CXADR-Like Membrane Protein Regulates Colonic Epithelial Cell Proliferation and Prevents Tumor Growth.

BACKGROUND & AIMS: CXADR-like membrane protein (CLMP) is structurally related to coxsackie and adenovirus receptor. Pathogenic variants in CLMP gene have been associated with congenital short bowel syndrome, implying a role for CLMP in intestinal development. However, the contribution of CLMP to regulating gut development and homeostasis is unknown. METHODS: In this study, we investigated CLMP function in the colonic epithelium using complementary in vivo and in vitro approaches, including mice with inducible intestinal epithelial cell (IEC)-specific deletion of CLMP (ClmpΔIEC), intestinal organoids, IECs with overexpression, or loss of CLMP and RNA sequencing data from individuals with colorectal cancer. RESULTS: Loss of CLMP enhanced IEC proliferation and, conversely, CLMP overexpression reduced proliferation. Xenograft experiments revealed increased tumor growth in mice implanted with CLMP-deficient colonic tumor cells, and poor engraftment was observed with CLMP-overexpressing cells. ClmpΔIEC mice showed exacerbated tumor burden in an azoxymethane and dextran sulfate sodium-induced colonic tumorigenesis model, and CLMP expression was reduced in human colorectal cancer samples. Mechanistic studies revealed that CLMP-dependent regulation of IEC proliferation is linked to signaling through mTOR-Akt-ß-catenin pathways. CONCLUSIONS: These results reveal novel insights into CLMP function in the colonic epithelium, highlighting an important role in regulating IEC proliferation, suggesting tumor suppressive function in colon cancer.

Unraveling the molecular mechanisms of selenite reduction: transcriptomic analysis of Bacillus reveals the key role of sulfur assimilation.

Selenite biotransformation by microorganisms is an effective detoxification and assimilation process. However, current knowledge of the molecular mechanisms of selenite reduction remains circumscribed. Here, the reduction of Se(IV) by a highly selenite-resistant Bacillus sp. SL (up to 50 mM) was systematically analyzed, and the molecular mechanisms of selenite reduction were investigated. Remarkably, 10 mM selenite was entirely transformed by the strain SL within 20 h, demonstrating a faster conversion rate compared to other microorganisms. Furthermore, glutathione (GSH) and exopolysaccharides (EPS) changes were also monitored during the process. Transcriptomic analysis revealed that the genes of ferredoxin-sulfite oxidoreductase (6.82) and sulfate adenylyltransferase (6.32) were significantly upregulated, indicating that the sulfur assimilation pathway is the primary reducing pathway involved in selenite reduction by strain SL. Moreover, key genes associated with NAD(P)/FAD-dependent oxidoreductases and thioredoxin were significantly upregulated. The reduction of Se(IV) was mediated by multiple pathways in strain SL. To our knowledge, this is the initial report to identify the involvement of sulfur assimilation pathway in selenite reduction for bacillus, which is rare in aerobic bacteria.

Indole metabolism by phenol-stimulated activated sludges: Performance, microbial communities and network analysis.

Indole and phenol often coexist in the coking wastewater, while the effects of phenol on microbial communities of indole metabolism were less explored. In this study, the microbial interactions within activated sludge microbial communities stimulated by indole (group A) or by indole and phenol (group B) were systematically investigated in sequencing batch reactors (SBRs). The results showed that the removal of indole was increased by adding phenol. By using high-throughput sequencing technology, it was found that α-diversity was reduced in both groups. According to the relative abundance analysis, the indole-degrading genus Comamonas was the core genus in both groups (33.94% and 61.40%). But another indole-degrading genus Pseudomonas was only enriched in group A with 12.22% relative abundance. Meanwhile, common aromatic degrading genus Dyella and Thermomonas were enriched only in group B. It was found that the relative abundance of cytochrome P450 and styrene degradation enzymes were increased in group B by PICRUSt analysis. Based on the phylogenetic molecular ecological networks (pMENs), module hub OTU_1149 (Burkholderia) was only detected in group B, and the positive interactions between the key functional genus Burkholderia and other bacteria were increased. This study provides new insights into our understanding of indole metabolism communities stimulated by phenol, which would provide useful information for practical coking wastewater treatment.

