Mitochondrial DNA Copy Number Variations and Serum Pepsinogen Levels for Risk Assessment in Gastric Cancer

Background: Variations in mitochondrial DNA copy number (mtDNA-CN) of peripheral blood leukocytes (PBLs), as a potential biomarker for gastric cancer (GC) screening has currently been subject to controversy. Herein, we have assessed its efficiency in GC screening, in parallel and in combination with serum pepsinogen (sPG) I/II ratio, as an established indicator of gastric atrophy. Methods: The study population included GC (n = 53) and non-GC (n = 207) dyspeptic patients. The non-GC group was histologically categorized into CG (n = 104) and NM (n = 103) subgroups. The MtDNA-CN of PBLs was measured by quantitative real-time PCR. The sPG I and II levels and anti-H. pylori serum IgG were measured by ELISA. Results: The mtDNA-CN was found significantly higher in GC vs. non-GC (OR = 3.0; 95% CI = 1.4, 6.4) subjects. Conversely, GC patients had significantly lower sPG I/II ratio than the non-GC (OR = 3.2; CI = 1.4, 7.2) subjects. The combination of these two biomarkers yielded a dramatic amplification of the odds of GC risk in double-positive (high mtDNA-CN-low sPGI/II) subjects, in reference to double-negatives (low mtDNA-CN-high sPGI/II), when assessed against non-GC (OR = 27.1; CI = 5.0, 147.3), CG (OR = 13.1; CI = 2.4, 72.6), or NM (OR = 49.5; CI = 7.9, 311.6) groups. Conclusion: The combination of these two biomarkers, namely mtDNA-CN in PBLs and serum PG I/II ratio, drastically enhanced the efficiency of GC risk assessment, which calls for further validations.

Immunohistochemical Analysis of LGR5 and TROY Expression in Gastric Carcinogenesis Demonstrates an Inverse Trend

Background: Two of the Wnt signaling pathway target genes, tumor necrosis factor receptor family member (TROY) and leucine-rich G-protein coupled receptor (LGR5), are involved in the generation and maintenance of gastrointestinal epithelium. A negative modulatory role has recently been assigned to TROY, in this pathway. Here, we have examined their simultaneous expression in gastric carcinogenesis. Methods: Tumor and paired adjacent tissues of intestinal-type gastric cancer (GC) patients (n = 30) were evaluated for LGR5 and TROY expression by immunohistochemistry. The combination of the percentage of positively¬ stained cells and the intensity of staining was defined as the composite score and compared between groups. The obtained findings were re-evaluated in a mouse model. Results: TROY expression in the tumor tissue was significantly lower than that of the adjacent tissue (2.5 ± 0.9 vs. 3.3 ± 0.9, p = 0.004), which was coincident with higher LGR5 expression (3.6 ± 1.1 vs. 2.7 ± 0.9, p = 0.001). This observation was prominent at stages II/III of GC, leading to a statistically significant mean difference of expression between these two molecules (p = 0.005). In the H. pylori infected-mouse model, this inverse expression was observed in transition from early (8-16 w) to late (26-50 w) time points, post treatment (p = 0.002). Conclusion: Our data demonstrates an inverse trend between TROY down-regulation and LGR5 up-regulation in GC tumors, as well as in response to H. pylori infection in mice. These findings support a potential negative modulatory role for TROY on LGR5 expression.

PAX2 expression is correlated with better survival in tamoxifen-treated breast carcinoma patients.

PAX2 (paired box gene 2) is a transcription factor, which is involved in both cell proliferation and carcinogenesis. This study aimed to investigate PAX2 expression in tamoxifen resistant (TAM-R) and tamoxifen sensitive (TAM-S) breast carcinoma patients and analyze its correlation with clinicopathological characteristics and survival. Immunohistochemical analysis was performed to evaluate PAX2 protein expression in 36 TAM-R and 36 TAM-S formalin-fixed paraffin-embedded (FFPE) breast tumor tissues. Data analysis indicated that PAX2 expression was significantly higher in TAM-S group in comparison to TAM-R (P = 0.014). Overexpression of PAX2 was significantly correlated with perineural invasion (PNI) (P = 0.025). Kaplan-Meier survival analysis showed significant association between high expression of PAX2 and better disease-free survival (P < 0.001) and also overall survival (P = 0.031). Multivariate cox regression analysis demonstrated that patients with increased expression of PAX2 have a trend toward improved disease free survival (OR = 0.065, 95% CI: 0.009-0.476; P = 0.007) and overall survival (OR = 0.147, 95% CI: 0.020-1.105; P = 0.062). Our data suggested that high expression of PAX2 could be associated with better survival in estrogen receptor positive tamoxifen-treated breast carcinoma patients.

