High expression of CASP1 induces atherosclerosis.

Atherosclerosis is a chronic, progressive vascular disease. The relationship between CASP1 gene expression and atherosclerosis remains unclear. The atherosclerosis dataset GSE132651 and GSE202625 profiles were downloaded from gene expression omnibus. Differentially expressed genes (DEGs) were screened. The construction and analysis of protein-protein interaction network, functional enrichment analysis, gene set enrichment analysis, and Comparative Toxicogenomics Database analysis were performed. Gene expression heatmap was drawn. TargetScan was used to screen miRNAs that regulate central DEG. 47 DEGs were identified. According to gene ontology analysis, they were mainly enriched in the regulation of stimulus response, response to organic matter, extracellular region, extracellular region, and the same protein binding. Kyoto Encyclopedia of Gene and Genome analysis results showed that the target cells were mainly enriched in the PI3K-Akt signaling pathway, Ras signaling pathway, and PPAR signaling pathway. In the enrichment project of Metascape, vascular development, regulation of body fluid levels, and positive regulation of cell motility can be seen in the gene ontology enrichment project. Eleven core genes (CASP1, NLRP3, MRC1, IRS1, PPARG, APOE, IL13, FGF2, CCR2, ICAM1, HIF1A) were obtained. IRS1, PPARG, APOE, FGF2, CCR2, and HIF1A genes are identified as core genes. Gene expression heatmap showed that CASP1 was highly expressed in atherosclerosis samples and low expressed in normal samples. NLRP3, MRC1, IRS1, PPARG, APOE, IL13, FGF2, CCR2, ICAM1, HIF1A were low expressed in atherosclerosis samples. CTD analysis showed that 5 genes (CASP1, NLRP3, CCR2, ICAM1, HIF1A) were found to be associated with pneumonia, inflammation, cardiac enlargement, and tumor invasiveness. CASP1 gene is highly expressed in atherosclerosis. The higher the CASP1 gene, the worse the prognosis.

Appraising the value of CircRNAs for the diagnosis and prognosis of esophageal squamous cell cancer: An updated meta-analysis.

OBJECTIVE: The purpose of this study was to thoroughly assess the relevance of circular RNAs (circRNAs) in the diagnosis and prognosis of esophageal squamous cell carcinoma (ESCC)， and design a systematic review and meta-analysis. METHODS: Using Stata 14.0 software, a meta-analysis was carried out by looking for pertinent studies up to February 20, 2023, in the online databases PubMed, Embase, Web of Science, and CNKI. The clinicopathologic and prognostic data were evaluated using the combined advantage ratio (OR) and combined hazard ratio (HR), respectively. The threshold effects and publication bias were quantified using Spearman's correlation and the Deeks funnel plot asymmetry tests, respectively. RESULTS: A total of 36 pertinent studies with a literature quality score of 7 or above were included in this study. Of them, 22 papers dealt with clinicopathological characterization, 15 dealt with prognostic analysis, and 13 dealt with diagnostic analysis. The findings demonstrated that high expression of upregulated circRNAs was associated with worse clinicopathological features (tumor size: OR=3.61, 95% CI:1.45-5.78; TNM stage: OR=2.12, 95% CI:1.41-2.83; lymph node metastasis: OR=2.87, 95% CI:1.67-4.07) and worse OS (HR=1.49, 95% CI:1.26-1.77). High downregulated circRNAs expression was linked to improved clinicopathologic characteristics (TNM staging: OR=0.35, 95% CI:0.13- 0.95) and longer survival (HR=0.48, 95% CI:0.27-0.84); combined sensitivity was 0.77 (95% CI: 0.71-0.82), specificity was 0.80 (95% CI:0.74-0.86), and area under the subject operating characteristic curve (AUC) was 0.86 (95% CI:0.82- 0.88). CONCLUSION: CircRNAs are useful for ESCC patient diagnosis and prognosis, and they are anticipated to be unique potential biomarkers for ESCC clinical diagnosis.

Comparing extracellular volume fraction with apparent diffusion coefficient for the characterization of breast tumors.

