of PTSD on COVID-19 exposed healthcare workers was apparent by shortened sleeping time, feeling of loneliness, poorer quality of life and intention to resign. CONCLUSIONS: Eight months after the end of the COVID-19 outbreak in Wuhan, the level of PTSD in healthcare workers exposed to COVID-19 was still high. Apart from the commonly recognized risk factors, comorbid chronic disease was identified as a new independent risk factor for developing PTSD. For countries where the pandemic is still ongoing or in case of future outbreaks of new communicable diseases, this study may contribute to preventing cases of PTSD in healthcare workers exposed to infectious diseases under such circumstances.

Tigecycline in the Treatment of Ventilator-Associated Pneumonia Due to Stenotrophomonas maltophilia: A Multicenter Retrospective Cohort Study.

INTRODUCTION: Tigecycline is a potential alternative to trimethoprim-sulfamethoxazole in treating Stenotrophomonas maltophilia infections due to its potent in vitro antimicrobial activity. Clinical evidence regarding the use of tigecycline in the treatment of S. maltophilia infections is scarce. In this study, we assessed the efficacy of tigecycline treating ventilator-associated pneumonia (VAP) due to S. maltophilia in comparison with fluoroquinolones. METHODS: This is a multicenter retrospective cohort study of patients admitted between January 2017 and December 2020 with the diagnosis of VAP caused by S. maltophilia receiving either tigecycline or fluoroquinolones as the definitive therapy ≥ 48 h. Clinical outcomes including 28-day mortality, clinical cure and microbiological cure were analyzed. RESULTS: Of 82 patients with S. maltophilia VAP included, 46 received tigecycline, and 36 received fluoroquinolones; 70.7% of patients had polymicrobial pneumonia, and the appropriate empiric therapy was applied to only 14.6% of patients. The overall 28-day mortality was 39%. Compared with patients receiving fluoroquinolones, tigecycline therapy resulted in worse clinical cure (32.6% vs. 63.9%, p = 0.009) and microbiological cure (28.6% vs. 59.1%, p = 0.045), while there was no statistical difference between 28-day mortality (47.8% vs. 27.8%, p = 0.105) in the two groups. Similar results were also shown in the inverse probability of treatment weighted univariable regression model and multivariable regression model. CONCLUSIONS: The standard dose of tigecycline therapy was associated with a lower clinical and microbiological cure rate but not associated with an increased 28-day mortality in patients with S. maltophilia VAP compared with fluoroquinolones. Considering the unfavorable clinical outcomes, we therefore recommend against using the standard dose of tigecycline in treating S. maltophilia VAP unless new clinical evidence emerges.

A new perspective of copper-iron effects on bloom-forming algae in a highly impacted environment.

Relatively little work has been done on the role of micronutrients in influencing development and progression of harmful algal blooms, yet micronutrients are ineluctably required for growth. Relatively small changes in micronutrient status have wide-ranging consequences. Here, we report results from mesocosm experiments with Microcystis and Desmodesmus spp., in mono- and mixed-cultures, to probe questions of how copper, iron, and copper-iron amendments affect growth, short-term assemblage progression, and production of siderophore, chalkophore, and microcystin in lake water from a large, hypereutrophic lake (Taihu, China). Our approach offers an entirely new perspective to understanding micronutrient dynamics in aqueous environments, as this is the first work to systematically screen for chalkophores and siderophores separately, as a function of copper/iron amendment, and using community trajectory analysis. Singular findings are summarized as follows: 1) unlike lab-based studies, in our work we observe neither dramatic copper-modulation of iron demand, nor evidence of an iron-protective effect from copper toxicity. 2) The interplay between chalkophore/siderophore production supports a concept model wherein Microcystis spp. varies behavior to individually and uniquely manage copper/iron requirements in a phased manner. In being able to specifically screen for chalkophores, we observe a previously unreported link between chalkophore and microcystin production that may relate to iron-limitation. 3) Regarding harmful algal bloom (HAB) persistance, the lake water itself influences mesocosm changes; differentiated effects for iron regarding growth indicators and/or reduction of Fe-limitation stress were found at an HAB-free field station (Xukou Bay), likely a consequence of low bioavailability of iron in this station as compared to HAB-impacted stations (half the initial dissolved iron concentration, persisting throughout experiments). The low dissolved iron accompanies more intense chalkophore/siderophore community trajectories.

ZIKV viral proteins and their roles in virus-host interactions.