Multiplex serology of Helicobacter pylori antigens in detection of current infection and atrophic gastritis - A simple and cost-efficient method.

INTRODUCTION: Helicobacter pylori express a large array of antigens, each of which is duly responsible for successful colonization and pathogenesis. Here, we have studied host serum antibody responses to four of its immunodominant antigens in association with the infection status and the resulting clinical outcomes. METHODS: For this purpose, four individual H. pylori proteins (UreB, CagA, Tip-α and HP0175) were produced in recombinant forms. Serum antibody responses of 246 (75 GC and 171 NUD) patients, against the above antigens, were evaluated by multiplex immunoblotting. The associations between the resulting data and the infection status, as well as clinical outcomes were evaluated using logistic regression models. RESULTS: Serum antibodies to all four recombinant antigens increased the chances of detecting screening ELISA-positive subjects, in an escalating dose-dependent manner, ranging from 2.6 (1.5-4.7) for HP0175 to 14.3 for UreB (4.3-50.7), exhibiting the lowest and highest odds ratios, respectively (PAdj ≤ 0.001), such that 98.2% of the subjects with antibodies to all four antigens, were also positive by the screening ELISA (P < 0.0001). Among the screening ELISA-positive subjects, the three antigens of CagA, Tip-α, and HP0175 were able to segregate current from past H. pylori infection (P < 0.05). Accordingly, subjects with antibodies to one or more antigen(s) were at 5.4 (95% CI: 1.8-16.4) folds increased chances of having current infection, as compared to triple negatives (PAdj = 0.003). In reference to the clinical outcomes, those with serum antibodies to CagA were more prevalent among gastric cancer, as compared to NUD patients (ORAdj: 5.4, 95% CI: 2.4-12.2, PAdj < 0.0001). When NUD patients were categorized according to their histopathologic status, multiple antigen analysis revealed that subjects with serum antibodies to one or more of the 3 current infection-positive antigens (CagA, Tip-α, and HP0175) were at 9.7 (95% CI: 2.1-44.9, P = 0.004) folds increased risk of atrophic gastritis, in reference to triple negatives. CONCLUSION: The non-invasive multiplex serology assay, presented here, was able to not only detect subjects with current H. pylori infection, it could also screen dyspeptic patients for the presence of gastric atrophy. This simple and cost-efficient method can supplement routine screening ELISAs, to increase the chances of detecting current infections as well as atrophic gastritis.

Prognostic Significance of Preoperative and Postoperative Plasma Levels of Ghrelin in Gastric Cancer: 3-Year Survival Study.

OBJECTIVES: We aimed to investigate prognostic effects of plasma levels of ghrelin before and after gastrectomy in gastric cancer (GC). METHODS: We followed 81 GC patients up to 3 years in this study. They were candidates for curative gastrectomy with or without neoadjuvant chemotherapy. Plasma levels of total and active ghrelins before and after the operation were assessed. Association of plasma levels of ghrelin with survival were assessed and adjusted for other potential prognostic factors using Cox regression analyses. RESULTS: Both total and active ghrelins dropped after gastrectomy (P<0.001 for both). Multiple Cox models revealed worse survival for patients with postoperative total ghrelins below median (hazards ratio (HR)=2.33, 95% confidence interval (CI): 1.01-5.41) or 25th percentile (HR=4.29, 95% CI: 1.48-12.44) compared with patients with higher ghrelin levels. In case of preoperative total ghrelin, patients with either second or third quartiles of plasma ghrelin showed worse survival compared with patients with the lowest quartile (HR=2.67, 95% CI: 1.11-6.38 for second quartile, and HR=2.32, 95% CI: 1.01-5.35 for third quartile vs. the lowest quartile). However, there was no difference between patients with the highest and lowest quartiles (HR=0.78, 95% CI: 0.22-2.73). Similar pattern was observed for preoperative active ghrelin (HR=4.92, 95% CI: 1.80-13.54 for second quartile, and HR=2.87, 95% CI: 1.11-7.38 for third quartile vs. the lowest quartile). Advanced TNM stage (HR=4.88, 95% CI: 1.10-21.77), cachexia (HR=2.99, 95% CI: 1.35-6.63), and receiving no neoadjuvant chemotherapy (HR=2.02, 95% CI: 1.04-3.92) were other poor prognostic factors. CONCLUSIONS: Preoperative and postoperative plasma levels of ghrelin could predict survival of GC patients with different patterns. This prognostic effect was independent of stage and cachexia. Measurement of plasma ghrelin in GC patients could complement conventional staging for more precise risk-stratification of the patients. Extrinsic admirations of ghrelin after total gastrectomy has potentials to improve survival of GC patients.