PURPOSE: To investigate the feasibility of dual-energy CT (DECT)-derived extracellular volume (ECV) fraction for characterization of breast tumors, compared to apparent diffusion coefficient (ADC) and validated against histopathological findings. MATERIAL AND METHODS: The ECV fraction and ADC were prospectively assessed in patients with breast tumors using chest DECT and breast MRI. The diagnostic performance of ECV fraction and ADC was accessed in predicting breast histopathological subtypes and pathological complete response (pCR) status. Histopathological sections were analyzed by digital image analysis. Pearson's correlation analysis was used to correlate between DECT and histopathological ECV fractions. RESULTS: This study included 271 patients, with 314 breast lesions (61 benign and 253 malignant). The ECV fraction and ADC showed comparable area under the curve (AUC) for distinguishing benign from malignant lesions (p = 0.123) and invasive carcinoma from ductal carcinoma in situ (p = 0.115). There were significant differences in ECV fraction between different hormone receptors and Ki67 states (p = 0.001 âˆ¼ 0.014), while ADC values only differed among various Ki67 states (p < 0.001). The ECV fraction was lower (p = 0.007), ADC was higher (p = 0.013) in pCR than in non-pCR group, with an AUC of 0.748 and 0.730 (p = 0.887), respectively. There was a positive correlation between DECT and histopathological ECV fractions (r = 0.615, p < 0.01). CONCLUSIONS: Routine chest DECT-derived ECV fraction is a viable quantitative imaging biomarker for predicting histopathological subtypes and pCR in patient with breast tumors, and correlated well with histopathology finding.

Quantitative parameters derived from dual-energy computed tomography for the preoperative prediction of early recurrence in patients with esophageal squamous cell carcinoma.

OBJECTIVES: This study aimed to assess the efficacy of quantitative parameters derived from dual-energy computed tomography (DECT) for the preoperative prediction of early recurrence (ER) in patients with esophageal squamous cell carcinoma (ESCC). METHODS: In total, 78 patients with ESCC who underwent radical esophagectomy and DECT from June 2019 to August 2020 were enrolled in this study. Normalized iodine concentration (NIC) and electron density (Rho) in tumors were measured using arterial and venous phase images, whereas unenhanced images were used to determine the effective atomic number (Zeff). Univariate and multivariate Cox proportional hazards models were used to identify independent risk predictors of ER. Receiver operating characteristic curve analysis was performed using the independent risk predictors. ER-free survival curves were constructed using the Kaplan-Meier method. RESULTS: NIC in the arterial phase (A-NIC; hazards ratio [HR], 3.91; 95% confidence interval [CI], 1.79-8.56; p = 0.001) and pathological grade (PG; HR, 2.69; 95% CI, 1.32-5.49; p = 0.007) were identified as significant risk predictors of ER. The area under the curve of A-NIC for predicting ER in patients with ESCC was not significantly higher than that of PG (0.72 vs. 0.66, p = 0.441). In a stratified survival analysis, patients with high A-NIC or poorly differentiated ESCC had a higher rate of ER than those with low A-NIC or highly/moderately differentiated ESCC. CONCLUSIONS: A-NIC derived from DECT can be used to noninvasively predict preoperative ER in patients with ESCC, with an efficacy comparable to that of pathological grade. CLINICAL RELEVANCE STATEMENT: Preoperative quantitative measurement of dual-energy CT parameters can predict the early recurrence of esophageal squamous cell carcinoma and serve as an independent prognostic factor to guide clinical designation of personalized treatment. KEY POINTS: • Normalized iodine concentration in the arterial phase and pathological grade were independent risk predictors of early recurrence in patients with esophageal squamous cell carcinoma. • Normalized iodine concentration in the arterial phase may be a noninvasive imaging marker for preoperatively predicting early recurrence in patients with esophageal squamous cell carcinoma. • The efficacy of normalized iodine concentration in the arterial phase derived from dual-energy computed tomography for predicting early recurrence is comparable to that of pathological grade.

[Regulation mechanism of resveratrol on odontogenic differentiation of human dental pulp stem cells].

