text:
MUC4 is a novel mediator in H. pylori infection‑related pancreatic cancer
Yu Gong
Shuai Chen
Yue Fu
Yu Liu
Yipeng Wang
Haojun Yang
Hanyang Liu
Liming Tang
Affiliations: Research Center of General Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu 213000, P.R. China,, Research Center of General Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu 213000, P.R. China, Charité‑University Medical Center, Department of Hematology, Oncology and Tumor Immunology, Virchow Campus, and Molecular Cancer Research Center, D‑13353 Berlin, Germany
Published online on: December 17, 2020 https://doi.org/10.3892/ol.2020.12384
Copyright: © Gong et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pancreatic cancer (PC) is a common malignant disease worldwide. Among the potential pathogenic factors, Helicobacter pylori (H. pylori) infection has been associated with the tumorigenesis of PC. The present study aimed to identify the differentially expressed genes (DEGs) of H. pylori infection‑associated PC and to investigate the key factors involved in PC tumorigenesis. Using bioinformatics methods, overlapping DEGs and key gene were identified from H. pylori‑infected gastric mucosa (GM) and H. pylori infection‑associated PC. Survival and tumor stage analyses were performed to assess the clinical associations. In addition, mucin 4 (MUC4) mRNA expression levels were measured in patient blood and tumor samples. According to the correlation analyses of four genes co‑expressed, potential biological processes were identified. MUC4 was identified to be associated with H. pylori infection, and its levels were significantly upregulated in PC samples compared with those in normal samples in TCGA dataset, the PC cell line and patient tissue samples. H. pylori infection was also associated with MUC4 expression in patients' blood and tissue samples. In conclusion, the results of the present study revealed a potentially pathogenic role of MUC4 in H. pylori infection‑associated PC. Thus, the tumorigenesis and metastasis of PC may be prevented by treating the H. pylori infection or using MUC4 antagonists.
Pancreatic cancer (PC) is one of most lethal types of malignancy worldwide, with equal incidence rates in men and women (~2.5%) (1). PC is a significant health burden worldwide, affecting >411,600 individuals and exhibiting a 5-year survival rate of <5% (2,3). Although medical technologies have improved in recent decades, no reliable diagnostic biomarkers are currently available for the detection of asymptomatic PC at an early stage. Consequently, PC is usually diagnosed at a late stage, when the majority of patients exhibit distant metastasis. Therefore, >80% of patients with PC are not eligible for surgical resection, which is currently considered to be the most effective treatment (4).
In humans, the incidence of PC is complex, and risk factors include a high-protein diet, high-fat diet, smoking and susceptibility to genetic mutations (5). However, another risk factor is gastric mucosal injury following Helicobacter pylori (HP) infection (6). Previous studies have reported that the pathophysiological actions of H. pylori colonization, alongside the gastric acidity, potentially modulate pancreatic tumorigenesis via N-nitrosamine intake-mediated hyperchlorhydria (6,7). Due to the crucial role that the tumor microenvironment serves in cancer progression, increasing attention has been paid to the extracellular matrix, stromal, immune and stem cells that predominate the PC microenvironment (8).
According to previous studies, H. pylori has developed mechanisms to co-exist in the harsh gastric microenvironment by inducing mucosal inflammation and immune activation (9–11). H. pylori may induce the secretion of inflammatory cytokines or neuroendocrine mediators by the infected gastric mucosa (GM) cells (12). In contrast to the direct carcinogenic impact of metabolites on GM or esophageal mucosa, H. pylori infection may promote neoplasia and metastasis of distant organs such as the pancreas by mediating the cellular microenvironment (13,14).
The results of the aforementioned studies have provided initial evidence of the incidence of H. pylori infection-associated PC; however, the biological mechanisms and key factors involved in this process remain poorly understood. In order to improve the current understanding, the present study aimed to identify the factors associated with the potential pathogenicity of H. pylori in PC tumorigenesis and metastasis.
The human PC cell line PANC-1 and the human normal ductal epithelial cell line HPDE6-C7 were purchased from the Cell Resource Center of Shanghai Institute of Biochemistry and Cell Biology, The Chinese Academy of Sciences. The cells were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and maintained at 37°C with 5% CO2.
Patient studies
Between June and December 2019, a total of 32 inpatient volunteers, among whom 20 patients had chronic gastritis without PC (10 HP− and 10 HP+) and 12 had PC (6 HP− and 6 HP+), were recruited at The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University (Changzhou, China) (Table I). The patients were diagnosed with H. pylori infection, PC or both. H. pylori infection was diagnosed by carbon 13 breath test and gastroscopy. GM injury was determined by gastroscopy. The diagnosis and tumor stage of PC were systemically assessed using imaging, surgery and biopsy. The study was approved by the Ethics Committee of the Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University [approval no: (2019) KYO 073-01], and all patients provided informed consent. PC and adjacent tissue samples (>5 cm from the tumor) were obtained during surgery. Blood samples were obtained after diagnosis. All samples (20 GM, 12 tumor, 12 adjacent tissue and 12 blood samples) were stored at −80°C and preprocessed for further research.
Clinicopathological characteristics of patients included in the present study.
HP+
HP−
Total, n 10 10 6 6
Sex, n
Male 6 5 4 4
Female 4 5 2 2
Age, years 39.56±4.64 40.32±5.03 53.87±4.23 56.42±8.53
GM injury, n (%) 8 (80%) 7 (70%) 2 (33%) 2 (33%)
Lymph node invasion, n (%) N/A N/A 2 (33%) 2 (33%)
Distant metastasis, n (%) N/A N/A 1 (16%) 0 (0%)
[i] GM, gastric mucosa; PC, pancreatic cancer; HP, Helicobacter pylori; N/A, not applicable.
Microarray datasets of H. pylori-infected GM [dataset nos. GSE6143 (15), GSE47797 (16), GSE70394 (17) and GSE5081 (18)] and PC [dataset nos. GSE85991 (19), GSE27890 and GSE55643 (20)] were downloaded from the Gene Expression Omnibus (GEO) database (http://www.ncbi.nlm.nih.gov/geo). By referring to the annotation information, the probe IDs were converted into the corresponding gene symbols. Basic information of the microarray datasets is displayed in Table II.
Table II.
Microarray datasets of patients with Helicobacter pylori infection and pancreatic cancer.
Sample character
(Refs.)
Wen et al, 2007 GSE6143 GPL193 Gastric mucosa H. pylori+ 9 (15)
Hanada et al, 2014 GSE47797 GPL13497 Gastric mucosa H. pylori+ 4 (57)
Costa et al, 2016 GSE70394 GPL6480 Gastric mucosa H. pylori+ 3 (17)
Galamb et al, 2008 GSE5081 GPL570 Injured gastric mucosa H. pylori+ 8 (18)
H. pylori− 8
Koutsioumpa et al, 2019 GSE85991 GPL570 Pancreatic cancer Tumor 2 (19)
Chinaranagari et al, GSE27890 GPL570 Pancreatic cancer Tumor 4 (58)
2015 Normal 4
Lunardi et al, 2014 GSE55643 GPL6480 Pancreatic cancer Tumor 45 (20)
Identification and overlap of differentially expressed genes (DEGs)
The DEGs between the tumor and control groups were identified using the GEO2R web tool (http://www.ncbi.nlm.nih.gov/geo/geo2r) by comparing the GEO microarray datasets. Probes without corresponding gene symbols or genes with redundant probe sets were eliminated or processed using the DAVID online tool (https://david.ncifcrf.gov/), respectively. A |log (fold-change)| >1 and P<0.01 were selected to identify statistically significant differences. The overlap analysis of DEGs from the H. pylori-infected GM and PC datasets was conducted and displayed in Venn diagrams (http://bioinformatics.psb.ugent.be/webtools/Venn/). In addition, the mRNA expression levels of the DEGs were presented in heatmaps.
Kyoto encyclopedia genes and genomes (KEGG) signaling pathway and gene ontology (GO) functional term enrichment analyses
