Campylobacter coli cultured from the stools of a patient with immunoproliferative small intestinal disease.

Campylobacter has been associated with immunoproliferative small intestinal disease (IPSID), on the basis of 16S rDNA sequencing, in situ hybridization, and immunohistochemistry. Here, for the first time, we have cultured Campylobacter from the stools of a patient with IPSID. Phenotypic analysis and whole genome sequencing identified Campylobacter coli. PCR on a IPSID tissue biopsy sample was positive for Campylobacter coli and negative for Campylobacter jejuni. These findings further support a causative role for Campylobacter in the development of IPSID.

Littoral cell angioma of the spleen--a surprising cause of anemia.

Littoral cell angioma is a rare tumor of the spleen, usually being considered benign and typically discovered incidentally. There are three different modalities of presentation: tumoral splenomegaly, long-standing iron deficient anemia or thrombocytopenia due to hypersplenism. However, some of its manifestations could generate the suspicion of a lymphoma or other more serious condition. We present the case of a 46-year-old man with splenomegaly and iron deficiency anemia. The tumor affected the whole spleen, which was surgically removed. The histopathological examination, together with immunophenotyping, established the diagnosis. Six months after the procedure, the patient is in very good condition. Several differential diagnoses were discussed, as well as the prognostic factors. The case illustrates a rare cause of anemia and the importance of pathology in uncovering such unusual causes for this.

K-ras gene mutation status in colorectal cancer: comparative analysis of pyrosequencing and PCR-RFLP.

BACKGROUND: In patients with high-stage colorectal carcinomas (CRC), anti-EGFR therapy is known to be effective only in cases with a wild-type K-ras gene status. Different procedures have been proposed for such evaluation. MATERIALS AND METHODS: The mutation status of K-ras gene, codons 12, 13 and 61 was determined in 250 CRC cases using the pyrosequencing assay. In addition, we compared the performance of the pyrosequencing procedure with that of PCR-RFLP in a subset (n=100) of the CRC samples the latter only in codons 12 and 13. RESULTS: Using pyrosequencing, 46.4% of the 250 CRC cases were found mutated. Most mutations were located in codon 12 (36.4% from all cases) and several were located in codon 61 (3.2%). All mutation identified by PCR-RFLP were confirmed by pyrosequencing and, in addition, one more mutated sample was identified in the subset of 100 samples. CONCLUSIONS: Both methods are highly specific and can profitably be used in the molecular diagnosis of colorectal cancer in order to establish the adequate therapy.

Foxp3 and IL17 expression in tumor infiltrating lymphocytes (TIL) and tumor cells - correlated or independent factors?

Tumor infiltrating lymphocytes (TIL), as a microenvironment component were studied in various epithelial tumors, with contradictory results. Recent data about regulatory T-cells (Treg) revealed new explanations for pro- and anti-tumor implications of TIL. Tregs immunoprofile was recently completed with Foxp3 expression. A T-cell fraction (Th) is producing cytokine IL17 and is now considered acting in tumor progression. Our study aimed to analyze immunohistochemically (IHC) Foxp3+ and IL17 expression in resected lung adenocarcinomas, since they could become possible targets in the antitumor immunotherapy. The studied material was represented by paraffin-embedded tumor fragments from 59 patients with TIL identified on HE staining. The antibodies used were Foxp3 and IL17. The statistical analysis used logistical regression on SPSS19 software (Chicago, IL, USA). TIL was usually mild or scarce. A positive statistic correlation resulted between the amounts of TIL in peritumoral and intratumoral location but without correlation to histopathological grading. Foxp3 and IL17 were present in TIL lymphocytes, tumor cells and fibroblasts; IL17 was expressed also in periendothelial cells (PEC). Foxp3 positivity was significantly correlated for lymphocytes÷tumor cells, lymphocytes÷fibroblasts and tumor cells÷fibroblasts, suggesting their concerted action. Tumor cells and lymphocytes Foxp3 expression was inversely correlated with the amount of TIL. Between lymphocytic Foxp3 and PEC IL17, we found a weak negative correlation. The TIL had a quite positive correlation with PEC IL17. In these conditions, Foxp3 could be a mediator of the tumor cells inhibitory aggression upon the immune system and could be used as a molecular target for biological antitumor therapy.