PURPOSE: To investigate whether resveratrol promotes odontogenic differentiation of human dental pulp stem cells(DPSCs) by up-regulating the expression of silent information regulator 1 (SIRT1) and activating ß-catenin signaling pathway. METHODS: Different concentrations of resveratrol(0, 10, 15, 20 and 50 µmol/L) were used to treat DPSCs for 7 days and 14 days, and cell proliferative activity was detected by CCK-8. After odontogenic differentiation induced by 15 µmol/L resveratrol for 7 days, alkaline phosphatase(ALP) staining was performed and real-time quantitative reverse transcription PCR(qRT-PCR) was used to detect the mRNA expression of Runt-related transcription factor 2 (Runx2), dentin sialophosphoprotein(DSPP) and dentin matrix protein-1(DMP-1) in DPSCs. Western blot was used to detect the expression of SIRT1 in DPSCs on a specific day (0, 3rd, 5th, 7th and 14th) after differentiation induction. Western blot was also used to detect the expression of SIRT1 and activated ß-catenin during odontogenic differentiation of DPSCs treated by 15 µmol/L resveratrol for 7 days. The experimental data was analyzed with GraphPad Prism 9 software package. RESULTS: 15 µmol/L resveratrol had no significant effect on proliferation of DPSCs on the 7th and 14th day; 15 µmol/L resveratrol promoted odontogenic differentiation of DPSCs and up-regulated mRNA expression of RUNX2, DSPP, and DMP-1 in DPSCs; the expression of SIRT1 was the highest on the 7th day during odontogenic differentiation induction. Resveratrol resulted in the increasing protein expressions of SIRT1 and activated ß-catenin when DPSCs was induced to odontogenic differentiation for 7 days. CONCLUSIONS: Resveratrol promotes odontogenic differentiation of human DPSCs by up-regulating the expression of SIRT1 protein and activating ß-catenin signaling pathway.

Multiple parameters from ultrafast dynamic contrast-enhanced magnetic resonance imaging to discriminate between benign and malignant breast lesions: Comparison with apparent diffusion coefficient.

PURPOSE: The purpose of this study was first to assess the diagnostic performance of ultrafast dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters compared to apparent diffusion coefficient (ADC) for distinguishing benign from malignant breast lesions and second to investigate the complementarity of ultrafast DCE-MRI with DWI in that task. MATERIALS AND METHODS: A total of 142 women (mean age, 48.42 ± 11.03 [SD]) years; range: 14-78 years) with 150 breast lesions who underwent breast ultrafast DCE-MRI were prospectively recruited. Ultrafast DCE-MRI semi-quantitative parameters (maximum slope [MS], time to peak [TTP], time to enhancement [TTE], and initial area under curve in 60 s [iAUC]), ultrafast DCE-MRI quantitative parameters (Kep, Ktrans, and Ve), and the ADC were estimated and compared between benign and malignant breast lesions. Classification performances were assessed using area under the receiver operating characteristic curve (AUC) and compared using Delong test. RESULTS: The ultrafast DCE-MRI semi-quantitative multiparameters (AUC, 0.913; 95% CI: 0.856-0.953) showed better classification performance than the quantitative multiparameters (AUC, 0.818; 95% CI: 0.747-0.876) (P = 0.022). No differences in AUC were found between ultrafast DCE-MRI semi-quantitative multiparameters and ADC (AUC, 0.912; 95% CI: 0.855-0.952) (P = 0.990). The combination of ultrafast DCE-MRI semi-quantitative multiparameters and ADC (AUC, 0.960; 95% CI: 0.915-0.985) showed better classification performance than the ultrafast DCE-MRI semi-quantitative multiparameters (P = 0.014) and quantitative multiparameters (P < 0.001). CONCLUSION: Ultrafast DCE-MRI can be used as an accurate method for discriminating benign from malignant breast lesions. The combination of ultrafast DCE-MRI and DWI significantly increases the diagnostic value of ultrafast DCE-MRI.

Development and application of a SFC-DAD-MS/MS method to determine carotenoids and vitamin A in egg yolks from laying hens supplemented with ß-carotene.

ß-Carotene, a provitamin A carotenoid, can be converted into vitamin A in animals' bodies, and can also be accumulated intactly in many animal products. In this study, supercritical fluid chromatography-tandem mass spectrometry was utilized to determine ß-carotene and different forms of vitamin A in eggs simultaneously. According to the results, ß-carotene contained in yolk reached a plateau after about 2 weeks of supplementation. With an increase in dietary supplement level, the amount of ß-carotene gradually increased, as well as slightly changing the yolk color. Moreover, the contents of retinoids including retinol, retinyl propionate, retinyl palmitate and retinyl stearate were also elevated in yolks with the ß-carotene additive levels; meanwhile, the lutein and zeaxanthin decreased. On the whole, ß-carotene in the diet of laying hens could be partially deposited in egg yolk, and the contents of vitamin A in yolk could be increased due to ß-carotene bioconversion.

Evaluation of optimal monoenergetic images acquired by dual-energy CT in the diagnosis of T staging of thoracic esophageal cancer.