The KEGG Orthology Based Annotation System (KOBAS 3.0; http://kobas.cbi.pku.edu.cn/kobas3) web server was used for gene/protein functional annotation (Annotation module) and functional set enrichment (Enrichment module) in the present study. GO functional term enrichment analysis bioinformatics annotation tool was used to determine gene functions and perform biological analysis (21). KEGG signaling pathway enrichment analysis was used to illustrate gene functions and biological pathways (22). P<0.01 was considered to indicate a statistically significant difference.
Data acquisition from The Cancer Genome Atlas (TCGA) database
Gene expression data of PC were acquired from TCGA (https://portal.gdc.cancer.gov) (23). Overall survival (OS) and disease-free survival (DFS) analyses of patients grouped into high/low mucin 4 (MUC4) mRNA expression groups based on the median expression levels were performed using the Gene Expression Profiling Interactive Analysis (GEPIA) online database (http://gepia.cancer-pku.cn) (24). The associations between the expression levels and tumor grades and the meta-analysis of the data from three previous studies (25–27) were analyzed using the Oncomine database (http://www.oncomine.com) (28).
Reverse transcription-quantitative PCR (RT-qPCR)
Total RNA was extracted from the PC cell line and tissues, as well as the corresponding controls, using TRIzol® reagent (Invitrogen; Thermo Fisher Scientific, Inc.). The total RNA was reverse-transcribed into cDNA using a PrimeScript™ RT reagent kit (Takara Biotechnology Co., Ltd.). qPCR was subsequently performed using a SYBR® Premix Ex Taq kit (Takara Biotechnology Co., Ltd.) on a 7500 Real-Time PCR system (Applied Biosystems; Thermo Fisher Scientific, Inc.). The reverse transcription was performed at 42°C for 1 h, followed by 95°C for 5 min. The thermocycling conditions were as follows: Initial denaturation step at 95°C for 10 min, followed by 40 cycles at 95°C for 15 sec and at 60°C for 1 min. The expression levels were calculated using the 2−ΔΔCq method (29). The primer pairs used were as follows: MUC4 forward, 5′-GACTTGGAGCTCTTTGAGAATGG-3′ and reverse, 5′-TGCAATGGCAGACCACAGTCC-3′; GAPDH forward, 5′-ATCATCCCTGCCTCTACTGG-3′ and reverse, 5′-GTCAGGTCCACCACTGACAC-3′.
Multilevel logistic regression analysis
Basic information of 22 patients (12 PC+HP+/− and 10 PC−HP−) was obtained from the medical records. The patients were grouped by age (>/<50 years), sex (male/female), MUC4 expression, HP infection (HP+/HP−) and PC diagnosis (PC+/PC−). Multilevel logistic regression analysis was made with grouped patients.
Data are presented as the mean ± SEM. SPSS 17.0 (SPSS statistics, IBM Inc.) was used for statistical analysis. GraphPad Prism 6.0 software (GraphPad Software, Inc.) was used to generate the receiver operating characteristic (ROC) curve and perform multilevel logistic regression analysis. Statistical differences were determined using one-way ANOVA with the Bonferroni post hoc test for multiple comparisons. Correlation analysis was performed using the Spearman correlation coefficient. P<0.05 was considered to indicate a statistically significant difference.
Identification of overlapping DEGs in H. pylori-infected GM and PC
A total of 32 patients and seven microarray datasets were included in the present study (Fig. 1). Following standardization and processing on the GEO database, DEGs were selected from three H. pylori-infected GM and three PC datasets. Using overlap analysis, six genes were selected among the 44 and 49 DEGs from H. pylori-infected GM and PC datasets, respectively, which were 6-phosphofructo-2-kinase (PFKFB3), sodium-dependent serotonin transporter (SLC6A4), claudin-1 (CLDN1), MUC4, prominin-2 (PROM2) and matrix metalloproteinase 9 (MMP9) (Fig. 2A). The functional annotation of these six genes is presented in Table III. The relative mRNA expression levels of the genes that were significantly upregulated are presented in heat maps in Fig. 2B. For further verification, gene expression data from patients with PC were downloaded from TCGA database. As presented in Fig. 2C, TCGA data analysis revealed that the expression levels PFKFB3, CLDN1, MUC4, PROM2 and MMP9 were significantly upregulated in the tumor tissues compared with those in the normal tissues (P<0.05). However, the expression levels of SLC6A4 were not significantly different between the tumor and normal tissues (P>0.05). These enriched genes were associated with a number of GO functional terms, such as 'biological regulation', 'regulation of biological process' and 'regulation of cellular process', and KEGG signaling pathways, including 'cytokine-cytokine receptor interaction', 'viral protein interaction with cytokine' and 'NF-kappa B signaling pathway' (Fig. 2D and E).
PRISMA flow of the present study. The research procedures of this study were classified into four parts: 'Identification', 'Screening', 'Eligibility' and 'Inclusion'.
Screening and enrichment of DEGs in H. pylori-infected GM and PC. (A) DEGs were selected with a fold-change >1 and P<0.01 among the mRNA expression profiling datasets of H. pylori-infected GM and PC. An overlap of six genes was identified among the H. pylori-infected GM and PC samples. (B and C) Detailed expression patterns of the six genes based on TCGA analysis are presented in (B) heatmaps and (C) boxplots. (D and E) The six genes were subjected to (D) GO functional term and (E) KEGG signaling pathway enrichment analysis. *P<0.01. Titles and percentages of each enriched term are represented. GM, gastric mucosa; PC, pancreatic cancer; DEGs, differentially expressed genes; H. pylori, Helicobacter pylori; TCGA, The Cancer Genome Atlas Gene Ontology Kyoto Encyclopedia of Genes and Genomes.
Table III.
Full names and functions of the six key differentially expressed genes identified in the present study.
PFKFB3 6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 Required for cell cycle progression and prevention of apoptosis; functions as a regulator of cyclin-dependent kinase 1, linking glucose metabolism to tumor cell proliferation and survival.
SLC6A4 Solute carrier family 6 member 4 A target of psychomotor stimulants, such as amphetamines and cocaine; a member of the sodium: neurotransmitter symporter family; a repeat length polymorphism in the promoter of this gene has been demonstrated to affect the rate of serotonin uptake.
CLDN1 Claudin 1 A member of the claudin family; an integral membrane protein; a component of tight junction strands, serving as a physical barrier to prevent solutes and water from passing freely through the paracellular space.
MUC4 Mucin 4, cell surface associated A major constituent of mucus; serves important roles in the protection of the epithelial cells; has been implicated in epithelial renewal and differentiation.
PROM2 Prominin 2 A member of the prominin family of pentaspan membrane glycoproteins; may be involved in the organization of plasma membrane microdomains.
MMP9 Matrix metallopeptidase 9 Involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction and tissue remodeling, as well as in disease processes, such as arthritis and tumor metastasis.
MUC4 contributes to H. pylori infection-associated PC
The GSE5081 dataset was used to compare the whole genome gene expression profile between patients with HP+ and HP− GM injury. Among the six selected genes, MUC4, MMP9 and CLDN1 were differentially expressed in the two groups. Notably, compared with normal GM tissues, MUC4 expression levels were upregulated in HP+ and downregulated in HP− GM injured tissues (Fig. 3A and B). These results indicated that MUC4 may be a potential factor involved in the biological processes of the H. pylori-infected GM. For further analysis, the expression profile of MUC4 in human tissues and the associated clinical information were produced using the GEPIA webtool. The human body maps illustrated that MUC4 expression levels were upregulated in pancreatic adenocarcinoma, cervical squamous cell carcinoma and endocervical adenocarcinoma, lung adenocarcinoma (LUAD) and stomach adenocarcinoma (STAD), but downregulated in prostate adenocarcinoma and head & neck squamous cell carcinoma compared with those the corresponding normal tissues (Fig. 3C and D). In addition, Oncomine-based meta-analysis indicated that the expression levels of MUC4 were significantly upregulated in cancer tissues compared with those in normal tissues in the three PC datasets (Fig. 3E). However, as presented in Fig. 3F, MUC4 was not associated with patient OS rates in PC. A weak association was identified between low MUC4 levels and DFS rates; however, this result was not significant (P>0.05; Fig. 3G). In addition, although MUC4 expression levels were upregulated at each tumor stage, no significant differences were identified among the four stages (P>0.05; Fig. 3H).
