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Reaction of the N-heterocyclic carbene-stabilized phosphinoaminosilylene ArN(SiMe3)Si(I(i)Pr)PPh2 (Ar = 2,6-(i)Pr2C6H3, I(i)Pr = 1,3-diisopropyl-4,5-dimethyl-imidazol-2-ylidene) (2) with dioxygen resulted in the formation of the two products ArN = Si(I(i)Pr)(OSiMe3)(OPPh2) (3) and ArN = Si(I(i)Pr)(OSiMe3)PPh2 (4), and the ratios of the two products can be controlled by the reaction temperatures and dioxygen concentrations.

Purpose To assess the value of semi-automated segmentation applied to diffusion MRI for predicting the therapeutic response of liver metastasis. Methods Conventional diffusion weighted magnetic resonance imaging (MRI) was performed using b-values of 0, 150, 300 and 450 s/mm2 at baseline and days 4, 11 and 39 following initiation of a new chemotherapy regimen in a pilot study with 18 women with 37 liver metastases from primary breast cancer. A semi-automated segmentation approach was used to identify liver metastases. Linear regression analysis was used to assess the relationship between baseline values of the apparent diffusion coefficient (ADC) and change in tumor size by day 39. Results A semi-automated segmentation scheme was critical for obtaining the most reliable ADC measurements. A statistically significant relationship between baseline ADC values and change in tumor size at day 39 was observed for minimally treated patients with metastatic liver lesions measuring 2–5 cm in size (p = 0.002), but not for heavily treated patients with the same tumor size range (p = 0.29), or for tumors of smaller or larger sizes. ROC analysis identified a baseline threshold ADC value of 1.33 μm2/ms as 75% sensitive and 83% specific for identifying non-responding metastases in minimally treated patients with 2–5 cm liver lesions. Conclusion Quantitative imaging can substantially benefit from a semi-automated segmentation scheme. Quantitative diffusion MRI results can be predictive of therapeutic outcome in selected patients with liver metastases, but not for all liver metastases, and therefore should be considered to be a restricted biomarker.

Persistent human papillomavirus (HPV) infection is the vital factor driving cervical carcinogenesis; however, other features of the local cervicovaginal microenvironment (CVM) may play a critical role in development of precancerous cervical dysplasia and progression to invasive cervical carcinoma (ICC). Here we investigated relationships between immunoproteomic and metabolic profiles and features of the cervicovaginal microenvironment, such as HPV status, vaginal microbiota (VMB), vaginal pH and genital inflammation, to better understand the complex interplay between host, virus and bacteria. In this multicenter study we enrolled 78 women with ICC, high- and low-grade squamous intraepithelial lesions, as well as HPV-positive and healthy HPV-negative controls. Vaginal swabs and cervicovaginal lavages (CVL) were collected for HPV genotyping, microbiome, metabolome and immunoproteome analyses. The VMB compositions were determined using 16S rRNA gene sequencing. Cervicovaginal metabolic fingerprints were profiled using liquid chromatography-mass spectrometry. Levels of immune mediators and other proteins in CVL samples were evaluated using multiplex cytometric bead arrays. Abnormal vaginal pH and dysbiotic non-Lactobacillus-dominated VMB were associated with Hispanic ethnicity and severity of cervical neoplasm. We also identified microbial signatures (e.g. Sneathia spp.) to be enriched in ICC and all precancerous groups. Notably, Sneathia abundance was also increased in patients with abnormal pH and those of Hispanic origin. Analyses of 62 protein targets in CVL samples revealed elevated levels of pro-inflammatory cytokines and chemokines, growth and angiogenic factors, apoptosis-related, immune checkpoint and other proteins in ICC patients. Levels of many of these proteins depended on the VMB structure and genital inflammation. These proteomic signatures positively correlated with dysbiotic non-Lactobacillus-dominated VMB and abnormal vaginal pH, both features associated with Hispanic ethnicity. Furthermore, metabolomic analysis also revealed that VMB, together with genital inflammation, are the major drivers of metabolic profiles in the local CVM. Finally, using hierarchical clustering analyses, we identified groups of patients who significantly varied in the levels of cancer-related proteins, genital inflammation, vaginal pH and VMB composition regardless of disease severity. These microenvironmental factors may impact the HPV persistence/progression and consequently increase the risk of cervical cancer. Our study demonstrated that the racial/ethnic differences in the VMB compositions may contribute to cervical cancer disparity in Hispanic women. In the future we are planning to expand our investigation of the VMB in Native American women, which will further illuminate the relationship between race/ethnicity, the VMB, and HPV. Citation Format: Pawel Laniewski, Zehra Esra Ilhan, Nicholas A. Bokulich, Haiyan Cui, Denise J. Roe, Dana M. Chase, J. Gregory Caporaso, Melissa M. Herbst-Kralovetz. Integrative multi-omics approach reveals complex interplay between HPV, host and microbiome during cervical carcinogenesis in Hispanic and non-Hispanic women [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr A094.

We investigated the cellulase-assisted extraction and anti-ultraviolet activity of water-soluble polysaccharides from the root of Flammulina velutipes on Caenorhabditis elegans. A Box-Behnken design experiment with three factors and three levels, including enzymolysis temperature, microwave time, and microwave power, was designed on the basis of the results of single-factor experiments. For improving the polysaccharide yield of F. velutipes root, the following optimal extraction conditions were used: 52.67°C enzymolysis temperature, 80s microwave time, and 144 W microwave power. Under optimal conditions, the actual measured value of the yield was 2.01% (w/w) and the predicted value was 2.06% (w/w). One fraction (FRP-2) was isolated and purified, and its characteristics were analyzed. The average mean molecular weight of FRP-2 was measured to be 2.60×10

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb), and remains a leading public health problem. Previous studies have identified host genetic factors that contribute to Mtb infection outcomes. However, much of the heritability in TB remains unaccounted for and additional susceptibility loci most likely exist. We perform a multistage genome-wide association study on 2949 pulmonary TB patients and 5090 healthy controls (833 cases and 1220 controls were genome-wide genotyped) from Han Chinese population. We discover two risk loci: 14q24.3 (rs12437118, Pcombined = 1.72 × 10−11, OR = 1.277, ESRRB) and 20p13 (rs6114027, Pcombined = 2.37 × 10−11, OR = 1.339, TGM6). Moreover, we determine that the rs6114027 risk allele is related to decreased TGM6 transcripts in PBMCs from pulmonary TB patients and severer pulmonary TB disease. Furthermore, we find that tgm6-deficient mice are more susceptible to Mtb infection. Our results provide new insights into the genetic etiology of TB., Genetic risk loci for tuberculosis (TB) have so far been identified in African and Russian populations. Here, the authors perform a three-stage GWAS for TB in Han Chinese populations and find two risk loci near ESRRB and TGM6 and further demonstrate that tgm6 protects mice from Mtb infection.