OBJECTIVES: The purpose of our study was to objectively and subjectively assess optimal monoenergetic image (MEI (+)) characteristics from dual-energy CT (DECT) and the diagnostic performance for the T staging in patients with thoracic esophageal cancer (EC). METHODS: In this retrospective study, patients with histopathologically confirmed EC who underwent DECT from September 2019 to December 2020 were enrolled. One standard polyenergetic image (PEI) and five MEI (+) were reconstructed. Two readers independently assessed the lesion conspicuity subjectively and calculated the contrast-to-noise ratio (CNR) and the signal-to-noise ratio (SNR) of EC. Two readers independently assessed the T stage on the optimal MEI (+) and PEI subjectively. Multiple quantitative parameters were measured to assess the diagnostic performance to identify T1-2 from T3-4 in EC patients. RESULTS: The study included 68 patients. Subjectively, primary tumor delineation received the highest ratings in MEI (+) 40 keV of the venous phase. Objectively, MEI (+) images showed significantly higher SNR compared with PEI (p < 0.05), peaking at MEI (+) 40 keV in the venous phase. CNR of tumor (MEI (+) 40 keV -80 keV) was all significantly higher than PEI in arterial and venous phases (p < 0.05), peaking at MEI (+) 40 keV in venous phases. The agreement between MEI (+) 40 keV and pathologic T categories was 81.63% (40/49). Rho values in venous phases had excellent diagnostic efficiency for identifying T1-2 from T3-4 (AUC = 0.84). CONCLUSIONS: MEI (+) reconstructions at low keV in the venous phase improved the assessment of lesion conspicuity and also have great potential for preoperative assessment of T staging in patients with EC.

Elbasvir/grazoprevir in children aged 3-18 years with chronic HCV genotype 1 or 4 infection: a pharmacokinetic modeling study.

BACKGROUND: Approximately 3.5 million children and adolescents worldwide are chronically infected with HCV. This study uses pharmacokinetic modeling to identify pediatric doses of elbasvir/grazoprevir (EBR/GZR) that achieve plasma concentrations similar to those seen in adults receiving the approved fixed-dose combination regimen of EBR/GZR. PATIENTS AND METHODS: We conducted a nonrandomized, single-arm, multicenter, open-label phase 2b trial in children and adolescents aged 3 to <18 years with chronic HCV genotype 1 or 4 infection (NCT03379506). Pharmacokinetic data were used to bridge efficacy and safety data from adults to children in a stepwise (oldest to youngest) manner. A total of 57 participants were enrolled: cohort 1 (aged 12 to <18 y), n=22; cohort 2 (aged 7 to <12 y), n=17; and cohort 3 (aged 3 to <7 y), n=18. RESULTS: Steady-state plasma exposures were achieved by week 4 for EBR and GZR in all cohorts and daily dosing achieved geometric mean steady-state area under the concentration-time curve at 0-24 hours that fell within comparability bounds established for adults. All participants achieved sustained virologic response 12 weeks after completing treatment (ie, undetectable HCV RNA 12 wk following completion of treatment). Headache (n=4), fatigue (n=4), and nausea (n=2) were the most common treatment-related adverse events (all mild or moderate); no participant discontinued because of an adverse event. CONCLUSIONS: Pediatric EBR/GZR pharmacokinetic models were successfully developed based on complex adult population pharmacokinetic models. At appropriate age-related doses, EBR/GZR is safe and effective in pediatric and adolescent participants with HCV infection.

XGBoost-based multiparameters from dual-energy computed tomography for the differentiation of multiple myeloma of the spine from vertebral osteolytic metastases.

