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PDZK1-interacting protein 1(PDZK1IP1), also known as MAP17, is encoded by the PDZK1IP1 gene and is a membrane-associated protein. PDZK1IP1 have been proven to be a potent regulator of cancer cell proliferation. However, the role of PDZK1IP1 in regulating goat subcutaneous preadipocyte proliferation is unknown. Here, we cloned the full-length coding sequence of PDZK1IP1 gene, investigated the potential functional of PDZK1IP1 in goat subcutaneous preadipocyte proliferation by gaining or losing function in vitro. Our results indicated that goat PDZK1IP1 gene consists of 345 bp, encoding a protein of 114 amino acids containing a typical PDZK1IP1 (MAP17) super family domain. Overexpression of PDZK1IP1 significantly increased the number of EdU-positive cells and cell viability, and also upregulated mRNA expression of cell proliferation-associated genes including CCND1 and CDK2 in vitro cultured cells. Conversely, knockdown of PDZK1IP1 mediated by siRNA technique significantly inhibited subcutaneous preadipocyte proliferation and downregulated mRNA expression of cell proliferation-associated genes including CCNE1, CCND1 and CDK2. Collectively, these results suggested that PDZK1IP1 can promote proliferation of goat subcutaneous preadipocyte.

Wenhao Ding,1,* Yanyan Li,1,* Yichun Bai,1 Yuhong Li,2 Lei Wang,3 Yongbin Wang1 1Department of Epidemiology and Health Statistics, School of Public Health, Xinxiang Medical University, Xinxiang, Henan Province, People’s Republic of China; 2National Center for Tuberculosis Control and Prevention, China Center for Disease Control and Prevention, Beijing, People’s Republic of China; 3Center for Musculoskeletal Surgery, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt–Universität Zu Berlin and Berlin Institute of Health, Berlin, Germany*These authors contributed equally to this workCorrespondence: Yongbin WangDepartment of Epidemiology and Health Statistics, School of Public Health, Xinxiang Medical University, Xinxiang, Henan Province, 453000, People’s Republic of ChinaEmail wybwho@163.comObjective: COVID-19 may have a demonstrable influence on disease patterns. However, it remained unknown how tuberculosis (TB) epidemics are impacted by the COVID-19 outbreak. The purposes of this study are to evaluate the impacts of the COVID-19 outbreak on the decreases in the TB case notifications and to forecast the epidemiological trends in China.Methods: The monthly TB incidents from January 2005 to December 2020 were taken. Then, we investigated the causal impacts of the COVID-19 pandemic on the TB case reductions using intervention analysis under the Bayesian structural time series (BSTS) method. Next, we split the observed values into different training and testing horizons to validate the forecasting performance of the BSTS method.Results: The TB incidence was falling during 2005– 2020, with an average annual percentage change of − 3.186 (95% confidence interval [CI] − 4.083 to − 2.281), and showed a peak in March–April and a trough in January–February per year. The BSTS method assessed a monthly average reduction of 14% (95% CI 3.8% to 24%) in the TB case notifications from January–December 2020 owing to COVID-19 (probability of causal effect=99.684%, P=0.003), and this method generated a highly accurate forecast for all the testing horizons considering the small forecasting error rates and estimated a continued downward trend from 2021 to 2035 (annual percentage change =− 2.869, 95% CI − 3.056 to − 2.681).Conclusion: COVID-19 can cause medium- and longer-term consequences for the TB epidemics and the BSTS model has the potential to forecast the epidemiological trends of the TB incidence, which can be recommended as an automated application for public health policymaking in China. Considering the slow downward trend in the TB incidence, additional measures are required to accelerate the progress of the End TB Strategy.Keywords: COVID-19, tuberculosis, BSTS, ARIMA, causal impact, nowcasting and forecasting

Skewness, as a proxy for extreme risks or losses, deserves more attention from risk management work of portfolio selection and futures hedging. We evaluate the hedging performance of strategies considering the skewness for two major benchmark international crude oil markets, Brent and WTI, with sample period ranging from June 11, 2018, to May 19, 2021. This paper contributes to the literature by accounting for futures basis and the skewness of the hedged portfolio return. Specifically, we first extend the existing literature of Lien (2010), whose study investigated the effect of skewness on optimal production and hedging decisions, to the case of a futures bias existing. Then, we propose minimum-risk hedging models wherein the return of the hedged portfolio return is assumed to follow a skew-normal distribution, which is a generalization of normality assumption. From the empirical results, we find that skewness cannot be ignored, otherwise it will lead to wrong hedging decision. Furthermore, hedging strategies under skew-normal distribution are outperformed than that under the normal distribution assumption. The research results of this paper have important implications for investors and decision makers to hedge the price risk of crude oil in extreme market conditions.

