Determination of the phylogenetic origins of the Árpád Dynasty based on Y chromosome sequencing of Béla the Third.

We set out to identify the origins of the Árpád Dynasty based on genome sequencing of DNA derived from the skeletal remains of Hungarian King Béla III (1172-1196) and eight additional individuals (six males, two females) originally interred at the Royal Basilica of Székesfehérvár. Y-chromosome analysis established that two individuals, Béla III and HU52 assign to haplogroups R-Z2125 whose distribution centres near South Central Asia with subsidiary expansions in the regions of modern Iran, the Volga Ural region and the Caucasus. Out of a cohort of 4340 individuals from these geographic areas, we acquired whole-genome data from 208 individuals derived for the R-Z2123 haplogroup. From these data we have established that the closest living kin of the Árpád Dynasty are R-SUR51 derived modern day Bashkirs predominantly from the Burzyansky and Abzelilovsky districts of Bashkortostan in the Russian Federation. Our analysis also reveals the existence of SNPs defining a novel Árpád Dynasty specific haplogroup R-ARP. Framed within the context of a high resolution R-Z2123 phylogeny, the ancestry of the first Hungarian royal dynasty traces to the region centering near Northern Afghanistan about 4500 years ago and identifies the Bashkirs as their closest kin, with a separation date between the two populations at the beginning of the first millennium CE.

DNA extraction from paraffin embedded colorectal carcinoma samples: A comparison study of manual vs automated methods, using four commercially kits.

BACKGROUND: Nucleic acid isolation from formalin-fixed, paraffin-embedded tissue (FFPET) samples is a daily routine in molecular pathology laboratories, but extraction from FFPET is not always easily achieved. Choosing the right extraction technique is key for further examinations. AIM: To compare the performance of four commercially available kits used for DNA extraction in routine practice. METHODS: DNA isolation was performed on 46 randomly selected formalin-fixed, paraffin-embedded (FFPE) colorectal adenocarcinoma (CRC) surgical specimens. Four commercially available extraction kits were used: two for manual DNA extraction (the PureLink Genomic DNA Mini Kit from Invitrogen and the High Pure FFPE DNA Isolation Kit from Roche) and two for automated DNA extraction (the iPrep Genomic DNA Kit from Invitrogen and the MagnaPure LC DNA Isolation Kit from Roche). The DNA concentration and quality (odds ratio) among the four systems were compared. The results were correlated with the clinicopathological aspects of CRC cases: age, gender, localization, macro- and microscopic features, lymph node metastases, and the lymph node ratio. RESULTS: The highest DNA concentration was obtained using the manual kits: 157.24 ± 62.99 ng/µL for the PureLink Genomic DNA Mini Kit and 86.64 ng/µL ± 43.84 for the High Pure FFPE DNA Isolation Kit (P < 0.0001). Lower concentrations were obtained with automated systems: 20.39 ± 21.19 ng/µL for the MagnaPure LC DNA Isolation Kit and 8.722 ± 6.408 ng/µL for the iPrep Genomic DNA Kit, with differences between the systems used (P < 0.0001). The comparison between age, gender, tumor localization, pT or pN stage and the lymph node ratio indicated no statistically significant difference in DNA concentration using any of the nucleic acid isolation kits. DNA concentration was influenced by the macroscopic features and grade of differentiation. A higher DNA concentration was obtained for well-differentiated polypoid colorectal adenocarcinomas (CRCs), compared with undifferentiated ulcero-infiltrative carcinomas, irrespective of the kit used. CONCLUSION: For research or diagnosis that needs high DNA concentrations, manual methods of DNA isolation should be used. A higher amount of DNA can be obtained from polypoid-type differentiated CRCs. Automated systems confer comfort and a lower amount of DNA that is, however, sufficient for classic polymerase chain reaction (PCR) and real-time quantitative PCR molecular examinations. All four commercially available kits can be successfully used in daily practice.

[Influence of HPV infection on cell cycle regulation in epithelial cell alterations of cervix uteri: Immunohistochemical and in situ hybridization observations]. / A HPV-fertõzés hatása a sejtciklus szabályozására a méhnyak laphámelváltozásaiban: immunhisztokémiai és in situ hibridizációs megfigyelések.

