Transcriptome analysis of Kluyveromyces marxianus under succinic acid stress and development of robust strains.

Kluyveromyces marxianus has become an attractive non-conventional yeast cell factory due to its advantageous properties such as high thermal tolerance and rapid growth. Succinic acid (SA) is an important platform molecule that has been applied in various industries such as food, material, cosmetics, and pharmaceuticals. SA bioproduction may be compromised by its toxicity. Besides, metabolite-responsive promoters are known to be important for dynamic control of gene transcription. Therefore, studies on global gene transcription under various SA concentrations are of great importance. Here, comparative transcriptome changes of K. marxianus exposed to various concentrations of SA were analyzed. Enrichment and analysis of gene clusters revealed repression of the tricarboxylic acid cycle and glyoxylate cycle, also activation of the glycolysis pathway and genes related to ergosterol synthesis. Based on the analyses, potential SA-responsive promoters were investigated, among which the promoter strength of IMTCP2 and KLMA_50231 increased 43.4% and 154.7% in response to 15 g/L SA. In addition, overexpression of the transcription factors Gcr1, Upc2, and Ndt80 significantly increased growth under SA stress. Our results benefit understanding SA toxicity mechanisms and the development of robust yeast for organic acid production. KEY POINTS: • Global gene transcription of K. marxianus is changed by succinic acid (SA) • Promoter activities of IMTCP2 and KLMA_50123 are regulated by SA • Overexpression of Gcr1, Upc2, and Ndt80 enhanced SA tolerance.

Solid basal adenoid cystic carcinoma of the breast: A case report and literature review.

RATIONALE: Adenoid cystic carcinoma (AdCC) is a rare malignancy of the breast with a low Ki-67 index and good prognosis. Owing to the rarity of breast AdCC, the misdiagnosis rate is as high as 50%, and there is no consensus or recognized guidelines for the treatment of this disease. Therefore, it is necessary to conduct a detailed clinical and pathological analysis in combination with a literature review to improve our understanding, diagnosis, and treatment of the disease. METHODS: A 68-year-old woman sought medical attention due to a recently increasing mass in the breast. The left breast mass was 1.3 cm × 1 cm in size. We analyzed the morphology, immunohistochemistry, and molecular characteristics of the tumor removed by surgery, and reviewed relevant literature. DIAGNOSES: Solid basal AdCC of the breast. INTERVENTIONS: We performed biopsy, immunohistochemistry and molecular testing on surgical resection specimens. OUTCOMES: Combining morphological and immunohistochemical features, it is consistent with solid basal AdCC of the breast, and Fish detected MYB gene break. LESSONS: Due to the high misdiagnosis rate of AdCC, accurate histopathological diagnosis is particularly important. At present, breast conserving surgery and local tumor resection are mainly used for the treatment of breast AdCC, and postoperative adjuvant radiotherapy is feasible.

Serum uric acid and outcome in hospitalized elderly patients with chronic heart failure through the whole spectrum of ejection fraction phenotypes.

INTRODUCTION: Elevated serum uric acid (SUA) levels have been associated with poor outcome in patients with heart failure (HF). Uric acid is associated with inflammation and microvascular dysfunction, which may differentially affect left ventricular ejection fraction (EF) phenotypes. We aimed to identify the role of SUA across EF phenotypes in hospitalized elderly patients with chronic HF. METHODS: We analyzed 1355 elderly patients who were diagnosed with chronic HF. All patients had SUA levels measured within the first 24 h following admission. Patients with left ventricle EF were categorized as having HF with reduced EF (HFrEF, EF < 40%), HF with mid-range EF (HFmrEF, 40%≦LVEF ≦ 49%) or HF with preserved EF (HFpEF, LVEF ≥ 50%). Endpoints were cardiovascular death, HF rehospitalization, and their composite. The median follow-up period was 18 months. RESULTS: Compared with the lowest SUA quartile, the highest SUA quartile was significantly associated with the endpoints (adjusted HR: 2.404, 95% CI: 1.178-4.906, P = 0.016; HR: 1.418, 95% CI: 1.021-1.971, P = 0.037; HR: 1.439, 95% CI: 1.049-1.972, P = 0.024, respectively). After model adjustment, a significant association of SUA with cardiovascular death and the composite endpoint persisted among HFrEF and HFmrEF patients in the highest SUA quartile (P < 0.05 for all). CONCLUSIONS: In hospitalized elderly patients with chronic HF, SUA is an independent predictor of adverse outcomes, which can be seen in HFrEF and HFmrEF patients.