Song Wen,1,* Congying Liu,1,* Yanyan Li,1 Junhong Pan,1 Thiquynhnga Nguyen,1 Ligang Zhou1,2 1Department of Endocrinology, Shanghai Pudong Hospital, Fudan University, Shanghai, 201399, People’s Republic of China; 2Shanghai Key Laboratory of Vascular Lesions Regulation and Remodeling, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai, 201399, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ligang ZhouDepartment of Endocrinology, Shanghai Pudong Hospital, Fudan University, Shanghai, 201399, People’s Republic of ChinaTel +8613611927616Email zhouligang@yahoo.comPurpose: Although psoriasis (PsO) is highly associated with insulin resistance (IR), the role of PsO on activity of insulin secretion or its action in diabetic patients has not been explored.Materials and Methods: In-patient data on type 2 diabetes (T2D) with or without PsO from 2016– 2019 in our hospital were analyzed. Data for 42 diabetic patients with PsO were compared with that of the control group (T2D only). Blood examinations with reference to the levels of fasting blood glucose, C-peptide, insulin, HbA1c, plasma lipids, lipoproteins, and kidney function were explored. HOMA-IR and HOMA-β models were established to explore IR and pancreatic β-cell function.Results: HOMA-IR level was significantly higher (P=0.0003< 0.05) in patients with PsO compared with the controls. Although the durations of diabetes in patients with PsO were significantly shorter compared with that of patients with diabetes only (P=0.012< 0.05), analysis of mean BMI, eGFR, plasma lipids, and lipoprotein showed no significant differences. Analysis of the level of fasting glucose and HOMA-β showed no statistical differences between the two groups. On the other hand, the levels of C-peptide of PsO group were significantly high in both fasting state (P=0.0182< 0.05) and after glucose challenge (P=0.0011< 0.01).Conclusion: The findings of this study show that under the same fasting conditions, patients with PsO may have relatively preserved pancreatic β-cell function, and PsO significantly increases IR.Keywords: psoriasis, type 2 diabetes mellitus, IR, β-cell function

As a member of the MicroRNA s (miRNAs) family, miR-421 has been widely studied in regulating the proliferation and apoptosis of cancer cells a. However, there are still no reports on miR-421 in regulating adipocyte differentiation and its related mechanisms. Accordingly, this study aimed to investigate the potential involvement of miR-421 in goat intramuscular preadipocytes (P_IMA). The expression level of miR-421 was measured via quantitative real-time PCR during goat P_IMA differentiation. And the effects of miR-421 on goat P_IMA differentiation were studied by liposome transfection, Oil red O staining and qRT-PCR. Furthermore, the miR-421 target was searched and the underlying mechanism was clarified by luciferase reporter assay and rescue experiment. Our results showed that inhibition of miR-421 could accumulation of lipid droplets by upregulation the expression level of AP2, LPL, C/EBPα and SREBP1. Further studies showed that fibroblast growth factor 13 (FGF13) was the direct target of miR-421. Knocking down of FGF13 expression could inhibit lipid droplet formation and down-regulated the expression of key adipogenic regulatory genes. In addition, the rescue experiment revealed that FGF13 is involved in miR-421-induced differentiation of goat P_IMA as a key factor. Overall, these findings indicate that miR-421 is a negative regulator in the progression of differentiation of goat P_IMA by inhibiting the expression of FGF13.

The learning analytics dashboard (LAD) has been recognised as a useful tool to facilitate teachers’ diagnosis and intervention in collaborative learning. However, little empirical evidence has been...

Aim ANGPTL8 is a cytokine expressed and secreted by liver and adipose tissue, and is involved in glucose, lipid, and energy metabolism. Although studies have shown that ANGPTL8 is elevated in type 2 diabetes mellitus (T2DM) and cardiovascular disease, few have examined the association between ANGPTL8 single-nucleotide polymorphisms and the risk of macrovascular complications in T2DM patients. This study aimed to explore the relationship between rs2278426 and carotid intima-media thickening (cIMT) in T2DM. Methods A total of 217 T2DM patients and 201 healthy control subjects with normal glucose tolerance were recruited in the study. T2DM patients were divided into two groups: T2DM patients without cIM thickening (cIMT

Aim We detected whether serum asprosin levels play a role in the occurrence and development of albuminuria in patients with type 2 diabetes mellitus (T2DM), which has not been previously discussed. Methods Based on urinary albumin/creatinine ratio (UACR), 207 T2DM patients were divided into T2DM patients with normoalbuminuria (UACR