Serum Antibodies against Helicobacter pylori Neutrophil Activating Protein in Carriers of IL-4 C-590T Genetic Polymorphism Amplify the Risk of Gastritis and Gastric Cancer.

BACKGROUND: Gastric cancer arises, mainly, on an inflammatory background. Helicobacter pylori neutrophil activating (HP-NAP) protein functions as a potent pro-inflammatory mediator. Similarly, IL-4 plays a critical role in the inflammation pathway, the levels of which are altered by C to T transition at position -590 in its promoter region. Here, we have aimed to assess the risk of gastritis and gastric cancer in the co-presence of these two inflammation modulating mediators. METHODS: Gastritis (n=58) and gastric cancer (n=31) patients were evaluated and compared with H. pylori-positive asymptomatic controls (n=46), for serum antibodies against recombinant HP-NAP and IL-4 C-590T single nucleotide polymorphism using immunoblotting and PCR-RFLP, respectively. Multivariable logistic regression, adjusting for age, gender and ethnicity, was used for data analysis. RESULTS: In terms of susceptibility to gastritis, seropositivity to HP-NAP projected a risk impact of 4.62 fold (OR=4.62, 95% CI=1.50-14.22), which when present in IL-4 -590 T carriers augmented the risk up to 9.7 fold (OR=9.70, 95% CI=2.06-45.69). A similar pattern, but of a stronger magnitude, occurred for the risk of gastric cancer, which was estimated at 9.07 fold (OR=9.07, 95% CI=1.99-42.0) for HP-NAP-seropositive subjects and was drastically amplified (OR=33.64, 95% CI=2.06-548.68), when double-positive (HP-NAP seropositive/IL-4 -590 T carrier) subjects were examined against double negatives (HP-NAP seronegative/IL-4 -590 CC). CONCLUSION: Our preliminary data indicate that serum antibodies against HP-NAP represent a state of risk, which is further exacerbated in IL-4 -590 T carriers. These biomarkers, if validated in larger prospective studies, can be used to screen for gastric cancer susceptibility.

Helicobacter pylori Peptidyl Prolyl Isomerase Expression Is Associated with the Severity of Gastritis.

PURPOSE: Helicobacter pylori secretory peptidyl prolyl isomerase, HP0175, is progressively identified as a pro-inflammatory and pro-carcinogenic protein, which serves to link H. pylori infection to its more severe clinical outcomes. Here, we have analyzed host HP0175-specific antibody responses in relation to the severity of gastritis. METHODS: The HP0175 gene fragment was PCR-amplified, cloned, expressed and purified by Ni-NTA affinity chromatography. Serum antigen-specific antibody responses of non-ulcer dyspeptic patients (N = 176) against recombinant HP0175 were detected by western blotting. The infection status of these subjects was determined by rapid urease test, culture, histology, and serology. The grade of inflammation and stage of atrophy were scored blindly according to the OLGA staging system. RESULTS: The recombinant HP0175 (rHP0175) was expressed as a ~35 kDa protein and its identity was confirmed by western blotting using anti-6X His tag antibody and pooled H. pylori-positive sera. Serum IgG antibodies against rHP0175 segregated our patients into two similar-sized groups of sero-positives (90/176, 51.1 %) and sero-negatives (86/176, 48.9 %). The former presented with higher grades of gastric inflammation (OR = 4.4, 95 % CI = 1.9-9.9, P = 0.001) and stages of gastric atrophy (OR = 18.3, 95 %CI = 1.4-246.6, P = 0.028). CONCLUSION: Our findings lend further support to the pro-inflammatory nature of H. pylori peptidyl prolyl isomerase (HP0175) and recommends this antigen as a non-invasive serum biomarker of the severity of H. pylori-associated gastritis.

CD44 and CD74: The promising candidates for molecular targeted therapy in oral squamous cell carcinoma.

