Diagnostic reliability of nested PCR depends on the primer design and threshold abundance of Helicobacter pylori in biopsy, stool, and saliva samples.

BACKGROUND: The aim of this work was to find a reliable nested PCR for the detection of Helicobacter pylori in biopsy, stool, and saliva specimens. MATERIALS AND METHODS: Novel nested PCR was elaborated and validated on 81 clinical biopsy, stool, and saliva samples from the same individual and compared to available H pylori assays: histology, rapid urease test (RUT), stool antigen test (SAT), 13 C-urea breath test (UBT). RESULTS: The efficiency and selectivity of 17 published nested polymerase chain reactions (PCR) available for Helicobacter pylori detection were re-evaluated. Most of them had serious limitations and mistakes in primer design. Hence, we elaborated a nested PCR for the unambiguous identification of H pylori in biopsy, stool, and saliva, using primers targeted to variable regions of the 16S ribosomal RNA (rRNA) gene. Moreover, we determined the detection limit by adding a known number of cells. This number was as low as 0.5 cells in a PCR vial, but due to the DNA isolation procedures, it required 1-5 × 103 cells/g or ml of specimen. The sensitivity for nested PCR from stomach biopsies was on the same scale as 13 C-UBT (93.8%), but it was much lower in amplifications from stool (31.3%). Sequencing of all obtained PCR products exclusively confirmed H pylori-specific DNA sequences. CONCLUSIONS: Elaborated nested PCR assay can serve as an auxiliary method for controversial samples (patients with bleeding or taking proton-pump inhibitor) in laboratories with basic equipment. The sensitivity and specificity for the amplification from gastric biopsies was almost like 13 C-UBT. Despite the good sensitivity, the threshold occurrence and the ability to survive in the oral cavity aside from and independent of the stomach is the reason why H pylori DNA cannot be reliably detected in saliva, stool, and some biopsy samples.

[Mesenterointestinocolonovascular arterial ischemic disease: guidelines of the angiology section of Slovak Medical Chamber (AS SMC, 2013)]. / Mezentériointestinokolonovaskulárna artériová ischemická choroba Odporúcania Sekcie angiológov Slovenskej lekárskej komory (2013).

Organovascular arterial ischemic diseases (cardiovascular, cerebrovascular, extremitovascular, renovascular, genitovascular, pulmovascular, mesenterovascular, dermovascular, oculovascular, otovascular, stomatovascular etc.) are an important manifestations of systemic atherosclerosis and other arterial diseases of vascular system (arteriolosclerosis/arteriolonecrosis; diabetic macroangiopathy; diabetic microangiopathy; Mönckeberg´s mediosclerosis/mediocalcinosis; arteritis - vasculitis; syndromes of arterial compression; fibromuscular dysplasia; cystic adventitial degeneration; arterial thrombosis; arterial embolism/thromboembolism; traumatic and posttraumatic arteriopathies; physical arteriopathies; chemical and toxic arteriopathies; iatrogenic arterial occlusions; dissection of aorta and of arteries; coiling; kinking; complicated arterial aneurysms; arteriovenous fistula, rare vascular diseases). Key clinical-etiology-anatomy-pathophysiology (CEAP) aspects of the mesenteriovascular arterial ischemic diseases are discussed in this article (project Vessels).

Interference of Known or Suspected Endometriosis in Reporting FDG PET/CT Performed in Another Indication.

INTRODUCTION: Endometriosis is a common gynecologic condition that may be visualized on 18F-FDG PET/CT and mimic lesions of malignancy. We analyzed the interference of known or suspected endometriosis in reporting 18F-FDG PET/CT performed in another indication. RESULTS: The PET/CT images of 18 women with known (n = 15) or suspected (n = 3) endometriosis were analyzed. Based on clinical follow-up and results of other imaging, biopsy, and/or postsurgical histology, the presence of lesions of endometriosis at the time of 18F-FDG PET/CT was confirmed in 13 of 18 patients (72%). The per-patient positivity rate of 18F-FDG PET/CT was 8/18 (44%; 95% confidence interval, 22%-69%). The patient-based detection rate of 18F-FDG PET/CT in patients with confirmed lesions of endometriosis was 8/13 (62%; confidence interval, 32%-86%). On per-lesion/site basis, 18F-FDG PET/CT detected 11 of 20 sites (55%) of endometriosis. The SUVmax of these lesions/sites ranged between 1.8 and 5.3 (median, 3.8). In 9 of 18 patients (50%), a total of 13 non-endometriosis-related lesions/sites were detected by 18F-FDG PET/CT; their SUVmax ranged between 2.7 and 23 (median, 9.4). CONCLUSION: The interference of known or suspected endometriosis in reporting 18F-FDG PET/CT performed in another indication was limited but possible and should be kept in mind, even in postmenopausal women, as the oldest patient with 18F-FDG-positive endometriosis was aged 63 years. The lesions of endometriosis showed inconstant 18F-FDG uptake with overlap of SUVmax with low-grade malignancies. In our series, the greatest SUVmax value of lesion of endometriosis was 5.3, somewhat higher than the threshold of 4 previously proposed for identification of malignant transformation of endometriosis.