Background Lysine demethylase 3A (KDM3A) has been increasingly recognized as an important epigenetic regulator involved in cancer development. This study aims to explore the relevance of KDM3A to cervical cancer (CC) progression and the molecules involved. Materials and methods Tumor and the adjacent tissues from CC patients were collected. KDM3A expression in tissues and CC cell lines and its correlation with the survival and prognosis of patients were determined. Malignant potentials of CC cells and the angiogenesis ability of HUVECs were measured to evaluate the function of KDM3A on CC progression. The interactions among KDM3A, H3K9me2 and ETS1, and the binding between ETS1 and KIF14 were validated through ChIP and luciferase assays. Altered expression of ETS1 and KIF14 was introduced to explore their roles in CC development. Results KDM3A was abundantly expressed in CC tissues and cells and linked to dismal prognosis of CC patients. Knockdown of KDM3A suppressed malignant behaviors of CC cells. KDM3A was found to increase ETS1 expression through the demethylation of H3K9me2. Overexpression of ETS1 blocked the inhibiting roles of sh-KDM3A. ETS1 could bind to the promoter region of KIF14 to trigger its transcription. Overexpression ofKIF14aggravated the malignant behaviors of CC cells and the angiogenesis ability of HUVECs, and it activated the Hedgehog signaling pathway. Artificial activation of Hedgehog by Sag1.5 diminished the effects of sh-KDM3A. These changes were reproduced in vivo. Conclusion This study evidenced that KDM3A promotes ETS1-mediated KIF14 transcription to promote CC progression with the involvement of the Hedgehog activation.

This pilot study aimed to potentially increase the consumption of cancer-preventative sulforaphane, available in broccoli, by introducing this vegetable in novel kimchi products. A panel of mostly ...

The demand for large-scale assessments in higher education, especially at an international scale, is growing. A major challenge of conducting these assessments, however, is that they require unders...

To attenuate positional errors of LiDAR-derived datasets for constructing digital elevation models (DEMs), a feature-preserving point denoising algorithm (F-PDA) is developed in this paper. F-PDA i...

Supply networks (SN) must maintain operations and connectedness under disruptions to remain competitive; this is referred to as SN resilience. Building a resilient SN is an underlying challenge in ...

Based on the knowledge-based view, this paper examines how firms can achieve an enhanced new product performance from the absorptive capacity under technological turbulence. By examining 165 Chines...

Manglietia yuyuanensis is an important afforestation and excellent broad-leaved tree species in southern China. In this study, we assembled the complete chloroplast genome of M. yuyuanensis based o...

In order to prevent spontaneous combustion of coal effectively and conveniently and achieve the simultaneous control of two main factors causing coal spontaneous combustion, namely, coal and oxygen...

In recent years, there has been an increasing interest among language educators in implementing and researching collaborative learning in wiki-based learning environments. However, little empirical...

We performed this study to observe the effects of different doses of ticagrelor and standard-dose clopidogrel on platelet reactivity and endothelial function in diabetic patients with stable coronary artery disease (CAD). Sixty type 2 diabetic patients were assigned to one-quarter standard-dose ticagrelor, half standard-dose ticagrelor, standard-dose ticagrelor and standard-dose clopidogrel groups. Light transmission aggregometry (LTA) and VerifyNow assay were used to measure platelet function. Endothelial function was assessed by measurement of flow-mediated vasodilation (FMD) and plasma von Willebrand factor (VWF) levels were detected. Enzyme-linked immunosorbent assay (ELISA) examined the Interleukin-8(IL-8) and IL-10. The results suggested that the one-quarter dose (34.0%± 14.7%), half-dose (26.9%± 11.6%) and standard-dose (17.3%± 10.3%) ticagrelor showed lower platelet aggregation rate than clopidogrel (52.8%± 18.3%; P ＜0.0001). PRU values in three ticagrelor groups were lower than that in cl...

This letter introduces an efficient bundle adjustment (BA) method based on the sparse Broyden-Fletcher-Goldfarb-Shanno (sparse BFGS, sBFGS) solution, which efficiently estimates camera poses and 3-...

Osteosarcoma (OS) is the most prevalent bone malignancy in childhood and adolescence, with highly aggressive and early systemic metastases. Here, we reported that celecoxib, a selective COX-2 inhibitor in the NSAID class, exhibits strong antitumor activity in dose dependent manner in two OS cell lines-143B and U2OS. We showed that celecoxib inhibits OS cell growth, causes G0/G1-phase arrest, modulates apoptosis and autophagy and reduces migration in OS cells. In addition, the results of fluorescent mitochondrial probe JC-1 test indicated that the mitochondrial pathway mediates celecoxib-induced apoptosis. Significantly, the autophagy inhibitor CQ combined with celecoxib causes greater cell proliferation inhibition and apoptosis. Pharmacologic inhibition of autophagy with another potent autophagy inhibitor SAR405 also enhances celecoxib-mediated suppression of cell viability. These results were confirmed with shRNAs targeting the autophagy-related gene Atg5. In OS tumor xenografts in vivo, celecoxib also presents antitumor activity. Taken together, our results shed light on the function and mechanism of antitumor action of celecoxib for treatment of OS patients.

