Design, synthesis, and evaluation of the in vitro activity of novel dual inhibitors of XOR and URAT1 containing a benzoic acid group.

Xanthine oxidoreductase (XOR) and uric acid transporter 1 (URAT1) are involved in the production and reabsorption of uric acid, respectively. However, the currently available individual XOR- or URAT1-targeted drugs have limited efficacy. Thus, strategies for combining XOR inhibitors with uricosuric drugs have been developed. Previous virtual screening identified Compounds 1-5 as hits for the potential dual inhibition of XOR/URAT1. Nevertheless, in vitro experiments yielded unsatisfactory results. The first round of optimization work on those hits was performed, and two series of compounds were designed and synthesized. Compounds of the A series exerted moderate inhibitory effects on URAT1, but extremely weak inhibitory effects on XOR. Compounds of the B series exerted strong inhibitory effects on both XOR and URAT1. B5 exhibited the greatest inhibitory activity, with similar inhibitory effects on XOR and URAT1. The half maximal inhibitory concentration (IC50 ) of XOR was 0.012 ± 0.001 µM, equivalent to that of febuxostat (IC50 = 0.010 ± 0.001 µM). The IC50 of URAT1 was 30.24 ± 3.46 µM, equivalent to that of benzbromarone (IC50 = 24.89 ± 7.53 µM). Through this optimization, the in vitro activity of most compounds of the A and B series against XOR and URAT1 was significantly improved versus that of the hits. Compound B5 should be further investigated.

Attenuation of Polysialic Acid Biosynthesis in Cells by the Small Molecule Inhibitor 8-Keto-sialic acid.

Sialic acids are key mediators of cell function, particularly with regard to cellular interactions with the surrounding environment. Reagents that modulate the display of specific sialyl glycoforms at the cell surface would be useful biochemical tools and potentially allow for therapeutic intervention in numerous challenging chronic diseases. While multiple strategies are being explored for the control of cell surface sialosides, none that shows high selectivity between sialyltransferases or that targets a specific sialyl glycoform has yet to emerge. Here, we describe a strategy to block the formation of α2,8-linked sialic acid chains (oligo- and polysialic acid) through the use of 8-keto-sialic acid as a chain-terminating metabolic inhibitor that, if incorporated, cannot be elongated. 8-Keto-sialic acid is nontoxic at effective concentrations and serves to block polysialic acid synthesis in cancer cell lines and primary immune cells, with minimal effects on other sialyl glycoforms.

Optomechanical switching of adsorption configurations of polar organic molecules by UV radiation pressure.

Using photoemission spectroscopy (PES), we have systematically investigated the behavior of polar organic molecule, chloroaluminum phthalocyanine (ClAlPc), adsorbed in the Cl-down configuration on the Ag(111) substrate at low temperature - 195 °C under UV irradiation with a range of different photon fluxes. Judging from the evolution of photoemission spectral line shapes of molecular energy states, we discovered that the Cl atoms are so robustly anchored at Ag(111) that the impinging photons cannot flip the ClAlPc molecules, but instead they crouch them down due to radiation pressure; we observe that the phthalocyanine (Pc) lobes bend down to interact with Ag atoms on the substrate and induce charge transfer from them. As photon flux is increased, radiation pressure on the Pc plane initiates tunneling of the Cl atom through the molecular plane to turn the adsorption configuration of ClAlPc from Cl-down to an upheld Cl-up configuration, elucidating an optomechanical way of manipulating the dipole direction of polar molecules. Finally, work function measurements provide a distinct signature of the resulting upheld Cl-up configuration as it leads to a large increase in vacuum level (VL), ~ 0.4 eV higher than that of a typical flat-on Cl-up configuration driven by thermal annealing.

Subjective and objective cognitive functioning among patients with breast cancer: effects of chemotherapy and mood symptoms.

