Molecular Pathology Research Papers - Academia.edu (original) (raw)

Techniques of molecular biology have improved diagnostic sensitivity, accuracy and validity in forensic medicine very much, especially in the field of identification (paternity testing, stain analysis). Since more than 10 years these techniques -meanwhile well established in clinical disciplines -are used also for other applications in forensic medicine: determination of cause and manner of death, tissue identification by mRNA and microRNA, examination of gene expression levels (survival time, time since death, cause of death), toxicogenetics.

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- Genetics, Algorithms, Forensic Science, Death

Hypersensitivity pneumonitis (HP) is a lung inflammatory disease caused by the inhalation of a variety of antigens. Previous studies support the role of the major histocompatibility complex (MHC) class II genes in the susceptibility to develop HP. However, the putative role of other MHC loci has not been elucidated. Transporters associated with antigen processing (TAP) genes are located within the MHC class II region and play an important role transporting peptides across the endoplasmic reticulum membrane for MHC class I molecules assembly. The distribution of single nucleotide polymorphisms (SNPs) in TAP1 genes was analyzed in 73 hypersensitivity pneumonitis (HP) patients and 58 normal subjects. We found a significant association of the allele Gly-637 GGC) ( p = 0.00004, OR = 27.30, CI = 3.87-548.04) and the genotypes Asp-637/Gly-637 ( p = 0.01, OR = 16.0, CI = 2.19-631.21), Pro-661/Pro-661 ( p = 0.006, OR = 11.30, CI = 2.28-75.77) with HP. A significant decrease in the frequency of the allele Pro-661 (CCA) ( p = 0.008, OR = 0.06, CI = 0-0.45), the genotype Asp-637/Asp-637 ( p = 0.01, OR = 0.17, 95% CI = 0.05-0.58) and the haplotype [Val-333 (GTC), Val-458 (GTG), Gly-637 (GGC), Pro-661 (CCA)] was detected in HP patients compared with controls ( p = 0.002, OR = 0.07, CI = 0.0-0.57). These findings suggest that TAP1 gene polymorphisms are related to HP risk, and highlight the importance of the MHC in the development of this disease.

Context.-Laboratories must validate all assays before they can be used to test patient specimens, but currently there are no evidence-based guidelines regarding validation of immunohistochemical assays. Objective.-To develop recommendations for initial analytic validation and revalidation of immunohistochemical assays. Design.-The College of American Pathologists Pathology and Laboratory Quality Center convened a panel of pathologists and histotechnologists with expertise in immunohistochemistry to develop validation recommendations. A systematic evidence review was conducted to address key questions. Electronic searches identified 1463 publications, of which 126 met inclusion criteria and were extracted. Individual publications were graded for quality, and the key question findings for strength of evidence. Recommendations were derived from strength of evidence, open comment feedback, and expert panel consensus. Results.-Fourteen guideline statements were established to help pathology laboratories comply with validation and revalidation requirements for immunohistochemical assays. Conclusions.-Laboratories must document successful analytic validation of all immunohistochemical tests before applying to patient specimens. The parameters for cases included in validation sets, including number, expression levels, fixative and processing methods, should take into account intended use and should be sufficient to ensure that the test accurately measures the analyte of interest in specimens tested in that laboratory. Recommendations are also provided for confirming assay performance when there are changes in test methods, reagents, or equipment.

- by Bruno Lomonte
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- Extracellular Matrix, Molecular Pathology, Adipose tissue, Basement Membrane

The Consortium for Blood Group Genes is a worldwide organization whose goal is to have a vehicle to interact, establish guidelines, operate a proficiency program, and provide education for laboratories involved in DNA and RNA testing for... more

- by Carlos De La Vega Elena and +2
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- Molecular Pathology, Program Development, Molecular plant pathology, United States

Phenylhydrazine (PHZ), a potent chemical causes toxicity on various tissues at various levels. Administration of phenylhydrazine mainly causes haematotoxicity which leads to the haemolytic anemia. In mammals PHZ induced anemia increased the iron absorption in spleen, liver and duodenum and finally iron metabolism was altered. Local demand and supply of Fe would increase erythropoeitic activity of the spleen so the size of spleen was increased that create the splenomegaly. PHZ induced anemia activate immune response which triggers phagocytosis in the spleen and liver. Apart from this administration of PHZ interfere the binding of erythropoietin (EPO) with erythropoietin receptors (EPOR) so that JAK-STAT would be affected.PHZ also showed genotoxic effect by creating single strand DNA damage.