OBJECTIVES: To evaluate the performance of extreme gradient boosting (XGBoost) combined with multiparameters from dual-energy computed tomography (mpDECT) to differentiate between multiple myeloma (MM) of the spine and vertebral osteolytic metastases (VOM). METHODS: For this retrospective study, 28 patients (83 lesions) with MM of the spine and 23 patients (54 lesions) with VOM who underwent DECT were included. The mpDECT for each lesion, including normalized effective atomic number, slope of the spectral Hounsfield unit curve, CT attenuation, and virtual noncalcium (VNCa), was obtained. Boruta was used to select the key parameters, and then subsequently merged with XGBoost to yield a prediction model. The lesions were divided into the training and testing group in a 3:1 ratio. The highest performance of the univariate analysis was compared with XGBoost using the Delong test. RESULTS: The mpDECT of MM was significantly lower than that of VOM (all p < 0.05). In univariate analysis, VNCa had the highest area under the receiver operating characteristic curve (AUC) in the training group (0.81) and testing group (0.87). Based on Boruta, 6 parameters of DECT were selected for XGBoost model construction. The XGBoost model achieved an excellent and stable diagnostic performance, as shown in the training group (AUC of 1.0) and testing group (AUC of 0.97), with a sensitivity of 80%, a specificity of 95%, and an accuracy of 88%, which was superior to VNCa (p < 0.05). CONCLUSIONS: XGBoost combined with mpDECT yielded promising performance in differentiating between MM of the spine and VOM. KEY POINTS: • The multiparameters obtained from dual-energy CT of multiple myeloma differed significantly from those of vertebral osteolytic metastases. • The virtual noncalcium offered the highest AUC in the univariate analysis to distinguish multiple myeloma from vertebral osteolytic metastases. • Extreme gradient boosting combined with multiparameters from dual-energy CT had a promising performance to distinguish multiple myeloma from vertebral osteolytic metastases.

In vivo Protein Interference: Oral Administration of Recombinant Yeast-Mediated Partial Leptin Reduction for Obesity Control.

Obesity-related diseases are always the major health problems that concern the whole world. Serial studies have reported that obesity development is closely related to the out-of-control leptin encoded by the obesity gene (ob). The latest report declaimed "Less Is More," a model explaining that partial leptin reduction triggers leptin sensitization and contributes to obesity control. Here, we came up with a novel concept, in vivo protein interference (iPRTi), an interesting protein knock-down strategy for in vivo partial leptin reduction. First, the specific immune response against leptin induced by the oral administration of ob recombinant yeast was confirmed. Subsequentally, leptin resistance was observed in diet-induced obese mice, and oral administration with ob recombinant yeast declined the circulating leptin and reduced significantly the body weight gain. To further investigate whether the iPRTi strategy is capable of obesity management, the diet-induced obese mice were administrated with ob recombinant yeast. All the indexes examined including the circulating leptin, triglyceride, and total cholesterol, as well as food intake and weight gain, demonstrated a positive effect of the iPRTi strategy on obesity control. In short, this study provides a novel strategy for the potential application of recombinant yeast for the therapy of obese individuals with leptin resistance.

Conjunctive Analyses of Bulk Segregant Analysis Sequencing and Bulk Segregant RNA Sequencing to Identify Candidate Genes Controlling Spikelet Sterility of Foxtail Millet.

Foxtail millet has gradually become a model gramineous C4 crop owing to its short growth period and small genome. Research on the development of its spikelets is not only directly related to the yield and economic value of foxtail millet but also can provide a reference for studying the fertility of other C4 crops. In this study, a hybrid population containing 200 offspring was constructed from the Xinong8852 and An15 parental lines, and two extreme trait populations were constructed from the F2 generation for analysis of the spikelet sterility. The F2 population conformed to a 3:1 Mendelian segregation ratio, and it was thus concluded that this trait is likely controlled by a single recessive gene. Bulk segregant analysis sequencing (BSA-Seq) was used to determine the candidate regions and candidate genes related to the development of foxtail millet spikelets. Additionally, the functional analysis of differentially expressed genes in populations with different traits was conducted by bulk segregant RNA sequencing (BSR-Seq). Finally, conjunctive analysis of BSA-Seq and BSR-Seq results, combined with biological information analysis, revealed six genes on chromosome VII that were ultimately identified as candidate genes controlling foxtail millet spikelet development. This study provides a new reference for research on foxtail millet sterility and lays a solid foundation for the examination of fertility in other gramineous crops.

Magnesium sulfate pharmacokinetics after intramuscular dosing in women with preeclampsia.