BACKGROUND: Previous findings regarding declines in cognitive functioning among patients with breast cancer (BC) before and after chemotherapy have been inconsistent. The present study explored the effect of BC and cancer-related chemotherapies on cognitive functioning. METHODS: A cross-sectional design was adopted to compare BC patients before their chemotherapy treatment, BC patients 3 ~ 9 months after the completion of chemotherapy, and noncancer controls. Evaluations of cognitive functioning included subjective and objective dimensions, with focus on memory, executive functioning, attention, and language. ANCOVA and Pearson's correlation analysis were used to examine the relationship among cancer, chemotherapy, cognitive performance, and psychological distress. RESULTS: After adjustment for intelligence quotient, anxiety, and depression, we found significant differences in the Semantic Association of Verbal Fluency between post-chemotherapy (C/T) patients and noncancer controls. Specifically, post-C/T patients scored lower than controls (p = 0.03, η2 = 0.07). No significant differences were found in other objective cognitive measures. However, both subjective and objective cognitive scores were significantly associated with depression, anxiety, and fatigue. In BC patients, levels of anxiety were positively correlated with measures of executive function. Among pre-C/T patients, self-perceived interference by fatigue was positively associated with better performances in some of the objective cognitive measures. CONCLUSION: Our findings suggest cognitive impairments in the domain of executive functioning among patients with BC who received chemotherapy. Providing relevant suggestions or strategies of managements for these negative consequences may help increase the long-term quality of life of patients with BC.

CUX2, BRAP and ALDH2 are associated with metabolic traits in people with excessive alcohol consumption.

Molecular mechanisms that prompt or mitigate excessive alcohol consumption could be partly explained by metabolic shifts. This genome-wide association study aims to identify the susceptibility gene loci for excessive alcohol consumption by jointly measuring weekly alcohol consumption and γ-GT levels. We analysed the Taiwan Biobank data of 18,363 Taiwanese people, including 1945 with excessive alcohol use. We found that one or two copies of the G allele in rs671 (ALDH2) increased the risk of excessive alcohol consumption, while one or two copies of the C allele in rs3782886 (BRAP) reduced the risk of excessive alcohol consumption. To minimize the influence of extensive regional linkage disequilibrium, we used the ridge regression. The ridge coefficients of rs7398833, rs671 and rs3782886 were unchanged across different values of the shrinkage parameter. The three variants corresponded to posttranscriptional activity, including cut-like homeobox 2 (a protein coded by CUX2), Glu504Lys of acetaldehyde dehydrogenase 2 (a protein encoded by ALDH2) and Glu4Gly of BRCA1-associated protein (a protein encoded by BRAP). We found that Glu504Lys of ALDH2 and Glu4Gly of BRAP are involved in the negative regulation of excessive alcohol consumption. The mechanism underlying the γ-GT-catalytic metabolic reaction in excessive alcohol consumption is associated with ALDH2, BRAP and CUX2. Further study is needed to clarify the roles of ALDH2, BRAP and CUX2 in the liver-brain endocrine axis connecting metabolic shifts with excessive alcohol consumption.

Correlations And Correlates Of Post-Traumatic Growth And Post-Traumatic Stress Symptoms In Patients With Breast Cancer.

PURPOSE: Although post-traumatic growth (PTG) and post-traumatic stress symptoms (PTSS) might develop and coexist after a major trauma, few studies have simultaneously examined them in patients with breast cancer. This study investigated the correlation between PTG and PTSS and their differential correlates in patients with breast cancer. PATIENTS AND METHODS: Overall, 145 patients with breast cancer were recruited. PTG and PTSS were assessed using the PTG inventory and the Chinese version of startle, physiological arousal, anger, and numbness, respectively. We investigated the effects of demographics, chemotherapy, depression, family support, alexithymia, and anxiety symptoms on PTG and PTSS. Multivariate linear regression analyses were performed to select the independent correlates of PTSS and PTG. RESULT: An association was observed between PTG and PTSS (r = 0.21). Based on multiple regression models, the common correlate of PTG (ß = 0.271) and PTSS (ß = 0.212) was anxiety symptoms. Differential independent correlates were years of education (ß = 0.272), receiving chemotherapy (ß = 0.248), and family support (ß = 0.259) for PTG, and chronic pain (ß = 0.316) and poor cognition (ß = -0.350) for PTSS. CONCLUSION: Differential correlates were observed for PTG and PTSS in patients with breast cancer. Possible mechanisms and relationships between PTG and PTSS were discussed.

Depression increases the risk of rotator cuff tear and rotator cuff repair surgery: A nationwide population-based study.