The re-emergence of Zika virus (ZIKV) and its associated neonatal microcephaly and Guillain-Barré syndrome have led the World Health Organization to declare a global health emergency. Until today, many related studies have successively reported the role of various viral proteins of ZIKV in the process of ZIKV infection and pathogenicity. These studies have provided significant insights for the treatment and prevention of ZIKV infection. Here we review the current research advances in the functional characterization of the interactions between each ZIKV viral protein and its host factors.

Differential O- and Glycosphingolipid Glycosylation in Human Pancreatic Adenocarcinoma Cells With Opposite Morphology and Metastatic Behavior.

Changes in the glycosylation profile of cancer cells have been strongly associated with cancer progression. To increase our insights into the role of glycosylation in human pancreatic ductal adenocarcinoma (PDAC), we performed a study on O-glycans and glycosphingolipid (GSL) glycans of the PDAC cell lines Pa-Tu-8988T (PaTu-T) and Pa-Tu-8988S (PaTu-S). These cell lines are derived from the same patient, but show an almost opposite phenotype, morphology and capacity to metastasize, and may thus provide an attractive model to study the role of glycosylation in progression of PDAC. Gene-array analysis revealed that 24% of the glycosylation-related genes showed a ≥ 1.5-fold difference in expression level between the two cell lines. Subsequent validation of the data by porous graphitized carbon nano-liquid chromatography coupled to a tandem ion trap mass spectrometry and flow cytometry established major differences in O-glycans and GSL-glycans between the cell lines, including lower levels of T and sialylated Tn (sTn) antigens, neoexpression of globosides (Gb3 and Gb4), and higher levels of gangliosides in the mesenchymal-like PaTu-T cells compared to the epithelial-like PaTu-S. In addition, PaTu-S cells demonstrated a significantly higher binding of the immune-lectins macrophage galactose-type lectin and galectin-4 compared to PaTu-T. In summary, our data provide a comprehensive and differential glycan profile of two PDAC cell lines with disparate phenotypes and metastatic behavior. This will allow approaches to modulate and monitor the glycosylation of these PDAC cell lines, which opens up avenues to study the biology and metastatic behavior of PDAC.

Advances in freshwater risk assessment: improved accuracy of dissolved organic matter-metal speciation prediction and rapid biological validation.

Speciation modeling of bioavailability has increasingly been used for environmental risk assessment (ERA). Heavy metal pollution is the most prevalent environmental pollution issue globally, and metal bioavailability is strongly affected by its chemical speciation. Dissolved organic matter (DOM) in freshwater will bind heavy metals thereby reducing bioavailability. While speciation modeling has been shown to be quite effective and is validated for use in ERA, there is an increasing body of literature reporting problems with the accuracy of metal-DOM binding in speciation models. In this study, we address this issue for a regional-scale field area (Lake Tai, with 2,400 km2 surface area and a watershed of 36,000 km2) where speciation models in common use are not highly accurate, and we tested alternative approaches to predict metal-DOM speciation/bioavailability for lead (Pb) in this first trial work. We tested five site-specific approaches to quantify Pb-DOM binding that involve varying assumptions about conditional stability constants, binding capacities, and different components in DOM, and we compare these to what we call a one-size-fits-all approach that is commonly in use. We compare model results to results for bioavailable Pb measured using a whole-cell bioreporter, which has been validated against speciation models and is extremely rapid compared to many biological methods. The results show that all of the site-specific approaches we use provide more accurate estimates of bioavailability than the default model tested, however, the variation of the conditional stability constant on a site-specific basis is the most important consideration. By quantitative metrics, up to an order of magnitude improvement in model accuracy results from modeling active DOM as a single organic ligand type with site-specific variations in Pb-DOM conditional stability constants. Because the biological method is rapid and parameters for site-specific tailoring of the model may be obtained via high-throughput analysis, the approach that we report here in this first regional-scale freshwater demonstration shows excellent potential for practical use in streamlined ERA.

Quantitative high-throughput approach to chalkophore screening in freshwaters.

There is an increasing need to study the effects of trace metal micronutrients on microorganisms in natural waters. For Fe, small Fe-binding ligands called siderophores, which are secreted from cells and bind Fe with high affinity, have been demonstrated to modulate bioavailability of this critical nutrient. Relatively little is known about secretion of strong Cu-binding ligands (chalkophores) that may help organisms navigate the divide between Cu nutrition and toxicity. A barrier to environmental chalkophore research is a lack of literature on chalkophore analysis. Here we report the development of a quantitative, high-throughput approach to chalkophore screening based on a popular competitive-ligand binding assay for siderophores wherein ligands compete for metal in a chromogenic ternary complex of chrome azurol sulfonate-metal-surfactant. We developed the assay for high-throughput analysis using a microplate reader. The method performance is slightly better than that of comparable screening approaches for siderophores. We find that levels of other metals in natural samples may be capable of causing matrix interferences (a neglected source of analytical uncertainty in siderophore screening) and that for our method this can be overcome by standard additions. In this respect the high-throughput nature of the technique is a distinct advantage. To demonstrate practical use, we tested samples from field mesocosm studies that were set up with and without Cu and Fe amendments; we find trends in results that are logical in the environmental context of our application. This approach will be useful in areas such as risk assessment for a rapid survey of metal speciation and bioavailability; investigators who perform structural studies might also benefit from this approach to rapidly screen and select samples with high Fe/Cu binding capacity for further study.