In this paper, a smart trash can based on the voice control is proposed. The garbage bin takes the Microcontroller as the control core and has a voice interaction function, that is, according to the speech recognition function instruction, the obstacle avoidance can be automatically realized during the movement process, and when the capacity reaches the preset value, it can prompt people to clean up. Meanwhile, the status of all trash cans can be displayed on the LCD screen. After debugging, the design has diversified functions and fully demonstrates practicality.

Four stable boron dipyrromethene (BODIPY)- and porphyrin-based bis-arylamine diradical dications were synthesized by two-electron oxidation of their neutral molecules. The two BODIPY-based dications have open-shell singlet ground states. UV/Vis absorption spectra of all four dications showed large redshifts in the NIR region compared to their neutral precursors with absorption maxima at 1274 and 1068 nm for the two BODIPY-based dications and 1746 and 2037 nm for the two porphyrin-based dications. Thus, two new types of NIR dyes with longer wavelengths are provided by the diradical-dication strategy, which can be applied for the generation of other NIR dyes with a range of different chromophores and auxochromes.

While high-risk human papillomavirus (HPV) infection is a well-established risk factor for cervical cancer, there are likely other factors within the local microenvironment that contribute to cervical carcinogenesis. Here we investigated relationships between HPV, vaginal pH, vaginal microbiota (VMB) composition, level of genital immune mediators and severity of cervical neoplasm. We enrolled women with low- and high-grade cervical dysplasia (LGD, HGD), invasive cervical carcinoma (ICC), and healthy controls. HPV16, HPV45, HPV58, and HPV31 were the most prevalent in our cohort with HPV16 and HPV31 genotypes more prevalent in Hispanics. Vaginal pH was associated with ethnicity and severity of cervical neoplasm. Lactobacillus dominance decreased with the severity of cervical neoplasm, which correlated with elevated vaginal pH. Hispanic ethnicity was also associated with decreased Lactobacillus dominance. Furthermore, Sneathia was enriched in all precancerous groups, ICC, abnormal pH and Hispanic origin. Patients with ICC, but not LGD and HGD, exhibited increased genital inflammatory scores and elevated specific immune mediators. Notably, IL-36γ was significantly associated with ICC. Our study revealed local, host immune and microbial signatures associated with cervical carcinogenesis and provides an initial step to understanding the complex interplay between mucosal inflammation, HPV persistence and the VMB.

Three bis(triarylamine) dications bridged with 2,2'-bithienyl group (12+), biphenyl group (22+) and [2,2]paracyclophane (32+) have been successfully isolated. The electronic structures of 12+ - 32+ show bridge dependence. Magnetic studies and DFT calculations show that dications 12+ - 32+ possess an open-shell singlet ground state with a thermally excited triplet state. Dication 32+ has rather small singlet-triplet energy gap and could basically be regarded as a nearly pure diradical. Diradical dications 12+ - 32+ represent rare examples of diradicaloids with intramolecular long-range π-conjugation or π-π coupling interaction.

A strain SMrs28 was isolated from the rhizosphere soil of a toxic plant Stellera chamaejasme and identified as Bacillus sp. on the basis of morphological and partial 16S rRNA gene sequence analysis. The crude extract of SMrs28 fermentation broth showed strong nematocidal activities in preliminary test. To define the active nematocidal metabolites of SMrs28, a novel compound (1), 4-oxabicyclo[3.2.2]nona-1(7), 5,8-triene, along with five known compounds (2-6), were isolated from the strain by various column chromatographic techniques and characterized on the basis of spectroscopic analysis. Results of the in vitro nematicidal tests showed that the metabolites presented different levels of activity at certain exposure conditions. Compounds (1-3) displayed LC50 values of 904.12, 451.26, 232.98 Rg/m1 and 1594.0, 366.62, 206.38 mu g/ml against Bursaphelenchus xylophilus and Ditylenchus destructor at 72 h, respectively. This is the first report of the nematicidal activity of the compounds as constituents of Bacillus sp.. Our findings help to find potential chemical structures to develop nematicides from microbial source for the management of nematode-infected plant diseases.

Populations of Stellera chamaejasme L. have been increasing constantly in recent years in some areas of the grassland in north China but why this toxic weed has become highly competitive is not clear. In order to determine if any potential allelochemicals are released into the soil environment by S. chamaejasme, we investigated the chemical composition of a water-washed solution of the living roots with rhizosphere soil. This led to the isolation and identification of seven compounds: umbelliferone (1), daphnoretin (2), chamaechromone (3), 7-methoxyneochamaejasmine A (4), mesoneochamaejasmin A (5), neochamaejasmin B (6), dihydrodaphnodorin B (7). All are secondary metabolites of S. chamaejasme. Bioassay showed that 1, 5 and 6 had a strong inhibitory effect on Festuca rubra L. and Medicago sativa seedlings. These compounds were quantified by high performance liquid chromatography in 25 root zone soil samples of S. chamaejasme collected at altitudes between 165 and 4741 m from the northeast to the Tibetan Plateau of China. All samples contained at least one of the phytotoxic compounds. Their content did not correlate with the altitude of the growing site. However, the level of chamaechromone negatively correlated with the soil pH. Principle components analysis indicated that the flavonoids might come from the same source. These potential allelochemicals from root release into the soil might play an important role in the highly competitive nature and broad ecological adaptability of S. chamaejasme in the wild.

Positive lymph node status in breast cancer is known to be an adverse prognostic factor, but the effect of lymph node (LN) status in inflammatory breast cancer (IBC) has not been evaluated. This study was designed to investigate the association between lymph node status and overall survival (OS) in individuals with IBC. Using the Surveillance, Epidemiology, and End Results (SEER) 18 registry, we collected data on 761 patients diagnosed with non-metastatic IBC from 2004 to 2008. Survival analysis was performed using the Kaplan–Meier method. Cox proportional hazard regression was performed to evaluate univariate and multivariate associations between estrogen and progesterone receptor (ER/PR) status, treatment, and OS. Positive nodal status was associated with a significant decrease in OS (p < 0.001). Five-year survival for LN-positive and LN-negative patients was 49 and 66 %, respectively. In node-positive patients, ER or PR positivity was associated with improved OS, (p = 0.025, p = 0.007). In node-positive patients, the combination of surgery and radiation therapy improved OS when compared with surgery alone (p = 0.002). Nearly 80 % of the patients in this study had nodal metastasis. Positive nodal status was found to be an adverse prognostic factor. ER/PR positivity and treatment with surgery and radiation in node-positive patients was found to improve outcomes. Further studies are required to characterize the biology of IBC and guide the optimal treatment of this disease.