Gene expression profiles of clinical samples and their associated predictive ability for patient survival. (A and B) Expression patterns of MUC4 are presented as a (A) heatmap and (B) scatter plot in injured GMs with or without H. pylori infection. (C) A body map and (D) histogram display the distribution of MUC4 in the human body. (E) Meta-analysis of MUC4 expression levels was performed. Red, upregulation and blue, downregulation in the tumor tissues compared with normal tissues. (F-H) MUC4-related (F) overall survival, (G) disease-free survival and (H) tumor stage analyses. *P<0.05. MUC4, mucin 4; GM, gastric mucosa; HP/H. pylori, Helicobacter pylori; TCGA, The Cancer Genome Atlas.
The human PC cell line PANC-1, 20 patients with chronic gastritis without PC and 12 patients with PC with or without H. pylori infection were used to verify the trends observed in MUC4 expression. Blood and tissue samples (GM, tumor and adjacent tissues) were collected and analyzed by RT-qPCR. The expression levels of MUC4 were upregulated in HP+ GM compared with those in HP− GM (P<0.05; Fig. 4A). In addition, compared with the normal tissues and cell lines, the expression levels of MUC4 were upregulated in both PC tissues and PANC-1 cells (P<0.05; Fig. 4B). The results of the blood tests revealed that patients in the HP+PC− group exhibited markedly upregulated MUC4 expression levels compared with those in the other groups. (P<0.05; Fig. 4C). These results indicated that the expression levels of MUC4 were significantly upregulated in H. pylori-infected GM and PC.
Verification of MUC4 expression levels in the GM, PC and blood. (A-C) mRNA expression levels of MUC4 in (A) injured GMs with or without H. pylori infection, (B) tumor tissues (relative to normal tissues) and PC cell lines (relative to a human normal ductal epithelial cell line), and (C) blood samples from patients with H. pylori infection or PC. (D) In a total of 26 patients, a ROC curve was generated to assess the sensitivity and specificity in HP-infection related PC of MUC4. (E) Multilevel logistic regression analysis of age, sex, high MUC4 levels and HP infection vs. PC status was conducted. *P<0.01. MUC4, mucin 4; GM, gastric mucosa; PC, pancreatic cancer; HP/H. pylori, Helicobacter pylori; ROC, receiver operating characteristic.
To assess the sensitivity of MUC4, 26 patients were divided into very low-risk (HP−PC−), low-risk (HP+PC−) and high-risk (HP+PC+) groups. A ROC curve was generated, and the results revealed that MUC4 exhibited moderate sensitivity for the identification of H. pylori-related PC (Fig. 4D). Multilevel logistic regression analysis of the effects of patient age, sex, MUC4 expression levels and H. pylori infection in PC was conducted, which revealed that high MUC4 expression levels and the presence of an H. pylori infection were positively associated with PC (Fig. 4E). Correlation analyses of multiple genes co-expressed with MUC4 were conducted using TCGA data, and four tumorigenic factors of PC, including receptor tyrosine-protein kinase erbB-2 (ErbB2), K-Ras, EGFR and SMAD2, were identified to be moderately positively correlated with MUC4 (Fig. 5A).
Correlation analyses and hypothetical mechanisms of the role of MUC4 in PC. (A) Correlation analyses between the expression levels of ErbB2, K-Ras, EGFR or SMAD2 and MUC4 were performed using The Cancer Genome Atlas. (B and C) Hypothetical mechanisms of (B) the secretion and transportation of MUC4 and (C) biological processes of MUC4 in PC. MUC4, mucin 4; PC, pancreatic cancer; ErbB2, receptor tyrosine-protein kinase erbB-2; HP, Helicobacter pylori.
The pancreas is an important retroperitoneal organ with exocrine and endocrine functions (30). Non-endocrine pancreatic tumors are usually malignant and have different histological features that can be classified as a ductal adenocarcinoma, cystadenocarcinoma or another type of malignant tumor, such as sarcoma and small cell carcinoma (31). In recent decades, the prognosis of PC has remained dismal, with limited improvements achieved for the diagnosis and treatment of the disease. Despite advances in surgery and comprehensive treatment regimens, the poor outcome of patients with PC remains unchanged (32). The results of epidemiological studies have indicated that age, obesity, smoking, long-term alcohol use, chronic pancreatitis and family history are all risk factors for PC (31). With the development of genomics and bioinformatics tools, several genetic mutations have been identified to be involved in PC development; for example, BRCA2 and K-Ras mutations, and loss of p16, SMAD4 or p53 function occur in the epithelium of precursor pancreatic diseases, which markedly accelerate the progression of tumorigenesis (33–35).
The stomach serves several exocrine functions that are similar to the pancreas in the gastrointestinal system, such as producing and secreting various polypeptides and proteins (36–38). H. pylori, recognized as the most common gastroduodenal infection, typically colonizes the human stomach (15). Chronic H. pylori infection has been associated with an increased risk of several types of disease or pathological lesions, including gastritis, peptic ulcers, dysplasia, neoplasia, mucosa-associated lymphoid tissue lymphoma and invasive gastric adenocarcinoma (39–41). In addition, a potential role of H. pylori infection in several extragastric diseases, such as neurodegenerative, cardiovascular, hepatobiliary, pancreatic and colorectal diseases, has been reported (7). Epidemiological and cohort studies in large numbers of PC cases have verified that H. pylori infection serves a role in the pathogenesis of chronic and autoimmune pancreatitis, diabetes and PC. Subsequent studies have suggested that H. pylori infection may affect the pancreatic physiology and contribute to the tumorigenesis of PC (17). A number of factors produced in response to infection, including ammonia, lipopolysaccharides and inflammatory cytokines, have been demonstrated to induce progressive damage in the pancreas (8). Maisonneuve and Lowenfels and Yadav amd Lowenfels (12,42) have reported that similar pathological manifestations to H. pylori-infected gastric tissues are also observed in PC. Other previous studies have indicated that increases in gastrin levels and decreases in somatostatin levels are involved in the mechanisms associated with H. pylori infection and PC (43). In addition, tissue inflammation, genomic DNA damage and various cytokines, including NF-κB, activator protein-1 and serum response elements, have been reported to contribute to the malignant transformation of pancreatic cells (5,44).
MUC4 has been identified to be associated with H. pylori infection. MUC4 is a major constituent of the mucus, the viscous secretion that covers epithelial surfaces, such as those in the trachea, colon and cervix, composed of highly glycosylated proteins called mucins (45). Multiple previous studies have reported a role of MUC4 in the aggressiveness of PC through its ability to enhance tumor growth and metastasis (46–49). The results of the present study demonstrated that the expression levels of MUC4 were also upregulated in LUAD and STAD compared with those in normal tissues, suggesting that the role of MUC4 may depend on the type of cancer.