The HPV16 E6 DNA incorporation into the cervical epithelial cell genome was determined. In addition, using p16, p21 and p27 protein immunohistochemistry, we intended to present how the normal cell cycle machinery was disturbed in cervical epithelial cells. In CIN1 the epithelial cells are transformed into koilocytes, p16 is not expressed. p21 is only visible over the parabasal cell layers, while p27 manifests in koilocytes and lymphocytes. In CIN2/3, one copy of HPV16 DNA is integrated in the epithelial cell genome. In CIN3, p16 is expressed in great quantities, while p21 can be seen in the upper 2/3 segment of cervical epithelium causing minimal cell differentiation. p27 is highly expressed in the basal cells of stratified epithelium blocking the autonomous proliferation phases of the cell cycle.

[Colorectal cancer susceptibility genetic variants in tumor free and colorectal carcinoma bearing Hungarian population. Individual predisposition to cancer]. / Vastagbélrák-hajlamosító DNS-szekvencia-variációk tumormentes és vastagbél-daganatos populációban Magyarországon. Az egyedi daganathajlam becslése.

Genome-wide association studies (GWAS) using population-based designs have identified many genetic loci, at which common variants can influence the risk of developing the sporadic colon cancers. These are single nucleotide polymorphisms (SNPs) located on different chromosomes, close to genes involved in cancer developing process, and the SNPs modify their functions, and as a consequence the cancer risk is increased. Our aim was to provide frequency distributions data of variable (risk) allele of six independent SNPs in patients with colorectal cancers and in control Hungarian population, predicting the increased risk effect of sequence variant of SNPs. We also investigated the frequency distribution of tumor localization between right or left half of large bowel as well as the RAS mutation status. 47 non-tumorous patients and 47 patients with colorectal cancer were given oral mucosa cells or blood samples for SNP analysis. After DNA isolation, an LC480 (Roche) type PCR instrument, asymmetric LATE PCR method and melting point analysis were used for detection of sequence variations, by the assistance of two SNP specific primers, unlabeled specific probe and intercalating fluorescent dye. Genomic frequency distribution of variable alleles of SNPs predisposed to tumor development have been investigated in colorectal cancer carrier patients and the results have been compared with the same allele frequency distribution data obtained from the non-tumorous control patients and from CEU population stored in SNPnexus data base. The homozygous risk alleles of SNPs showed a 1.5-2.3-time increase in colorectal cancer carrier patients then in control and CEU patients, but the heterozygous risk allele distribution was identical in tumorous and control population. The frequency distribution of homozygous risk alleles of six SNPs was also investigated in the same time and some patients. Among 47 patients with colorectal cancer, in 3 patients carrying 3 SNPs with homozygous risk alleles, in additional 5 tumor samples two and 24 samples contain only one SNP's homozygous risk alleles, and in 15 patients, SNPs with homozygous risk alleles do not occur. In 47 control patients, only 3 samples contain two SNPs with homozygous risk alleles and 17 samples contain only one SNP with homozygous risk alleles. Significant differences of the tumorous and the control population can be seen detected. NRAS mutation was not found. Our results showed a real increased risk effect of several newly recognized low-penetrance colorectal cancer susceptibility genetic variants by influence of the regulation of neighboring genes, however, the degree of cancer risk is individual, and influenced by others environmental factors, such as dietary factors. Orv Hetil. 2018; 159(40): 1614-1623.

Paradoxical expression pattern of the epithelial mesenchymal transition-related biomarkers CD44, SLUG, N-cadherin and VSIG1/Glycoprotein A34 in gastrointestinal stromal tumors.