The displaced polymer clamp entered the duodenum causing an abscess: A case report.

RATIONALE: The Hem-o-lok clip, made from a nonabsorbable polymer, and its predecessor the metal ligation clip have been used widely for laparoscopic or robot-assisted surgery to ligate the cystic duct after a cholecystectomy, to ligate the appendix after an appendectomy, or control hemorrhage or on occasion to occlude a fistula or enterotomy. Displacement of these ligation clips to distant sites is an extremely rare complication in clinical practice. PATIENT CONCERNS: The patient is a 67-year old female who sought medical attention for 3 days due to worsening intermittent upper abdominal pain and poor appetite. Gastroscopy showed both an ulcer and the presence of a foreign object embedded in the anterior wall of the duodenal bulb, consistent with what looked like a polymer-based ligation clip. After removal of the foreign body, which turned out to be a remnant of the polymer clip, no further pus was seen, but fresh granulomatous tissues were seen at the base. DIAGNOSES: a polymer-based clip-induced duodenal bulb erosion with a local contained enterically draining abscess. INTERVENTIONS: The patient recovered after removing foreign bodies under gastroscopy and receiving anti infection treatment. OUTCOMES: The patient recovered after removing foreign bodies under gastroscopy and receiving anti infection treatment. LESSONS: In laparoscopic cholecystectomy, attention should be paid to the correct surgical techniques, possibly by decreasing the number of such clips used or considering use of absorbable clips, ligature wires, ligation with absorbable suture material, or ultrasonic resection, all of which can be used for clipless cholecystectomy.

Changes in the incidence and prevalence of ischemic stroke and associations with natural disasters: an ecological study in 193 countries.

Epidemiological studies have indicated that natural disasters have important impacts on ischemic stroke. This study determined the associations between natural disasters and the incidence and prevalence of ischemic stroke at the global level. A 28-year ecological trend study was performed to estimate worldwide changes in the incidence and prevalence of ischemic stroke and their associations with natural disasters by analyzing data from 193 countries. Quantum geographic information system-based visualization and multivariable linear regression were used. Changes in the incidence and prevalence of ischemic stroke, as well as disaster occurrence, varied among the different regions over the past 28 years (p < 0.001). Multiple linear regression revealed an independent and positive association between disaster occurrence and the incidence of ischemic stroke in males, females and both sexes combined (standardized coefficients = 0.515, 0.470 and 0.483, p < 0.001); similar associations were found for the prevalence of ischemic stroke (standardized coefficients = 0.471, 0.417 and 0.438, p < 0.001). The incidence and prevalence of ischemic stroke changed significantly at the global level and were independently associated with natural disasters. Both males and females show common but different vulnerabilities to natural disasters. This evidence supports policy making and resource allocation for disaster response and disease burden reduction.

[Optimization of Tidal-Combined Flow Constructed Wetland System and Its Removal Effect on Antibiotic Resistance Genes].

Antibiotic resistance genes (ARGs) pose a serious threat to environmental biology and public health, along with the discharge and spread of wastewater. The advanced treatment of ARGs in wastewater therefore deserves special attention. In our previous study, we found that tidal flow constructed wetlands can effectively remove multiple ammonia from wastewater. In this study, we further optimized tidal flow constructed wetland systems by adding baffles and cultivating plants; we investigated the influence of process optimization on the removal of ARGs and the influence of functional microorganism distribution on nitrogen removal. The results show that the addition of baffles and plants can effectively improve the removal efficiency of ARGs, with the maximum removal rate of 21 resistance genes, in 7 categories, reaching 83.82%-100.0% with the simultaneous addition of baffles and plants. These removal rates were significantly higher than the increase resulting from a single baffle or plant group. From the comparison of the absolute abundance of ARGs in the substrate and plants, it is clear that the baffles can promote the enrichment of ARGs in the wetland substrate, while uptake by plants is also a way of removing ARGs. Combined with the results of nitrogen-cycle functional gene sequencing, system optimization can increase the diversity and richness of nitrification and denitrification functional microorganisms in the substrate, which is consistent with the higher removal rate of nitrification and total nitrogen in wastewater.