Antibacterial properties and mechanism of selenium nanoparticles synthesized by Providencia sp. DCX.

Selenium nanoparticles (SeNPs) have attracted great interest as a potential antimicrobial agent. However, there is limited research on the antibacterial activity and possible mechanisms of biosynthesized SeNPs. In this study, spherical bio-SeNPs with an average size of 120 nm were synthesized by strain Providencia sp. DCX. The SeNPs were further applied to investigate the antibacterial properties of model bacteria, including Gram-positive (Staphylococcus aureus, Bacillus cereus and Bacillus subtilis) and Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli and Vibrio parahemolyticus). The biosynthesized SeNPs demonstrated strong inhibition activity against the growth of these pathogens. When treated with 500 mg/L SeNPs, most of the tested bacteria were destructed within 12 h, among which the mortality rates of Gram-negative bacteria were much better. The leakage tests illustrated that there existed more proteins and polysaccharides outside the cells after reacted with bio-SeNPs. It was indicated that the leakages of proteins and polysaccharides were caused by permeability changes of membranes and the disruption of cell walls. And the change of reactive oxygen species (ROS) intensity indicated that oxidative damage may play the significant role in the antibacterial processes. The results showed that several bacteria could be effectively inhibited and destructed, suggesting the potential of using the biosynthesized SeNPs as antibacterial agents for bacterial infectious diseases.

Treatment Outcome of Combined Continuous Venovenous Hemofiltration and Hemoperfusion in Acute Paraquat Poisoning: A Prospective Controlled Trial.

OBJECTIVES: To investigate whether combined continuous venovenous hemofiltration and hemoperfusion among paraquat-poisoned patients would improve survival. DESIGN: Prospective, controlled interventional study over 4 years. SETTING: Single, tertiary, academic medical center. PATIENTS: We recruited patients admitted to Shanghai Tenth People's Hospital within 48 hours after paraquat ingestion. Exclusions were under 14 years old, ingestion of paraquat with other toxicants, pregnant, a history of chronic pulmonary disease, psychosis, hyperthyroidism, or diabetes with impaired liver or renal function. INTERVENTIONS: All patients were assigned to receive continuous venovenous hemofiltration with hemoperfusion therapy (continuous venovenous hemofiltration group) and to receive conventional therapy (conventional group). The study outcomes were death from any cause within 90 days after paraquat ingestion and the frequencies of hypoxia, acute kidney injury, or adverse events. MEASUREMENTS AND MAIN RESULTS: Of the 110 enrolled patients, 59 were assigned to continuous venovenous hemofiltration group and 51 to conventional group. The two groups had similar baseline demographics and clinical features. At 90 days after paraquat ingestion, 19 of 59 patients (32.2%) in the continuous venovenous hemofiltration group and 29 of 51 patients (56.9%) in the conventional group had died (hazard ratio, 0.43; 95% CI, 0.24-0.76; p = 0.004). In multivariable Cox proportional hazard models controlling for baseline characteristics, combined continuous venovenous hemofiltration and hemoperfusion was independently associated with reduced risk of death compared with conventional therapy (adjusted hazard ratio, 0.35; 95% CI, 0.19-0.64; p = 0.001). Patients in the continuous venovenous hemofiltration group, as compared to the conventional group, had a reduced occurrence rate of hypoxia (40.7% vs 72.5%; p = 0.001) and of acute kidney injury (59.3% vs 78.4%; p = 0.03). Hypophosphatemia and thrombocytopenia were more common in the continuous venovenous hemofiltration group (p < 0.05). CONCLUSIONS: In patients with paraquat poisoning, treatment with combined continuous venovenous hemofiltration and hemoperfusion significantly improved 90-day survival rates.

Association of Biotin Ingestion With Performance of Hormone and Nonhormone Assays in Healthy Adults.