A panel of nine hypoxia regulated genes, selected from a previously published fifty gene panel, was investigated for its ability to predict hypoxic ovarian cancer phenotypes. All nine genes including vascular endothelial growth factor A, glucose transporter 1, phosphoglycerate mutase 1, lactate dehydrogenase A, prolyl 4-hydroxylase, alpha-polypeptide 1, adrenomedullin, N-myc downstream regulated 1, aldolase A, and carbonic anhydrase 9 were upregulated in the HEY and OVCAR-3 human ovarian cell lines cultured in vitro under hypoxic compared to normoxic conditions as measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The gene panel was also elevated in HEY xenograft tumor tissue compared to HEY cells cultured in normoxia. The HEY xenograft tissue demonstrated heterogeneous positive immunohistochemical staining for the exogenous hypoxia biomarker pimonidazole, and the hypoxia regulated protein carbonic anhydrase IX. A quantitative nuclease protection assay (qNPA) was developed which included the nine hypoxia regulated genes. The qNPA assay provided similar results to those obtained using qRT-PCR for cultured cell lines. The qNPA assay was also evaluated using paraffin embedded fixed tissues including a set of five patient matched primary and metastatic serous cancers and four normal ovaries. In this small sample set the average gene expression was higher in primary and metastatic cancer tissue compared to normal ovaries for the majority of genes investigated. This study supports further evaluation by qNPA of this gene panel as an alternative or complimentary method to existing protein biomarkers to identify ovarian cancers with a hypoxic phenotype.

Reaction of the N-heterocyclic carbene (NHC)-stabilized silylene ArN(SiMe3)Si(IiPr)Cl (1, Ar = 2,6-iPr2C6H3, IiPr = 1,3-diisopropyl-4,5-dimethyl-imidazol-2-ylidene) with SiCl4 resulted in the formation of three different products NHC-stabilized dichlorosilaimine ArN=Si(IiPr)Cl2 (2), aminotrichlorosilane ArN(SiMe3)SiCl3 (3) and a silaimine dimer (ArNSiCl2)2 (4) under different conditions. The products can be controlled by reaction conditions. Compound 2 is the first example of a less bulky NHC-supported silaimine via the reaction of a silylene with SiCl4.

Bone metastasis (BM) is the advanced complication of breast cancer, while bone marrow-derived mesenchymal stem cells (BMSCs) in the microenvironment unclearly contribute to cancer metastasis. This study investigated potential roles of transforming growth factor- (TGF-) α in the interaction between breast cancer and BMSCs in BM. Clinical cases of breast cancer with bone metastasis (BMBC), breast cancer without bone metastasis (Non-BM-BC), and benign fibroadenoma (Benign) were enlisted in a retrospective study. TGF-α was found obviously overexpressed in BM lesion of BMBC compared to primary lesion of both BMBC and Non-BM-BC (P < 0.01), and TGF-α was higher in primary lesion of both BMBC and Non-BM-BC (P < 0.01) than Benign group. Interestingly, TGF-α in nontumor tissues of both BMBC and Non-BM-BC was at a higher level than Benign group (P < 0.01), and numbers of macrophages in nontumor tissues of both BMBC and Non-BM-BC (P < 0.01) were higher than Benign group. Furthermore, in cultured human BMSCs, TGF-α stimulated production of procancer cytokines including IL-6, VEGF, FGF10, FGF17, and TGF-β1 in a dose-dependent manner. Thus, TGF-α in BC could potentially be an important signal of carcinogenesis and metastasis. Macrophages in the nontumor tissue of BC may not be protective but could promote cancer metastasis.

Diisodecyl phthalate (DIDP) is considered to be one of the less toxic phthalates. However epidemiological studies suggest that DIDP is associated with the occurrence of asthma. The effect of DIDP exposure on allergic asthma and the underlying mechanism have not been fully elucidated. Here, mice were exposed to DIDP and sensitization with OVA. The results demonstrated that DIDP exposure aggravated allergic asthma. Exposure to 15 mg/kg/day DIDP markedly exacerbated airway remodeling and promoted airway hyperresponsiveness (AhR). The study suggests that exposure to DIDP not only promotes a predominant Th2 response, but also induces Th17-type immunity. The induced allergic asthma was accompanied by elevation of IgE, an increase in TSLP expression and exacerbation of oxidative stress. Inhibition of oxidative stress by Vitamin E effectively alleviated the airway remodeling and AhR induced by DIDP and OVA sensitization. Treatment with Vitamin E inhibited the Th2 response and the production of TSLP. Blocking the activation of p38 MAPK by SB203580 prevented elevation of IL-1β and IL-17A induced by DIDP and OVA sensitization and effectively alleviated Th17 type asthmatic lesions. These results suggest that exposure to DIDP exacerbates the Th2 and Th17 response through aggravating oxidative stress and activation of the p38 MAPK pathway.

Owing to the complicated and time-consuming regenerative process, the repair of injured peripheral nerves depends largely on ongoing stem-cell therapy. Decades ago, researchers successfully isolated and identified muscle-derived stem cells (MDSCs) and discovered their potential for multidifferentiation. MDSCs play an important role in trauma repair associated with neuromuscular and vascular injury by simultaneously promoting tissue regrowth via direct differentiation and systematic secretion under physiological conditions. However, the isolation, culture, induction and application of MDSCs require further methodological analysis before clinical application. In this review, we comprehensively discuss the challenges associated with neural regeneration and reviewed the progress of stem cell based regenerative medicine, in an effort to realize the potential of MDSCs in nerve regeneration.