Although MUC4 levels were upregulated in H. pylori infection-associated GM and PC samples compared with those in normal tissues, there is currently lack of evidence of an association between the two in existing studies. Following the analysis of MUC4 expression levels in patient blood and tissues, cytokine transportation and biological networks were hypothetically constructed and visualized in the present study (Fig. 5B and C). The results of the present study revealed that MUC4 expression levels were upregulated in patients with H. pylori infection regardless of GM damage compared with those in normal tissues. Thus, H. pylori infection potentially promotes the intracellular transcription and the extracellular secretion of MUC4 in GM cells. The circulatory and immune systems and the digestive tract appear to be main routes for MUC4 transportation (46,50). Generally, the microenvironment of PC cells consists of vessels, immune cells, stroma, stem cells, stellate cells and fibroblasts (8,51,52). MUC4 is transported to the surrounding microenvironment, where it potentially participates in the cellular biological processes of PC. MUC4 has been reported to serve as an intramembrane ligand and co-mediate with ErbB2 (53,54). Correlation analyses in the present study revealed that the expression levels of PC tumorigenic factors (ErbB2, K-Ras, EGFR and SMAD2) were associated with those of MUC4 in PC tissues. A previous study has concluded that the MUC4/ErbB2 complex contributes to processes such as differentiation, proliferation, cell cycle, angiogenesis and migration by regulating the EGF, K-Ras, PI3K/AKT and TGF-β signaling pathways (48,55). Therefore, MUC4 produced by the H. pylori-infected GM may be involved in mediating cell survival and migration by regulating the PC cell microenvironment.
MUC4 is considered to serve a crucial role in the tumorigenesis and metastasis of PC (47,56). In the present study, MUC4 was found to be upregulated in H. pylori-infected GM. Nevertheless, the underlying mechanisms of MUC4 between H. pylori infection and PC have received limited attention, especially regarding the cellular transport mechanism. Thus, further studies are required. However, limited samples of H. pylori-infected GM are currently available in public databases, which is a limitation of the present study. Additional representative datasets with large samples are needed in future studies.
In conclusion, the results of the present study revealed that MUC4 may be a cytokine involved in the pathogenesis of PC and may represent a novel treatment target. Based on these results, it may be speculated that silencing MUC4 may reduce the tumorigenic risk of H. pylori infection in PC.
This study was supported by the Jiangsu Natural Science Foundation (grant no. BK20181155), the Nanjing Medical University (grant no. 2017NJMU043), the Changzhou Department of Health (grant no. QN201711) and the Changzhou No. 2 People's Hospital (grant no. 2018K003).
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
YG, LT and HL designed the study. HY and YF collected tissue samples and basic information from the patients and carried out statistical analyses. YL and YW carried out the bioinformatics analysis. SC performed the experiments. HL wrote and edited the manuscript. All authors read and approved the final manuscript.
This study was approved by the Ethics Committee of the Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University [No: (2019) KYO 073-01]. All blood samples and tissues involved in this study were collected with informed consent from the patients.
gastric mucosa
Gene Expression Omnibus
DEGs
differentially expressed genes
Kyoto Encyclopedia of Genes and Genomes
overall survival
LUAD
stomach adenocarcinoma
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text:
Sahyadrica
of the mountains
Central Indian Journal
Barefoot Notes
The Cogito
Log of the Woods
By the campfire
Every season is marked by an event or a phenomenon that defines that season, and is so skillfully woven onto a timeline that it forms a periodic rhythm – the beauty of which lies in a meshwork of colours, scents, songs, and something that cannot be seen, smelled, or heard – a purpose, which I think comes close to what we humans call love. The purpose is but the only force that drives every plant or animal to display colours, release pheromones, and sing melodies.
What's special about man is perhaps his way of appreciating nature's mysteries and sharing it with others of his kind, and not in building bridges and airplanes; those feats were long conquered by nature.
What's special is this: no bird can sing of an autumn sunrise or of sound of the crashing waves, although we and they equally feel it, and our lives depend upon it. Our greatest strength perhaps lies in understanding what gave birth to us, and to them – indeed to all of us – and in respecting that wisdom than manipulating i…
It is pitch dark outside as I sit and write this. The tree-line has dissolved into the empty space above, with only a few stars gleaming down upon the darkened earth. I am sitting by an incandescent light I switched on at the click of a button, and I plugged in my computer, turned on some music, and began to think.
There in this darkness that has set in at six o'clock in the evening is a village devoid of all that I possess right now. This village probably lies within a hundred kilometers around me, is devoid of electricity, is devoid of any form of artificial light save for the kerosene lamps hung from corrugated ceilings. This village is older than you and I, and dates back to the bygone era of our forefathers. Its houses have been surrounded by woods for eons, and the darkness that falls on this village tonight is no different from the one that engulfed it last year on this day. Every morning since the harvest moon hung low in the sky, men, women, and children alike have been hacki…
The Last Wave by Pankaj Sekhsaria: a review
I have always wanted to visit the Andaman Islands, and I had written to Pankaj nearly half a decade ago about it, but I didn't intend to visit the islands as a tourist, especially after the tsunami. The trip however never materialized and years later, today, Pankaj helped me visit the great islands through "The Last Wave", and I learnt much more about the island than I would have if I were to visit as a mere tourist. Simply put, the book is relevant to those who have been to the islands as it is to those who wish to. The Last Wave's cover is outlaid on a mellow shade of green, with sights and signs distinctive of the Andamans: a Jarawa standing on what appears to be the Andaman Trunk Road, a fish and a dinghy signifying the basic livelihood of the island, a mugger basking near the base of the book, and between the title and the name of the author rests the flower of Papilionanthe teres. The back-cover, in continuation of the front, shows a Leatherback turtle along the sea, a Narconda…
Sewri: the perfect picture?
by Vishal Rasal
I have visited Sewri only twice, and on both occasions I've asked myself: what on Earth are these birds doing here? There are answers that are quite startling. In this article, Vishal talks about Sewri and its renowned visitors, Lesser and Greater Flamingos, the plausible reasons why they chose this place, and what we can deduce about their future from recent updates on the conservation-versus-development debate on Sewri. In Vishal's words… The city of Mumbai is never short of surprises. From skyscrapers cluttering the precarious coasts of the Arabian Sea to the forest fringes of Sanjay Gandhi National Park, this city is one of the handful few to harbour great biodiversity sandwiched between a large human population. And just as Mumbai proudly boasts of whales off its coasts and leopards in its forests, it is also peculiarly famous for its purple-winged migrants – flamingos. Not long ago Sewri was just a bleak and drab railway station on Mumbai's harbor-line rail ro…
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License. Contact me if you need to use any of my works.
© 2008-2019 Sahyadrica | Aniruddha Dhamorikar
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A Field Guide to Insects and Spiders of Kanha Tiger Reserve (India) is out! Click the image for more information.
The Modern Day Falconer
Growing Amidst Nature
The Legend of Sahyadri
In God's Garden
A Penchant for the Wild
Of Mahua and Her People
To be a Fish
Decoys, Shotguns and Conservation
To each his own fear
The Shared Waters
The Sweet One and the Warm One
In Search of the Summer Angel
Place I Left Behind
The Lizard that Lost its Way
The Plight of India's Vultures
Story of the Yellow Crazy Ant
The Crinum Chance
The Earth-Lion of India
The Purple Winged
On Konkan Tides
In Conversation with Sandhill Cranes
Dandeli-Anshi Tiger Reserve
Mother Wasp II
Mother Wasp
Toby Garter: A Video Documentary
Chasing Tiger Beetles
Common Spider Families of Mumbai
Tracing Monsoon
Following the Plants
Following the Insects
Order in Chaos
An Ode to Graminoids
How to Point-and-Shoot
Read List
Biodiversity in Focus Blog
Bug Eric
Butterflies of Singapore
Catching Flies
Musings from the north-east
nature of a man