BACKGROUND: Considering molecular target therapy concept in the treatment of oral squamous cell carcinoma (OSCC), many attempts have been performed to introduce an effective molecular marker during recent years. Several investigations have emphasized on the role of CD44 in various cancers and few studies have mentioned CD24 and CD74. The purpose of this study was to investigate the relationship between CD44, CD24 and CD74 expressions and several clinical or histopathological factors in OSCC patients. MATERIALS AND METHODS: In our analytical cross-sectional study, forty primary OSCC specimens were immunohistochemically stained for CD44, CD24, and CD74 proteins. Then, the relationship between their expressions and age, sex, lymph node metastasis, and histopathologic grading was statistically analyzed using Mann-Whitney nonparametric and t-test. Furthermore, P < 0.05 was considered as significant. RESULTS: CD44 and CD74 proteins were significantly over-expressed in OSCC patients with high grade (P = 0.001 and P = 0.001) as compared to those with low grade. Furthermore, CD74 immunoreactivity showed significantly higher expression in patients with lower age (P = 0.039). Considering lymph node metastasis, we observed significant overexpression of CD74 in patients with no lymph node involvement (P = 0.033). CONCLUSION: Our observations support the significant role of membranous CD44 protein in progression of OSCC and also introduce CD74 protein as a probable interfering factor in different aspects of OSCC.

Age-Specific Gastric Cancer Risk Indicated by the Combination of Helicobacter pylori Sero-Status and Serum Pepsinogen Levels.

BACKGROUND: Serologic screening of gastric cancer (GC) by serum pepsinogens (sPG) levels and Helicobacter pylori (Hp) sero-status, though highly informative, has provided heterogeneous results. Here, we have evaluated the modifying effects of demographic factors on the risk impact of Hp sero-status/sPG levels in gastric cancer, with particular emphasis on age. METHODS: A cross-sectional study was carried out on 1341 individuals (GC = 578, healthy = 763), who were stratified into two age groups: 35-59 years (middle-aged, n = 830) and ≥ 60 years (60 years-plus, n = 511). Demographic factors and serological states (Hp sero-staus and sPG levels) were recorded by subject interview and serum ELISAs, respectively. Covariate-specific odds ratios were calculated by multivariable logistic regression. RESULTS: Hp infection was consistently associated with increased sPGI and sPGII levels in the 60 year-plus, but not the middle-aged group. The joint examination of the variable states of the three serum biomarkers (Hp serology, sPGI, and sPGI/II ratio), in the 60 year-plus age group, demonstrated a stepwise escalation of risk from the single (sPGI low; OR = 2.6), to double (sPGI low/sPGI/II low; OR = 3.55, and Hp positive/sPGI low; OR = 5.0) and ultimately triple (Hp positive/PGI low/PGI/II low; OR = 10.48) positive states, in reference to the triple negatives. However, this pattern was not exhibited in the middle-aged subjects. CONCLUSION: Age was clearly identified as a modifying factor on the risk projection of the combined states of Hp serology and sPG levels in gastric cancer screening, reflected by the augmented (~10.5 fold) risk of GC in the triple positive (Hp positive/sPGI low/sPGI/II low) 60 year-plus subjects, which was not evident in the middle-aged group.

Lack of association between human papillomavirus infection and colorectal cancer.

INTRODUCTION: Colorectal cancer is the third leading cause of cancer-related deaths worldwide, with nearly one million new cases identified annually. Different factors might cause colorectal cancer, one of the most prevalent cancers among both men and women. Viral aetiology in cancerous malignancies is a very important issue and so far a number of viral strains have been identified as tumour oncogene viruses. Viral infections, such as human papillomavirus (HPV), have recently been suggested as a risk factor for colorectal cancer. However, the aetiology of the disease is still unknown. AIM: To assessed the association between HPV infection and colorectal cancer. MATERIAL AND METHODS: In this study, 50 cancer tissue samples and 50 samples without colon cancer were studied in order to identify HPV through polymerase chain reaction (PCR). Of 42 adenocarcinomas, 10 were well differentiated, 30 moderated differentiated, and 2 were poorly differentiated. DNA extraction was verified by beta globin gene amplification; specific PCR was carried out based on HPV L1 consensus primers MY09/MY11. RESULTS: HPV DNA was not identified in any of the normal, adenocarcinoma, or adenoma samples. CONCLUSIONS: In contrast with previous studies, the current research failed to establish a relationship between HPV infection and the incidence of colon cancer. Considering the existing inconsistencies, it is recommended that further studies be conducted with larger sample size.

Clinicopathologic features predicting involvement of non- sentinel axillary lymph nodes in Iranian women with breast cancer.

