High purity of human secreted phospholipase A2 group IIE in Pichia pastoris using basal salts medium comparison with YPD medium.

Secreted phospholipase A2s (sPLA2s) are a group of enzymes with 6-8 disulfide bonds that participate in numerous physiological processes by catalyzing the hydrolysis of phospholipids at the sn-2 position. Due to their high content of disulfide bonds and hydrolytic activity toward cell membranes, obtaining the protein of sPLA2s in the soluble and active form is challenging, which hampers their functional study. In this study, one member of recombinant human sPLA2s, tag-free group IIE (GIIE), was expressed in Pichia pastoris. The protein GIIE was purified from the crude culture supernatant by a two-step chromatography procedure, a combination of cation exchange and size-exclusion chromatography. In the shake flask fermentation, Protein of GIIE with higher purity was successfully obtained, using basal salts medium (BSM) instead of YPD medium. In the large-scale fermentation, each liter of BSM produced a final yield of 1.2 mg pure protein GIIE. This protocol will facilitate further research of GIIE and provide references for the production of other sPLA2 members.

Variations in the structural and functional properties of flaxseed gum from six different flaxseed cultivars.

Although flaxseed gum (FG) has been widely studied, the differences in its structure and function with respect to various flaxseed cultivars remain unclear. In this study, our objective was to examine the differences between FG samples obtained from different flaxseed cultivars based on their structural and functional properties. Specifically, FG samples from the different cultivars were extracted via hot water extraction followed by ethanol precipitation. Thereafter, they were analyzed via zeta potential measurements, Fourier-transform infrared (FTIR) spectroscopy, X-ray diffractometry (XRD), and scanning electron microscopy (SEM). The results demonstrated that the different cultivars showed significantly different FG yields (p < .05; range, 5.83%-7.36%). Further, the FTIR spectra of the FG samples were slightly different but showed typical polysaccharide absorption peaks. Furthermore, the XRD patterns obtained predominantly showed an amorphous region and a small crystalline region, while the SEM images obtained at 1,000× magnification revealed that the samples had smooth and irregular surfaces, with a scaly structure. However, at 20,000× magnification, the FG samples showed slight structural differences. Additionally, the zeta potentials of the FG samples (range, -19.4 to -30.6 mV; p < .05) were cultivar-dependent and indicated the presence of negatively charged macromolecules. This implies that the FG samples from the different cultivars show diverse structural properties. Our findings not only provide useful information regarding FG samples extracted from different cultivars but also serve as a theoretical basis for the application of FG in food processing.

Quantifying the response of surface urban heat island to urban greening in global north megacities.

Urban heat island, a phenomenon that urban temperature is higher than the rural area nearby, affects directly citizens' human health and well-being. However, the cooling effect from urban green space (UGS) and the attribution of the different land processes to surface urban heat island intensity (SUHI) under different background climates remains unclear. The coarse-grained model was used to estimate summer SUHI in three different background climatic zones and for seven agglomerations (BTH, JP, LD, NAAC, NAGL, YZ, UQ). Results indicate that (1) the temperate zone had the highest daytime SUHI (0-10 °C), while the arid zone has the lowest daytime SUHI (-1-2 °C). In both temperate and cold zone, the daytime SUHI was higher than the nighttime SUHI. The SUHI in downtown was higher (more than 2 °C) than in the suburbs. (2) The increasing precipitation can enhance daytime SUHI while can weaken nighttime SUHI in all three climatic zones. The increasing temperature tends to enhance SUHI in both daytime and nighttime (exclude UQ). (3) The cooling effects of UGS in daytime SUHI were highly dependent on the background climate (cold > temperate > arid). (4) The nighttime SUHI could be effectively offset when UGSFs were greater than 0.48, 0.82, 0.97, 0.95 in NAAC, NAGL, YZ, and UQ. This article highlights the different feedback of urban green space to UHII and supports green infrastructure intervention as an effective means of reducing urban heat stress at urban agglomeration scales.

Effects of mobile phone application combined with or without self-monitoring of blood glucose on glycemic control in patients with diabetes: A randomized controlled trial.