redpajama_set_name: RedPajamaCommonCrawl


Home MMA News Al Iaquinta Conor McGregor claims he is "back", Al Iaquinta responds!
Perennial UFC lightweight contender Al Iaquinta continues to troll former two-division UFC champion Conor McGregor on social media.
Yesterday afternoon the UFC unveiled it's new "Legacy Belts" on Twitter.
The flashy new belt design prompted the promotions first ever 'champ champ', Conor McGregor, to announce that he is officially back.
"Ok I'm back." announced the Irish superstar in response to the promotions newly revealed 'Legacy Belts'.
McGregor's statement triggered the following response from Al Iaquinta.
"Suuure you are, Conor." replied Iaquinta accompanied by a picture of McGregor in a humbled state following his UFC 229 loss to Khabib Nurmagomedov.
"Raging Al" continued to attack the Irish superstar by posting the following.
This is not the firs time that Al Iaquinta and Conor McGregor have jostled on social media.
The top lightweight contenders went at it just last week, see that here.
Al Iaquinta has won five of his past six octagon appearances, his latest being a unanimous decision victory over Kevin Lee last month in Milwaukee.
As for Conor McGregor, the Irish superstar was most recently seen in action at September's UFC 229 event, where he suffered a fourth-round submission loss to the aforementioned Khabib Nurmagomedov.
"Mystic Mac" is hoping to rematch "The Eagle" later this year. However, UFC President Dana White would prefer that Conor McGregor rematch Dustin Poirier at lighweight in his next octagon appearance.
Would you like to see the UFC book a lightweight bout between Al Iaquinta and Conor McGregor? Who do you think would win? Sound off in the comments section PENN Nation!