Importance: Biotinylated antibodies and analogues, with their strong binding to streptavidin, are used in many clinical laboratory tests. Excess biotin in blood due to supplemental biotin ingestion may affect biotin-streptavidin binding, leading to potential clinical misinterpretation. However, the degree of interference remains undefined in healthy adults. Objective: To assess performance of specific biotinylated immunoassays after 7 days of ingesting 10 mg/d of biotin, a dose common in over-the-counter supplements for healthy adults. Design, Setting, and Participants: Nonrandomized crossover trial involving 6 healthy adults who were treated at an academic medical center research laboratory. Exposure: Administration of 10 mg/d of biotin supplementation for 7 days. Main Outcomes and Measures: Analyte concentrations were compared with baseline (day 0) measures on the seventh day of biotin treatment and 7 days after treatment had stopped (day 14). The 11 analytes included 9 hormones (ie, thyroid-stimulating hormone, total thyroxine, total triiodothyronine, free thyroxine, free triiodothyronine, parathyroid hormone, prolactin, N-terminal pro-brain natriuretic peptide, 25-hydroxyvitamin D) and 2 nonhormones (prostate-specific antigen and ferritin). A total of 37 immunoassays for the 11 analytes were evaluated on 4 diagnostic systems, including 23 assays that incorporated biotin and streptavidin components and 14 assays that did not include biotin and streptavidin components and served as negative controls. Results: Among the 2 women and 4 men (mean age, 38 years [range, 31-45 years]) who took 10 mg/d of biotin for 7 days, biotin ingestion-associated interference was found in 9 of the 23 (39%) biotinylated assays compared with none of the 14 nonbiotinylated assays (P = .007). Results from 5 of 8 biotinylated (63%) competitive immunoassays tested falsely high and results from 4 out of 15 (27%) biotinylated sandwich immunoassays tested falsely low. Conclusions and Relevance: In this preliminary study of 6 healthy adult participants and 11 hormone and nonhormone analytes measured by 37 immunoassays, ingesting 10 mg/d of biotin for 1 week was associated with potentially clinically important assay interference in some but not all biotinylated assays studied. These findings should be considered for patients taking biotin supplements before ordering blood tests or when interpreting results. Trial Registration: clinicaltrials.gov Identifier: NCT03034707.

ZMYND10 is mutated in primary ciliary dyskinesia and interacts with LRRC6.

Defects of motile cilia cause primary ciliary dyskinesia (PCD), characterized by recurrent respiratory infections and male infertility. Using whole-exome resequencing and high-throughput mutation analysis, we identified recessive biallelic mutations in ZMYND10 in 14 families and mutations in the recently identified LRRC6 in 13 families. We show that ZMYND10 and LRRC6 interact and that certain ZMYND10 and LRRC6 mutations abrogate the interaction between the LRRC6 CS domain and the ZMYND10 C-terminal domain. Additionally, ZMYND10 and LRRC6 colocalize with the centriole markers SAS6 and PCM1. Mutations in ZMYND10 result in the absence of the axonemal protein components DNAH5 and DNALI1 from respiratory cilia. Animal models support the association between ZMYND10 and human PCD, given that zmynd10 knockdown in zebrafish caused ciliary paralysis leading to cystic kidneys and otolith defects and that knockdown in Xenopus interfered with ciliogenesis. Our findings suggest that a cytoplasmic protein complex containing ZMYND10 and LRRC6 is necessary for motile ciliary function.

Nephrocystin-5, a ciliary IQ domain protein, is mutated in Senior-Loken syndrome and interacts with RPGR and calmodulin.

Nephronophthisis (NPHP) is the most frequent genetic cause of chronic renal failure in children. Identification of four genes mutated in NPHP subtypes 1-4 (refs. 4-9) has linked the pathogenesis of NPHP to ciliary functions. Ten percent of affected individuals have retinitis pigmentosa, constituting the renal-retinal Senior-Loken syndrome (SLSN). Here we identify, by positional cloning, mutations in an evolutionarily conserved gene, IQCB1 (also called NPHP5), as the most frequent cause of SLSN. IQCB1 encodes an IQ-domain protein, nephrocystin-5. All individuals with IQCB1 mutations have retinitis pigmentosa. Hence, we examined the interaction of nephrocystin-5 with RPGR (retinitis pigmentosa GTPase regulator), which is expressed in photoreceptor cilia and associated with 10-20% of retinitis pigmentosa. We show that nephrocystin-5, RPGR and calmodulin can be coimmunoprecipitated from retinal extracts, and that these proteins localize to connecting cilia of photoreceptors and to primary cilia of renal epithelial cells. Our studies emphasize the central role of ciliary dysfunction in the pathogenesis of SLSN.