AIM: To evaluate the immunohistochemical (IHC) expression of five biomarkers, commonly involved in epithelial mesenchymal/mesenchymal epithelial transition (EMT/MET), in gastrointestinal stromal tumors (GISTs). METHODS: In 80 consecutive GISTs the IHC examinations were performed using the EMT-related antibodies E-cadherin, N-cadherin, SLUG, V-set and immunoglobulin domain containing 1 (VSIG1) and CD44. RESULTS: The positivity rate was 88.75% for SLUG, 83.75% for VSIG1, 36.25% for CD44 and 10% for N-cadherin. No correlation was noted between the examined markers and clinicopathological parameters. Nuclear positivity for SLUG and VSIG1 was observed in all cases with distant metastasis. The extra-gastrointestinal stromal tumors (e-GISTs) expressed nuclear positivity for VSIG1 and SLUG, with infrequent positivity for N-cadherin and CD44. The low overall survival was mainly dependent on VSIG1 negativity (P = 0.01) and nuclear positivity for SLUG and/or CD44. CONCLUSION: GIST aggressivity may be induced by nuclear up-regulation of SLUG and loss or cytoplasm-to-nuclear translocation of VSIG1. SLUG and VSIG1 may act as activated nuclear transcription factors. The CD44, but not N-cadherin, might also have an independent prognostic value in these tumors. The role of the EMT/MET-related transcription factors in the evolution of GISTs, should be revisited with a larger dataset. This is the first study exploring the IHC pattern of VSIG1 in GISTs.

[Proposal for the modernization of cervical screening procedure in Hungary]. / Javaslat a hazai méhnyakszurési eljárásrend korszerusítésére.

Two main considerations played roles in creation of new cervical screening system. One was the proven fact that high-risk human papilloma virus infection plays a role in the development of cervical cancer and pre-cancerous lesions. The other was the implementation of the HPV infection's biological behavior in the new screening strategy. The new screening procedure faithfully reflects the cervical carcinogenesis. An organised, population-based and age differentiated screening method could increase attendance of screening and could decrease the possibility of interval cancer rate due to increased sensitivity. Orv Hetil. 2017; 158(52): 2062-2067.

Immunohistochemical features and staging of early gastric cancer.

INTRODUCTION: The aim of this study was to analyze the particularities of early gastric cancer (EGC) and their importance for staging, prognosis, and therapy. MATERIAL AND METHODS: A total of 338 GCs diagnosed and surgically removed in three medical institutes from Eastern Europe were retrospectively examined, and the EGCs were further examined. Besides the demographic factors and tumor characteristics, immunostains were performed with E-cadherin, HER-2, p53, Ki67, MLH-1, MSH-2, COX-2, VEGF-A, CD31, and CD105. RESULTS: From the 338 GCs, 29 were EGCs, the average being similar in Poland and Hungary (12.37% and 13.33% respectively) but lower in Romania (5.61%). The rate of lymph node metastases was 20.69% (n = 6). Two of the cases presented liver metastases, both of them having a multifocal aspect. In 1 of these cases, limited to the mucosa, intramural carcinomatosis of the lymph vessels was seen in submucosa, muscularis propria, and subserosa. COX-2 positivity was observed in 14 (48.72%) cases. COX-2 was directly correlated with microvessel density determined with CD31 (p < 0.001) and CD105 (p = 0.03). Same correlation with CD31 and CD105 was seen for HER-2 (p = 0.03 and p = 0.0007). The only negative independent prognostic factors for overall survival were tumor localization at the proximal stomach and male gender, regardless of age. CONCLUSIONS: In EGCs, intramural carcinomatosis of the lymph vessels and multifocality should be separately described in every surgical pathology report, as indicators of aggressiveness. Microsatellite status, E-cadherin, HER-2, p53, and Ki67 do not have prognostic value in EGC, but the highly angiogenic pattern is a possible therapeutic target.

PKCθ utility in diagnosing c-KIT/DOG-1 double negative gastrointestinal stromal tumors.