Berchemiella wilsonii (Rhamnaceae) is an endangered endemic tree to China. Here, the chloroplast genome was characterized by next-generation Illumina paired-end sequencing. The complete genome comprised 160,076 bp in length, with an inverted repeat region of 26,514 bp, a small single copy (SSC) region of 18,717 bp, and a large single copy region of 88,331 bp. A total of 132 genes were identified, including 83 protein-coding genes, 37 transfer RNA genes, and eight rRNA genes. The overall GC content of the whole genome was 37.23%. Phylogenetic analysis showed that B. wilsonii was most closely related to Berchemiella berchemiifolia.

Zhanzhan Li,1 Yanyan Li,2 Shipeng Yan,3 Jun Fu,1 Qin Zhou,1 Xinqiong Huang,1 Liangfang Shen1 1Department of Oncology, Xiangya Hospital, Central South University, Changsha, China; 2Department of Nursing, Xiangya Hospital, Central South University, Changsha, China; 3Office of Tumor Prevention and Control, Hunan Provincial Tumor Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, China Background: The potential benefits and possible risks associated with combined nimotuzumab and concurrent chemoradiotherapy in patients with advanced nasopharyngeal carcinoma (NPC) have yet to be determined.Methods: The databases PubMed, Web of Science, China National Knowledge Infrastructure, and Wanfang were systematically searched through February 2017 for studies comparing combined nimotuzumab and chemoradiotherapy versus chemoradiotherapy alone in the treatment of NPC. Primary outcomes were complete and partial responses, and the secondary outcome was adverse reactions. The random-effect model was used to pool relative risks (RRs) and 95% confidence intervals (CIs).Results: Nine randomized control trials and six cohort studies were included in the final analysis (n=1,015 patients). Compared with chemoradiotherapy alone, chemoradiotherapy combined with nimotuzumab was associated with an increased response rate (RR=1.11, 95% CI: 1.01–1.22). Combined treatment further reduced the occurrence rate of erythropenia (RR=0.11, 95% CI: 0.05–0.28) and neutropenia (RR=0.12, 95% CI: 0.05–0.27). The differences in the rates of other complications were not significant.Conclusion: Nimotuzumab combined with concurrent chemoradiotherapy is more effective in patients with advanced NPC than chemoradiotherapy alone. Patients receiving combination therapy did not have a higher rate of adverse reactions. Nimotuzumab can thus be recommended as an adjunct therapy in patients with advanced NPC.Keywords: randomized controlled trial, cohort studies, nasopharyngeal carcinoma, relative risk, radiotherapy, chemoradiotherapy

The classification of airborne laser scanning ALS point clouds is a necessary step in the process of digital elevation model DEM construction. Motivated by the surface interpolation filtering scheme, a multi-resolution hierarchical classification MHC method was developed to perform classification in our previous research. However, similar to almost all filtering methods, MHC suffers from the problem of misclassifying off-terrain and ground points in complex landscapes with steep slopes and large discontinuities. To improve the classification accuracy in the above landscapes, the original MHC is improved in this article by adopting a region growing-based segmentation method to maximally capture the initial ground seeds of MHC. Furthermore, considering the high sensitivity of principle component analysis PCA to outliers, a robust PCA approach based on least trimmed squares LTS is proposed to estimate point normals. Fifteen groups of data sets provided by the International Society for Photogrammetry and Remote Sensing ISPRS Working Group III/3 are utilized to compare the performances of the improved and original MHCs. Results indicate that the improved MHC is more accurate than the original MHC, irrespective of accuracy measures. In addition, we also compare the results of our method with three new filtering methods developed in the years 2013 and 2014. It is found that regardless of landscapes i.e. urban and rural samples, the improved MHC is averagely more accurate than the three filtering methods in terms of type II error, and irrespective of accuracy measures i.e. types I, II and total errors, our method outperforms the three filtering methods for the rural samples. In short, the robust segmentation-based MHC demonstrates great potential to ALS-derived DEM construction in rural areas.

The complete chloroplast genome of Picea neoveitchii was assembled in this study. The genome comprised 124,234 bp in length. The GC content was 38.77%. A total of 116 genes are successfully annotat...

A role for transforming growth factor-β1gene has been suggested in the etiology of IgA nephropathy. However, results have been inconsistent. In this study, a meta-analysis was performed to further clarify the association between transforming growth factor-β1-509C/T gene polymorphism and the susceptibility of IgA nephropathy. PubMed, EMBASE, Web of Science, CNKI, WanFang, and VIP Data were searched for eligible studies. Pooled odds ratios (ORs) with 95% confidence intervals were calculated using a fixed-effects model or random-effects model. A total of eight publications involving 1355 IgA nephropathy patients and 1464 controls met the inclusion and were analyzed. The pooled ORs for the association between TGF-β1gene-509C/T polymorphism and IgA nephropathy risk were not statistically significant under all genetic models (for CT+TT vs. CC: OR = 1.09; 95% CI = 0.92-1.29, p = 0.490; for TT vs. CT+CC: OR = 1.14; 95% CI = 0.94-1.38, p = 0.081; for CC vs. TT: OR = 0.87; 95% CI = 0.69-1.08, p = 0.195; for C allele vs. T allele: OR = 0.92; 95% CI = 0.83-1.03, p = 0.149). In the stratified analysis by ethnicity, results also showed no significant association between TGF-β1 gene-509C/T polymorphism and IgA nephropathy risk in both European and Asian populations. This meta-analysis does not support the hypothesis that TGF-β1 gene-509C/T polymorphism is a risk factor for the development of IgA nephropathy.