Establishment and application of a reverse dot blot assay for 13 mutations of hearing-loss genes in primary hospitals in China.

Background: Hearing loss is a common sensorineural dysfunction with a high incidence in China. Although genetic factors are important causes of hearing loss, hearing-related gene detection has not been widely adopted in China. Objective: Establishing a rapid and efficient method to simultaneously detect hotspot hearing loss gene mutations. Methods: A reverse dot blot assay combined with a flow-through hybridization technique was developed for the simultaneous detection of 13 hotspot mutations of 4 hearing loss-related genes including GJB2, GJB3, SLC26A4, and the mitochondrial gene MT-RNR1. This method involved PCR amplification systems and a hybridization platform. Results: The technique can detect 13 hotspot mutations of 4 hearing loss-related genes. And a total of 213 blood samples were used to evaluate the availability of this method. Discussion: Our reverse dot blot assay was a simple, rapid, accurate, and cost-effective method to identify hotspot mutations of 4 hearing loss-related genes in a Chinese population.

Critical role of thrombospondin-1 in promoting intestinal mucosal wound repair.

Thrombospondin-1 (TSP1) is a matricellular protein associated with the regulation of cell migration through direct binding interactions with integrin proteins and by associating with other receptors known to regulate integrin function, including CD47 and CD36. We previously demonstrated that deletion of an epithelial TSP1 receptor CD47 attenuates epithelial wound repair following intestinal mucosal injury. However, the mechanisms by which TSP1 contributes to intestinal mucosal repair remains poorly understood. Our results show upregulated TSP1 expression in colonic mucosal wounds and impaired intestinal mucosal wound healing in vivo upon intestinal epithelial specific loss of TSP1 (VillinCre/+Thbs1f/f or Thbs1ΔIEC). We report that exposure to exogenous TSP1 enhanced migration of IECs in a CD47- and TGFß1-dependent manner, and that deficiency of TSP1 in primary murine colonic epithelial cells resulted in impaired wound healing. Mechanistically, TSP1 modulated epithelial actin cytoskeletal dynamics by suppression of RhoA activity, activation of Rac1, and changes in F-actin bundling. Overall, TSP1 was found to regulate intestinal mucosal wound healing via CD47 and TGFß1, coordinate integrin-containing cell-matrix adhesion dynamics and remodel the actin cytoskeleton in migrating epithelial cells to enhance cell motility and promote wound repair.

Localization of retinitis pigmentosa 2 to cilia is regulated by Importin beta2.

Ciliopathies represent a newly emerging group of human diseases that share a common etiology resulting from dysfunction of the cilium or centrosome. The gene encoding the retinitis pigmentosa 2 protein (RP2) is mutated in X-linked retinitis pigmentosa. RP2 localizes to the ciliary base and this requires the dual acylation of the N-terminus, but the precise mechanism by which RP2 is trafficked to the cilia is unknown. Here we have characterized an interaction between RP2 and Importin ß2 (transportin-1), a member of the Importin-ß family that regulates nuclear-cytoplasmic shuttling. We demonstrate that Importin ß2 is necessary for localization of RP2 to the primary cilium because ablation of Importin ß2 by shRNA blocks entry both of endogenous and exogenous RP2 to the cilium. Furthermore, we identify two distinct binding sites of RP2, which interact independently with Importin ß2. One binding site is a nuclear localization signal (NLS)-like sequence that is located at the N-terminus of RP2 and the other is an M9-like sequence within the tubulin folding cofactor C (TBCC) domain. Mutation of the NLS-like consensus sequence did not abolish localization of RP2 to cilia, suggesting that the sequence is not essential for RP2 ciliary targeting. Interestingly, we found that several missense mutations that cause human disease fall within the M9-like sequence of RP2 and these mutations block entry of RP2 into the cilium, as well as its interaction with Importin ß2. Together, this work further highlights a role of Importin ß2 in regulation of the entry of RP2 and other proteins into the ciliary compartment.

Application of Abbott ID NOW in the emergency department for SARS-CoV-2 detection: A medical center's perspective.