BACKGROUND: Chronic inflammation is known to be associated with both rotator cuff tears (RCTs) and depression. However, no epidemiological studies with a longitudinal follow-up have been performed to prove this association. We aimed to investigate whether depressed patients had an elevated risk of RCT and subsequent repair surgery compared with those without depression. METHODS: This retrospective cohort study comprised of patients diagnosed with depression between 2000 and 2010 (depression cohort) and patients without depression (non-depression cohort, 1:2 age and sex matched). The risk of RCT and rotator cuff repair surgery were determined during a 13-year follow-up (2000-2013) between these two cohorts. RESULTS: This study included 26,868 patients with depression and 53,736 patients without depression. The incidence of RCT was 648 and 438 per 100,000 person-years in the depression and non-depression cohorts, respectively. The adjusted hazard ratio (HR) was 1.46 (95% confidence interval [CI], 1.36-1.57) for depressed patients. The incidence of rotator cuff repair surgery was 28 and 18 per 100,000 person-years in the depression and non-depression cohorts, respectively. Depressed patients also had a significantly increased risk of subsequent rotator cuff repair surgery (adjusted HR = 1.46; 95% CI, 1.04-2.06). CONCLUSION: The present study showed that depression was associated with an increased risk of rotator cuff tear and rotator cuff repair surgery.

Prospecting for microbial α-N-acetylgalactosaminidases yields a new class of GH31 O-glycanase.

α-Linked GalNAc (α-GalNAc) is most notably found at the nonreducing terminus of the blood type-determining A-antigen and as the initial point of attachment to the peptide backbone in mucin-type O-glycans. However, despite their ubiquity in saccharolytic microbe-rich environments such as the human gut, relatively few α-N-acetylgalactosaminidases are known. Here, to discover and characterize novel microbial enzymes that hydrolyze α-GalNAc, we screened small-insert libraries containing metagenomic DNA from the human gut microbiome. Using a simple fluorogenic glycoside substrate, we identified and characterized a glycoside hydrolase 109 (GH109) that is active on blood type A-antigen, along with a new subfamily of glycoside hydrolase 31 (GH31) that specifically cleaves the initial α-GalNAc from mucin-type O-glycans. This represents a new activity in this GH family and a potentially useful new enzyme class for analysis or modification of O-glycans on protein or cell surfaces.

Facile Formation of ß-thioGlcNAc Linkages to Thiol-Containing Sugars, Peptides, and Proteins using a Mutant GH20 Hexosaminidase.

Thioglycosides are hydrolase-resistant mimics of O-linked glycosides that can serve as valuable probes for studying the role of glycosides in biological processes. The development of an efficient, enzyme-mediated synthesis of thioglycosides, including S-GlcNAcylated proteins, is reported, using a thioglycoligase derived from a GH20 hexosaminidase from Streptomyces plicatus in which the catalytic acid/base glutamate has been mutated to an alanine (SpHex E314A). This robust, easily-prepared, engineered enzyme uses GlcNAc and GalNAc donors and couples them to a remarkably diverse set of thiol acceptors. Thioglycoligation using 3-, 4-, and 6-thiosugar acceptors from a variety of sugar families produces S-linked disaccharides in nearly quantitative yields. The set of possible thiol acceptors also includes cysteine-containing peptides and proteins, rendering this mutant enzyme a promising catalyst for the production of thio analogues of biologically important GlcNAcylated peptides and proteins.

Expression of Bmi-1 and PAI-1 in esophageal squamous cell carcinoma.

AIM: To determine the correlation between invasiveness, migration and prognosis in esophageal squamous cell carcinoma (ESCC) and expression of the B-cell-specific Moloney leukemia virus insert site 1 (Bmi-1) and plasminogen activator inhibitor-1 (PAI-1). METHODS: Eighty previously untreated patients who underwent surgical excision of ESCC were included. The expression of Bmi-1 and PAI-1 was examined immunohistochemically in formalin-fixed paraffin-embedded primary tissue specimens. The relationships between the expression of Bmi-1 and PAI-1, the clinicopathologic features of ESCC, and the survival rate of ESCC patients were also discussed. The correlation between Bmi-1 and PAI-1 protein expression in ESCC was analyzed. The relationship between Bmi-1 and PAI-1 expression and ESCC prognosis was evaluated using a Cox regression model and Kaplan-Meier survival curve analysis. RESULTS: The rates of positive Bmi-1 and PAI-1 expression in ESCC were higher than those in normal esophageal tissue (P < 0.05). The expression of Bmi-1 and PAI-1 was correlated with depth of invasion and lymph node metastasis (P < 0.05), but not with patient age, tumor size or nationality (P > 0.05). The expression of Bmi-1 was positively correlated with that of PAI-1 (P < 0.05). The 10-year overall survival rate for all patients was 20% (16/80). Univariate Kaplan-Meier survival analysis showed that patients with high expression of esophageal PAI-1 and Bmi-1 had lower survival, however, the difference was not statistically significant. Cox multivariate analysis showed that PAI-1 and Bmi-1 were not independent factors for survival rate, while the depth of tumor invasion and metastasis were independent factors affecting patient survival. CONCLUSION: The expression of Bmi-1 and PAI-1 plays a role in ESCC progression, and may be used as a prognostic marker in ESCC.