An ethyl acetate-assisted ionic liquid dispersive liquid–liquid microextraction method was developed for the determination of tetracyclines in water samples with high-performance liquid chromatography using a variable wavelength detector. A mixture of ethyl acetate and 1-butyl-3-methylimidazolium hexafluorophosphate was used to isolate lanthanum(III) chelates of tetracyclines. The addition of ethyl acetate significantly enhanced the extraction efficiencies. Experimental parameters affecting the efficiency such as the identity and volume of ionic liquid, the volume of ethyl acetate, chelating time, centrifuge time, pH, and ionic strength were evaluated. Under the optimum conditions, the calibration curves were linear and the correlation coefficients (r 2 ) ranged from 0.9985 to 0.9992 over concentration levels of 0.01–0.50 µg mL−1. The limits of detection (signal-to-noise ratio = 3) of the method ranged from 0.97 to 1.14 ng mL−1, whereas inter- and intra-day relative standard deviations (n = 5) were betwee...

A novel, practical, and environmentally friendly technique, termed concentrated sulfuric acid cleanup and ultrasound-assisted surfactant-enhanced emulsification microextraction (CSAC-UASEM), was combined with HPLC for the preconcentration and determination of five phenolics in water and Arthrospira (Spirulina) platensis samples. The main advantages include that the concentrated sulfuric acid is used to decrease macromolecular interferences prior to microextraction and, unlike dispersive liquid–liquid microextraction procedures, no dispersive organic solvent is required. Chloroform and sodium dodecyl sulfate were used as the extraction solvent and emulsifier, respectively. The algal cell preparation and CSAC-UASEM procedure parameters, including selection of cleanup method, ultrasound power, cell cytocylasis time, type and volume of extraction solvent, extraction temperature, ultrasound-extracted time, and sample pH, were optimized. At the fortification levels of 1.0 and 10.0 µg/L, the enrichment factors o...

A sequential adjustment (SA) method is employed to decrease the computational cost of high-accuracy surface modeling (HASM), and the SA of HASM (HASM-SA) is being developed. A mathematical surface was used to comparatively analyze the computing speed of SA and the classical iterative solvers provided by MATLAB 7.7 for solving the system of linear equations of HASM. Results indicate that SA is much faster than the classical iterative solvers. The computing time of HASM-SA is determined by not only the total number of grid cells but also the number of sampling points in the computational domain. A real-world example of surface modeling of digital elevation models (DEMs) with various resolutions shows that HASM-SA is averagely more accurate and much faster than the commonly used interpolation methods, such as inverse distance weighting (IDW), kriging, and three versions of spline, namely regularized spline (RSpline), thin-plate spline (TPS), and ANUDEM in terms of root mean square error (RMSE), mean absolute...

Low testosterone levels may be a signal of poor health. This study aimed to investigate the effects of age and abnormal metabolism on sex hormones in Chinese male.Three hundred and thirty-seven elder men were enrolled into this single-center, cross-sectional study, and their sex hormone levels and metabolic parameters were assessed.Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and sex-hormone-binding globulin (SHBG) concentrations increased with age, while free testosterone index (FTI), testosterone secretion index (TSI), estradiol (E2)/SHBG and progestin (PROG) decreased. Abnormal metabolisms were related to androgen indices (TT, FT, BT, FTI, TSI, T/E2), SHBG and E2/SHBG even after adjusting by age and macrovascular disease. Obesity and overweight, hyperglycemia and dyslipidemia were the most important abnormal metabolism that related to decreased androgen indices. Including SHBG in the stepwise regression increased the explanation effect of TT and BT by 32.7% and 28.5%, respectively, and all metabolic indices were excluded. Abnormal metabolism indies (BMI and PBG) were correlated to the decrease in SHBG levels, while age and LH was positively correlated to SHBG levels.Age and abnormal metabolism were independently important factors associated with the sex hormone levels in elderly Chinese men, which were all mediated by SHBG.