The mandate for a proper preservation in histopathological tissues.

A sequence of technically reproducible procedures is mandatory to guarantee a proper preservation of tissues and to build up the basis for sound diagnoses. However, while the goal of these procedures was, until recently, to assure only structural (histological and cytological) preservation, an appropriate preservation of antigenic properties and of nucleic acid integrity is now additionally requested, in order to permit pathologists to provide the biological information necessary for the adoption of personalized therapies. The present review analyses the sequence of technical steps open to critical variations. Passages such as dehydration, paraffin embedding, sectioning and staining are relatively well standardized and allow adoption of dedicated (automatic) apparatuses, while other pre-analytical steps, i.e. time and modalities of transfer of surgical specimens from the surgical theatre to the pathology laboratory (s.c. "ischemia time") and the type and length of fixation are not standardized and are a potential cause of discrepancies in diagnostic results. Our group is involved in European-funded projects tackling these problems with the concrete objective of implementing a model of effective tumors investigations by high performance genetic and molecular methodologies. The problem of the discrepant quality level of histopathological and cytological preparations involved five European countries and exploiting the potential of "virtual slide technology". Concrete issues, techniques and pitfalls, as well as proposed guidelines for processing the tissues are shown in this presentation.

Cellular immunophenotypes in human embryonic, fetal and adult heart.

The cellular immunoprofile of cardiac dysfunctions and lesions of ischemic etiology are insufficiently studied to date, especially regarding the contribution of non-cardiomyocytic structures. Aiming to explore this immunoprofile, we used immunohistochemistry applied on embryonic, fetal and adult normal or ischemic myocardium. We observed a decrease of smooth muscle alpha-actin expression in fetal vs. embryonic cardiomyocytes, its absence in normal adult myocardium and its intense expression in the fibrotic scars of ischemic myocardium. DDR2 and vimentin, which are present in the interstitial cells and cardiomyocytes of the embryo, fetus and normal adult heart, are absent in the fibrotic scar tissue and cicatricial infarction, the latter expressing smooth muscle alpha-actin and CD34. This suggested that myofibroblasts and not local fibroblasts that participate in ischemic remodeling. An EGFR-positive vascular network was better represented in the ischemic heart than in the adult normal one, a fact possibly related to EGFR implication in cardiac ischemic pre- and post-conditioning. Therefore, cardiomyocytes and non-cardiomyocytic cells have an undulating immunoprofile according to the intrauterine life stage or age after birth, and a variable contribution in cardiac lesions, mostly in ischemic ones.

Nestin and caveolin-1 in the diagnosis of GISTs.

Gastrointestinal stromal tumors (GISTs) are the most common primary mesenchymal neoplasias of the gastrointestinal tract, typically expressing c-kit (CD117) and CD34. Recently, it was reported that nestin and caveolin-1 are also expressed in some human sarcomas, GISTs included. We performed a retrospective study on formalin fixed, paraffin embedded samples from 81 cases of confirmed GISTs, aiming to characterize their immunohistochemical profile, including nestin and caveolin-1 expressions. Tissue samples were evaluated immunohistochemically for CD117, CD34, nestin and caveolin-1. The patients (M:F 36:45), aged 46 to 84 years, had spindle cell type GISTs in 56.7% of cases, epithelioid in 30.8% and mixed pattern in 12.3%. Immunohistochemically, CD117 was positive in 88.9% of GISTs, CD34 in 85.1%, nestin in 77.7% and caveolin-1 in 71.6% of the tumors. Of nine c-kit negative GISTs, 66.7% expressed nestin, the same as caveolin-1 and 44.5% expressed both nestin and caveolin-1. Statistical analysis using Kendall's and Spearman's tests revealed significant correlations between nestin and caveolin-1 expressions (p=0.024). Our results suggest that nestin and caveolin-1 could be considered sensitive markers in GISTs, together with CD117 and CD34, for diagnostic purposes. Their significant expression in CD117 negative GISTs could represent an immunohistochemical alternative in establishing the diagnosis of these tumors.