AIMS/INTRODUCTION: There is potential for mobile applications to deliver new self-management interventions for chronic disease, especially in diabetes. The aim of the present study was to evaluate the effects of a mobile phone application (MPA) combined with or without self-monitoring of blood glucose (SMBG) on glycemic control in patients with diabetes. MATERIALS AND METHODS: The study was a 24-week period, four-arm parallel group, non-blinded, randomized trial. A total of 185 patients with mean age of 52 years were randomized to group A (no MPA and no SMBG), group B (SMBG only), group C (MPA only) and group D (both MPA and SMBG were used). Changes in glycated hemoglobin (HbA1c), fasting plasma glucose and 1,5-anhydroglucitol from baseline to week 24 were analyzed. RESULTS: At 24 weeks, the HbA1c levels in patients of all groups decreased significantly from baseline. There were significant differences in the proportions of patients that achieved HbA1c <7% between groups, especially in group C and group D, compared with group A at week 24 (60.4%, 62.2% vs 25.5%, all P < 0.05). 1,5-Anhydroglucitol changes were obvious in group A and group C at week 24 from baseline (all P < 0.05 within groups). Factorial analysis of anova showed that MPA intervention was the main effective factor for HbA1c change (F = 4.59, P = 0.034), and there was no effect on HbA1c change for SMBG intervention (P = 0.975). CONCLUSIONS: Implementation of the MPA, Diabetes-Carer, is effective in improving the proportion of HbA1c <7% in patients with type 2 diabetes.

Effect of Cognitive Behavioral Therapy on Improving the Cognitive Function in Major and Minor Depression.

The aim of this study was to investigate the effectiveness of cognitive behavioral therapy (CBT) on improving the cognitive function in minor depression (MiD) and major depression (MaD). The study will constitute a placebo-controlled single-blind parallel-group randomized controlled trial. The selected participants will be randomly allocated into one of two parallel groups with a 1:1 ratio: the CBT-based group and the general health education group. CBT significantly alleviated depressive symptoms of MiD and MaD at 12 weeks (p < 0.001), and the treatment effect was maintained for at least 12 months (p < 0.001). Interestingly, CBT significantly promotes more cognitive function of MiD and partial cognitive function of MaD at 12 weeks in the intervention group than in the control group (p < 0.01). CBT can alleviate depressive symptoms of both minor and MaDs. The effectiveness of CBT is different on improving the cognitive function in MiD and MaD.

FBP2 and Talin-1 are potential protein markers for Mongolian medicine symptom evaluation in viral infectious diseases.

BACKGROUND: Influenza, measles, and mumps are common viral infectious diseases in Mongolia. The traditional Mongolian medicine (TMM) classified them as warm disease, and still plays a major role in the diagnoses and treatments. METHODS: To interpret the connotation of the complex theoretical system in TMM with scientific technique, in this study, a high throughput mass spectrometry was used to identify potential protein markers of TMM symptom types. Fifty venous blood samples were drawn from influenza, measles and mumps patients. Differential proteins between samples of patients diagnosed as immature and mature heat in TMM were detected by mass spectrometry. RESULTS: After proteomics analysis, 1500 proteins and 7619 polypeptides were identified and 1323 in total showed differential expression between those 2 symptom types; then enrichment analysis of the differentially expressed proteins revealed the significant biological functions related to the differentially expressed proteins, including cardiomyopathy, several bacterial and parasitic infections, bacterial invasion of epithelial cells, insulin signaling pathway, and regulation of actin cytoskeleton. The network analysis showed that FBP2 and Talin-1 were critical points and might determine the evolution directions of TMM warm disease symptom. CONCLUSIONS: This study suggests that the identified core differential proteins may be regarded as potential biomarkers, and benefit to evaluate the evolutionary tendency of TMM warm disease symptoms.

Predictors of neurocognitive impairment at 2years after a first-episode major depressive disorder.