BACKGROUND: Almost half of the breast cancer patients with positive sentinel lymph nodes have no additional disease in the remaining axillary lymph nodes. This group of patients do not benefit from complete axillary lymph node dissection. This study was designed to assess the clinicopathologic factors that predict non-sentinel lymph node metastasis in Iranian breast cancer patients with positive sentinel lymph nodes. MATERIALS AND METHODS: The records of patients who underwent sentinel lymph node biopsy, between 2003 and 2012, were reviewed. Patients with at least one positive sentinel lymph node who underwent completion axillary lymph node dissection were enrolled in the present study. Demographic and clinicopathologic characteristics including age, primary tumor size, histological and nuclear grade, lymphovascular invasion, perineural invasion, extracapsular invasion, and number of harvested lymph nodes, were evaluated. RESULTS: The data of 167 patients were analyzed. A total of 92 (55.1%) had non-sentinel lymph node metastasis. Univariate analysis of data revealed that age, primary tumor size, histological grade, lymphovascular invasion, perineural invasion, extracapsular invasion, and the number of positive sentinel lymph nodes to the total number of harvested sentinel lymph nodes ratio, were associated with non-sentinel lymph node metastasis. After logistic regression analysis, age (OR=0.13; 95% CI, 0.02-0.8), primary tumor size (OR=7.7; 95% CI, 1.4-42.2), lymphovascular invasion (OR=19.4; 95% CI, 1.4- 268.6), extracapsular invasion (OR=13.3; 95% CI, 2.3-76), and the number of positive sentinel lymph nodes to the total number of harvested sentinel lymph nodes ratio (OR=20.2; 95% CI, 3.4-121.9), were significantly associated with non-sentinel lymph node metastasis. CONCLUSIONS: According to this study, age, primary tumor size, lymphovascular invasion, extracapsular invasion, and the ratio of positive sentinel lymph nodes to the total number of harvested sentinel lymph nodes, were found to be independent predictors of non-sentinel lymph node metastasis.

Prognostic significance of matrix metalloproteinase-7 in gastric cancer survival: a meta-analysis.

The prognostic role of matrix metalloproteinase-7 in gastric cancer survival has been widely evaluated. However, the results are controversial. We aimed to set up a meta-analysis to reach a conclusion on the prognostic significance of metalloproteinase-7 in gastric cancer survival as well as its association with clinicopathological parameters. We searched popular databases from 1988 until October 2014 to gather eligible peer-reviewed papers addressing the prognostic effect of matrix metalloproteinase-7 in gastric cancer patients' survival. The CASP check list was used for quality appraisal. Pooled hazard ratio (HR) for survival and odds ratio (OR) for association with their 95% confidence interval (CI) were considered as summary measurements. Finally, 1208 gastric cancer patients from nine studies were included in the meta-analysis. Pooled HR estimate for survival was 2.01 (95% CI = 1.62 - 2.50, P < 0.001), which indicated a significant poor prognostic effect for matrix metalloproteinase-7. Sensitivity analysis detected no dominancy for any study. No publication bias was detected according to Egger's and Begg's tests. Clinicopathological assessment revealed that higher matrix metalloproteinase-7 expression is associated with deeper invasion (pooled OR = 3.20; 95% CI = 1.14 - 8.96; P = 0.026), higher TNM stage (pooled OR = 3.67; 95% CI = 2.281-5.99; P<0.001), lymph node metastasis (pooled OR = 2.84; 95% CI = 1.89 - 4.25; P<0.001), and distant metastasis (pooled OR = 3.68; 95% CI = 1.85 - 7.29; P<0.001), but not with histological grade. This meta-analysis indicated a significant poor prognostic effect of matrix metalloproteinase-7 in gastric cancer survival. Additionally it was associated with aggressive tumor phenotype.

Gastrointestinal tumor board: an evolving experience in Tehran Cancer Institute.

Gastrointestinal (GI) cancers are a significant source of morbidity and mortality in Iran, with stomach adenocarcinoma as the most common cancer in men and the second common cancer in women. Also, some parts of Northern Iran have one of the highest incidences of esophageal cancer in the world. Multi-disciplinary organ-based joint clinics and tumor boards are a well-recognized necessity for modern treatment of cancer and are routinely utilized in developed countries, especially in major academic centres. But this concept is relatively new in developing countries, where cancer treatment centres are burdened by huge loads of patients and have to cope with a suboptimum availability of resources and facilities. Cancer Institute of Tehran University of Medical Sciences is the oldest and the only comprehensive cancer treatment centre in Iran, with a long tradition of a general tumor board for all cancers. But with the requirements of modern oncology, there has been a very welcome attention to sub-specialized organ-based tumor boards and joint clinics here in the past few years. Considering this, we started a multi-disciplinary tumor board for GI cancers in our institute in early 2010 as the first such endeavor here. We hereby review this 2-year evolving experience. The process of establishment of a GI tumor board, participations from different oncology disciplines and related specialties, the cancers presented and discussed in the 2 years of this tumor board, the general intents of treatment for the decisions made and the development of interest in this tumor board among the Tehran oncology community will be reviewed. The GI tumor board of Tehran Cancer Institute started its work in January 2010, with routine weekly sessions. A core group of 2 physicians from each surgical, radiation and medical oncology departments plus one gastroenterologist, GI pathologist and radiologist was formed, but participation from all interested physicians was encouraged. An electronic database was kept from the beginning. The number of patients presented in the tumor board increased from 4 in January 2010 to 16 in December 2011. Most patients were presented by radiation oncology department (38%) and then surgical (36%) and medical oncology (20%) departments. Physicians' participation also grew from an average of 8 each session to 12 in the same months, with a number of cancer specialists taking part from other university hospitals in Tehran. A total number of 225 patients were presented with a treatment decision made in this 2-year period. The majority of cases were colorectal (32%), stomach (23%), and esophageal (17%) cancers. The number of pancreatic (7%) and hepatobiliary (6%) cancers were much smaller. Most decisions were for a primary treatment (surgery or radiochemotherapy) and then a neoadjuvant approach.  Tehran Cancer Institute's GI tumor board is one of the first multi-disciplinary organ-based tumor boards in Iran, and as such has made a successful start, establishing itself as a recognized body for clinical decisions and consultations in GI oncology. This experience is growing and evolving, with newer presentation and discussion formats and adapted guidelines for treatment of GI cancers in Iran sought.