C677T and A1298C methylenetetrahydropholate reductase (MTHFR) polymorphisms as factors involved in ischemic stroke.

BACKGROUND: Ischemic stroke is a major health problem. Data regarding the possible association between ischemic stroke and the polymorphism of methylenetetrahydropholate reductase (MTHFR) C677T and A1298C are still conflictual. AIM: The study tried to assess the association of the two MTHFR polymorphisms with ischemic stroke in a series of patients from a unique hospital center. MATERIALS AND METHODS: The study comprised a total of 127 patients (67 with non-cardioembolic ischemic stroke diagnosed by computed tomography or magnetic resonance imaging) and 60 control cases. The method we used was reverse hybridization performed on peripheral blood for C677T and A1298C polymorphisms. In all patients a careful clinical examination, laboratory analyses of cholesterol, glucose amount and triglycerides, as well as their medical history were available. RESULTS: The mean age of stroke patients was 68.73 years, and 55.2% were males. Gene analysis for C677T disclosed the presence of TT genotype in more control subjects than in stroke series (15% and 7.46% respectively). Also, the overall T allele (CT+TT cases) was present in 71.6% of control cases, as compared with 44.7% stroke patients. 1298C allele was almost equally distributed among the two series. No statistically significant correlations of the two genotypes with infarct localization and dimensions ant with other potential risk factors (hypertension, lipids, diabetes mellitus) were observed. CONCLUSIONS: The two MTHFR polymorphisms, C677T and A1298C, seemed not related to the onset of ischemic stroke in our study. However, they could be rather involved in hemorrhagic stroke, as seen in our control patients. Further evaluation on larger series is mandatory since homocysteine activity (related to MTHFR activity) could be easily influenced by folate or cobalamin derivatives.

Clinico-pathological correlations in fatal ischemic stroke. An immunohistochemical study of human brain penumbra.

Ischemic stroke is one of the most frequent pathologies with high invalidating potential and a leading cause of death. The brain tissue adjacent to the central necrotic core, defined as penumbra, was extensively characterized mostly by imaging techniques and in animal models. Our goal was to identify a large panel of molecules in this particular area on human brains harvested at autopsy. Twenty-one patients with ischemic stroke and seven control cases were taken into study. We used immunohistochemistry to characterize necrotic lesions. Metalloproteinases, mostly MMP-9, seem to be involved in brain ischemia, but as a protective and not as a deleterious factor. Apoptotic molecules are not increasingly expressed in stroke compared to control cases. Mast cell enzymes chymase and tryptase are described for the first time in neurons and glia, even with unclear significance. Microglia appears active in stroke and stimulating methods directed to it could be useful. Nitric oxide synthases and cyclooxygenase-2 were also involved in stroke cases but not in control ones. Other factors as VEGF and its receptors, PDGF, b-FGF or TNF-alpha showed no significant expression related to ischemic brain injury. Animal study of penumbra and human tissue findings are distinct and research should be focused on the latter approach in order to find valuable and safe therapeutic methods.

Neurodegenerative changes in human aging brain. An autopsy study.