Concerns about the risk of titanium dioxide nanoparticles (TiO2 NPs) to human health and environment are gradually increasing due to their wide range of applications. In this study, cytotoxicity, DNA damage, and apoptosis induced by TiO2 NPs (5 nm) in A549 cells were investigated. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays revealed the time- and concentration-dependent cytotoxic effects of TiO2 NPs in a concentration range of 50 to 200 μg/mL. A statistically significant (p

Allelopathy, the negative effect on plants of chemicals released to the surroundings by a neighboring plant, is an important factor which contributes to the spread of some weeds in plant communities. In this field, Stellera chamaejasme L. (Thymelaeaceae) is one of the most toxic and ecologically-threatening weeds in some of the grasslands of north and west China. Bioassay-guided fractionation of root extracts of this plant led to the isolation of eight flavonoids 1-8, whose structures were elucidated by spectroscopic analysis. All compounds obtained, except 7-methoxylneochaejasmin A (4) and (+)-epiafzelechin (5), showed strong phytotoxic activity against Arabidopsis thaliana seedlings. Seedling growth was reduced by neochamaejasmin B (1), mesoneochamaejasmin A (2), chamaejasmenin C (3), genkwanol A (6), daphnodorin B (7) and dihydrodaphnodorin B (8) with IC50 values of 6.9, 12.1, 43.2, 74.8, 7.1 and 27.3μg/mL, respectively, and all of these compounds disrupted root development. Endogenous auxin levels at the root tips of the A. thaliana DR5::GUS transgenic line were largely reduced by compounds 1, 2 and 6-8, and were increased by compound 4. Moreover, the inhibition rate of A. thaliana auxin transport mutants pin2 and aux1-7 by compounds 1-8 were all lower than the wild type (Col-0). The influence of these compounds on endogenous auxin distribution is thus proposed as a critical factor for the phytotoxic effect. Compounds 1, 2, 4 and 8 were found in soils associated with S. chamaejasme, and these flavonoids also showed phytotoxicity to Clinelymus nutans L., an associated weed of S. chamaejasme. These results indicated that some phytotoxic compounds from roots of S. chamaejasme may be involved in the potential allelopathic behavior of this widespread weed.

Polysaccharides derived from edible fungi inhibit the proliferation of tumor cells. In this study, we investigated the effects of a polysaccharide (PN50G) from Pleurotus nebrodensis on A549 cell proliferation and apoptosis. MTT assay showed that PN50G induced apoptosis in the A549 cells in a dose-dependent. However, PN50G did not affect the proliferation viability of human fetal lung fibroblast cells MRC-5. Scanning electro microscopy (SEM) results indicate that PN50G induced a typical apoptotic morphological feature in A549. DNA accumulation and fragmentation were determined by acridine orange/ethidium bromide (AO/EB) staining. Flow cytometric analysis demonstrated that PN50G caused A549 cell apoptosis via cell arrest at the S phase. PN50G also extended the comet tail length in single-cell gel electrophoresis and disrupted the mitochondrial membrane potential as determined by Rdamine-123 staining. Further analysis by qRT-PCR showed that the expression of caspase-3 and caspase-9 mRNA increased. These findings suggest that PN50G can inhibit A549 cell proliferation and induce apoptosis mainly by activating the intrinsic mitochondrial pathway.

Background and Objectives This study was performed to investigate recent trends and factors associated with immediate breast reconstruction (IBR) using a large population-based registry. We hypothesized that rates of IBR have increased since passage of the Women's Health and Cancer Rights Act of 1998. Methods The SEER (surveillance, epidemiology and end results) database was used to evaluate Stage I–III breast cancer (BC) patients who underwent total mastectomy from 1998 to 2008. Univariate and multivariate analyses were performed to study predictors of IBR. Results Of 112,348 patients with BC treated by mastectomy 18,001 (16%) had IBR. Rates of IBR increased significantly from 1998 to 2008 (P

The reaction of the ene-diamine LH2 (L = [ArNC(Me)C(Me)NAr]2–, Ar = 2,6-iPr2C6H3) with AlMe3 and AlEt3 yielded the aluminum dimethyl complex (LH)AlMe2 (1) and the radical (L•)AlEt2 (2; L• = [ArNC(Me)C(Me)NAr]•–), respectively. Treatment of 2 with O2 and TEMPO (2,2,6,6-tetramethylpiperidin-1-oxyl) led to hydrogen abstraction from the methyl group at the ligand backbone to give the diamagnetic species (L*)AlEt2 (3; L* = [ArN═C(Me)C(CH2)NAr]−) and (L*)Al(TEMPO)(Et) (4). In contrast, the reaction of 2 with iodine resulted in the iodination of the Al–C bonds and yielded the radical (L•)AlI2 (5). Reaction of 2 with BCl3 led to electron transfer between the α-diimine radicals with the formation of the aluminum cation [(L0)AlCl2]+[AlCl4]− (6; L0 = ArN═C(Me)(Me)C═NAr) and the diazaborole LBCl (7). The structures of compounds 2, 3, and 7 have been determined by X-ray single-crystal diffraction, and the paramagnetic species 2 has been characterized by EPR measurements.

Stellera chamaejasme, a perennial weed which is an ecological threat, is widely distributed in some grasslands of Central and Eastern Asia. Our previous studies have identified several allelochemicals including two coumarins (umbelliferone and daphnoretin), from S. chamaejasme, and confirmed that allelopathy contributed to the competitive behavior of this weed. In this study, the inhibitory effects of umbelliferone and daphnoretin on lettuce seedlings and the mechanisms of their phytotoxicity were investigated. Results showed that shoot and root elongation and fresh weight of lettuce seedlings were effectively inhibited by umbelliferone in a concentration-dependent manner. Daphnoretin showed a weaker phytotoxicity. Both of the coumarins arrested the mitosis process in lettuce root tips and induced proline overproduction. Additionally, loss of cell viability and overproduction of reactive oxygen species in lettuce root cells were found after treatments with umbelliferone. Moreover, umbelliferone caused lipid peroxidation. These results suggested that umbelliferone displayed stronger phytotoxicity than daphnoretin on lettuce growth, and that the two coumarins had different mechanisms of phytotoxicity. That of daphnoretin was mainly dependent on its inhibitory effects on mitosis. Umbelliferone caused membrane lipid peroxide formation and cell death by inducing ROS overproduction, and impacted cell division, which resulted in growth inhibition of the receptor plant.

Magnetic Ba3(PO4)2/Fe3O4-nanoparticle (called BPFN) was prepared, characterized, and developed as a low-cost adsorbent for malachite green (MG) from aqueous solution. Factors such as adsorption temperature, pH of solution, dosage of adsorbent, adsorption kinetics and isotherms were investigated. The maximum adsorption capacity obtained in this work was 1639mgg-1 at 45°C and pH6. The adsorption process fitted the pseudo-first-order kinetic model and Langmuir isotherm model. Evidences from zeta potential, Fourier transform infrared spectroscopy (FT-IR), and X-ray photoelectron spectroscopy (XPS) data revealed that the adsorption process was driven by electrostatic attraction, the interaction between Lewis base N(CH3)2 in MG and Lewis acid Ba sites of BPFN. In addition, the BPFN could be easily regenerated by a magnet and the adsorption capacity maintained at 70% after five cycles. The present study suggests that the BPFN had high potential of removing MG from wastewater.