Disability-Adjusted Life Years (DALYs) Due to Ischemic Heart Disease (IHD) Associated with Natural Disasters: A Worldwide Population-Based Ecological Study.

Background: Recent studies have reported an association between natural disasters of various kinds and ischemic heart disease (IHD). We investigated the association between Disability-adjusted life years (DALYs) due to IHD and natural disasters and aimed to assess DALYs as a quantification of the burden of IHD related to natural disasters at the global level. Methods: Country-specific data of natural disaster impacts DALYs due to IHD and socioeconomic variables were obtained from open sources over the period of 1990-2013 and 2014-2017. A population-based trend ecological design was conducted to estimate the association between trends in DALYs and natural disasters (occurrence, casualties and total damage), adjusting for socioeconomic variables. Results: Most countries have experienced increases in natural disaster occurrences and decreases in DALYs during this study period. The unadjusted correlation analysis demonstrated a positive and significant correlation between DALYs and natural disasters for females and for both sexes (R = 0.163 and 0.146, p = 0.024 and 0.043), and a marginally significant correlation for males (R = 0.128, p = 0.076). After adjusting for socioeconomic variables, multiple linear regression demonstrated independent associations between the occurrence and DALYs due to IHD for males, females and both sexes (standardized coefficients = 0.192, 0.23 and 0.187, p = 0.016, 0.004 and 0.022). Conclusions: A weak but significantly positive association between natural disaster and IHD was confirmed and quantified at the global level by this DALY metric analysis. Adaptation strategies for natural disaster responses and IHD disease burden reduction need to be developed.

Velocity mismatch effect on the dynamic MTF of a TDI image sensor.

By analyzing the process of time delay integration dynamic imaging, we establish a model of velocity mismatch. Based on this model, we analyze the influence of different factors on the dynamic imaging process, and a modulation transfer function (MTF) is used to evaluate imaging quality. According to the simulation, the velocity mismatch and scan stage are the main factors for image quality. The MTF of the image sensor decreases with the velocity mismatch, and the scan stage increases. In addition, an image with higher contrast can be obtained in a short integration time. However, a shorter integration time leads to insufficient sampling. Furthermore, we establish a dynamic MTF testing system, and evaluate the experiment at different imaging modes. Through data comparison, the experimental data are consistent with theoretical data.

Highly enhanced biodegradation of pharmaceutical and personal care products in a novel tidal flow constructed wetland with baffle and plants.

Research on decentralized wastewaters deserves special focus due to the potential abundance of emerging organic pollutants including pharmaceutical and personal care products (PPCPs), which might pose serious threats to the local water bodies and even to human health. Constructed wetland (CW) is a common decentralized wastewater treatment technology, with a certain ability to eliminate PPCPs. Nonetheless, PPCPs removal in common CWs is frequently challenging, besides, the removal mechanism remains elusive. Based on our previous study, tidal flow constructed wetlands (TFCWs) is effective in nitrogen removal. Here, 3 TFCWs with different modifications (baffle, plants, both baffle and plants) were constructed to treat raw domestic sewage and specifically to evaluate the removal efficiencies and mechanism of PPCPs. 24 PPCPs including 7 antibiotics, 8 steroid hormones and 9 biocides were detected in the level of 1.10 ± 0.29 ng/L-799 ± 10.6 ng/L in the influents. Consequently, we found that modification with both baffle and plants significantly influenced the removal of PPCPs. Moreover, the highest removal rates of biocides (97.1 ± 0.29%), steroid hormones (99.8 ± 0.02%), and antibiotics (90.2 ± 1.60%) were achieved via both baffles and plants in TFCWs. Based on the mass balance analysis, microbial degradation dominated the removal of PPCPs with a percentage higher than 85.7%, followed by substrate adsorption (5.22 × 10-2-14.3%) and plant uptake (1.66 × 10-3-0.44%). Further, 16S rDNA sequencing analysis revealed that the presence of baffle and plants improve the removal efficiency of PPCPs by means of enhancing microbial diversity and changing dominant microorganisms. Moreover, Thaumarchaeota was potentially the key microorganism in the phylum level for PPCPs elimination by TFCWs through LEfSe (linear discriminant analysis (LDA) effect size) analysis. These findings provide new insights into the removal of PPCPs in CWs.