A plate-based high-throughput activity assay for polysialyltransferase from Neisseria meningitidis.

Polysialyltransferases (PSTs) assemble polysialic acid (PSA) and have been implicated in many biological processes. For example, certain bacteria such as neuroinvasive Neisseria meningitidis decorate themselves in a PSA capsule to evade the innate immune system. Identifying inhibitors of PSTs therefore represents an attractive therapeutic goal and herein we describe a high-throughput, robust, and sensitive microtiter-plate-based activity assay for PST from N. meningitidis. A trisialyl lactoside (GT3) serving as the acceptor substrate was immobilized on a 384-well plate by click chemistry. Incubation with PST and CMP-sialic acid for 30min resulted in polysialylation. The immobilized PSA was then directly detected using a green fluorescent protein (GFP)-fused PSA-binding protein consisting of the catalytically inactive double mutant of an endosialidase (GFP-EndoNF DM). We report very good agreement between kinetic and inhibition parameters obtained with our on-plate assay versus our in-solution validation assay. In addition we prove our assay is robust and reliable with a Z' score of 0.79. All aspects of our assay are easily scalable owing to optimization trials that allowed immobilization of acceptor substrates prepared from crude reaction mixtures and the use of cell lysates. This assay methodology enables large-scale PST inhibitor screens and can be harnessed for directed evolution screens.

Use of compact digital cervicography: an adjuvant screening tool for precancerous cervical lesions.

OBJECTIVE: In an attempt to develop a new model to screen for cervical cancer, we set up a digital camera mounted with a macro lens for cervicography. This digitalized compact form of cervicography with the Reid colposcopic index (RCI) can be applied as an adjuvant tool to screen for abnormal Papanicolaou (Pap) smears. MATERIALS AND METHODS: Among 21,532 cases screened with a Pap smear, we enrolled 125 cases with precancerous smears for this prospective study. An 8-megapixel Canon digital camera with a macro lens and a light source comprised the compact digital cervicographic system. The results of compact digital cervicography were reported using the RCI scoring system. Cases with high-grade squamous intraepithelial lesion or atypical glandular cells of undetermined significance were transferred to medical centers for further confirmation and management. In total, 119 cases underwent compact digital cervicography. The data were analyzed with the McNemar test. RESULTS: With the digitized results of compact digital cervicography, we were able to show patients with precancerous Pap smears their cervix and help them understand its status. For patients with an abnormal Pap smear, the results of performing compact digital cervicography combined with the RCI scoring system were statistically significant (p < 0.05). CONCLUSION: The use of compact digital cervicography with the RCI scoring system is a significant device for screening abnormal Pap cases. It helps to improve patient care. Further research is required to understand the discrepancies among diagnostic results of Pap smears (the Bethesda system), cervicography with RCI, and histopathology.

[A dynamic observation on serum cytokine and immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer].