MicroRNAs (miRNAs) may function as tumor suppressor or onco‑miRNAs and have critical roles in the pathogenesis of gastric cancer (GC). The exact function and mechanism of miRNA (miR)‑320a in GC remains to be elucidated. The present study performed gain‑ and loss‑of‑function analyses by transfecting cells with mimics or inhibitors and subsequently performing colony formation, proliferation and cisplatin‑sensitivity assays. Additionally, in vivo xenograft models were also performed. Bioinformatics algorithms, luciferase reporter activity assay and western blotting were used to predict the potential target of miR‑320a. Additionally, the effect of knockdown or overexpression of ADAM metallopeptidase domain 10 (ADAM10) on cell growth and chemosensitivity was examined. The expression of miR‑320a and ADAM10 was also determined in primary tumors. The present study revealed that the expression of miR‑320a was reduced in GC cells and ectopic miR‑320a expression significantly inhibited cell growth in vitro and in vivo and enhanced the sensitivity of GC cells to cisplatin. ADAM10 was a direct target of miR‑320a in GC. Knockdown of ADAM10 attenuated the proliferative ability of GC cells, and increased the sensitivity of GC cells to cisplatin. The upregulated ADAM10 accelerated cell growth rate and reduced the cisplatin‑sensitivity of cells. Clinically, a significantly negative correlation was identified between the expression of miR‑320a and mRNA levels of ADAM10 in tumors. The findings of the present study suggested that miR‑320a may function as a tumor suppressor in GC progression and potential therapeutic strategies for GC may be based on the miR‑320a/ADAM10 axis.

Since the strong antineoplastic potential against A549 cells of Pleurotus nebrodensis polysaccharide (PN50G) in vitro has been proven previously, the definitive mechanism of PN50G-induced apoptosis in A549 cells in vivo was further investigated. All the results indicated that PN50G significantly suppressed tumor growth in A549 tumor-bearing mice. Tumor cells treated with PN50G were arrested in the G0/G1 phase, and marked changes in the expression of cell cycle-related proteins, including cyclin D1, cyclin A and cyclin B1, were observed. Moreover, western blotting analysis indicated that PN50G triggered the mitochondrial apoptotic pathway, for an increased Bax/Bcl-2 ratio, release of cytochrome c, cleavage of caspase-3 and PRPP in A549 tumor cells were observed. And the decrease in the expression of the translation related protein P70S6K was observed, because PN50G activated AMPK phosphorylation, but inhibited PI3K/AKT phosphorylation and suppressed the activation of the mammalian target of rapamycin (mTOR) induced by PN50G. In vivo imaging was performed on tumor-bearing mice, and the results indicated that PN50G significantly increased the intracellular levels of reactive oxygen species (ROS). Furthermore, it indicated that PN50G promoted the protein expression of Beclin 1 and LC-3 in a dose-dependent manner. All the results suggested that PN50G-mediated apoptosis and autophagy of A549 tumor cells in vivo mainly involved in the mitochondrial pathway and the AMPK/PI3K/mTOR pathway.

The Database of Individuals at High Risk for Breast, Ovarian, or Other Hereditary Cancers at the Arizona Cancer Center in Tucson, Arizona, assesses cancer risk factors and outcomes in patients with a family history of cancer or a known genetic mutation. We analyzed the subset of clinic probands who carry deleterious BRCA gene mutations to identify factors that could explain why mutations in BRCA2 out number those in BRCA1. Medical, family, social, ethnic and genetic mutation histories were collected from consenting patients’ electronic medical records. Differences between BRCA1 and BRCA2 probands from this database were analyzed for statistical significance and compared to published analyses.. A significantly higher proportion of our clinic probands carry mutations in BRCA2 than BRCA1, compared with previous reports of mutation prevalence. This also holds true for the Hispanic sub-group. Probands with BRCA2 mutations were significantly more likely than their BRCA1 counterparts to present to the high risk clinic without adiagnosis of cancer. Other differences between the groups were not significant. Six previously unreported BRCA2 mutations appear in our clinic population. The increased proportion of probands carrying deleterious BRCA2 mutations is likely multifactorial, but may reflect aspects of Southern Arizona’s unique ethnic heritage.

The NHC-supported silylaminosilylene Ar(SiMe3)N(Cl)Si(IiPr) (1; Ar = 2,6-iPr2C6H3, IiPr = 1,3-diisopropyl-4,5-dimethylimidazol-2-ylidene) is an efficient and stereospecific bis-silylation reagent for a range of functionalized alkynes to yield cis-1,2-bis-silylated alkenes via a 1,4-silyl migration from the amino nitrogen atom to an alkyne carbon atom. The reaction is regio- and stereospecific for terminal alkynes with an electron-withdrawing substituent, thus providing a facile access to 1,2-bis-silylated alkenes with two different silyl groups.

The first 2-aminophosphasilene, [Ar(Me3Si)N]HSi═PAr′ (4, Ar = 2,6-iPr2C6H3, Ar′ = 2,6-Mes2C6H3), bearing a hydride ligand on the three-coordinate silicon atom has been synthesized and structurally characterized. Both X-ray data of 4 and DFT calculations on the model compound (H2N)HSi═PH (4′) disclosed that the amino group on the silicon atom results in significant N–Si–P π conjugation.

Background and objective: Pulmonary tuberculosis and diabetes mellitus (DM) are closely associated. The objective of this study was to determine whether the expression of CD4+CD25+CD127− T-cells (regulatory T-cells (Treg)) is associated with diabetic pulmonary tuberculosis. Methods: Flow cytometry was used to determine the frequencies of CD4+CD25+ and CD4+CD25+CD127− T-cells in peripheral blood, bronchoalveolar lavage fluid (BALF) and pleural effusions from 120 patients (30 with pulmonary tuberculosis and DM (TBDM), 30 with pulmonary tuberculosis without DM (TB), 30 with tuberculous pleurisy without DM (TBP) and 30 healthy volunteers). The concentrations of interferon (IFN)-γ and interleukin (IL)-10 in BALF and pleural effusions were determined by enzyme-linked immunosorbent assay. Results: Treg frequencies in peripheral blood were significantly higher in patients with TBDM, TB and TBP than in the control group, with the frequency in TBDM being the highest (P