BACKGROUND: The aim of this study was to evaluate the diagnosis value of an immunohistochemical (IHC) panel of three antibodies for the diagnosis of gastrointestinal stromal tumors (GISTs). MATERIAL AND METHODS: In 80 consecutive GISTs without lymph node metastases, the IHC examinations were performed using the antibodies CD117 (c-KIT), DOG-1 and c-theta (PKCθ) protein. The diagnostic value of PKCθ in c-KIT/DOG-1 negative GISTs has been explored in fewer than 10 Medline-indexed papers. RESULTS: The c-KIT, PKCθ and DOG-1 positivity was noted in 92.50% (n = 74), 90% (n = 72) and 76.25% (n = 61) of the cases, respectively. All of the C-KIT negative cases (n = 6) were also DOG-1 negative but displayed PKCθ positivity. All of the DOG-1 positive cases (n = 61) also expressed c-KIT. No correlation between the examined markers and clinicopathological parameters was noted. CONCLUSIONS: The PKCθ sensitivity is similar to c-KIT and superior to DOG-1 sensitivity. All of the c-KIT/DOG-1 negative GISTs seem to express PKCθ. For a proper diagnosis of GIST, the c-KIT/DOG-1/PKCθ panel should be used, with possible therapeutic but not prognostic value.

[Human papillomavirus associated cervix uteri morbidity in Hungary: epidemiology and correlation with the HPV types and the simultaneous cytological diagnosis]. / Humán papillomavírus asszociált méhnyak-megbetegedések Magyarországon: epidemiológia és a HPV-típusok összefüggése a párhuzamosan végzett citológiai diagnózissal.

INTRODUCTION: Persistent infection of human papillomavirus is known to cause cervical intraepithelial neoplasia or cancer in the cervix uteri and other HPV-associated cancers in different localization. Based on epidemiological and biological data, principally the high risk HPV is responsible for development of cervical these cancers. However, we have no information about the frequently distribution of different HPV types and what is the correlation between the HPV types and cytological diagnosis in cervical intraepithelial neoplasia (CIN). AIM: In this paper, we are going to present new data involving incidence and mortality of HPV-associated cancers during the period of 2009-2015 in Hungary. We are also going to investigate the correlation of cervical cytological diagnosis and HPV typing, and the preventive effect of HPV vaccination. METHOD: The epidemiological data spring from the National Cancer Registry. HPV typing was performed by Linear Array HPV Genotyping Test. Simultaneous cytological diagnosis and HPV typing was carried out on 2048 cytological samples collected in period of 2009-2016. RESULTS: According to the epidemiologic data, the most frequently occurring HPV-associated cancer is the laryngeal carcinoma in man, and the cervical cancer in woman in Hungary. During the 2009-2015 time intervals, the frequency distribution of head and neck cancers was not changed in man, but the incidence of tongue root squamous cell carcinomas was gradually increasing in woman. We have defined the clinical significance of single and simultaneously multiple HPV infection and have investigated the correlation of the HPV frequency distribution and cytological diagnosis in CIN. It was found that in the cytological negativity of probably/possibly carcinogen pHR-HPV group classified by IACR was much more frequent as in HR-HPV group (56% versus 47%). The presence of simultaneous multiplex HPV infection betokens an increased cancer risk. According to the international publications, the ratio of HPV16 just twice as big as in cervical cancer, what we found in CIN (60% versus 30%). The frequency order of the HPV18 is 2nd in cancer, and 9th in CIN. Comparing the frequency distribution of HR/pHR-HPVs in cervical cancer and CIN, the HR-HPV35 is very rarely occurring in CIN, the pHR-HPV56, 66, and 73 is more frequently seen in CIN as in carcinoma. Appreciated the preventive value of anti-HPV vaccines, we have found a significant differences in group with 1 HPV/sample and in group with more than 1 HPV/sample. CONCLUSION: The frequency distribution of tongue root squamous cell carcinoma and cervical cancer was gradually increasing in woman. The overall preventive effect of 9-valent vaccine is 80.3%. This preventive value should be higher because of the transformation ability of the different HPV types is not same. Out of consideration for HPV incidence in cancer, the preventive effect of 9-valent or 4-valent vaccines might reach to 93% or 73%. However, the pHR-HPVs are biologically active, it is not sufficient for the inclusion of these HPV types into population-wide HPV-DNA based cervical screening programs. Orv Hetil. 2017; 158(31): 1213-1221.

Proposal of a Dukes-MAC-like staging system for gastric cancer.