BACKGROUND: Neurocognitive impairment is a contributor to major depressive disorder (MDD). However, MDD patients show great variability in the level and course of deficits. The present longitudinal study was to identify predictors of neurocognitive impairment in first-episode MDD patients. METHODS: Neurocognitive performance was analyzed in a cohort of 100 patients at 2years after a first-episode MDD. Subgroups, deficit type vs. non-deficit type, were compared on baseline clinical, neuropsychological, premorbid and sociodemographic characteristics. The analysis was performed using the multivariate logistic regression to obtain a model for neurocognitive impairment determination. The predicted probabilities of multivariate logistic regression were analyzed using receiver operating characteristic (ROC) curve. RESULTS: Fifty-two percent of MDD participants presented general neurocognitive impairment. The regression analyses demonstrated that clinical and sociodemographic characteristics were not predictive variables. A model composed of processing speed, executive function, and attention, dexterity correctly classified 85.8% of the MDD patients with deficit type. ROC curve indicated that the changes of these three cognitions could identify MDD with deficit type from MDD with non-deficit type. In addition, ROC curve also indicated that processing speed and executive function could identify MDD from CN subjects. Finally, processing speed performance was negatively correlated with Hamilton Depression Scale scores in both MDD with deficit and non-deficit type. CONCLUSION: The present study provides novel insights on frequency and neurocognitive profile of subtypes of patients showing impairment. Our results suggest that processing speed impairment is a trait dimension of the disorder related to specific cognitive dysfunctions and the severity of depression.

Design, synthesis, and structure-activity relationship studies of conformationally restricted mutilin 14-carbamates.

We report herein the design, synthesis, and structure-activity relationship studies of conformationally restricted mutilin 14-carbamates based on the structure of SB-222734. The antibacterial activities of these newly synthesized compounds were also evaluated and compared with linezolid and retapamulin. Results showed that most of the target compounds exhibit good potency in inhibiting the growth of Gram-positive bacteria including Methicillin-susceptible Staphylococcus aureus MSSA (MIC: 0.0625-2µg/mL), Methicillin-resistant S. aureus MRSA (MIC: 0.0625-2µg/mL), Methicillin-susceptible Staphylococcus epidermidis MSSE (MIC: 0.0625-2µg/mL), Methicillin-resistant S. epidermidis MRSE (MIC: 0.0625-2µg/mL), and Streptococcus pneumonia (MIC: 0.0625-4µg/mL). In particular, three remarkable compounds of this series (12l, 12m, and 21l) exhibited comparable in vitro antibacterial profiles to that of retapamulin.

Novel 4-azasteroidal N-glycoside analogues bearing sugar-like D ring: synthesis and anticancer activities.

A series of novel N-glycoside analogues with 4-azasteroid moiety bearing sugar-like D ring were conveniently synthesized by constructing the core dihydropyran ring embedded in 4-azasteroidal skeleton which was prepared from 4-aza-5α-androst-3,17-dione 1 in four steps. The structure of 6b were unambiguously proved by the appropriate X-ray structural analysis. Anticancer activity was found for all of the analogues with purinyl moiety against breast cancer (MCF-7), human neuroblastoma (SK-N-SH), cervical cancer cell (HeLa) and prostatic cancer (PC-3), while the analogue 7 containing 1,2,4-triazole heterocycle as the nucleobase was inactive against all of the tested cancer cell lines. The biology results showed the purinyl moiety attached to the pyran ring of 6a-d, substituent at 6'-position of purine base and introduction of a halogen atom at 2'-position of 6'-chloropurine had obviously effect on the evaluated anticancer activity.

Design, synthesis, and structure-activity relationship studies of highly potent novel benzoxazinyl-oxazolidinone antibacterial agents.

A series of novel benzoxazinyl-oxazolidinones bearing nonaromatic heterocycle or aryl groups were designed and synthesized. Their in vitro and in vivo antibacterial activities were investigated. Most of the (3S, 3aS) biaryl benzoxazinyl-oxazolidinones exhibited potent activity against Gram-positive pathogens. SAR trends were observed; a pyridyl C ring was preferable to other 5- or 6-member aryl C rings, while fluorine substitution on the B ring generated derivatives with reduced activity. Various substituent group positions on the pyridyl ring were also evaluated. The resulting compounds displayed excellent activity against linezolid-resistant strains. Compound 45 exhibited excellent in vitro activity, with a MIC value of 0.25-0.5 µg/mL against MRSA and an activity against linezolid-resistant strains of 8-16-fold higher potency than linezolid. In a MRSA systemic infection model, compound 45 displayed an ED(50) < 5.0 mg/kg, a potency that is nearly 3-fold better than that of linezolid. This compound also showed excellent pharmacokinetic profiles, with a half-life of more than 5 h as well as an oral bioavailability of 81% in rats.