Purpose: This phase I trial evaluated intraperitoneal (i.p.) pemetrexed, cisplatin, and paclitaxel in optimally debulked ovarian cancer. Experimental Design: Dose escalation of day 1 i.p. pemetrexed accrued three patients to each of five dose levels (60–1,000 mg/m2), along with day 2 i.p. cisplatin (75 mg/m2) and day 8 i.p. paclitaxel (60 mg/m2). The goals were to determine maximum tolerated dose (MTD), 18-month progression-free survival (PFS), and pharmacokinetics of i.p. pemetrexed. Results: Cycles, given every 21 days, had an 80% 6-cycle completion rate. There was minimal grade III toxicity in the first 4 dose levels and remarkably an almost complete absence of peripheral neuropathy and alopecia. At the highest dose level, two of three patients experienced ≥grade III and dose-limiting toxicity (DLT; hematologic, infection, gastrointestinal). There was a pharmacokinetic advantage for i.p. pemetrexed with an intraperitoneal:plasma area under the concentration–time curve ratio of 13-fold. Neither analysis of pharmacokinetic nor homocysteine levels explains the unexpected severity of toxicity in those two patients. On the basis of plasma C24h levels, the 42 cycles at ≥500 mg/m2 i.p. pemetrexed without DLT, the MTD appears to be 500 mg/m2. Median PFS is 30.1 months; 18-month PFS is 78.6% (median follow-up 22.4 months). Conclusions: This i.p.-only regimen in front-line ovarian cancer is feasible with PFS in line with recent literature. We suggest phase II trials of this regimen in this population with i.p. pemetrexed at 500 mg/m2. The favorable toxicity profile at doses

The 14-3-3 proteins are a set of seven highly conserved proteins that have recently been implicated in having a role in human tumorigenesis. However, the mechanism by which 14-3-3 proteins may act in this capacity is not well understood. In this study, we examined the expression of one of the 14-3-3 family members, 14-3-3σ, since it was shown previously to be aberrantly altered in human tumors. Using quantitative rtPCR and immunohistochemistry, we found that the expression levels of 14-3-3σ were elevated in the majority of human non-small cell lung cancers (NSCLC) we examined. Surprisingly, we found that the 14-3-3σ gene was hypomethylated in lung tumors relative to normal lung tissue suggesting that decreased DNA methylation resulted in increased expression of 14-3-3σ in NSCLC. We also determined the gene copy number for 14-3-3σ in tumor samples and found no significant correlation with elevated mRNA expression. And also no mutations were found in 14-3-3σ gene. Overall, our data suggest that misregulated expression of 14-3-3σ gene may be due to altered methylation status. © 2011 Wiley-Liss, Inc.

Tuberculous pleurisy, which is characterized by an extensive inflammatory exudate in the pleural space, is a common manifestation of extrapulmonary tuberculosis caused by Mycobacterium tuberculosis infection but is difficult to diagnose. This study compared concentrations of three cytokines (interferon-γ and interleukin [IL]−2 and −4) and soluble Fas ligand in serum and pleural effusions from 36 patients with tuberculous pleurisy, 30 with malignant pleurisy and 30 healthy volunteers. Interferon-γ concentration was significantly higher in serum and pleural effusions, and the concentration of soluble Fas ligand was significantly higher in pleural effusion, from patients with tuberculous pleurisy than in those with malignant pleurisy. Levels of IL-2 and IL-4 were also raised in serum, but failed to reach statistical significance. Interferon-γ and soluble Fas ligand levels were significantly higher in pleural effusion than in serum. Pleural soluble Fas ligand concentrations correlated linearly with those of interferon-γ in patients with tuberculous pleurisy. These findings suggest a predominant T-helper-1 response to M. tuberculosis infection.

T cell-mediated immunity can produce durable anti-melanoma responses resulting in improved survival of metastatic melanoma patients. Gamma-interferon-inducible lysosomal thiol reductase (GILT) is critical for MHC class II-restricted presentation of multiple melanoma antigens to CD4+ T cells and facilitates cross-presentation on MHC class I for activation of CD8+ T cells. Our prior study found that GILT expression is increased in melanocytes of malignant melanoma specimens compared with benign nevi. To determine whether GILT expression is associated with inflammation, expression in halo nevi specimens was compared to nevi without lymphocytic infiltrate by immunohistochemistry. A halo nevus is a benign nevus with a lymphocytic infiltrate which leads to regression of the nevus. GILT, but not MHC class II, expression was increased in melanocytes of halo nevi compared to nevi without lymphocytic infiltrate. Analysis of a publicly available gene expression profiling cohort of 457 cutaneous melanoma specimens revealed that GILT expression was associated with IFN-γ, TNF-α, and IL-1β expression in human melanoma. In vitro exposure of human melanoma cell lines to IFN-γ or inflammatory cytokines, such as TNF-α, induced GILT expression in melanoma cell lines, which lacked GILT expression at baseline. Vemurafenib, a BRAF inhibitor used in the treatment of metastatic melanoma, enhanced IFN-γ-induced GILT and MHC class II expression in a melanoma cell line. Together these data demonstrate that inflammation, alone or in combination with a current therapeutic agent, induces high levels of GILT expression in human melanoma.

INTRODUCTION Of about 20% of patients with relapsed/refractory Hodgkin lymphoma (HL), approximately half achieve long-term remissions after high-dose chemoradiotherapy and autologous stem cell transplantation (HDT/ASCT). Treatment with a comprehensive program using second-line chemotherapy with ICE (ifosfamide, carboplatin, etoposide) before HDT/ASCT identified a clinical prognostic model, but prognostic biologic markers in relapsed/refractory HL remain unclear. We sought to determine if we could identify such markers, and if our comprehensive second-line program could overcome their significance. METHODS Pre-ICE biopsy specimens of 191 patients enrolled on 1 of 2 Institutional Review Board-approved clinical trials of HDT/ASCT. We performed immunohistochemistry staining for Bcl-2, Bax, Bim, p53, and interleukin-6. Samples were considered positive if more than 10% of Hodgkin Reed-Sternberg cells stained at any intensity. RESULTS Ninety-one patients had sufficient tissue available. Forty-eight patients (53%) had an event and 36 (40%) died. Median event-free survival (EFS) was 8.5 years, median overall survival (OS) was not reached, and median follow-up was 8.8 years. Bcl-2 was overexpressed in 37/91 (41%), Bax in 28/65 (43%), Bim in 9/72 (13%), p53 in 38/89 (43%), and interleukin-6 in 58/84 (69%) patients. Overexpression of these biomarkers had no statistically significant association with EFS or OS, except for association of Bim overexpression with inferior OS (P = 0.0385). The 3-factor clinical model (B symptoms at relapse, extranodal disease, and complete remission duration of < 1 y) remained highly significant (0/1 vs. 2/3 factors) for EFS and OS (P = 0.0008 and P = 0.0001, respectively). CONCLUSIONS Despite the evidence that p53 and Bcl-2 overexpression may predict a worse prognosis with initial treatment, it seems that at relapse such overexpression is either not prognostically significant, or that the treatment with ICE and HDT/ASCT overcomes its significance. Subsequent studies should further address the role of Bim in both initial and relapsed/refractory settings.