text:
Project Origin Best of El Salvador auction crowns inaugural winners
Project Origin has celebrated the best of El Salvador coffee in its inaugural event, which took place from 2 – 8 May.
With a score of 88.75, Ever Leonel Díaz Pérez from the Milaydi farm in the Chalatenango region took top honours in the Washed Category.
The top bidder for this coffee at US$10.30 per pound was Sydney Coffee Company, who outbid 19 other contenders for the washed-processed Pacamara varietal, paying a total value of US$12,535.10.
The Honey/Natural Category winner, with a score of 88.25, was Raúl Rivera from the Santa Rosa farm in the Chalatenango region. Project Origin secured this honey processed Pacamara coffee, paying US$9.10 per pound. This equates to a total value of US$10,155.60.
Other roasters to show their support for El Salvador coffee included Dramanti Artisan Roaster, Knockbox Coffee Company, Hong Coffee, Beijing HMC, Lighthouse Coffee Snd, and many more.
"An absolute highlight of the event was announcing the winners Jorge, and Ever," said Project Origin's Habib Maarbani. "Raul has been a previous Cup Of Excellence (COE) winner and reminded us why we love honey and natural Pacamaras from El Salvador. Ever was a stranger to the spotlight and his story really proved why events such as these can help change the specialty coffee industry."
The Milaydi farm is only one manzana (about 2 acres). As such, Ever can only produce six to fifteen 60-kilogram bags of coffee per year. Habib says Ever has had trouble entering other competitions given the minimum lot size requirements.
"In the past he has sold his coffee for about US$2 per pound to other producers, and his coffee would get combined with other lots then get entered into competitions, often winning and being sold for quite high prices," Habib said. "This year he submitted six bags into the Project Origin: Best of El Salvador Auction and won. He earned himself an auction price of US$9.10 per pound and direct credit for the quality of that coffee – two things he has never had before."
World Barista Champion (WBC) Sasa Sestic created the new auction program to help develop coffee growing communities and advance quality coffee globally.
"My objective is to continue supporting the specialty coffee industry by connecting coffee communities all over the world," Sasa said. "I truly think this auction is the most humble way to connect coffee buyers with consumers, and also to educate both farmers and roasters on how we can improve the specialty coffee industry."
The Best of El Salvador supported coffee producers who have previously placed in the top 15 of COE in El Salvador, including Gilberto Baraona, Jorge Raul Rivera and Mauricio Salaverria. In 2015, the Alliance for Coffee Excellence decided to "temporarily scale back" COE competitions, which included the El Salvador COE.
To conduct the Best of El Salvador auction, a national jury first put coffee entries through a pre-selection process and shortlisted coffees scoring above 85 points. The international jury then cupped 36 lots and selected the top eight coffees in the Washed Category and Honey/Natural Category.
"Those 16 coffees became our finalists and our auction lots and they were incredibly tasty coffees," Habib said.
On the last day of the auction, coffees that scored 86+ points were auctioned off. The auction included an online bidding system for those who could not physically attend.
To see the full results visit: hub.cropster.com/store/auctions/best-of-el-salvador
The next Best of Project Origin auction will take place in Santa Barbara, Honduras from 4 July – 10 July.
To join the judging panel or obtain samples of the coffees that make the International Judging round please email habib@onacoffee.com.au


text:
package org.apache.cassandra.cql.jdbc;

import java.net.InetAddress;
import java.net.UnknownHostException;
import java.nio.ByteBuffer;
import java.sql.Types;

import org.apache.cassandra.utils.ByteBufferUtil;

public class JdbcInetAddress extends AbstractJdbcType<InetAddress>
{
    public static final JdbcInetAddress instance = new JdbcInetAddress();

    JdbcInetAddress()
    {
    }

    public boolean isCaseSensitive()
    {
        return false;
    }

    public int getScale(InetAddress obj)
    {
        return 0;
    }

    public int getPrecision(InetAddress obj)
    {
        return obj.toString().length();
    }

    public boolean isCurrency()
    {
        return false;
    }

    public boolean isSigned()
    {
        return true;
    }

    public String toString(InetAddress obj)
    {
        return obj.getHostAddress();
    }

    public boolean needsQuotes()
    {
        return false;
    }

    public String getString(ByteBuffer bytes)
    {
        return compose(bytes).getHostAddress();
    }

    public Class<InetAddress> getType()
    {
        return InetAddress.class;
    }

    public int getJdbcType()
    {
        return Types.OTHER;
    }

    public InetAddress compose(ByteBuffer bytes)
    {
        try
        {
            return InetAddress.getByAddress(ByteBufferUtil.getArray(bytes));
        }
        catch (UnknownHostException e)
        {
            throw new AssertionError(e);
        }
    }

    public ByteBuffer decompose(InetAddress value)
    {
        return ByteBuffer.wrap(value.getAddress());
    }
}

redpajama_set_name: RedPajamaGithub


Habit: slow growing evergreen tree reaching great heights if given enough time. Open-grown trees have a broad, rounded crown and straight trunk, while those growing in shadier sites have a narrow, conical crown. The bark is thick and fissured and mature trunks have a large burl at the trunks base. Leaves and stems on younger shoots are covered in small brownish yellow hairs, as the leaves age they become smooth and gray-green above and only fuzzy below. They are oblong reaching 5 in (12 cm) long with sharply toothed margins, and a stiff leathery texture. Flowers are produced on both male and female catkins from April -August, and the fruit is an acorn taking two years to fully develop.
Ecology: endemic to a small area from the Southern Oregon Coast to the Southern California Coast as well as on the foothills of the Sierra Nevada Mountain range on dry wooded slopes at elevations from 140-5000 ft (42-1525 m).
Growing Conditions: full sun to partial shade, deep well drained moist to dry soil. Tan oak is best adapted to a cool-moist coastal climate.
Lithocarpus densiflorus is susceptible to sudden oak death.


text:
Q: NFS inbound adapter configuration for multiple directory using single inbound adapter In spring integration, I want to poll files from different source directories (each interface configured have different source directories) which is configured in as sourcePath in yml file (dynamically) like below. N number of interfaces can be added by user.
 interfaces:
      -
        sourceType: NFS
        sourcePath: /Interface-1/Inbound/text
        target: Interface-1
        targetType: S3
        targetPath: test-bucket-1
      -
        sourceType: NFS
        sourcePath: /Interface-2/Inbound/text
        target: Interface-2
        targetType: S3
        targetPath: test-bucket-2