BACKGROUND: Current intramuscular magnesium dosing regimens in low and middle-income countries are based on indirect absorption parameters to inform pharmacokinetic and pharmacodynamic response. OBJECTIVE: To determine if therapeutic serum magnesium levels are obtained in women with severe preeclampsia receiving intramuscular administration of magnesium sulfate using the Pritchard regimen and to compare the key pharmacokinetic variables to those previously published. STUDY DESIGN: Serum magnesium levels were obtained at multiple time points at baseline and after magnesium sulfate administration from women with severe preeclampsia receiving the standard Pritchard regimen for seizure prophylaxis at Bayero University, Kano, Nigeria. The pharmacokinetic profiles were constructed for the study cohort and the updated pharmacokinetic model was compared with the one that was previously published. RESULTS: A total of 80 blood samples were collected from 20 women with severe preeclampsia (45 collected before childbirth and 35 collected after childbirth). After 11.5 hours of magnesium sulfate administration, 63% of women in the cohort had serum magnesium levels of ≥2.0 mmol/L. Data from women receiving the Pritchard regimen combined with data from women previously modeled after the receipt of intravenous magnesium sulfate were adequately described using a 2-compartment model with first-order absorption and linear elimination from the central compartment. All structural pharmacokinetic parameters including clearance, central volume of distribution, peripheral volume of distribution, and intercompartment clearance were adjusted for maternal weight, and the clearance was further adjusted for serum creatinine level and antepartum or postpartum status. The simulated pharmacokinetic profiles of the updated pharmacokinetic model and the previously published pharmacokinetic model are similar. In previously published pharmacokinetic modeling, absorption rate constant=0.32 and absolute bioavailability=0.86. In the updated pharmacokinetic model, absorption rate constant=0.45 and absolute bioavailability=0.91. CONCLUSION: These data support the use of the Pritchard regimen as acceptable to achieve therapeutic serum magnesium levels and support the reported simulation of serum magnesium levels and eclampsia response associated with different intramuscular regimens.

Alternative Magnesium Sulfate Dosing Regimens for Women With Preeclampsia: A Population Pharmacokinetic Exposure-Response Modeling and Simulation Study.

Magnesium sulfate is the anticonvulsant of choice for eclampsia prophylaxis and treatment; however, the recommended dosing regimens are costly and cumbersome and can be administered only by skilled health professionals. The objectives of this study were to develop a robust exposure-response model for the relationship between serum magnesium exposure and eclampsia using data from large studies of women with preeclampsia who received magnesium sulfate, and to predict eclampsia probabilities for standard and alternative (shorter treatment duration and/or fewer intramuscular injections) regimens. Exposure-response modeling and simulation were applied to existing data. A total of 10 280 women with preeclampsia who received magnesium sulfate or placebo were evaluated. An existing population pharmacokinetic model was used to estimate individual serum magnesium exposure. Logistic regression was applied to quantify the serum magnesium area under the curve-eclampsia rate relationship. Our exposure-response model-estimated eclampsia rates were comparable to observed rates. Several alternative regimens predicted magnesium peak concentration < 3.5 mmol/L (empiric safety threshold) and eclampsia rate ≤ 0.7% (observed response threshold), including 4 g intravenously plus 10 g intramuscularly followed by either 8 g intramuscularly every 6 hours × 3 doses or 10 g intramuscularly every 8 hours × 2 doses and 10 g intramuscularly every 8 hours × 3 doses. Several alternative magnesium sulfate regimens with comparable model-predicted efficacy and safety were identified that merit evaluation in confirmatory clinical trials.

Central metabolic pathway modification to improve L-tryptophan production in Escherichia coli.

Tryptophan, an aromatic amino acid, has been widely used in food industry because it participates in the regulation of protein synthesis and metabolic network in vivo. In this study, we obtained a strain named TRP03 by enhancing the tryptophan synthesis pathway, which could accumulate tryptophan at approximately 35 g/L in a 5 L bioreactor. We then modified the central metabolic pathway of TRP03, to increase the supply of the precursor phosphoenolpyruvate (PEP), the genes related to PEP were modified. Furthermore, citric acid transport system and TCA were upregulated to effectively increase cell growth. We observed that strain TRP07 that could accumulate tryptophan at approximately 49 g/L with a yield of 0.186 g tryptophan/g glucose in a 5 L bioreactor. By-products such as glutamate and acetic acid were reduced to 0.8 g/L and 2.2 g/L, respectively.

Pharmacokinetic Interactions between the Hepatitis C Virus Inhibitors Elbasvir and Grazoprevir and HIV Protease Inhibitors Ritonavir, Atazanavir, Lopinavir, and Darunavir in Healthy Volunteers.