Reaction of cyclic diaminochlorosilanes with 1,3-bis(tert-butyl)imidazol-2-ylidene resulted in the facile formation of the corresponding stable and transient diaminosilylenes. This novel dehydrochlorination route could be applied for the generation of four- and five-membered N-heterocyclic silylenes with a range of different substituents under very mild conditions. Activation of an olefinic C−H bond and reduction of a cyclic diaminochlorosilane have been observed for these new transient silylenes.

A bigenic MUC1.Tg/MIN mouse model was developed by crossing Apc/(MIN/+) (MIN) mice with human MUC1 transgenic mice to evaluate MUC1 antigen-specific immunotherapy of intestinal adenomas. The MUC1.Tg/MIN mice developed adenomas at a rate comparable to that of MIN mice and had similar levels of serum MUC1 antigen. A MUC1-based vaccine consisting of MHC class I-restricted MUC1 peptides, a MHC class II-restricted pan-helper peptide, unmethylated CpG oligodeoxynucleotide and GM-CSF caused flattening of adenomas and significantly reduced the number of large adenomas. Immunization was successful in generating a MUC1-directed immune response evidenced by increased MUC1 peptide-specific anti-tumor cytotoxicity and IFN-gamma secretion by lymphocytes.

A novel alkali extractable polysaccharide (designated as PNA-2) was purified from Pleurotus nebrodensis and the effects of purified PNA-2 on the proliferation and apoptosis of human hepatic cancer cells (HepG2) were investigated in this study. The results of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay indicated that PNA-2 inhibited the proliferation of HepG2 cells by apoptosis induction, which was also characterized using scanning electron microscopy (SEM). Moreover, the expression of apoptosis-associated mRNA, proteins and the cell-cycle arrest at the G0/G1 phase was determined using RT-qPCR, Western blot and flow cytometry, respectively. A notable inhibition of the migration rate of PNA-2-treated HepG2 cells was observed using a cell scratch assay. DNA damage was observed using a comet assay and AO/EB staining in HepG2 cells, which were exposed to PNA-2. Induction of the mitochondria-mediated intrinsic apoptotic pathway by PNA-2 was indicated by the loss of mitochondrial membrane potential (ΔΨm), Bcl-2 dysregulation and cytochrome c release. All the results suggested that the mitochondria-mediated intrinsic apoptotic pathway could be involved in PNA-2-mediated apoptosis of human liver carcinoma cells HepG2. Finally, the results indicated that PNA-2 significantly suppressed tumor growth in HepG2 tumor-bearing mice, indicating that PNA-2 may be developed as a candidate drug or functional food factor to prevent or treat liver cancer.

This study is the first to describe the composition and characteristics of culturable bacterial isolates from the rhizosphere and bulk soil of the medicinal plant Stellera chamaejasme L. at different growth stages. Using a cultivation-dependent approach, a total of 148 isolates showing different phenotypic properties were obtained from the rhizosphere and bulk soil. Firmicutes and Actinobacteria were the major bacterial groups in both the rhizosphere and bulk soil at all 4 growth stages of S. chamaejasme. The diversity of the bacterial community in the rhizosphere was higher than that in bulk soil in flowering and fruiting stages. The abundance of bacterial communities in the rhizosphere changed with the growth stages and had a major shift at the fruiting stage. Dynamic changes of bacterial abundance and many bacterial groups in the rhizosphere were similar to those in bulk soil. Furthermore, most bacterial isolates exhibited single or multiple biochemical activities associated with S. chamaejasme growth, which revealed that bacteria with multiple physiological functions were abundant and widespread in the rhizosphere and bulk soil. These results are essential (i) for understanding the ecological roles of bacteria in the rhizosphere and bulk soil and (ii) as a foundation for further evaluating their efficacy as effective S. chamaejasme growth-promoting rhizobacteria.

Background: A high-fat diet is a risk factor for the development of inflammatory bowel disease (IBD) in humans. Deoxycholate (DOC) is increased in the colonic contents in response to a high-fat diet. Thus, an elevated level of DOC in the colonic lumen may play a role in the natural course of development of IBD. Methods: Wild-type B6.129 mice were fed an AIN-93G diet, either supplemented with 0.2% DOC or unsupplemented and sacrificed at 1 week, 1 month, 3 months, 4 months, and 8 months. Colon samples were assessed by histopathological, immunohistochemical, and cDNA microarray analyses. Results: Mice fed the DOC-supplemented diet developed focal areas of colonic inflammation associated with increases in angiogenesis, nitrosative stress, DNA/RNA damage, and proliferation. Genes that play a central role in inflammation and angiogenesis and other related processes such as epithelial barrier function, oxidative stress, apoptosis, cell proliferation/cell cycle/DNA repair, membrane transport, and the ubiquitin-proteasome pathway showed altered expression in the DOC-fed mice compared with the control mice. Changes in expression of individual genes (increases or reductions) correlated over time. These changes were greatest 1 month after the start of DOC feeding. Conclusions: The results suggest that exposure of the colonic mucosa to DOC may be a key etiologic factor in IBD. The DOC-fed mouse model may reflect the natural course of development of colitis/IBD in humans, and thus may be useful for determining new preventive strategies and lifestyle changes in affected individuals.

Background: Off-pump coronary artery bypass (OPCAB) grafting is gaining popularity; however, decreased mid-term graft patency and increased coronary reintervention rates are reported. Study Design: Five hundred and ninty-one consecutive unselected patients underwent OPCAB grafting from February 2000 to April 2004 (mean follow-up 38.54 ± 0.54 months). One hundred and thirteen patients had ≤2 grafts, and four hundred and seventy-eight had ≥3 grafts. At least one radial artery graft was present in 398 patients, 328 received postoperative Clopidogrel, and 391 received postoperative statins. History of at least one percutaneous coronary intervention (PCI) was present in 192 patients. Results: Twenty-nine patients developed recurrent angina, nine had myocardial infraction, and twenty underwent coronary reintervention. Five patients died of sudden cardiac death. Overall mortality was 4.9% during follow-up (29 patients). Postoperative Clopidogrel and statins as well as intraoperative shunt use significantly decreased symptom recurrence and adverse cardiac events. Diabetes, chronic obstructive pulmonary disease, prior history of PCI, and utilization of radial artery grafts were positive predictors of symptom recurrence and adverse cardiac events. Utilization of radial artery grafts, history of PCI as well as low preoperative ejection fraction increased mortality. Number of bypass grafts, type of conduit, grafted territory, hyperlipidemia, or prior coronary artery bypass graft surgery (CABG) did not influence symptom recurrence, adverse cardiac events or mortality. Conclusions: OPCAB grafting can be performed with low symptom recurrence, adverse cardiac events, and mortality rates. Modification of intra- and postoperative management strategies may improve outcomes.