Is it possible to poll the files from different source folders using single inbound adapter (using atomic reference) or need more than one inbound adapter?
Currently application polls files from base directory.
   <file:inbound-channel-adapter id="filesInboundChannel"
    directory="file:${base.path}" auto-startup="false" scanner="scanner" auto-create-directory="true">

    <integration:poller id="poller" max-messages-per-poll="${max.messages.per.poll}" fixed-rate="${message.read.frequency}" task-executor="pollingExecutor">
        <integration:transactional transaction-manager="transactionManager" />
    </integration:poller>
  </file:inbound-channel-adapter>

Can someone give an advice on this or is there any other way can also achieve the same goal

A: Yes, you can use a single <file:inbound-channel-adapter> for this task. To make it rotate over a list of directories for scanning you need to configure an AbstractMessageSourceAdvice implementation for the <poller> of that adapter to change a directory when afterReceive(boolean messageReceived, MessageSource<?> source) gets a false for the receive operation. So, this way the next poll will get already a new directory for scanning.
As a sample you can take a look into the recently introduced a RotatingServerAdvice: https://github.com/spring-projects/spring-integration/blob/master/spring-integration-file/src/main/java/org/springframework/integration/file/remote/aop/RotatingServerAdvice.java
https://docs.spring.io/spring-integration/docs/current/reference/html/messaging-channels-section.html#conditional-pollers

redpajama_set_name: RedPajamaStackExchange


Size: 4.0 - 5.0 m2 | Whole hides | Leather goods, Clothing, Orthopedics| Thickness : from 0.7 mm. to 0.8 mm.
Often when you opt for a certain property, you deliver on another property. In the case of Ross Nappa, this is really different. It is a supple nappa leather with a pronounced firmness. You do not just tear it apart. Due to the elasticity that the leather has, it can be used very well for upholstery and clothing. The form will not be lost.
We see it as a solid leather which does its job very well.
Thickness; 0.7 mm. – 0.8 mm.
Whole hides; 4.0 m2 to 5.5 m2.


text:
Wayfinders: Optimism Will Lead the Way
Photo: Annabelle Armstrong-Temple
Last year was a year like no other. As we continue to navigate the pandemic in 2021, we must consider the possible long-term consequences to how young people hope and plan for the future. The good news is this: research shows that young people continue to be optimistic despite having to navigate significant disruptions to their lives over the past year. Working together, we can nurture that feeling of optimism and help guide them to a path that leads them to the vibrant futures they envision for themselves.
Inside this month's Wayfinders:
A newly released report about how young people in New York City came together to save a summer employment program,
A recent analysis on race and gender equity gaps in access to both AP and dual enrollment coursework across states and school districts,
Important research findings released in December highlighting that as Black and Latino young people adjust to the realities of COVID-19, they are also paying attention and engaged on racial justice issues,
Bill & Melinda Gates' annual letter which highlights the importance of student supports, like mentoring and advising, especially in the COVID context,
And finally, a collection of articles we're reading and events we're looking forward to in February.
Youth Voice is Not a Box You Check Off
"This work helped me find my voice. I have always wanted to be a doctor, but this experience has made me wonder whether I should be a public servant and political leader. I saw the ability that public officials have to help their community, and I've started wondering if that's the path for me. My community needs the help, and I now know that I have a powerful voice." – La'Toya Beechum, student organizer and lead author of Youth Voice is Not a Box You Check Off
During the summer of 2020, following New York City's cancellation of the Summer Youth Employment Program, young people came together and worked with other youth advocates to save the program for thousands of students. This report tells the story of that advocacy, showcasing the work of young people through the lens of the mediums they used to organize amidst the continued threat of the Covid pandemic — texts, tik toks, zoom calls, and online petitions among other digital tools and assets. This student-led narrative was developed jointly by Teens Take Charge and HERE to HERE, affirming the importance of SYEP, the value of real-world learning experiences, and the vital role youth voice must play in systems change. Read the full report here.
How Equitable Is Access to AP and Dual Enrollment?
A recent analysis from Community College Research Center (CCRC) reveals race and gender equity gaps in access to both AP and dual enrollment coursework across states and school districts. However, while equity gaps in access are widespread, about one in five districts nationally have closed such gaps in access to dual enrollment. Differences across states and districts in access to dual enrollment raise important questions about how educators can design these programs to advance equity. In fall 2020, CCRC and the Aspen Institute released the Dual Enrollment Playbook: A Guide to Equitable Acceleration for Students, which details the practices of nine dual enrollment programs in Florida, Ohio, and Washington that have narrowed or closed equity gaps — both in access to dual enrollment and in post–high school college outcomes — for Black, Latino, Indigenous, and Pacific Islander students.
Young People Remain Engaged and Hopeful on Racial Justice
The pandemic has caused many young people to reconsider their expectations related to education and work. At the same time, as young people adjust to the impacts of COVID-19, they are paying attention to and participating in conversations and events happening around them related to policing and racial justice — these experiences are also shaping how they think about their futures. To them, being able to be engaged in promoting change on these issues feels more like an opportunity than a burden. This sense of possibility feels connected to young people's strong sense of personal agency, observed in earlier research conducted by Equitable Futures, and their belief that they are the most important changemakers in their own lives.
Engaging and empowering young people with the support they need to navigate their educational and career pathways has been a challenge; however, as youth development professionals, educators, policy makers, grantmakers and program designers, we must know that young people are still present and engaged on issues that matter to them — even while navigating the pandemic. Review the full research findings here.
Bill & Melinda Gates released their annual letter yesterday, shedding light on the ways COVID-19 has disrupted our daily lives, how it's exacerbated inequality, and the work the foundation has done to help support the recovery, including for students.
Here's what else we're reading:
A case study on how the mismatch between the skills employers need and the training young job seekers receive is being tackled in New Orleans
An education and employment promise to prevent the forces that relegate millions of young adults to the economic margins
A look into how a 13-year-old is thinking about the future and whether college will be free in 4 years
Another example showing advising may be key to getting more young people to and through college