Testing for SARS-CoV-2 is crucial to tracking and controlling the pandemic. In particular, rapid testing in settings such as the emergency department (ED) could improve time to diagnosis and promote proper infection control measures. Early in the COVID-19 pandemic, we implemented the Abbott ID NOW COVID-19 method for screening symptomatic ED patients. However, due to concerns of suboptimal sensitivity, samples with a negative result were reflexed to the lab for confirmatory testing by the TaqPath COVID-19 Combo RT-PCR method. This study analyzed 6773 ID NOW results from April 2020 to September 2020 in the ED, of which 10% (n = 673) were positive and reported directly. The rest 90% (n = 6100) were negative and reflexed to RT-PCR. Among them, 3% (n = 175) turned positive on RT-PCR while 97% (n = 5925) of the results were consistently negative. The cycle threshold (Ct) values of the false-negative samples (n = 175) showed 90% (n = 158) of them with relatively low viral loads (Ct ≥ 30) with median Ct value at 35, while a number of samples (n = 17) had low Ct values (Ct < 30) and no clear explanation for false-negative results. Our study demonstrates that the Abbott ID NOW, despite it's sensitivity limitations, was capable of providing near real-time results for 10% of symptomatic patients presenting to the ED allowing for improved management and workflow. However, our study findings emphasize the need to reflex negative specimens to a higher sensitivity method when prevalence is high and false-negative results are intolerable.

A novel Crumbs3 isoform regulates cell division and ciliogenesis via importin beta interactions.

The Crumbs family of apical transmembrane proteins regulates apicobasal polarity via protein interactions with a conserved C-terminal sequence, ERLI. However, one of the mammalian Crumbs proteins, Crumbs3 (CRB3) has an alternate splice form with a novel C-terminal sequence ending in CLPI (CRB3-CLPI). We report that CRB3-CLPI localizes to the cilia membrane and a membrane compartment at the mitotic spindle poles. Knockdown of CRB3-CLPI leads to both a loss of cilia and a multinuclear phenotype associated with centrosomal and spindle abnormalities. Using protein purification, we find that CRB3-CLPI interacts with importin beta-1 in a Ran-regulated fashion. Importin beta-1 colocalizes with CRB3-CLPI during mitosis, and a dominant-negative form of importin beta-1 closely phenocopies CRB3-CLPI knockdown. Knockdown of importin beta-1 blocks targeting of CRB3-CLPI to the spindle poles. Our data suggest an expanded role for Crumbs proteins in polarized membrane targeting and cell division via unique interactions with importin proteins.

Different toxicity to liver and gill of zebrafish by selenium nanoparticles derived from bio/chemical methods.

With the wide application of selenium nanoparticles (SeNPs) in pharmaceutical fields, the toxicity assessment is of great significance. In this study, zebrafish were selected as model organisms to compare the toxicity of SeNPs derived from biological and chemical methods. The results showed that the size of bio-SeNPs was about 5-fold bigger than chem-SeNPs. When exposed to SeNPs for 96 h, LC50 of bio-SeNPs and chem-SeNPs was 1.668 mg/L and 0.699 mg/L, respectively. Compared with the control, the results showed a significant increase in oxidative toxicity index (P < 0.05), such as glutathione (GSH), superoxide dismutase (SOD) of the liver, and gill in SeNPs-treated group. The neurotoxicity index, such as acetylcholinesterase (AchE) and Na+-K+-ATP enzyme activity, was significantly decreased both in the liver and gill (P < 0.05). It was found that the toxicity of bio-SeNPs to the liver and gill of zebrafish was lower than chem-SeNPs and the toxicity to the liver was higher than gill. In this study, the toxicity of chem-SeNPs and bio-SeNPs to the target organs of zebrafish were systematically evaluated, which provided the basis for the safe application of SeNPs synthesized by different pathways.

JAM-A signals through the Hippo pathway to regulate intestinal epithelial proliferation.

JAM-A is a tight-junction-associated protein that contributes to regulation of intestinal homeostasis. We report that JAM-A interacts with NF2 and LATS1, functioning as an initiator of the Hippo signaling pathway, well-known for regulation of proliferation. Consistent with these findings, we observed increased YAP activity in JAM-A-deficient intestinal epithelial cells (IEC). Furthermore, overexpression of a dimerization-deficient mutant, JAM-A-DL1, failed to initiate Hippo signaling, phenocopying JAM-A-deficient IEC, whereas overexpression of JAM-A-WT activated Hippo signaling and suppressed proliferation. Lastly, we identify EVI1, a transcription factor reported to promote cellular proliferation, as a contributor to the pro-proliferative phenotype in JAM-A-DL1 overexpressing IEC downstream of YAP. Collectively, our findings establish a new role for JAM-A as a cell-cell contact sensor, raising implications for understanding the contribution(s) of JAM-A to IEC proliferation in the mammalian epithelium.