The aim of this study was to present an epidemiological update regarding the classical prognostic parameters of gastric cancer (GC) in 3 countries from Eastern Europe and to suggest a modification of the pTNM staging system. In 333 consecutive cases which were diagnosed between 2003 and 2012 in 3 departments of pathology from Romania, Hungary, and Poland, the following parameters were analyzed: age and gender of patients, tumor localization, macroscopic and microscopic aspects including the degree of discohesivity, depth of tumor infiltration, and pTNM stage. From all of the studied parameters, the following proved to have independent prognostic value, indicating a lower survival rate: presence of distant metastases (p=0.001), lymph node positivity (p=0.0009), depth of tumor infiltration (p=0.04), age over 50 (p=0.02), proximally located tumors (p=0.03), and ulceroinfiltrative or diffusely infiltrative macroscopic aspect (p=0.0002). The pT2N1-3 staged cases showed a worse prognosis compared with the pT3N0 ones (p=0.02). Regardless of depth of invasion, the lymph node status remains the strongest indicator of the survival rate in GC. The pTN staging system should be adapted and a Dukes-MAC-like staging system should include the following groups: stage A1-T1N0, stage A2-T1N1-3, stage B1-T2N0, stage B2-T2N1-3, stage C1-T3N0, stage C2-T3N1-3, and stage D-T4N0-3. The grade of discohesivity/budding is not a prognostic factor in GC.

Human papilloma virus status evaluation and survival description in selected oropharyngeal and laryngeal squamous cell carcinoma patients from Hungary.

PURPOSE: Many patients with oropharyngeal squamous cell carcinomas do not have any of the traditional risk factors associated with head and neck squamous cell cancers (HNSCC). Epidemiologic and molecular studies have identified human papillomavirus (HPV) as a causative agent, viral tumors presenting a better survival and being important risk factors together with the long established ones, tobacco and alcohol consumption, in head and neck cancers. The purpose of this study was to establish the incidence of HPV-associated HNSCCs, to identify the most frequent HPV type and to evaluate the overall survival and recurrence rates of HPV-positive cases in comparison with HPV-negative HNSCCs. METHODS: A retrospective analysis from the database of the National Institute of Oncology from Budapest was performed and the following parameters were analyzed: age, age at diagnosis, gender, primary tumor location, tumor histopathology, TNM stage, HPV status, date of recurrence, last visit and date of death. RESULTS: Out of 81 patients with HNSCCs 55 (67.9%) were male and 26 (32.1%) female. HNSCCs were more frequent in men (2.11:1) and the majority of the patients (81.7%) were diagnosed in advanced stages (TNM III and IV). HPV status was evaluated in nearly half (48.14%) of the patients and HNSCCs were positive for HPV in 43.6% of the cases. These were more frequent in patients over 50 years (76.66%), in men (76.47%) and in oropharyngeal location (94.1%). HPV-16 type was associated with malignancy in 82.35% of the cases. Disease recurrence was more frequent in HPV-negative (31.81%) vs HPV-positive cases (29.41%) and mortality rate was inferior in HPV-positive 33.33% vs negative (38.09%) tumors (p=0.52). CONCLUSIONS: In Hungary HNSCCs are more frequent in men than in women. HPV positivity is higher in men vs women and in oropharyngeal vs laryngeal location. Overall survival rate was superior in HPV-positive vs HPV-negative cases. Disease recurrence was more frequent in HPV-negative vs HPV-positive cases.

Geographic particularities in incidence and etiopathogenesis of sporadic gastric cancer.

It is known that geographical differences in the prevalence and etiopathogenesis of gastric cancer exist across the world. Eastern Europe and East Asia are two of the largest endemic areas of gastric cancer in the world, yet there are few studies comparing its features in these two regions. Based on our experience and literature data, we performed a review that is mainly focused on the etiology and pathogenesis of sporadic gastric cancer and its geographic particularities. Geographic prevalence of specific Helicobacter pylori strains is also synthesized. The pathogenesis of gastric cancer in patients from countries of the authors, respectively Japan, Romania, Hungary and Poland, is particularly examined.

[Pregnancy and delivery with transfer of vitrified blastocysts following trophectoderm biopsy]. / Terhesség és szülés trophectoderma-biopsziát követoen vitrifikált blasztociszták beültetésével.