This study was to investigate whether polymerized human placenta hemoglobin (PolyPHb) given before ischemia protects in vivo rat heart function against ischemia/reperfusion (I/R) injury. Forty-five male Sprague-Dawley rats were randomly divided (n = 15 per group) into a sham group, control group (pretreatment with Lactated Ringer's solution), or PolyPHb group (pretreatment with 0.1 gHb/kg PolyPHb). Rat hearts were subjected to 30-min ischemia by occlusion of left anterior descending, followed by 2-hr reperfusion. As compared to the control group, PolyPHb preserved cardiac function and reduced cardiac troponin-I release and histopathological changes. Therefore, PolyPHb pretreatment provided a profound cardioprotective effect on the in vivo rat heart.

Heat shock protein 90 (Hsp90), an essential molecular chaperone that regulates the stability of a wide range of oncogenic proteins, is a promising target for cancer therapeutics. We investigated the combination efficacy and potential mechanisms of sulforaphane, a dietary component from broccoli and broccoli sprouts, and 17-allylamino 17-demethoxygeldanamycin (17-AAG), an Hsp90 inhibitor, in pancreatic cancer. MTS assay demonstrated that sulforaphane sensitized pancreatic cancer cells to 17-AAG in vitro. Caspase-3 was activated to 6.4-fold in response to simultaneous treatment with sulforaphane and 17-AAG, whereas 17-AAG alone induced caspase-3 activity to 2-fold compared to control. ATP binding assay and coimmunoprecipitation revealed that sulforaphane disrupted Hsp90-p50(Cdc37) interaction, whereas 17-AAG inhibited ATP binding to Hsp90. Concomitant use of sulforaphane and 17-AAG synergistically downregulated Hsp90 client proteins in Mia Paca-2 cells. Co-administration of sulforaphane and 17-AAG in pancreatic cancer xenograft model led to more than 70% inhibition of the tumor growth, whereas 17-AAG alone only suppressed the tumor growth by 50%. Our data suggest that sulforaphane potentiates the efficacy of 17-AAG against pancreatic cancer through enhanced abrogation of Hsp90 function. These findings provide a rationale for further evaluation of broccoli/broccoli sprout preparations combined with 17-AAG for better efficacy and lower dose-limiting toxicity in pancreatic cancer.

A MME-HPLC (UV) method was developed for the determination of chlorogenic acid, rutin, and quercetin; nonionic surfactant Triton X-100 was used to extract and preconcentrate active ingredients from Honeysuckle prior to their determination by HPLC-UV. The experimental conditions were optimized by a L16 (45) orthogonal array design (OAD) with five factors such as concentration and kind of TX, liquid/solid ratio, extraction temperature, and time at four levels using statistical analysis. Under optimum conditions, that is, 5% Triton X-100 (w/v), liquid/solid ratio of 400:1 (mL · g−1), ultrasonic-assisted extraction for 30 min at 30°C, the extraction content of the active ingredients reached the highest value. When compared with commonly used solvents, 5% Triton X-100 yielded, in a way, higher than the extraction efficiency of methanol. For the preconcentration of compounds by cloud-point extraction, sodium chloride was added to the solution to facilitate the phase separation. The preconcentration factor for e...

The aim of this study was to use silica nanoparticles as the carrier for controlled release of tilmicosin. Tilmicosin was selected as a drug model molecule because it has a lengthy elimination half-life and a high concentration in milk after subcutaneous administration. Three samples of tilmicosin-loaded silica nanoparticles were prepared with different drug-loading weight. The drug-loading weight in three samples, as measured by thermal gravimetric analysis, was 29%, 42%, and 64%, respectively. With increased drug-loading weight, the average diameter of the drug-loaded silica nanoparticles was increased from 13.4 to 25.7 nm, and the zeta potential changed from-30.62 to-6.78 mV, indicating that the stability of the drug-loaded particles in the aqueous solution decreases as drug-loading weight increases. In vitro release studies in phosphate-buffered saline showed the sample with 29% drug loading had a slow and sustained drug release, reaching 44% after 72 h. The release rate rose with increased drug-loading weight; therefore, the release of tilmicosin from silica nanoparticles was well-controlled by adjusting the drug loading. Finally, kinetics analysis suggested that drug released from silica nanoparticles was mainly a diffusion-controlled process.

In this paper, we study the perfect and the insulated conductivity problems with multiple inclusions imbedded in a bounded domain in $\mathbb{R}^n, n\ge 2$. For these two extreme cases of the conductivity problems, the gradients of their solutions may blow up as two inclusions approach each other. We establish the gradient estimates for the perfect conductivity problems and an upper bound of the gradients for the insulated conductivity problems in terms of the distances between any two closely spaced inclusions.