Design and synthesis of pyridine-substituted itraconazole analogues with improved antifungal activities, water solubility and bioavailability.

To improve antifungal activities, water solubility and bioavailability, a series of novel analogues of itraconazole-containing pyridine rings were designed and synthesized. Their antifungal activities were evaluated in vitro against six clinically important fungi by measuring the minimal inhibitory concentrations (MICs). Most of the compounds showed more potent antifungal activities than that of itraconazole. In particular, the analogues 30d, 30c, 31c, and 36d exhibited much higher solubility and bioavailability than that of itraconazole. The bioavailability of 36d (42.2%) was five times higher than that of itraconazole (8%) and was negative for genetic toxicology in the Ames test.

Water-soluble phosphate prodrugs of pleuromutilin analogues with potent in vivo antibacterial activity against Gram-positive pathogens.

A phosphate prodrug strategy was investigated to address the problem of poor aqueous solubility of pleuromutilin analogues. Water-soluble phosphate prodrugs 6a, 6b and 6c of pleuromutilin analogues were designed and synthesized. Three compounds all exhibited excellent aqueous solubility (>50mg/mL) at near-neutral pH and sufficient stability in buffer solution. In particular, the phenol pleuromutilin prodrug 6c displayed favourable pharmacokinetic profiles and comparable potency with vancomycin against MSSA and MRSA strains in vivo.

MCT-1 protein interacts with the cap complex and modulates messenger RNA translational profiles.

MCT-1 is an oncogene that was initially identified in a human T cell lymphoma and has been shown to induce cell proliferation as well as activate survival-related pathways. MCT-1 contains the PUA domain, a recently described RNA-binding domain that is found in several tRNA and rRNA modification enzymes. Here, we established that MCT-1 protein interacts with the cap complex through its PUA domain and recruits the density-regulated protein (DENR/DRP), containing the SUI1 translation initiation domain. Through the use of microarray analysis on polysome-associated mRNAs, we showed that up-regulation of MCT-1 was able to modulate the translation profiles of BCL2L2, TFDP1, MRE11A, cyclin D1, and E2F1 mRNAs, despite equivalent levels of mRNAs in the cytoplasm. Our data establish a role for MCT-1 in translational regulation, and support a linkage between translational control and oncogenesis.

Substitution of the autophosphorylation site Thr516 with a negatively charged residue confers constitutive activity to mouse 3-phosphoinositide-dependent protein kinase-1 in cells.

3-Phosphoinositide-dependent protein kinase-1 (PDK-1)is a serine/threonine kinase that has been found to phosphorylate and activate several members of the AGC protein kinase family including protein kinase B (Akt), p70 S6 kinase, and protein kinase Czeta. However, the mechanism(s) by which PDK-1 is regulated remains unclear. Here we show that mouse PDK-1 (mPDK-1) undergoes autophosphorylation in vitro on both serine and threonine residues. In addition, we have identified Ser(399) and Thr(516) as the major mPDK-1 autophosphorylation sites in vitro. Furthermore, we have found that these two residues, as well as Ser(244) in the activation loop, are phosphorylated in cells and demonstrated that Ser(244) is a major in vivo phosphorylation site. Abolishment of phosphorylation at Ser(244), but not at Ser(399) or Thr(516), led to a significant decrease of mPDK-1 autophosphorylation and kinase activity in vitro, indicating that autophosphorylation at Ser(399) or Thr(516) is not essential for mPDK-1 autokinase activity. However, overexpression of mPDK-1(T516E), but not of mPDK-1(S244E) or mPDK-1(S399D), in Chinese hamster ovary and HEK293 cells was sufficient to induce Akt phosphorylation at Thr(308) to a level similar to that of insulin stimulation. Furthermore, this increase in phosphorylation was independent of the Pleckstrin homology domain of Akt. Taken together, our results suggest that mPDK-1 undergoes autophosphorylation at multiple sites and that this phosphorylation may be essential for PDK-1 to interact with and phosphorylate its downstream substrates in vivo.