Corticosteroid treatment of patients with coronavirus disease 2019 (COVID-19).

OBJECTIVES: To assess the efficacy of corticosteroid treatment of patients with coronavirus disease 2019 (COVID-19). DESIGN, SETTING: Observational study in the two COVID-19-designated hospitals in Wuhu, Anhui province, China, 24 January - 24 February 2020. PARTICIPANTS: Thirty-one patients infected with the severe acute respiratory coronavirus 2 (SARS-CoV-2) treated at the two designated hospitals. MAIN OUTCOME MEASURES: Virus clearance time, length of hospital stay, and duration of symptoms, by treatment type (including or not including corticosteroid therapy). RESULTS: Eleven of 31 patients with COVID-19 received corticosteroid treatment. Cox proportional hazards regression analysis indicated no association between corticosteroid treatment and virus clearance time (hazard ratio [HR], 1.26; 95% CI, 0.58-2.74), hospital length of stay (HR, 0.77; 95% CI, 0.33-1.78), or duration of symptoms (HR, 0.86; 95% CI, 0.40-1.83). Univariate analysis indicated that virus clearance was slower in two patients with chronic hepatitis B infections (mean difference, 10.6 days; 95% CI, 6.2-15.1 days). CONCLUSIONS: Corticosteroids are widely used when treating patients with COVID-19, but we found no association between therapy and outcomes in patients without acute respiratory distress syndrome. An existing HBV infection may delay SARS-CoV-2 clearance, and this association should be further investigated.

Effectiveness and Safety of High Dose Tigecycline for the Treatment of Severe Infections: A Systematic Review and Meta-Analysis.

BACKGROUND: Studies assessing the effect of high dose tigecycline on severe infections are limited and remain controversial. OBJECTIVES: To assess systematically the effectiveness and safety of high dose tigecycline in the treatment of severe infections. METHODS: Pubmed, Web of Science, Embase, MEDLINE, Cochrane Library and ClinicalTrials were searched up to February 20, 2019 for studies that compared the effectiveness and safety of high dose tigecycline with standard dose tigecycline or other non-tigecycline-containing regimens in the treatment of severe infections. Rates for all-cause mortality, clinical cure, microbiological eradication and adverse events were analysed. RESULTS: Ten studies with 593 patients were included. The results indicated that using high dose tigecycline resulted in better outcomes compared with controls with lower all-cause mortality (OR 0.44, 95% CI 0.30-0.66, p < 0.0001), higher clinical cure (OR 3.43, 95% CI 2.09-5.63, p < 0.00001), higher microbiological eradication (OR 2.25, 95% CI 1.44-3.50, p = 0.0003), and without increasing adverse events rates. Subgroup analysis showed that high dose tigecycline reduced all-cause mortality in nosocomial acquired pneumonia (OR 0.39, 95% CI 0.22-0.70, p = 0.002), bloodstream infections (OR 0.19, 95% CI 0.06-0.58, p = 0.004) and mixed infections (OR 0.20, 95% CI 0.07-0.59, p = 0.003), with no statistical differences in complicated intra-abdominal infections (OR 2.04, 95% CI 0.80-5.23, p = 0.14). In carbapenem-resistant pathogens, the microbiological eradication rate in those given high dose tigecycline did not differ from controls (OR 1.07, 95% CI 0.44-2.60, p = 0.87), although mortality was reduced (OR 0.20, 95% CI 0.09-0.45, p = 0.0001). The main limitation of the review is that most of the included studies are observational studies with small sample sizes and high risks of bias. CONCLUSIONS: High dose tigecycline treatment is effective and safe for severe infections owing to its lower all-cause mortality, higher clinical cure, microbiological eradication and comparable adverse events. However, as a result of the high risks of bias of the included studies, well-designed randomised clinical trials are warranted to establish the effectiveness and safety of high dose tigecycline compared with standard dose tigecycline and other commonly used antibiotics.