The combination of the hepatitis C virus (HCV) nonstructural protein 5A (NS5A) inhibitor elbasvir and the NS3/4A protease inhibitor grazoprevir is a potent, once-daily therapy indicated for the treatment of chronic HCV infection in individuals coinfected with human immunodeficiency virus (HIV). We explored the pharmacokinetic interactions of elbasvir and grazoprevir with ritonavir and ritonavir-boosted HIV protease inhibitors in three phase 1 trials. Drug-drug interaction trials with healthy participants were conducted to evaluate the effect of ritonavir on the pharmacokinetics of grazoprevir (n = 10) and the potential two-way pharmacokinetic interactions of elbasvir (n = 30) or grazoprevir (n = 39) when coadministered with ritonavir-boosted atazanavir, lopinavir, or darunavir. Coadministration of ritonavir with grazoprevir increased grazoprevir exposure; the geometric mean ratio (GMR) for grazoprevir plus ritonavir versus grazoprevir alone area under the concentration-time curve from 0 to 24 h (AUC0-24) was 1.91 (90% confidence interval [CI]; 1.31 to 2.79). Grazoprevir exposure was markedly increased with coadministration of atazanavir-ritonavir, lopinavir-ritonavir, and darunavir-ritonavir, with GMRs for grazoprevir AUC0-24 of 10.58 (90% CI, 7.78 to 14.39), 12.86 (90% CI, 10.25 to 16.13), and 7.50 (90% CI, 5.92 to 9.51), respectively. Elbasvir exposure was increased with coadministration of atazanavir-ritonavir, lopinavir-ritonavir, and darunavir-ritonavir, with GMRs for elbasvir AUC0-24 of 4.76 (90% CI, 4.07 to 5.56), 3.71 (90% CI, 3.05 to 4.53), and 1.66 (90% CI, 1.35 to 2.05), respectively. Grazoprevir and elbasvir had little effect on atazanavir, lopinavir, and darunavir pharmacokinetics. Coadministration of elbasvir-grazoprevir with atazanavir-ritonavir, lopinavir-ritonavir, or darunavir-ritonavir is contraindicated, owing to an increase in grazoprevir exposure. Therefore, HIV treatment regimens without HIV protease inhibitors should be considered for HCV/HIV-coinfected individuals who are being treated with elbasvir-grazoprevir.

Pharmacokinetics of elbasvir and grazoprevir in subjects with end-stage renal disease or severe renal impairment.

PURPOSE: To describe the phase 1 and population pharmacokinetic investigations that support dosing recommendations for elbasvir/grazoprevir (EBR/GZR) in hepatitis C virus-infected people with advanced chronic kidney disease. METHODS: This was an open-label, two-part, multiple-dose trial (MK-5172 PN050; NCT01937975) in 24 non-HCV-infected participants with end-stage renal disease (ESRD) or severe renal impairment who received once-daily EBR 50 mg and GZR 100 mg for 10 days. Population pharmacokinetic analyses from the phase 3 C-SURFER study (PN052, NCT02092350) were also conducted. RESULTS: When comparing haemodialysis (HD) and non-HD days in participants with ESRD, geometric mean ratios (GMRs) (90% confidence intervals [CIs]) for EBR and GZR AUC0-24 were 1.14 (1.08-1.21) and 0.97 (0.87-1.09). When comparing ESRD and healthy participants, GMRs (90% CIs) for EBR and GZR AUC0-24 were 0.99 (0.75-1.30) and 0.83 (0.56-1.22) on HD days, and 0.86 (0.65-1.14) and 0.85 (0.58-1.25) on non-HD days. GMRs (90% CIs) for AUC0-24 in participants with severe renal impairment relative to healthy controls were 1.65 (1.09-2.49) for GZR and 1.86 (1.38-2.51) for EBR. In population modelling of data from C-SURFER, absolute geometric means of steady-state EBR AUC0-24 were 2.78 and 3.07 µM*h (HD and non-HD recipients) and GZR AUC0-24 were 1.80 and 2.34 µM*h (HD and non-HD recipients). CONCLUSIONS: EBR/GZR represents an important treatment option for HCV infection in people with severe renal impairment and those with ESRD. No dosage adjustment of EBR/GZR is required in people with any degree of renal impairment, including those receiving dialysis.

Population Pharmacokinetic Modeling to Evaluate Standard Magnesium Sulfate Treatments and Alternative Dosing Regimens for Women With Preeclampsia.