Epithelial JAM-A is fundamental for intestinal wound repair in vivo.

Junctional adhesion molecule-A (JAM-A) is expressed in several cell types, including epithelial and endothelial cells, as well as some leukocytes. In intestinal epithelial cells (IEC), JAM-A localizes to cell junctions and plays a role in regulating barrier function. In vitro studies with model cell lines have shown that JAM-A contributes to IEC migration; however, in vivo studies investigating the role of JAM-A in cell migration-dependent processes such as mucosal wound repair have not been performed. In this study, we developed an inducible intestinal epithelial-specific JAM-A-knockdown mouse model (Jam-aERΔIEC). While acute induction of IEC-specific loss of JAM-A did not result in spontaneous colitis, such mice had significantly impaired mucosal healing after chemically induced colitis and after biopsy colonic wounding. In vitro primary cultures of JAM-A-deficient IEC demonstrated impaired migration in wound healing assays. Mechanistic studies revealed that JAM-A stabilizes formation of protein signaling complexes containing Rap1A/Talin/ß1 integrin at focal adhesions of migrating IECs. Loss of JAM-A in primary IEC led to decreased Rap1A activity and protein levels of Talin and ß1 integrin, and it led to a reduction in focal adhesion structures. These findings suggest that epithelial JAM-A plays a critical role in controlling mucosal repair in vivo through dynamic regulation of focal adhesions.

Selenium nanoparticles with photocatalytic properties synthesized by residual activated sludge.

The treatment and disposal of residual activated sludge is a worldwide problem and the research on its reuse is still only in the earliest stages. Selenite is a toxic pollutant, while selenium nanoparticles (SeNPs) are environmentally friendly and have promising application prospects. At present, the reduction mechanism of selenite under the complex system is still poorly understood. In order to explore the mechanism of SeNPs synthesis by activated sludge resource utilization, SeNPs were synthesized by activated sludge extracts of domestic sewage (DSeNPs) and coking sewage (CSeNPs), respectively. The synthesis process, zeta potential and morphology size of SeNPs were changed by pH value, extract concentration and extract composition. Under the same synthesis conditions, the morphologies of DSeNPs and CSeNPs were mainly spherical and pseudo-spherical, while CSeNPs also contained pseudo-rod shape particles. The sizes and crystal grains of CSeNPs were smaller than those of DSeNPs. Compared with DSeNPs, a specific protein (~35 kDa) was found on the surface of CSeNPs using SDS-PAGE. By analyzing the fluorescence images of the two SeNPs, it was found that the relative contents of proteins, α-d-glucopyranose polysaccharides, and ß-d-glucopyranose polysaccharides on their surfaces were obviously different (P < 0.05). The present study demonstrated that proteins, polysaccharides, humic-like and fulvic acid-like substances cooperated in the formation and stabilization of SeNPs. Furthermore, CSeNPs (bandgap: 1.68 eV) had more desirable photocatalytic performance than DSeNPs (bandgap: 1.84 eV). Under the light condition, CSeNPs could degrade Rhodamine B faster without adding hydrogen peroxide. This experiment provided a new insight into the resource utilization of activated sludge and a reference for the synthesis of nanometer selenium with excellent performance.

Non-prescription Fentanyl Positive Toxicology: Prevalence, Positive Predictive Value of Fentanyl Immunoassay Screening, and Description of Co-substance Use.