LIN28B-AS1-IGF2BP1 binding promotes hepatocellular carcinoma cell progression.

IGF2BP1 overexpression promotes hepatocellular carcinoma (HCC) progression. Long non-coding RNA LIN28B-AS1 directly binds to IGF2BP1. In the present study, LIN28B-AS1 and IGF2BP1 expression and their potential functions in HCC cells were tested. Genetic strategies were applied to interfere their expression, and cell survival, proliferation and apoptosis were analyzed. We show that LIN28B-AS1 is expressed in established/primary human HCC cells and HCC tissues. RNA-immunoprecipitation (RIP) and RNA pull-down results confirmed that LIN28B-AS1 directly associated with IGF2BP1 protein in HCC cells. LIN28B-AS1 silencing (by targeted siRNAs) or knockout (KO, by CRISPR-Cas9 method) depleted IGF2BP1-dependent mRNAs (IGF2, Gli1, and Myc), inhibiting HCC cell growth, proliferation, migration, and invasion. Conversely, ectopic overexpression of LIN28B-AS1 upregulated IGF2BP1-dependent mRNAs and promoted HCC cell progression in vitro. Importantly, ectopic IGF2BP1 overexpression failed to rescue LIN28B-AS1-KO HepG2 cells. LIN28B-AS1 siRNA and overexpression were ineffective in IGF2BP1-KO HepG2 cells. In vivo, LIN28B-AS1 KO-HepG2 xenograft tumors grew significantly slower than the control tumors in the nude mice. Taken together, we conclude that LIN28B-AS1 associates with IGF2BP1 to promote human HCC cell progression in vitro and in vivo.

I-BET726 suppresses human skin squamous cell carcinoma cell growth in vitro and in vivo.

Bromodomain-containing protein 4 (BRD4) is a potential therapeutic target of skin squamous cell carcinoma (SCC). I-BET726 is a novel BRD4 inhibitor. Its potential effect in skin SCC cells was tested in the present study. We show that I-BET726 potently inhibited survival, proliferation, cell cycle progression, and migration in established (A431/SCC-9/SCC-12/SCC-13 lines) and primary human skin SCC cells. I-BET726 induced significant apoptosis activation in skin SCC cells. It was more efficient in inhibiting skin SCC cells than known BRD4 inhibitors (JQ1, CPI203, and AZD5153). I-BET726 not only downregulated BRD4-regulated proteins (c-Myc, Bcl-2, and cyclin D1), but also inhibited sphingosine kinase 1 (SphK1) and Akt signalings in SCC cells. Restoring Akt activation, by a constitutively active S473D mutant Akt1 ("caAkt1"), partially inhibited I-BET726-induced cytotoxicity in A431 cells. In vivo, I-BET726 oral administration potently inhibited A431 xenograft growth in severe combined immunodeficient mice. Downregulation of BRD4-regulated proteins and inhibition of the SphK1-Akt signaling were detected in I-BET726-treated A431 xenograft tumor tissues. Together, I-BET726 inhibits skin SCC cell growth in vitro and in vivo.

Structural stability of a polyetheretherketone femoral component-A 3D finite element simulation.

BACKGROUND: Because its mechanical properties are similar to cortical bones of the knee, polyetheretherketone (PEEK) material has been used to make total knee arthroplasty (TKA) components. This study investigated the PEEK femoral component deformation of a TKA system and compared the data with that of a cobalt-chromium (CoCr) component. METHODS: A 3D finite element knee model was constructed using CT images of a normal subject. A knee prosthesis was installed on the model to simulate a TKA knee. The material properties of the bone were assumed linear and transverse isotropic. The femoral component was modeled using a PEEK or CoCr material. A compressive load was applied to the knee at full extension. Tibiofemoral contact stresses and femoral component deformations were analyzed. FINDINGS: Under a 3 kN load, the maximal Von-Mises stresses in the femoral component were 14.39 MPa and 30.05 MPa for the PEEK and CoCr components, respectively. At the tibial polyethylene surface, the CoCr femoral component caused higher contact stresses (>2.2%) than the PEEK component. The deformation of the PEEK component was over 3 times larger than that of the CoCr component (0.65 × 10-3 mm vs 0.2 × 10-3 mm). INTERPRETATION: The PEEK femoral component could result in lower contact stresses, but larger deformations in the TKA knee compared to the CoCr component. An increased deformation of the PEEK component indicates a reduction in its structural strength. Future investigation should examine if the reduced structural strength will affect the in-vivo component-bone interface integration and affect the component fatigue life.