Neurodegenerative pathological changes are known as occurring in human brain, in some way paralleling aging. We characterized prospectively the occurrence of cortical senile plaques and neurofibrillary tangles in 55 adult human subjects, by post-mortem examination. We tried to determine if aging is associated with greater senile plaque and neurofibrillary tangles burden and what is the cortical distribution of lesions, regardless the mental status of the patient. The series comprised a large spectrum of ages, from 30 to 97-year-old. Immunohistochemistry for amyloid-beta (Abeta) and tau protein was the technique we used. ApoE genotyping was performed in 33 cases by polymerase chain reaction. In our series brain Abeta deposition as senile plaques occurred only after 65-year-old. These accumulations were strongly associated with the occurrence of neurofibrillary tangles. However, several very old patients were lacking both beta-amyloid and tau-positive lesions. As a result, even though Abeta and tau protein show a certain predilection for brain deposition in elder people, their relationship with aging still needs further investigation, mostly in human subjects.

Pancreatic cancer: incidence, treatment and survival trends--1175 cases in Calvados (France) from 1978 to 2002.

AIM: To assess the trends in incidence, therapeutic modalities and survival of pancreatic cancer between 1978 and 2002 in a well-defined population, as recorded in the Calvados digestive cancer registry database. PATIENTS AND METHODS: All patients living in Calvados with a diagnosis of pancreatic cancer were registered. Clinical data and treatment modalities were prospectively recorded. This 25-year database was divided into five 5-year periods. Data were compared using log-rank tests and the Cox model. RESULTS: A total of 1175 cases of pancreatic cancer (617 men, 558 women) were registered. Its incidence increased with an average annual coefficient of +2.8% in men and +5.1% in women. Therapeutic modalities changed over the five time periods: surgical resection increased from 6.8 to 13.4% (median survival 15 months) while radiation therapy and/or chemotherapy also increased from 5.5 to 13.2%. Palliative surgery decreased from 54.6 to 32.0% and favored interventional endoscopic techniques. Postoperative mortality decreased significantly. Survival increased significantly over the five time periods, although the median survival time remained stable (4 months). CONCLUSION: From 1978 to 2002, pancreatic cancer incidence increased in Calvados (France). Therapeutic modalities changed, with endoscopic treatments preferred over palliative surgery. The improvement in survival could be explained by the decrease in postoperative mortality.

Georges Marinesco and the early research in neuropathology.

OBJECTIVE: To present the scientific contributions of Georges Marinesco (1863-1938) and place his achievements within the context of early neuropathology research. BACKGROUND: Neuropathology is a relatively recent medical field, its origins dating to the late 19th century. RESULTS: One of the most important neuroscientists of that period was the Romanian-born Georges Marinesco. He became a neurologist under Charcot's guidance at the Salpêtrière Hospital, in Paris. In 1892, Paul Blocq and Marinesco gave a first account of senile plaques, having used their pathologic skills in the examination of nine deceased epileptic patients. They did not, however, relate the plaques to dementia. Marinesco made discoveries in neuropathology which he described from a histopathologic perspective, and introduced new medical terms such as neuronophagia, chromatolysis, and medullomyoblastoma. He also drew correlations between clinical neurologic findings and morphology, for example in congenital cerebellar ataxia, syringomyelia, and parkinsonism. From 1899 he used cinematography as a medical research tool. CONCLUSION: Marinesco was a prolific researcher in the field of neuropathology, especially neurodegeneration but also in clinical neurology. He is now considered the founder of the modern Romanian school of neurology.

Endoscopic ultrasonography is an independent predictive factor of prognosis in locally advanced esophageal cancer. Results from the randomized FFCD 9102 study from the Fédération Francophone de Cancérologie Digestive.