Case Report: Skeletal Muscle Lymphoma as a Result of Slow Centrifugal Migration of Untreated Primary Neurolymphomatosis?

Introduction: Fludeoxyglucose (18F) (FDG) hybrid positron emission tomography/computed tomography (PET/CT) is currently a well-documented tool for diagnosis, staging, and therapeutic follow-up of lymphoma with significant impact on therapeutic decisions. Patient Concerns and Interventions: We reported a case of a 71-year-old woman with diffuse large B-cell lymphoma (DLBCL) of the left gluteal muscles as a possible result of slow centrifugal migration of untreated neurolymphomatosis (NL) of the lumbosacral plexus suggested on FDG PET/CT 4 years ago, when the patient was complaining for weakness and numbness of the left leg, but the proposed biopsy of peripheral nerve was not performed. Four years later, no pathological FDG uptake was present in nerves and lymph nodes, but PET/CT detected multiple FDG-positive infiltrates in the left gluteal muscles, appearing as a continuation of previously involved nerves. Diagnosis: The biopsy of muscular infiltrates confirmed DLBCL. Outcomes: The therapy was started, and a complete remission was achieved after three lines of treatment. Conclusion: This case contributes to limited knowledge on development of skeletal muscle lymphoma (SML): It suggests the macroscopically isolated, FDG-positive SML involving more than one muscular compartment as a possible consequence of natural course of untreated primary NL previously revealed by peripheral neuropathy and suspected on FDG PET/CT. This observation further justifies the consideration of implementation of FDG PET/CT into diagnostic algorithm while evaluating the peripheral neuropathy, in which the NL, albeit rare, is a part of differential diagnosis.

Identification of Whole-Serum Glycobiomarkers for Colorectal Carcinoma Using Reverse-Phase Lectin Microarray.

Colorectal cancer (CRC) is one of the most common types of cancer among men and women worldwide. Efforts are currently underway to find novel and more cancer-specific biomarkers that could be detected in a non-invasive way. The analysis of aberrant glycosylation of serum glycoproteins is a way to discover novel diagnostic and prognostic CRC biomarkers. The present study investigated a whole-serum glycome with a panel of 16 different lectins in search for age-independent and CRC-specific glycomarkers using receiver operating characteristic (ROC) curve analyses and glycan heat matrices. Glycosylation changes present in the whole serum were identified, which could lead to the discovery of novel biomarkers for CRC diagnostics. In particular, the change in the bisecting glycans (recognized by Phaseolus vulgaris erythroagglutinin) had the highest discrimination potential for CRC diagnostics in combination with human L selectin providing area under the ROC curve (AUC) of 0.989 (95% CI 0.950-1.000), specificity of 1.000, sensitivity of 0.900, and accuracy of 0.960. We also implemented novel tools for identification of lectins with strong discrimination power.

Novel MLH1 and MSH2 germline mutations in the first HNPCC families identified in Slovakia.

Hereditary nonpolyposis colorectal cancer (HNPCC) is a dominantly-inherited cancer predisposition syndrome, in which the susceptibility to cancer of the colon, endometrium and ovary is linked to germline mutations in DNA mismatch repair (MMR) genes. We have recently initiated a cancer prevention program in suspected HNPCC families in the Slovak Republic. The first ten families fulfilling Amsterdam criteria or Bethesda guidelines were screened for germline mutations in MLH1 and MSH2, two MMR genes most frequently mutated in HNPCC families. Six mutations were identified, five of which have not been reported previously. Two of the three new mutations in MLH1 (c.380+2T>A; c.307-2A>C) were absent from 100 chromosomes of healthy controls and probably cause a splicing defect, while the third was a 1 bp deletion (c.1261delA). In the MSH2 gene, one new nonsense (c.1030C>T [p.Q344X]) and one missense (c.524T>C [p.L175P]) mutation were identified. This latter variant was not found in 104 alleles of healthy control individuals. Moreover, a previously-reported pathogenic mutation (c.677G>T [p.R226L]) was found in one kindred. The clinical data and the genotypic and phenotypic evaluation of the tumors indicate that all the new alterations are pathogenic HNPCC mutations.

Collagen type IV in epithelial tumours of colon.

Collagen type IV in the lamina propria mucosae is one of the main components of the basement membrane of normal and transitional colon mucosa. The aim of the present study was to assess the use of anti-collagen type IV antibodies in the evaluation of biological activity of epithelial tumours of the colon. Formalin-fixed and paraffin-embedded specimens of polyps and carcinomas of the colon from 14 patients were analyzed. In transitional mucosa around epithelial tumours, only minor deformities of the evenly thick collagen type IV-containing basement membranes were found. This pattern was different in polyps where collagen type IV-positive basement membrane components extended between basolateral membranes of epithelial cells. Local changes of collagen type IV positivity in basement membranes of polyps were observed. Positivity of epithelial basement membranes disappeared in adenocarcinomas but there was an increased positivity in fibrillar components of stroma. Basement membranes of microvessels in lamina propria mucosae were also positive for collagen type IV. Similar observations were made in the stroma of polyps. Our results indicate that loss of collagen type IV in basement membranes of adenocarcinomas is related to loss of differentiation and the malignant potential of epithelial tumours of colon.