Two liquid-phase microextraction (LPME) approaches, static direct-immersed single-drop microextraction (DI-SDME) and continuous-flow microextraction (CFME), were used to extract methomyl in water samples and their respective extraction efficiencies were compared. Several important parameters affecting extraction efficiency such as the type of extraction solvent, solvent drop volume, stirring speed or flow rate, extraction time and salt concentration were optimised. The optimised conditions were as follows: 3.0-µL tetrachloroethane (C2H2Cl4) as the extraction solvent, 15% NaCl (w/v), 15 min extraction time and stirring speed at 600 rpm for DI-SDME; 3.5-µL C2H2Cl4 as the extraction solvent, 15% NaCl (w/v), 21 min extraction time and flowing rate at 0.8 mL min−1 for CFME. Under the previous optimal conditions, the linear range, detection limit (S/N = 3) and precision (RSD, n = 6) were 5.0-5000 ng mL−1, 1.5 ng mL−1, 6.9% for DI-SDME, and 4.0–10000 ng mL−1, 2.5 ng mL−1, 4.6% for CFME, respectively. Lake and ri...

The effects of two farm manures, bovine manure (BM) and chicken manure (CM), on imazaquin degradation in a sandy loam soil were studied under laboratory conditions. The degradation of imazaquin in BM and CM-amended soil (10% by w/w) was about 2.4 and 1.5 times, respectively, faster than that in unamended soil. Different amendment ratios of BM to soil lead to different degradation rates of imazaquin. However, higher mix ratio did not necessarily increase the degradation, and the optimal active degradation of imazaquin was observed at the approximate mix ratio of 10:1 of soil to BM. Imazaquin degradation rate was enhanced 2∼ 4 times in non-sterilized soil over that in SGI (sterilization by gamma irradiation) soil. In non-sterilized soil, biological mechanisms accounted for 48.7% to 74.6% of imazaquin degradation, which suggested that microbial transformation was the primary pathway for imazaquin degradation in soil. The degradation rate of imazaquin for SSA (sterilization by sodium azide) treatment was 1.33...