BACKGROUND: No multivariate study has assessed the independent prognostic role of endoscopic ultrasonography (EUS) in esophageal cancer, even when considering computed tomography (CT). OBJECTIVE: To evaluate the prognostic value of EUS in esophageal cancer before exclusive or preoperative radiochemotherapy. METHODS: From 1993 to 1999, the FFCD 9102 study enrolled 444 patients who had cancer of the thoracic esophagus, stages T3-4, N0-1 and M0 on CT. The patients received two sessions of chemotherapy in addition to radiotherapy. The 259 patients with objective response and no contraindications for further treatment were randomized to undergo surgery or to continue with radiochemotherapy. EUS was performed in 174 patients enrolled in the trial (mean age: 59 years). Tumor characteristics and lymph node status were prospectively recorded. A Cox statistical model was used to identify any predictive prognostic factors among the clinical, EUS and CT data. RESULTS: In the multivariate analysis, three factors were associated with a poor prognosis: inability to ingest solid food (OR: 1.98; P=0.0008); more than three neoplastic subdiaphragmatic lymph nodes (LN) on EUS (OR: 2.41; P<0.0045) and age>65 (OR: 1.53; P<0.056). Their prognostic value persisted after adjustment for type of treatment given. Two- and five- year survival rates were 21.5 and 10.5%, respectively, in the presence of three neoplastic subdiaphragmatic LN, and 43 and 30%, respectively, in all other cases. CONCLUSION: Degree of dysphagia, age and presence of neoplastic subdiaphragmatic LN on EUS were independently predictive of the prognosis for locally advanced esophageal cancer. EUS results should be taken into account in future trials.

Giant intracranial endolymphatic sac tumor (ELST). Case presentation and histogenetic considerations.

We present a giant tumor of the skull base compressing the brain in a 40-years-old man. The tumor was policystic at imaging. Its histopathology, immunohistochemical profile and long evolution suggest an endolymphatic sac tumor (ELST), a rare case of neoplasia. Since the patient had multiple otolaryngological procedures in his medical history, a possible traumatic pathogenesis could be suspected. On the other way, some immunohistochemical aspects found in our case may imply a histogenesis divergent from that currently accepted. This could be from either the organ of Corti or some local cells that generate a resemblance with a systemic tumor, the so-called benign mesothelioma. Further studies are needed in order to clarify this topic.

Angiocentric glioma: presentation of two cases with dissimilar histology.

OBJECTIVE AND IMPORTANCE: Angiocentric glioma (AG) is a recently described tumor of the brain which was included as a distinct entity in the 2007 WHO classification. To date only 26 cases have been reported in the literature. We describe two additional cases of this possibly confusing lesion of the brain. Emphasis is put on variations in the histopathological picture. CLINICAL PRESENTATION: The patients (20- and 55-year-old males) presented with seizures and headaches, respectively. Imaging examination showed a small cortical-subcortical tumor in each case. Both tumors were totally removed. MATERIAL AND METHODS: Paraffin blocks from the two cases were examined with classical histopathology stainings and immunohistochemistry for GFAP, vimentin, EMA, neurofilament protein, synaptophysin, S100 protein, CD31, CD34, FVIII, smooth muscle actin and Ki67. RESULTS: The tumor proliferation was restricted around small intraparenchymal vessels. Immunohistochemistry demonstrated positivity for glial and negativity for vascular or neuronal markers. The cell shape and arrangement was different in the two cases. CONCLUSIONS: AG is a peculiar tumor of uncertain histogenesis but with certain glial reactivity. Histopathology is variable but restricted, for unknown reasons, to perivascular areas. Apparently, a better prognosis than for other gliomas is distinctive. Further studies are needed in order to expand the information regarding the clinical behavior and therapeutic approach of this tumor type.

An intrajugular paraganglioma. Unusual presentation of a classical tumor.

Paragangliomas arise from the extraadrenal neuroendocrine system. They are locally aggressive tumors, causing adjacent invasion, bone destruction and compression related symptoms. We present a 35-years-old woman with a peculiar paraganglioma lacking all these features, and strictly located within the jugular vein. Differential diagnosis is detailed since other entities could have dissimilar clinical behavior. To the best of our knowledge, this is a very unusual site of occurrence for paragangliomas, and only two other comparable cases have been described.

Meningeal melanocytosis in a young patient--an autopsy diagnosis.