Seroreactivity to Helicobacter pylori antigens as a risk indicator of gastric cancer.

BACKGROUND: Multiple etiologic factors are suspected to cause gastric cancer, the most important of which is infection with virulent types of Helicobacter pylori. MATERIALS AND METHODS: We have compared 102 gastric cancer patients with 122 non-ulcer, non-cancer dyspeptic patients. Gastric specimens were evaluated for H. pylori infection by tissue-based detection methods. Patient sera underwent antigen-specific ELISA and western blotting using a Helicoblot 2.1 kit and antibody responses to various H. pylori antigens were assessed. RESULTS: The absolute majority (97-100%) of both groups were H. pylori seropositive. Multivariate regression analysis demonstrated serum antibodies to the low molecular weight 35kDa protein to be protective and reduce the risk of gastric cancer by 60% (OR:0.4; 95%CI:0.1-0.9). Conversely, seroreactivity to the 89kDa (VacA) protein was significantly higher in gastric cancer patients (OR:2.7; 95%CI:1.0-7.1). There was a highly significant association (p<0.001) between seroreactivity to the 116kDa (CagA) and 89kDa (VacA) proteins, and double positive subjects were found at nearly five fold (OR:4.9; 95%CI:1.0-24.4) enhanced risk of gastric cancer as compared to double negative subjects. CONCLUSIONS: Seroreactivity to H. pylori low (35kDa) and high (116kDa/89kDa) molecular weight antigens were respectively revealed as protective and risk indicators for gastric cancer.

Amelioration by chicory seed extract of diabetes- and oleic acid-induced non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) via modulation of PPARα and SREBP-1.

We evaluated the effect of chicory (Cichorium intybus L.) seed extract (CI) on hepatic steatosis caused by early and late stage diabetes in rats (in vivo), and induced in HepG2 cells (in vitro) by BSA-oleic acid complex (OA). Different dosages of CI (1.25, 2.5 and 5 mg/ml) were applied along with OA (1 mM) to HepG2 cells, simultaneously and non-simultaneously; and without OA to ordinary non-steatotic cells. Cellular lipid accumulation and glycerol release, and hepatic triglyceride (TG) content were measured. The expression levels of sterol regulatory element-binding protein-1c (SREBP-1c) and peroxisome proliferator-activated receptor alpha (PPARα) were determined. Liver samples were stained with hematoxylin and eosin (H&E). Significant histological damage (steatosis-inflammation-fibrosis) to the cells and tissues and down-regulation of SREBP-1c and PPARα genes that followed steatosis induction were prevented by CI in simultaneous treatment. In non-simultaneous treatment, CI up-regulated the expression of both genes and restored the normal levels of the corresponding proteins; with a greater stimulating effect on PPARα, CI acted as a PPARα agonist. CI released glycerol from HepG2 cells, and targeted the first and the second hit phases of hepatic steatosis. A preliminary attempt to characterize CI showed caffeic acid, chlorogenic acid, and chicoric acid, among the constituents.

Impact of methylenetetrahydrofolate reductase C677T polymorphism on the risk of gastric cancer and its interaction with Helicobacter pylori infection.