The ability of microbes to adapt in response to an environmental change, such as the infection of a warm-blooded host is an important mechanism in pathogenesis. One major mechanism by which microbes respond to an environmental change is through the modification or remodeling of their membranes. Lipid A, or endotoxin, the bioactive component of lipopolysaccharide (LPS) is the major component of the outer leaflet of the outer membrane of Gram-negative bacteria. Lipid A, the membrane anchor of LPS is a β-(1′-6)-linked glucosamine disaccharide backbone with amide linked fatty acids at the 2 and 2′ positions, ester linked fatty acids at the 3 and 3′ positions and terminal phosphate moieties at the 1 and 4′ positions. Early steps in lipid A synthesis are conserved across Gram-negative species (Raetz pathway) though species-specific structural modifications are also observed. Modification of lipid A in response to environmental stressors such as pH, cation concentration, oxygen saturation and osmolarity have been shown in a number of bacterial species to contribute to condition-specific membrane phenotypes. Temperature has also been shown to play an important role in lipid A structure supported by the presence of specific genes for growth at high temperatures in Escherichia coli (htrB/lpxM) and temperature dependent hypoacylation in Yersinia pestis. The ability of bacterial species to alter or modify their lipid A in response to specific environmental cues is important for pathogenesis. In our recent manuscript, we demonstrated the importance of temperature as an environmental cue in outer membrane remodeling of the intracellular pathogen, Francisella novicida. Francisella has been identified in many environmental niches and a wide array of hosts and vectors including humans, lagomorphs, arthropods and amoebae. Throughout its complex and varied life cycle, Francisella adapts to a range of temperatures that alter the composition of its lipid A. Specifically, after growth at lower temperatures normally encountered in arthropods or water environments (~25°C), Francisella synthesizes a lipid A with predominately shorter fatty acids (16 carbons in length). At higher temperatures, like those encountered in warm-blooded hosts (37°C), lipid A is composed primarily of longer chain fatty acids (18 carbons in length). Recent work from our lab determined this shift in lipid A fatty acid composition was regulated at the genetic and enzymatic level utilizing two N-linked acyltransferases, LpxD1 and LpxD2 (Fig. 1). Figure 1. Overview of lpxD control of pathogenesis. Francisella has a unique ability to adapt upon entering various environmental niches. Of particular interest is the activity of the acyltransferases LpxD1 and LpxD2. Upon entry of warm temperatures ... lpxD is conserved in most Gram-negative organisms and is present in an operon containing the essential genes fabZ, lpxB and lpxA. LpxD adds N-linked fatty acids to the glucosamine backbone at the “lipid X” stage present on the inner leaflet of the inner membrane and prior to MsbB transport to the outer leaflet of the inner membrane. Activity of LpxD has been shown to be required for bacterial viability. Interestingly, sequence comparisons of lpxD1 and lpxD2 from Francisella show low amino acid sequence identity (34%) suggesting distinct genetic origins, as opposed to gene duplication. lpxD1 is similar to the lpxDs found in enteric bacteria, whereas lpxD2 is more closely related to lpxDs from anaerobic bacteria, such as Desulfovibrio and Fusobacterium. The close relationship to the anaerobic species is striking, as Francisella is a strict aerobe. lpxD1 and lpxD2 are highly conserved across all Francisella subspecies (> 98%) indicating a critical role in remodeling the bacterial membrane. Also, the duplication of lpxD in the relatively small genome of Francisella (1,782 genes) highlights a potentially critical role for both genes in bacterial virulence. Regulation of both lpxD1 and lpxD2 is tightly controlled in Francisella. Microarray data showed that lpxD1 was upregulated at 37°C, in contrast to lpxD2 which was upregulated at 25°C. Not only was the expression of these genes temperature regulated, but their abilities to catalyze acylation was also linked to temperature. The LpxD1 enzyme catalyzed the addition of longer chain acyl groups at 37°C, while the LpxD2 enzyme added shorter chain acyl groups at 25°C. Genetic deletion of these genes allowed further insight into their role in pathogenesis, leading to distinct phenotypes. Deletion of lpxD1 resulted in a smaller lipid A structure similar to that seen after growth of wild-type Francisella at lower temperatures, whereas deletion of lpxD2 “locked” the lipid A into a larger configuration similar to wild-type samples grown at warm temperatures. These mutants, having different lipid A phenotypes and lacking the ability to alter their membrane composition, were tested in a variety of assays to determine their role in virulence. Francisella is lethal to mice at extremely low doses (LD100 ~10 CFU) regardless of infection route. To assess if the overall lethality of the mutants was affected, mice were infected with increasing amounts of each mutant, as well as WT Francisella. The ΔlpxD2 mutant maintained full virulence at amounts similar to that of the WT strain. Interestingly, the ΔlpxD1 mutant, which is locked in the smaller, cold phenotype, showed a total lack of virulence even at the highest doses tested (5 × 106 CFU, 500,000 × LD100). As the ΔlpxD1 mutant was completely attenuated after a single primary infection dose, we sought to determine if the ΔlpxD1 mutant could induce protective immunity against a lethal wild-type challenge. Our results showed that protection was dependent on the primary infection dose. When mice were vaccinated using a prime-boost model they were completely protected, regardless of the immunizing dose. To determine if specific membrane alterations were responsible for attenuation in the lpxD mutant strains, a variety of lipid A/membrane functions were examined. As is seen in lipid A isolated from WT strains of Francisella, neither mutant produced a lipid A that resulted in NFκB-mediated proinflammatory responses through TLR4, suggesting that the attenuation was not due to increased recognition by the host innate immune sensors. Another function of lipid A is to resist killing by host peptides by modulating both the charge and permeability of the outer membrane. The ΔlpxD2 mutant, with the larger lipid A was shown to be more resistant to cationic antimicrobial peptides (CAMPs) presumably due to decreased permeability in the outer membrane when compared with WT Francisella. Decreased permeability was confirmed by measuring the uptake of ethidium bromide into cells. Conversely, the ΔlpxD1 mutant showed increased permeability and sensitivity to CAMPs. Temperature regulation of virulence factors is common in many bacterial species. Generally, temperature regulates the production of classical virulence factors such as toxins in C. difficile, or master regulators of virulence, such as prfA in L. monocytogenes. Here we have shown that a single, temperature regulated, non-classical virulence gene is important for pathogenesis of Francisella. The inability to alter its lipid A and thus remodel its membrane upon infection of warm-blooded hosts is an important mechanism in the overall pathogenesis of Francisella. Though the loss of lpxD2 showed no effect on murine pathogenesis, it may be required for growth in the environment or arthropod vectors. These questions are currently being pursued in alternative models. The importance of maintaining proper membrane function and organization upon adaptation to varied environmental conditions is essential. In Francisella, the tight control of lipid A biosynthetic enzymes occurring at multiple levels; transcriptional and enzymatic mark a critical role in pathogenesis. Controlled regulation of the lpxD1 gene potentially works as a “virulence switch” and alters not only the membrane permeability and resistance to host defenses but plays a critical role in turning on or off pathogenesis. The consequences of this seemingly simple acyl-chain alteration, addition of two carbons to lipid A remain to be explored. Additionally modulation of lipid A structure has a profound impact on the protein constituency in the outer membrane, suggesting a role for specific outer-membrane proteins to be present or absent during pathogenesis. This new cadre of membrane proteins may have far-reaching effects on signaling both from extracellular stimuli and other partners in the membrane. These initial observations in Francisella invite examination of the temperature controls placed on lipid A in other bacterial species that are found at multiple temperatures and how these may affect virulence.

We introduce, along the lines of [4], an integer valued degree for second order fully nonlinear elliptic operators which is invariant under homotopy within elliptic operators. We also give some applications to the bifurcation problem for nonlinear elliptic equations. Applications to the existence of solutions of certain fully nonlinear elliptic equations on compact manifolds can be found in [7].