Application of preimplantation genetic diagnosis makes it possible to transfer only embryos unaffected by a certain genetic disorder. The authors have applied the combination of trophectoderm biopsy and vitrification in order to detect a monogenic disorder. Previously diagnosed type 1 neurofibromatosis of the woman was the indication for genetic examination. In vitro fertilisation and embryo culture was performed using sequential culture mediums. Seven blastocysts could be sampled on the fifth day and were vitrified subsequently. Two blastocysts turned out to be genetically normal based on the result of genetic examination using polimerase chain reaction. A healthy boy was delivered following the transfer of warmed blastocysts and an uneventful singleton pregnancy.

New Insights in Histogenetic Pathways of Gastric Cancer.

The aim of this paper was to describe 3 possible histogenetic pathways for poorly cohesive (diffuse) carcinomas and 2 for intestinal-type gastric carcinomas (GCs), which might influence the behavior of GC. In the present observational study, 102 patients with early (n = 50) and advanced GCs (n = 52) were evaluated, and the histogenetic background was analyzed. All of the cases were sporadic GCs. For particular aspects, Maspin, E-cadherin, and SLUG immunostains were performed. For our final conclusions, the results were correlated with literature data. In early stages, poorly cohesive carcinomas can display 3 histogenetic pathways, with particular molecular behaviors: "carcinoma with intraepithelial pagetoid onset" (with or without a switch from E-cadherin to SLUG positivity), "carcinoma with early lymphatic invasion" (carcinoma limited to mucosa but with carcinomatosis of the lymph vessels from subjacent layers), and "microglandular-type poorly cohesive carcinoma" (the onset is similar with adenocarcinoma but abrupt dedifferentiation can be seen in the submucosa, with persistence of a dual component in the deep layers). The intestinal type carcinoma can be developed on the background of superficially located dysplasia ("classic adenocarcinoma") or in the submucosal heterotopic mucosa ("adenocarcinoma arising from the mucosal infolding in the submucosa"). Based on personal observations correlated with literature data, 5 histopathogenetic pathways are proposed with specific denominations. Each of them can partially explain the aberrant behavior of early gastric cancer.

[Liquid-based cervical cytology ThinPrep screening in Hungary]. / A folyadékalapú méhnyakrákszurés eredménye Magyarországon.

INTRODUCTION: Liquid-based cervical cytology has been developed as an alternative for conventional Papanicolaou cervical cytology. AIM: The aim of this study was to determine the quality assurance of liquid-based cervical cytology. METHOD: 4573 cervical cytology smears were classified according to the Bethesda (2001) system. Human papilloma virus infection was detected and subtyped from reflex test using real-time polymerase chain reaction. RESULTS: 4573 smears were classified according to the Bethesda (2001) system. Negative diagnosis was made in 2323 cases (50.8%), non neoplastic in 2017 cases (44.1%), and positive for intraepithelial lesions or malignancy in 233 cases (5.1%). Unsatisfactory smear for diagnosis was found in 43 cases (0.9%), low-grade squamous intraepithelial lesion in 87 cases (1.9%), high-grade squamous intraepithelial lesion in 24 cases (0.5%), and carcinoma in 23 cases (0.5%). Fifty-nine of the cases were histologically verified and 4 falsely negative cases were detected. The sensitivity, specificity and the positive predictive value were 93.2%, 100% and 100%, respectively. Compared to an identical time periods of the previous three years, the low- and high-grade squamous intraepithelial lesion increased from 0.82% to 2.51%. Eighty one human papilloma virus tests were performed with a positive predictive value of 99%. CONCLUSIONS: The auditing values of the liquid-bases cervical cytology results meet the proposed threshold values. Liquid-bases cervical cytology is an alternative cervical cytology and it seems to be significantly better than conventional Papanicolaou cervical cytology in all parameters.

Molecular and immunohistochemical profile of a basaloid (cloacogenic) carcinoma of the sigmoid colon: possible predictive value for clinical outcomes.