BACKGROUND: Attempts for early detection of gastric cancer have recently focused on host's genetic susceptibility factors and gene-environment interactions. We have, herein, studied the association of MTHFR C677T single nucleotide polymorphism (SNP) and its interaction with Helicobacter pylori infection, smoking, age and gender on the risk of gastric cancer among an Iranian population. METHODS: Gastric cancer patients (n = 450) and cancer-free controls (n = 780) were studied for serum H. pylori-specific IgG antibodies by ELISA and MTHFR C677T polymorphism (SNP) by PCR-RFLP. Demographic and life style data were collected through patient interviews. Unconditional logistic regression model estimated odds ratio (OR) and the corresponding 95% confidence intervals (CI). RESULTS: The interactions of MTHFR genotype with H. pylori infection (P = 0.03), age (P = 0.049) and gender (P = 0.007) were statistically significant. Accordingly, MTHFR C677T carriers who were also positive for H. pylori infection exhibited 80% (OR = 1.8, 95% CI = 1.0-2.9) significant excess risk of non-cardia gastric cancer. Furthermore, subjects over the age of 50 or female subjects carrying MTHFR C677T SNP showed 40 (OR = 1.4, 95% CI = 1.0-2.0) and 100 (OR = 2.0, 95% CI = 1.2-3.2) percent increased risk of gastric cancer, respectively. CONCLUSION: MTHFR C677T SNP seems to increase the risk of gastric cancer and the effect is significantly inflated by interactions with H. pylori infection, age and gender.

Human papillomavirus detected in esophageal squamous cell carcinoma in Iran.

INTRODUCTION: Considering the different results obtained regarding the association of HPV with the development of esophageal squamous cell carcinoma (ESCC) in different populations, we aimed to determine the frequency of HPV infection and its subtypes in ESCC in Iranian patients. METHOD: A total of 100 paraffin-embedded tissue samples of ESCC diagnosed during 1991 and 2005 in the Institute of Cancer affiliated to Tehran University of Medical Sciences were selected. Seven out of 100 samples were excluded due to low quality of extracted DNA from paraffin-embedded specimens. Thus, 93 samples were included for HPV genotyping. RESULT: All samples were examined using SPF10 primers for HPV detection. HPV DNA was positive in 8 out of 93 (8.6%) ESCC specimens. Using INNO-LiPA genotyping system we detected the genotypes of 5 out of 8 HPV-positive samples. Both HPV types 16 and 6 were detected in 3 specimens; one sample was positive for HPV type 18 and 2 samples were co-infected with two HPV types. There were no statistically significant differences between HPV-positive and HPV-negative cases with regard to clinical and pathologic findings. Three samples were positive for SPF10 indicating HPV infection; however, the exact HPV type could not be clarified using INNO-LiPA genotyping . CONCLUSION: In conclusion, the present study showed that a small proportion of ESCC specimens from Iran harbor HPV16, 18 genome using a highly sensitive method. As different rates have been reported from Iran, a more widespread study with more precise definition of geographical differences could delineate the potential involvement of HPV in the development of ESCC in Iranian population.

El fenotipo de las mucinas en el esófago de Barrett / Mucin phenotypes in Barrett's esophagus

AntecedentesEl esófago de Barrett es una reconocida lesión precursora de adenocarcinoma esofágico. Aunque generalmente asociada al reflujo gastroesofágico, los mecanismos patogénicos de la enfermedad no son bien conocidos. El objetivo del presente estudio es explorar la historia natural e identificar marcadores de progreso del proceso precanceroso.Material y métodosSe utilizaron cortes histológicos de 67 especímenes de esófago correspondientes a 14 pacientes con esófago de Barrett, a los que se siguió entre 1–9 años. Se clasificaron las lesiones en: esófago de Barrett sin displasia, indefinido para displasia o con displasia. Se evaluó la expresión de diferentes mucinas en las células caliciformes y en las columnares usando técnicas de histoquímica e inmunohistoquímica.ResultadosEn todos los casos se comprobó la presencia de metaplasia intestinal incompleta. Las células columnares dentro del epitelio metaplásico contenían mucinas neutras. A mayor severidad de la lesión se encontró significativamente menor expresión de sialomucinas en las células columnares (p de tendencia igual a 0,03). En sujetos con lesiones indefinidas para displasia se observó un mayor contenido de sulfomucinas en las células caliciformes (p=0,034) y de MUC2 en las células columnares (p=0,029) que en sujetos con esófago de Barrett sin displasia. Se observó expresión de la mucina intestinal MUC2 y de la mucina gástrica MUC5AC en todas las muestras. MUC6, una mucina de las glándulas profundas gástricas, se presentó ocasionalmente.ConclusiónLa evaluación de los perfiles de mucinas en el esófago de Barrett sugiere una transición gradual del fenotipo del epitelio metaplásico a medida que la lesión avanza en el tiempo(AU)