Purpose: Barrett’s esophagus (BE) is a common premalignant lesion of the distal part of the esophagus that arises as a consequence of chronic duodenogastroesophageal reflux. Interleukin (IL)-6 is a pleiotropic cytokine that regulates immune defense mechanisms and hematopoiesis. In addition, IL-6 may also be involved in malignant transformation and tumor progression. IL-6 has been shown to inhibit apoptosis. The major aim of this study was to evaluate expression of IL-6 in BE at the protein and mRNA levels. In addition, we tested whether proteins that are associated with IL-6 signaling, phosphorylated signal transducer and activator of transcription 3 and two antiapoptotic proteins, Bcl-xL and Mcl-1, are also expressed in the same tissues.Experimental Design: Biopsies of duodenum, BE, and squamous epithelium were evaluated by using a human cytokine protein array, ELISA, real-time PCR, and immunohistochemistry.Results: Increased IL-6 levels were found to be secreted from BE tissue compared with duodenum or squamous epithelium from sites adjacent or 5 cm away from the BE lesion. IL-6 mRNA was also elevated in BE compared with duodenum or squamous epithelium in five of seven patients. Immunohistochemical studies confirmed IL-6 expression in intestinal glandular epithelium in BE tissue. Activated signal transducer and activator of transcription 3, Mcl-1, and Bcl-xL are present at higher levels in BE glands, with lower levels being found in duodenum or squamous epitheliumConclusions: These data, taken together, suggest that elevated IL-6 levels in BE may contribute to the development of apoptosis resistance, thereby placing this epithelium at higher risk of developing malignancy.

Thioredoxin-1 is a redox protein that, when overexpressed, causes increased cancer-cell growth and inhibited apoptosis. Thioredoxin-1 expression has been reported to be increased in several human primary tumors, but its relationship to tumor progression and patient survival has not been established. We studied the expression of thioredoxin-1 as measured with immunohistochemical staining in paraffin-embedded human normal colonic mucosa, adenomatous polyps, and primary and metastatic colorectal cancer. Thioredoxin-1 expression was not increased in 12 colorectal adenomatous polyps, compared with 8 samples of normal colonic mucosa, but was significantly increased in 12 primary colorectal cancers (P

OBJECTIVE: Gastroesophageal reflux disease (GERD) plays a major role in the development of Barrett’s esophagus. Recently, we demonstrated that duration of esophageal acid exposure in the distal esophagus correlates with the length of Barrett’s mucosa. The aim of this study was to determine whether there is a relationship between the rate of the change in acid exposure along the esophagus and the length of Barrett’s esophagus. METHODS: A total of 17 patients (16 men and one woman; mean age 66 ± 15 yr, range 41–83 yr) with varying lengths of biopsy-proven Barrett’s esophagus were recruited prospectively into the study. Ambulatory 24-h esophageal pH monitoring was performed using a commercially available pH probe with four sensors located 5 cm apart. For each patient, a least squares regression line of the fraction of the study that the pH was

The distribution of α6/α3 integrin in adhesion complexes at the basal membrane in human normal and cancer prostate glands was analyzed in 135 biopsies from 61 patients. The levels of the polarized α6/α3 integrin expression at the basal membrane of prostate tumor glands were determined by quantitative immunohistochemistry. The α6/α3 integrin expression was compared with Gleason sum score, pathological stage, and preoperative serum prostate-specific antigen (PSA). The associations were assessed by statistical methods. Eighty percent of the tumors expressed the α6 or α3 integrin and 20% was integrin-negative. Gleason sum score, but not serum PSA, was associated with the integrin expression. Low Gleason sum score correlated with increased integrin expression, high Gleason sum score with low and negative integrin expression. Three prostate tumor phenotypes were distinguished based on differential integrin expression. Type I coexpressed both α6 and α3 subunits, type II exclusively expressed a6 integrin, and type III expressed α3 integrin only. Fifteen cases were further examined for the codistribution of vinculin, paxillin, and CD 151 on frozen serial sections using confocal laser scanning microscopy. The α6/α3 integrins, CD151, paxillin, and vinculin were present within normal glands. In prostate carcinoma, α6 integrin was colocalized with CD 151, but not with vinculin or paxillin. In tumor phenotype I, the α6 subunit did not colocalize with the α3 subunit indicating the existence of two different adhesion complexes. Human prostate tumors display on their cell surface the α6β1 and/or α3β1 integrins. Three tumor phenotypes associated with two different adhesion complexes were identified, suggesting a reorganization of cell adhesion structures in prostate cancer.

Background: Kallikrein-related peptidase 6 (KLK6) belongs to the family of human tissue kallikrein genes, majority of which are shown to be differentially expressed in cancers. Clinical studies have demonstrated that upregulation of KLK6 in primary colorectal tumors and lymph nodes correlates with serosal invasion, liver metastasis and indicative of poor prognosis in patients. It has been reported that KLK6 protein is involved in regulation of the epithelial-mesenchymal transition (EMT) program in an organ-specific context. The aim in this study was to investigate contribution of KLK6 enzyme in the EMT during neoplastic transformation in the colon. Results: We expressed enzymatically active or inactive KLK6, using pcDNA3.1(+)preproKLK6 and pcDNA3.1(+)preproKLK6 Ser197Ala mutant plasmids, in Caco-2 colon cancer cell line, which has been characterized before as a very low KLK6 expresser with an undetectable secreted KLK6. Stable isogenic clones were selected and further evaluated for their ability to migrate and invade using in vitro assays and to metastasize in vivo using SCID orthotopic mouse model. We found no effect of KLK6 enzyme activity on migration of Caco-2 cells, expressing the empty vector (Caco-2 mock), and Caco-2 cells, expressing an enzymatically active KLK6 (Caco-2 KLK6 wt) or inactive KLK6 (Caco-2 KLK6 mut). But Caco-2 KLK6 wt cells demonstrated the invasive phenotype in Matrigel invasion assays (p Conclusion. These findings demonstrate that KLK6 enzyme activity is required for colon cancer progression via induction of the EMT program. We identified the TGF-β- signaling pathway as a mechanism driving the EMT in colon cancer cells expressing KLK6 enzyme. Citation Format: Hwudaurw Chen, Earlphia Sells, Haiyan Cui, Ritu Pandey, George Pampalakis, Georgia Sotiropoulou, Thomas Doetschman, Natalia A. Ignatenko. Human tissue Kallikrein 6 enzyme activity regulates epithelial-mesenchymal transition in colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 854. doi:10.1158/1538-7445.AM2017-854