OBJECTIVE: To study the changes of serum interleukin-2 (IL-2), interleukin-8 (IL-8) and immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer, and to probe the relationship between the levels of IL-2, IL-8, IgG, IgA and IgM and the progress of cancer. METHODS: The serum levels of IL-2 and IL-8 were detected by enzyme-linked immunosorbent assay for 72 case of primary esophageal cancer, 68 advanced esophageal cancer and 120 healthy controls, and the level of immunoglobulin (IgG, IgA, IgM) in patients with esophageal cancer was dynamically observed. RESULTS: The IL-2 level in patients with early esophageal cancer [(1.69 +/- 0.53) ng/ml] or late esophageal cancer [(1.11 +/- 0.60) ng/ml] was lower than the control group [(2.78 +/- 0.51) ng/ml] (P < 0.01), the late esophageal cancer group was lower than early esophageal cancer group (P < 0.05). The level of IL-8 in patients with early esophageal cancer [(85.48 +/- 6.14) ng/L] or late esophageal cancer [(121.41 +/- 6.22) ng/L] was much higher than the control group [(54.48 +/- 12.20) ng/L] (P < 0.01), the late esophageal cancer group was much higher than early esophageal cancer group (P < 0.01); There was correlation between the levels of IL-2 and IL-8 and the worsen-extent of the tumour in patients with early esophageal cancer or late esophageal cancer. But the level of IgG [(12.23 +/- 2.50) g/L], IgM [(1.60 +/- 0.80) g/L] in the patients with esophageal cancer compared with the level of IgG [(11.65 +/- 3.70) g/L], IgM [(1.46 +/- 0.71) g/L] in the health control group have no significant difference (P > 0.05), the level of IgA [(3.50 +/- 1.10) g/L] in patients with esophageal cancer Compared with the control group [(1.88 +/- 1.08) g/L] has significant difference (P < 0.01), and along with the worsen-extent of the tumor in patients the level of IgA has the increased tendency. CONCLUSION: The IL-8 might accelerate the pathogenesis of esophageal cancer, and the IL-2 might restrain. The positive correlation between the level of IgA and the patients with esophageal cancer is observed in this study; the immune maladjustment of IL-2, IL-8 and IgA might be correlative to esophageal cancer, and the IL-2, IL-8 and IgA levels might be an available index for the severity of esophageal cancer, Which may be of some help for clinic practitioners to judge the progress, curative effect and prognosis of the cancer.

[Serum level and genotype of interleukin-6 in patients with esophageal cancer].

OBJECTIVE: To investigate the polymorphism of the interleukin-6 (IL-6) gene promoter-572C/G and -634C/G in patients with esophageal cancer, and to study the relation between the serum level and genotype of interleukin-6 and esophageal cancer. METHODS: Peripheral blood samples were collected from 118 patients with esophageal cancer and 130 healthy persons as controls. The polymorphism of IL-6 was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The serum level of IL-6 was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The frequency of the genotype GG of the IL-6 (-634) site in the cancer group was 12.7%, significantly higher that that of the control group (3.8%, P<0.05). The risk of esophageal cancer of the G allele carriers was 1.759 times that of the C allele carriers (OR=1.759, 95% CI=1.150-2.691). The serum level of IL-6 of the esophageal cancer group was (16.9+/-5.3) ng/L, significantly higher than that of the control group [(4.6+/-2.6) ng/L, P<0.01]. The serum level of IL-6 of the esophageal cancer with the G allele carriers was (18.8+/-6.1) ng/L, significantly higher than that of the esophageal cancer without the G allele carriers [(13.2+/-6.0) ng/L, P<0.01]. There was no significant difference in the distribution of the IL-6 gene-572C/G polymorphism between the 2 groups (P>0.05). CONCLUSION: IL-6 gene-634C/G polymorphism is associated with the esophageal cancer. The IL-6 allele G carriers may be at increased risk of the esophageal cancer because of the increase of the IL-6 expression.

Effect of electroacupuncture on neurotrophin expression in cat spinal cord after partial dorsal rhizotomy.

Neuroplasticity of the spinal cord following electroacupuncture (EA) has been demonstrated although little is known about the possible underlying mechanism. This study evaluated the effect of EA on expression of neurotrophins in the lamina II of the spinal cord, in cats subjected to dorsal rhizotomy. Cats received bilateral removal of L1-L5 and L7-S2 dorsal root ganglia (DRG, L6 DRG spared) and unilateral EA. They were sacrificed 7 days after surgery, and the L6 spinal segment removed and processed by immunohistochemistry and in situ hybridization histochemistry, to demonstrate the expression of neurotrophins. Significantly greater numbers of nerve growth factor (NGF) and neurotrophin-3 (NT-3) positive neurons, brain-derived neurotrophic factor (BDNF) immunoreactive varicosities and NT-3 positive neurons and glial cells were observed in lamina II on the acupunctured (left) side, compared to the non-acupunctured, contralateral side. Greater number of neurons expressing NGF mRNA was also observed on the acupunctured side. No signal for mRNA to BDNF and NT-3 was detected. The above findings demonstrate that EA can increase the expression of endogenous NGF at both the mRNA and protein level, and BDNF and NT-3 at the protein level. It is postulated that EA may promote the plasticity of the spinal cord by inducing increased expression of neurotrophins.