OBJECTIVES: Opioid overdose deaths in Massachusetts linked to illicitly-manufactured fentanyl have increased dramatically. In response, an urban safety-net hospital added urine fentanyl testing with reflex confirmation testing to its standard urine toxicology panel. The goals of this study were to describe fentanyl toxicology test results, identify the positive predictive value of presumptive fentanyl immunoassay, and describe co-substance use among those with unexpected fentanyl positive results. METHODS: We included urine toxicology tests from January through June 2016 analyzed at an urban safety-net hospital. We excluded tests from individuals prescribed or administered fentanyl within the preceding 72 hours. Positive fentanyl immunoassay tests underwent reflex chromatography confirmation testing. Samples that confirmed positive for acetyl fentanyl and/or fentanyl and/or norfentanyl were considered true positives. RESULTS: Of 11,873 urine samples, 10.4% of samples screened fentanyl positive and 8.8% were confirmed fentanyl positive. The positive predictive value of a positive urine fentanyl screen was 85.7%. Of 4398 unique patients, 13.2% had at least 1 test confirmed positive for nonprescription fentanyl. Patients with a confirmed fentanyl positive drug test were more likely to have positive urine drug test for barbiturates, benzodiazepines, cocaine, methadone, and opiates, and less likely to have oxycodone or buprenorphine. CONCLUSIONS: At an urban safety-net hospital, nonprescription fentanyl use was common and was associated with greater use of other substances favoring routine fentanyl testing. Although the positive predictive value of the screening test was high, confirmation testing detected substantial numbers of false positives, especially in older patients. Therefore, fentanyl confirmation testing should be used when results will change treatment approach and patient education.

[Characteristics of selenium nanoparticles synthesized by cell-free supernatant Cupriavidus sp. SHE].

In recent years, selenium nanoparticles (SeNPs) have been widely used in many fields such as nanotechnology, biomedicine and environmental remediation due to their good electrical conductivity, photothermal properties and anticancer properties. In this study, the cell-free supernatant, whole cell and the cell-free extracts of the strain Cupriavidus sp. SHE were used to synthesize SeNPs, and several methods were applied to analyze the crystal structure and surface functional groups of the nanoparticles. Finally, Pseudomonas sp. PI1 (G⁺) and Escherichia coli BL21 (G⁻) were selected to investigate the antibacterial properties of SeNPs. Cell-free supernatant, whole cell and cell-free extracts of the strain could synthesize SeNPs. As for the cell-free supernatant, selenite concentration of 5 mmol/L and pH=7 were favorable for the synthesis of SeNPs. TEM images show that the average size of nanospheres synthesized by the supernatant was 196 nm. XRD analysis indicates the hexagonal crystals structure of SeNPs. FTIR and SDS-PAGE confirmed the proteins bound to the surfaces of SeNPs. SeNPs synthesized by cell-free supernatant showed no antimicrobial activities against Pseudomonas sp. PI1 and Escherichia coli BL21 (DE3). These results suggest that proteins played an important role in biotransformation of SeNPs in an eco-friendly process, and SeNPs synthesized in this study were non-toxic and biologically compatible, which might be applied in other fields in the future.

A retrospective analysis reveals a predictor of survival for the patient with paraquat intoxication.

Feasible and accurate predictors are urgently needed to evaluate the survival for patients with paraquat poisoning since the high mortality of paraquat poisoning always resulted in the loss of both life and money. Multiple predictors have been developed to predict prognosis of the patients with PQ poisoning, which however heavily depend on the time of admission to hospitals. Here we reported a feasible and accurate prognosis predictor for patients with paraquat poisoning that is independent of the time of admission to hospitals. Patients with paraquat poisoning were enrolled in this study according to the inclusion and exclusion criteria, which were grouped into survivors and non-survivors based on the 90-days follow-up investigation. The concentration of paraquat in serum and urine, and the baseline clinical parameters associated with the injuries of the liver, kidney, and lung were evaluated to predict the survival of these patients by using receiver operating characteristic curve (ROC) analysis, univariate and multivariate cox regression analyses. A total of 114 patients was included in this study with a survival rate of 54.4%. The median survival days of non-survivors were 6.0 (95%Cl: 4.0-7.8). A new predictor, namely paraquat concentration-associated multiorgan injury index (PCAMII), was established by integrating serum and urine paraquat concentration, serum creatinine, alanine aminotransferase, aspartate transaminase, total and direct bilirubin, at different weighting coefficients, with the accuracy of about 90%. The model to predict the survival probability by PCAMII was established with good fitness (R2 = 0.9325), providing the simulated survival rates comparable to the clinical data. PCAMII, which is independent of hospital admission time, is a feasible and accurate marker to predict the survival rate of patients with PQ poisoning.