Magnesium sulfate is the standard therapy for prevention and treatment of eclampsia. Two standard dosing regimens require either continuous intravenous infusion or frequent, large-volume intramuscular injections, which may preclude patients from receiving optimal care. This project sought to identify alternative, potentially more convenient, but similarly effective dosing regimens that could be used in restrictive clinical settings. A 2-compartment population pharmacokinetic (PK) model was developed to characterize serial PK data from 92 pregnant women with preeclampsia who received magnesium sulfate. Body weight and serum creatinine concentration had a significant impact on magnesium PK. The final PK model was used to simulate magnesium concentration profiles for the 2 standard regimens and several simplified alternative dosing regimens. The simulations suggest that intravenous regimens with loading doses of 8 g over 60 minutes followed by 2 g/h for 10 hours and 12 g over 120 minutes followed by 2 g/h for 8 hours (same total dose as the standard intravenous regimen but shorter treatment duration) would result in magnesium concentrations below the toxic range. For the intramuscular regimens, higher maintenance doses given less frequently (4 g intravenously + 10-g intramuscular loading doses with maintenance doses of 8 g every 6 hours or 10 g every 8 hours for 24 hours) or removal of the intravenous loading dose (eg, 10 g intramusculary every 8 hours for 24 hours) may be reasonable alternatives. In addition, individualized dose adjustments based on body weight and serum creatinine were proposed for the standard regimens.

Comparative Genomic and Genetic Functional Analysis of Industrial L-Leucine- and L-Valine-Producing Corynebacterium glutamicum Strains.

Corynebacterium glutamicum is an excellent platform for the production of amino acids, and is widely used in the fermentation industry. Most industrial strains are traditionally obtained by repeated processes of random mutation and selection, but the genotype of these strains is often unclear owing to the absence of genomic information. As such, it is difficult to improve the growth and amino acid production of these strains via metabolic engineering. In this study, we generated a complete genome map of an industrial L-valine-producing strain, C. glutamicum XV. In order to establish the relationship between genotypes and physiological characteristics, a comparative genomic analysis was performed to explore the core genome, structural variations, and gene mutations referring to an industrial L-leucine-producing strain, C. glutamicum CP, and the widely used C. glutamicum ATCC 13032. The results indicate that a 36,349 bp repeat sequence in the CP genome contained an additional copy each of lrp and brnFE genes, which benefited the export of L-leucine. However, in XV, the kgd and panB genes were disrupted by nucleotide insertion, which increase the availability of precursors to synthesize L-valine. Moreover, the specific amino acid substitutions in key enzymes increased their activities. Additionally, a novel strategy is proposed to remodel central carbon metabolism and reduce pyruvate consumption without having a negative impact on cell growth by introducing the CP-derived mutant H+/citrate symporter. These results further our understanding regarding the metabolic networks in these strains and help to elucidate the influence of different genotypes on these processes.

Identification and application of a growth-regulated promoter for improving L-valine production in Corynebacterium glutamicum.

BACKGROUND: Promoters are commonly used to regulate the expression of specific target genes or operons. Although a series of promoters have been developed in Corynebacterium glutamicum, more precise and unique expression patterns are needed that the current selection of promoters cannot produce. RNA-Seq technology is a powerful tool for helping us to screen out promoters with expected transcriptional strengths. RESULTS: The promoter PCP_2836 of an aldehyde dehydrogenase coding gene from Corynebacterium glutamicum CP was identified via RNA-seq and RT-PCR as a growth-regulated promoter. Comparing with the strong constitutive promoter Ptuf, the transcriptional strength of PCP_2836 showed a significant decrease that from about 75 to 8% in the stationary phase. By replacing the native promoters of the aceE and gltA genes with PCP_2836 in the C. glutamicum ATCC 13032-derived L-valine-producing strain AN02, the relative transcriptional levels of the aceE and gltA genes decreased from 1.2 and 1.1 to 0.35 and 0.3, and the activity of their translation products decreased to 43% and 35%, respectively. After 28 h flask fermentation, the final cell density of the obtained strains, GRaceE and GRgltA, exhibited a 7-10% decrease. However, L-valine production increased by 23.9% and 27.3%, and the yield of substrate to product increased 43.8% and 62.5%, respectively. In addition, in the stationary phase, the intracellular citrate levels in GRaceE and GRgltA decreased to 27.0% and 33.6% of AN02, and their intracellular oxaloacetate levels increased to 2.7 and 3.0 times that of AN02, respectively. CONCLUSIONS: The PCP_2836 promoter displayed a significant difference on its transcriptional strength in different cell growth phases. With using PCP_2836 to replace the native promoters of aceE and gltA genes, both the transcriptional levels of the aceE and gltA genes and the activity of their translation products demonstrated a significant decrease in the stationary phase. Thus, the availability of pyruvate was significantly increased for the synthesis of L-valine without any apparent irreversible negative impacts on cell growth. Use of this promoter can enhance the selectivity and control of gene expression and could serve as a useful research tool for metabolic engineering.