Sensitive and rapid detection of Salmonella enterica serovar Indiana by cross-priming amplification.

Salmonella enterica serovar Indiana (S. Indiana) was the most frequently reported foodborne pathogen, which has a broad host range including poultry, swine, and humans. Traditional methods used for the detection of S. Indiana from contaminated food products are time-consuming and labor-intensive. Therefore, rapid detection methods with high sensitivity and specificity are vitally important to prevent the spread of S. Indiana. In this study, we developed a nearly instrument-free, simple molecular method which incorporates cross-priming amplification (CPA) combined with a nucleic acid detection strip (NADS) for sensitive detection of S. Indiana. A set of CPA primers was designed based on S. Indiana specific nucleotide sequences and the specificity of CPA-NADS was tested against 42 bacterial strains. The results showed that this method was highly specific for detection of S. Indiana. The sensitivity of CPA-NADS was evaluated and compared with that of the serovar-specific PCR method and the real-time PCR method. The limit of detection of the CPA method was 8.997 fg/µL for genomic DNA and 6.2 × 101 CFU/mL for bacteria in pure culture. An application of the CPA assay was conducted with 90 inoculated specimens by S. Indiana. The accuracy of CPA-NADS was consistent with the results of the traditional culture-based methods in inoculated specimens. This method showed a higher sensitivity than the serovar-specific PCR method did and was more convenient to perform. In conclusion, we demonstrated that the CPA-NADS system offers high specificity, sensitivity, rapidity, and a simple detection tool for screening S. Indiana.

Novel plasmid-mediated colistin resistance gene mcr-7.1 in Klebsiella pneumoniae.

Objectives: To identify a novel plasmid-mediated colistin resistance gene in Klebsiella pneumoniae isolated from chickens in China. Methods: WGS was used to identify a novel colistin resistance gene. The transferability of plasmids carrying mcr-7.1 was investigated by conjugation experiments. The expression of the mcr-7.1 gene was examined using an expression vector. Results: A novel plasmid-mediated colistin resistance gene mcr-7.1, sharing 70% amino acid identity with the mcr-3 gene, was identified in three K. pneumoniae strains isolated from chickens in China. The mcr-7.1 gene was found in an IncI2-type plasmid (pSC20141012) that co-harboured the blaCTX-M-55 gene in one isolate. pSC20141012 can be transferred from K. pneumoniae SC20141012 to Escherichia coli J53Azr, exhibiting a ≥8-fold increase in colistin MIC compared with the recipient E. coli J53Azr. Conclusions: We identified a novel plasmid-mediated colistin resistance gene named mcr-7.1 in K. pneumoniae in China. The prevalence of mcr-7.1 in various species of human and animal origin needs to be investigated immediately.

Characterization of pHeBE7, an IncFII-type virulence-resistance plasmid carrying blaCTX-M-98b, blaTEM-1, and rmtB genes, detected in Escherichia coli from a chicken isolate in China.

Recently, a novel variant of the CTX-M enzyme, CTX-M-98, was detected in Escherichia coli isolates from food animals. However, few plasmids carrying blaCTX-M-98 have been fully characterized. In this study, we sequenced the complete pHeBE7 plasmid, an 86,015-bp plasmid that contains the blaCTX-M-98b, blaTEM-1, rmtB, and traT genes, using whole-genome sequencing. The backbone of pHeBE7 shows a high similarity (>99%) to pMC-NDM, which carries the blaNDM-1 gene, however its mosaic regions remain relatively unique among sequenced plasmids. We discovered that a typical ISEcp1-blaCTX-M-IS903 element in the mosaic region harbors the blaCTX-M-98b gene. Conjugation and growth competition assays indicate that pHeBE7 can be easily transmitted and that it confers a limited fitness cost to the recipient cell. The genetic characterization of pHeBE7 may improve our knowledge of how antibiotic resistance disseminates in enterobacteria.