Primary diffuse leptomeningeal melanocytosis is a very rare form of brain tumor. We report on a rapidly fatal case in an 18-year-old man presenting with symptoms and imaging features suggestive for subarachnoid hemorrhage or meningitis. The laboratory findings and imaging examination were still confusing and the diagnosis remained unclear during the patient's life. Autopsy was the cornerstone in disclosing the lesion, confirming its usefulness in the assessment of such unusual cases. The complete profile of the tumor was obtained only by histology and immunohistochemistry. Clinicians and pathologists must be aware of diagnosis difficulties in this rare disease which can represent a serious challenge in clinical practice.

Comparison of a guaiac based and an immunochemical faecal occult blood test in screening for colorectal cancer in a general average risk population.

BACKGROUND: The guaiac faecal occult blood test (G-FOBT) is recommended as a screening test for colorectal cancer but its low sensitivity has prevented its use throughout the world. METHODS: We compared the performances of the reference G-FOBT (non-rehydrated Hemoccult II test) and the immunochemical faecal occult blood test (I-FOBT) using different positivity cut-off values in an average risk population sample of 10,673 patients who completed the two tests. Patients with at least one test positive were asked to undergo colonoscopy. RESULTS: Using the usual cut-off point of 20 ng/ml haemoglobin, the gain in sensitivity associated with the use of I-FOBT (50% increase for cancer and 256% increase for high risk adenoma) was balanced by a decrease in specificity. The number of extra false positive results associated with the detection of one extra advanced neoplasia (cancer or high risk adenoma) was 2.17 (95% confidence interval 1.65-2.85). With a threshold of 50 ng/ml, I-FOBT detected more than twice as many advanced neoplasias as the G-FOBT (ratio of sensitivity = 2.33) without any loss in specificity (ratio of false positive rate = 0.99). With a threshold of 75 ng/ml, associated with a similar positivity rate to G-FOBT (2.4%), the use of I-FOBT allowed a gain in sensitivity of 90% and a decrease in the false positive rate of 33% for advanced neoplasia. CONCLUSIONS: Evidence in favour of the substitution of G-FOBT by I-FOBT is increasing, the gain being more important for high risk adenomas than for cancers. The automated reading technology allows choice of the positivity rate associated with an ideal balance between sensitivity and specificity.

Vascular convolutes in the brain: a peculiar entity.

The present study intends to systematically assess a lesion with potential clinical expression - the vascular convolutes of the brain parenchyma. Those, firstly described by Gertz and Frydl in 1987, are frequently reported in various studies as being associated to aging. Their mechanism seems to be an arteriolar tortuosity. The association with pathologically important manifestations, as brain infarct or hemorrhage could be of clinical importance. The study was done on 70 consequently autopsied cases, on paraffin-embedded material, using the classical histological staining methods: Hematoxylin-Eosin and Masson's Trichrome, as well as the Gömöri silver method for reticulum. In our series, only two cases had specific features suggesting vascular convolutes. More frequently were encountered vascular tortuosities, which seems unrelated to the former. We conclude that vascular convolutes and vascular tortuosities are dissimilar lesions. Both are less represented in reality than reported in the literature. Possible mechanisms of the two changes and their pathogenic significance are discussed.

Skull base tumor invading both cavernous sinuses. Adenoid cystic carcinoma mimicking a meningioma.

We present a case of an anterior skull base tumor invading both cavernous sinuses and extending into the right orbit in a 55 years old female. The radiological aspect was confusing, being highly suggestive for an extensively invasive meningioma. However, the orbital portion of the tumor, which was surgically removed, proved to be an adenoid cystic carcinoma. Some peculiar immunohistochemical findings were obvious, as well as a lack of continuity of the tumor with the local lacrimal gland. This is an unusual situation, when a lacrimal gland tumor spread along the cavernous sinus, cross the midline and approaches the contralateral orbit. Such local extension should be considered in the differential diagnosis of anterior skull base tumors.