A 61-year-old woman was hospitalized with a 5-week history of abdominal discomfort, change in bowel habits, and weight loss. Colonoscopy showed a protruded tumor of the sigmoid colon first diagnosed as undifferentiated carcinoma. Surgical resection of the sigmoid colon was performed. Histological examination of the surgical specimen showed a proliferation of basaloid cells arranged in tumor clusters with central comedonecrosis and peripheral palisading of the nuclei. The tumor invaded the subserosa and presented liver metastasis without lymph node metastases. The tumor cells were marked by keratin AE1/AE3, keratin 5/6, epithelial membrane antigen, bcl-2, vascular endothelial growth factor, CD105, neuron-specific enolase, MLH-1, MSH-2, and p53, and were negative for keratin 7/20, chromogranin, synaptophysin, carcinoembryonic antigen, p63, c-KIT, and maspin. A high p53 nuclear index was also detected. On the basis of these characteristics and molecular examinations, the final diagnosis was microsatellite stable/human papilloma virus-negative/K-ras mutated/BRAF wild-type basaloid carcinoma (BC). Only seven BCs of the colon were reported in the literature, this being the eighth one and the first case that reports new molecular findings about microsatellite instability, K-ras/BRAF mutations, angiogenesis, and maspin expression in BC, with direct involvement in targeted therapy.

Serrated pathway adenocarcinomas: molecular and immunohistochemical insights into their recognition.

INTRODUCTION: Colorectal adenocarcinomas (CRC) developed through serrated pathway seem to present particular behavior compared with the non-serrated ones, but recognition of them is difficult to do. The aim of our paper was to establish some criteria to facilitate their identification. MATERIALS AND METHODS: In 170 consecutive CRCs, we performed immunohistochemical staining with Cytokeratin 7 (CK7) and Cytokeratin 20 (CK20) and also with p53 and MLH-1. At the same time, we analyzed BRAF and K-ras mutations and the microsatellite status of CRC. RESULTS: 26.47% of cases expressed CK7, and 16.47% were CK20-negative. Diffuse positivity for CK7 was associated in the proximal colon with CK20 negativity or weak positivity, BRAF mutations, lack of K-ras mutations, and p53 and MLH-1 negativity. All these cases were microsatellite-unstable and were diagnosed in stage II. Those cases from the distal colon and rectum that expressed CK7 were K-ras-mutated and had low p53 index and MLH-1 positivity, independent of the CK20 expression. CONCLUSIONS: CK7, associated with MLH-1 and p53 expression, and also with the microsatellite status, BRAF and K-ras pattern, might be used to identify the CRC potentially going through serrated pathway. The serrated pathway adenocarcinomas of the proximal colon that do not display the morphological features of this pattern are more frequent CK7+/p53-/MLH-1-/BRAF-mutated/K-ras-wt/MSI cases, but those located in the distal colorectal segments seem to be CK7+/CK20+/p53-/MLH-1+/BRAF wt/K-ras-mutated/MSS cases.

Non-Epstein-Barr virus associated lymphoepithelioma-like carcinoma of the esophagogastric junction with microsatellite instability, K-ras wild type.

A 60-year-old male was admitted to our hospital for gastric cancer. Considering his general condition, total gastrectomy and dissection of regional lymph nodes were performed. Macroscopically, a 45 mm × 20 mm × 10 mm-sized, ulcero-infiltrative tumor located in the esophagogastric junction was described. Microscopically, the tumor consisted of a poorly differentiated adenocarcinoma intermingled with dense lymphoid infiltration predominantly composed of T-cell lymphocytes. The tumor cells infiltrated the submucosa, muscularis and subserosal layers of the stomach, respectively the esophageal adventitia. No metastases were noticed in the 58 regional lymph nodes. Based on the histopathological features, the diagnosis was lymphoepithelioma-like carcinoma, pT3N0 stage. In situ hybridization for Epstein-Barr virus showed no nuclear signal in tumor cells. The p53 expression was observed in fewer than 10% of the tumor cells. Real-time PCR analysis showed microsatellite instability without K-ras mutation in codon 12. No recurrences or metastases were reported 6 months after surgical intervention. No adjuvant therapy was performed.