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text:
Home Brief Encounters
Chögyam Trungpa Rinpoche in Quick Charcoal
William Gilkerson
Charcoal sketch by William Gilkerson
I met Rinpoche in 1971, becoming a student of his soon after, and for life. During the course of it, he assigned to me an eclectic array of different tasks and challenges. One of these was to make portrait sketches of him as he went about his business of the moment. Typically, I would get a summons, sometimes when he was delivering a talk nearby and it was easy to get to him, but he didn't hesitate to telephone from, for instance, Colorado, putting me on an airplane at short notice. Military venues were costumed with uniforms, many kinds; lectures were mostly suit and tie; for a bedroom event, messing with my mind, he wore nothing at all; for a session during which he made calligraphy, he wore Zen robes and his hair in a Japanese topknot.
This was his costume when I sketched my favourite of all the drawings that I made of him. I discarded a lot more elaborate attempts than I kept, but this particular little two-minute, unfinished toss-off was a happy accident and a keeper for sure. At the time, I couldn't wait to show it to him. His usual reaction to my work was to examine it piece by piece in stony silence, acknowledging things only with an inscrutable "Hmmm," nothing else.
Handing him my proud little drawing, I hoped for a less lethargic response this time, maybe even a tickle of praise, although I was braced for any kind of criticism. Line work, style, composition, all were open to it, hopefully something—anything. He studied the picture for a while.
"Hmmm" came his inevitable comment.
Only much later did his reviews start to come in, and they were all carom shots bounced off sangha chums of old, never directly to me. "He told me you're the only artist who ever captured his essence," reported a grinning old student who had written down the quote. Then the same message trickled in from other sources, bathing me in warm, molten honey in a way that no doubt would have wrinkled my teacher's nose.
Thereafter, I framed my remaining pictures—those I hadn't given to friends or discarded for lack of merit—and hung them through the house. Most prominent was the quick calligraphic little sketch that I had liked from the beginning. Quite shamelessly, I had fallen in love with a piece of my own work. For one thing, it reminded me that whatever expected art lessons I hadn't got were in my dreams, and my teacher's intended instructions were in modelling, not drawing. Or so it seemed. He never told me that, but he demonstrated it throughout every meeting, talk, whatever the session, by taking a comfortable pose at the outset, and returning to it, like the out-breath after every diversion. While he was gesticulating, talking, shifting, I had time to fill in the static details while waiting for the foundational pose again. Unlike the unstirring model of a figure-drawing class, or the rigid subject of a formal portrait, this model was always animated, never static, yet reliably back where wanted for the portrait's sake. Lessons to be sure, I reflected, looking at my enshrined little drawing over the years.
What's your own favourite of your Rinpoche pictures?" asked the Vajra Regent Osel Tendzin, during a visit to Martin's River, and I told him. "Will you give it to me—" he asked at once, and I did, after some anguished hesitation. "Thank you," he said. It was a raucous dinner party, and I trusted he would forget about the picture as an evening of emptied bottles played out. Finally departing, during farewells at the door, he turned to an attendant: "Mr. Gilkerson will give you the picture that he gave me earlier." And that was that.
Or so it seemed at the time. During my period of mourning for the lost icon, I fretted that I had never made a reproduction and asked to borrow it back for that purpose, whereon it was returned. After making the photocopies, I felt no great rush to return the original. By then, the Regent had got quite a lot of other things to deal with; also he was preparing to move and it seemed very unlikely that during his last days in Nova Scotia he would have any thought to spare for an insignificant little picture.
This was mistaken. "The Regent wants to know when you're going to return his drawing of the Vidyadhara" came the voice of a secretary on the phone. So, back it went again, as trucks were being loaded with his possessions, bound for California where he planned a permanent retreat. I later heard that the truck with most of his pictures was burned in a road accident with full loss. And that, seemingly, was the end of it at last.
But it wasn't. Some months after the Regent's death, my little picture showed up by complete surprise, via courier, in fine condition, like Lassie Come Home, with a note from his main caregiver at the end, to whom he had left it. "I thought you might like to have this back," she commented.
Recounted at the request of Walter Fordham for the Chronicles of CTR.
© 2009 William Gilkerson
Bill's award winning Novel, PIRATE'S PASSAGE, was just re released with a new cover. Check it out. PIRATE'S PASSAGE is being produced as an animated, made-for-television film. Produced and co-written by Donald Sutherland along with co-writer, Brad Peyton. Sutherland also stars as the voice of the lead character, Captain Johnson. For more on William Gilkerson and his work, visit GilkersonArt.com.
Richard Arthure on Meeting Chogyam Trungpa
Kasung Log: Bringing Them Around
Chögyam Trungpa Rinpoche Retreat in Massachusetts
May I never be separated from you
Light of Blessings: Supplication to the Eleventh Trungpa Chökyi Gyatso
Tulku Thondup Rinpoche
Francesca Fremantle: A Poem to Trungpa Rinpoche


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Secret Money System Review - Does it Really Work?
Ronnie M. and Juan Gabriel's Secret Money System review. Does the program really work or is it another make money online scam? Let's find out.
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Upon the successful completion of the Simon Skjodt Assembly Hall addition project, the Indiana University Athletic Department discovered an opportunity to enhance an area of underutilized space within the building. The space located under the lower bowl was being utilized for storage, back of house restrooms, and a various ancillary needs. With direct access to the basketball court, it made sense to reclaim this space as game day locker rooms, a players lounge, shower/restrooms, and coaching conference spaces, all of which did not previously exist within the facility.
After early discussions with the University it became apparent this space needed to be on par with many of the universities IU competes with for talent. The current coaching staff indicated that, while the space needed to be improved, there was also a strong desire to have it be modest. Above all requirements, the space needed to speak to the rich history of Indiana University Basketball with an eye to the future.
In order to capture this aesthetic the design team incorporated wood, limestone, and back painted glass as the foundation of the finish palette. These finishes would serve as a clean backdrop to the iconic branding components found in the furnishings and implemented throughout the design. The overall composition created the motivational and energetic space required to attract new athletes, while reminding them of the honor involved in representing IU.


John "Stinker" Clarke, Di Morrissey, MC Craig Hamilton ABC, Kate Washington MP, Port Stephens.
Stinker has many talents! He's published four books and I was honoured to launch his latest children's book – "The True Adventures of Clarabelle". It's a true story based on the 1955 floods when a cow was washed down the river and out to sea to be rescued and taken to Broughton Island. It's a charming story with lovely illustrations by Illena Clarke.
Also present was Leonard Connelly who actually met Clarabelle when he was 15, back in 1955!
Interestingly, at the book launch at Port Stephens, I met Leonnard O'Connell, who remembers as a young nipper meeting the real Clarabelle in a paddock after her big adventure back in '55!
It was a lovely day, and Stinker and his family are just lovely!