Removal of antibiotics from piggery wastewater by biological aerated filter system: Treatment efficiency and biodegradation kinetics.

This study aimed to investigate the removal efficiency and mechanism for antibiotics in swine wastewater by a biological aerated filter system (BAF system) in combination with laboratory aerobic and anaerobic incubation experiments. Nine antibiotics including sulfamonomethoxine, sulfachloropyridazine, sulfamethazine, trimethoprim, norfloxacin, ofloxacin, lincomycin, leucomycin and oxytetracycline were detected in the wastewater with concentrations up to 192,000ng/L. The results from this pilot study showed efficient removals (>82%) of the conventional wastewater pollutants (BOD5, COD, TN and NH3-N) and the detected nine antibiotics by the BAF system. Laboratory simulation experiment showed first-order dissipation kinetics for the nine antibiotics in the wastewater under aerobic and anaerobic conditions. The biodegradation kinetic parameters successfully predicted the fate of the nine antibiotics in the BAF system. This suggests that biodegradation was the dominant process for antibiotic removal in the BAF system.

Nitrogen removal and its relationship with the nitrogen-cycle genes and microorganisms in the horizontal subsurface flow constructed wetlands with different design parameters.

This study aims to investigate nitrogen removal and its relationship with the nitrogen-cycle genes and microorganisms in the horizontal subsurface flow constructed wetlands (CWs) with different design parameters. Twelve mesocosm-scale CWs with four substrates and three hydraulic loading rates were set up in the outdoor. The result showed the CWs with zeolite as substrate and HLR of 20 cm/d were selected as the best choice for the TN and NH3-N removal. It was found that the single-stage mesocosm-scale CWs were incapable to achieve high removals of TN and NH3-N due to inefficient nitrification process in the systems. This was demonstrated by the lower abundance of the nitrification genes (AOA and AOB) than the denitrification genes (nirK and nirS), and the less diverse nitrification microorganisms than the denitrification microorganisms in the CWs. The results also show that microorganism community structure including nitrogen-cycle microorganisms in the constructed wetland systems was affected by the design parameters especially the substrate type. These findings show that nitrification is a limiting factor for the nitrogen removal by CWs.

Comparison of Iodixanol and Iopromide in Patients With Renal Insufficiency and Congestive Heart Failure Undergoing Coronary Angiography by Hemodynamic Monitoring.

We investigated the impact of contrast media (CM) with different osmolality on cardiac preload in patients with chronic kidney disease (CKD) and congestive heart failure (CHF). Patients with CKD and CHF were equally randomized to receive either iso-osmolar contrast media (IOCM) iodixanol or low-osmolar contrast media iopromide. We measured cardiac preload indexes by invasive hemodynamic monitoring before and after CM injection. Major adverse cardiac events postprocedures were recorded. Increase in extravascular lung water index was only seen in the iopromide group ( P < .001), while global end diastolic index and central venous pressure were all significantly increased from baseline in the both groups ( P < .001, respectively), and the increase in cardiac preload indexes was significantly greater in the iopromide group than in the iodixanol group ( P < 0.001). The overall incidence of acute heart failure was more frequently observed in the iopromide group ( P = 0.027). Low-osmolar contrast media iopromide significantly increased cardiac preload in patients with CKD and CHF undergoing cardiac catheterization procedures compared with IOCM iodixanol.

Colistin Resistance Gene mcr-1 and Its Variant in Escherichia coli Isolates from Chickens in China.

The mcr-1 gene was detected in 5.11% (58/1136) of Escherichia coli isolates of chicken origin from 13 provinces in China. A novel mcr-1 variant, named mcr-1.3, encoding an Ile-to-Val functional variant of MCR-1 was identified in a sequence type 155 (ST155) strain. An mcr-1.3-containing IncI2 plasmid, pHeN867 (60,757 bp), was identified. The transfer of pHeN867 led to a 32-fold increase in the MIC of colistin in the recipient, exhibiting an effect on colistin resistance that was similar to that of mcr-1.