Background: Barrett’s esophagus is a known precursor lesion of esophageal adenocarcinoma. Although itis generally associated with gastroesophageal reflux, the pathogenic mechanisms of the disease are notwell understood. The aim of this study was to explore the natural history and to identify markers ofprogression of the precancerous process.Material andmethods: Histological sections of 67 esophageal specimens were used in this study. Theywere obtained from 14 subjects with Barrett’s esophagus who were followed from 1 to 9 years. The lesionswere histologically classified as: Barrett’s esophagus without dysplasia,indefinitefordysplasia,ordysplasia. Expression of various mucins in goblet and columnar cells was assessed by histochemistry andimmunohistochemistry.Results: Incomplete intestinal metaplasia was observed in all the specimens. Columnar cells with in themetaplastic epithelium expressed neutral mucins. Sialo mucins were significantly less expressed incolumnar cells as the lesions increased in severity(p trend=0.03). Subjects with indefinite dysplasia lesionshad significantly higher expression of sulphomucins in goblet cells(p=0.034)and of MUC2 in columnarcells (p=0.029)than subjects with Barrett’s esophagus without dysplasia. Expression of the intestinalmucin MUC2 and gastricmucin MUC5AC was observed in all specimens. MUC6, a mucin of the deep gastricglands, was occasionally expressed.Conclusion: The evaluation of the mucin profiles in Barrett’s esophagus suggests a gradual transition of themetaplastic epithelium phenotype as the lesion advances in time(AU)

El fenotipo de las mucinas en el esófago de Barrett: Mucin phenotypes in Barrett's esophagus.

ANTECEDENTES: El esófago de Barrett es una reconocida lesión precursora de adenocarcinoma esofágico. Aunque generalmente asociada al reflujo gastroesofágico, los mecanismos patogénicos de la enfermedad no son bien conocidos. El objetivo del presente estudio es explorar la historia natural e identificar marcadores de progreso del proceso precanceroso. MATERIAL Y M#ENTITYSTARTX000E9;TODOS: Se utilizaron cortes histológicos de 67 especímenes de esófago correspondientes a 14 pacientes con esófago de Barrett, a los que se siguió entre 1 - 9 años. Se clasificaron las lesiones en: esófago de Barrett sin displasia, indefinido para displasia o con displasia. Se evaluó la expresión de diferentes mucinas en las células caliciformes y en las columnares usando técnicas de histoquímica e inmunohistoquímica. RESULTADOS: En todos los casos se comprobó la presencia de metaplasia intestinal incompleta. Las células columnares dentro del epitelio metaplásico contenían mucinas neutras. A mayor severidad de la lesión se encontró significativamente menor expresión de sialomucinas en las células columnares (p de tendencia igual a 0,03). En sujetos con lesiones indefinidas para displasia se observó un mayor contenido de sulfomucinas en las células caliciformes (p=0,034) y de MUC2 en las células columnares (p=0,029) que en sujetos con esófago de Barrett sin displasia. Se observó expresión de la mucina intestinal MUC2 y de la mucina gástrica MUC5AC en todas las muestras. MUC6, una mucina de las glándulas profundas gástricas, se presentó ocasionalmente. CONCLUSI#ENTITYSTARTX000F3;N: La evaluación de los perfiles de mucinas en el esófago de Barrett sugiere una transición gradual del fenotipo del epitelio metaplásico a medida que la lesión avanza en el tiempo.

Interobserver variations in histopathological assessment of gastric pathology.

AIM: Considering the fact that histology and its grading systems are accepted gold standards in diagnosis of diverse clinical disorders, assessing the accuracy and reliability of this method of diagnosis is of utmost importance. Thus, this study was performed to measure the agreement values between four independent histopathology readings for identical indices under one scoring guideline using three different approaches. METHODS: Four independent pathologists participated in this study and were blinded to the clinical diagnosis of patients. Various histological features were examined on gastric tissue specimens according to the updated Sydney system. RESULTS: Statistical analysis revealed that our null hypothesis about the existing agreement between different histopathological observations is rejected for chronic gastritis, the presence of inflammatory activity, atrophy and Helicobacter pylori, whereas there were significant inter-observation agreements in regard to the presence of lymphoid follicles, intestinal metaplasia and dysplasia. Pairwise analysis showed that different grading scales resulted in different kappa values ranging from poor to excellent agreements. The best kappa values were observed for the evaluation of dysplasia between two independent pathologists. CONCLUSIONS: This assessment has demonstrated that standardisation of less quantitative grading scales resulting in consistent readings is essential for affirmative clinical diagnosis and devising effective treatment strategies.