- by Antonio Oliva
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- Genetics, Algorithms, Forensic Science, Death

T his article focuses on the challenges of generating comprehensive diagnostic reports in hematopathology. In particular, two main challenges that diagnosticians face are (1) interpreting and understanding the rapid advances in molecular and genetic pathology, which have gained increasing importance in classifications of hematopoietic neoplasms, and (2) managing the logistics of reporting ancillary studies and incorporating them effectively into a final synthesized report. This article summarizes many important genetic findings in hematopoietic neoplasms, which are required for accurate diagnoses, and discusses practical issues to generating accurate and complete hematopathology reports.

- by Robert Ohgami
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- Molecular Pathology

Object: Mutations of the gene encoding isocitrate dehydrogenase (IDH) have been shown in a significant proportion of diffuse gliomas. These mutations are specific to gliomas and their utility for diagnosis and prognostication of these tumors is being proclaimed. The present study was conducted with the aim of assessing frequency of IDH1 mutations in gliomas, their correlation with other molecular alterations along with a comprehensive review of available literature. Methods: A total of 100 gliomas of various grades and subtypes from Indian patients were screened for assessing frequency of IDH1 mutations. The findings were correlated with TP53 mutations, 1p/19q deletion, EGFR amplification and PTEN deletion status. The detailed comprehensive review of literature was performed comparing all studies available till date. Results: IDH1 mutations in codon 132 were observed in 46% cases. The frequency was 68.8% in grade II, 85.7% in grade III and 12.8% in GBMs. R132H mutation was most frequent (84.8%). Overall frequency of these mutations was relatively higher in oligodendroglial tumours as compared to astrocytic phenotype (66.7% versus 38.4%; p = 0.06). Primary GBMs showed IDH1 mutation in only 4.4% cases. In contrast, 66.7% of secondary GBMs harboured this alteration. Patients with IDH1 mutations were significantly younger as compared to those without mutation (p = 0.001). There was a significant correlation between IDH1 mutation and TP53 mutation (p = 0.004). Although IDH1 mutation showed a positive correlation with 1p/19q deletion, the association was not statistically significant (p = 0.653). There was no correlation with EGFR amplification or PTEN deletion. Conclusion: IDH1 mutations are present in large proportion of Indian patients with diffuse astrocytic and oligodendroglial neoplasms similar to the reported literature form west. The frequency is lower in primary GBMs and as compared to secondary GBMs. Association with younger age and positive correlation with TP53 mutation and 1p/19q loss is observed. More importantly it is emerging as an independent prognostic marker. Hence the greatest challenge now is establishing a reliable user friendly test for incorporating this novel genetic alteration to routine clinical practice.

- by vikash chand
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- Adolescent, Molecular Pathology, Child, Fluorescence in situ hybridization
- by Mary Sheppard
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- Molecular Biology, Cardiovascular, Biopsy, Molecular Pathology

The myelodysplastic syndromes (MDS) are a heterogeneous group of myeloid disorders characterised by impaired peripheral blood cell production due to bone marrow dysplasia affecting one or more of the major myeloid cell lines. MDS are one of Þ ve major categories of myeloid neoplasms according to the World Health Organization (WHO) classiÞ cation system for haematological cancers. Given their cytological and cytogenetic heterogeneity, these diseases probably constitute a group of molecularly distinct entities with variable degrees of ineffective haematopoiesis and susceptibility to leukaemic transformation. Recent studies provide some insights into the physiopathology of MDS. In the early stages, one mechanism contributing to hypercellular marrow and peripheral blood cytopenia is a signiÞ cant increase in programmed cell death (apoptosis) in haematopoietic cells. Furthermore, altered responses in relation to cytokines, the immune system and bone marrow stroma also contribute to the disease phenotype. Deletions of chromosome 5q31-q32 are the most common recurring cytogenetic abnormalities detected in MDS. The 5q-syndrome is a new entity recognised in the WHO classiÞ cation since 2001 and is associated with a good prognosis. Haplo-insufÞ ciency of multiple genes mapping to the common deleted region at 5q31-32 may contribute to the pathogenesis of 5q-syndrome and other MDS with 5q-deletion. Many studies have demonstrated that altered DNA methylation and histone acetylation can alter gene transcription. Abnormal methylation of transcription promoter sites is universal in patients with MDS, and the number of involved loci is increased in high-risk disease and secondary leukaemias. A better understanding of the pathogenesis of MDS can contribute to the development of new treatments such as hypomethylating drugs, immunomodulatory agents such as lenalidomide, and immunosuppressive drugs aimed at reversing the speciÞ c alteration that results in improvement in patients with MDS.

- by Marisa Calabuig
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- Nutrition and Dietetics, Molecular Pathology, Gene Mapping, Cell line

Bone tumors are considered by most pathologists difficult to diagnose as they are rare, have overlapping morphology, need radiological correlation, and the usefulness of immunohistochemistry is limited, making conventional morphology the cornerstone of the diagnosis. Over the past decade, more and more has become known of the molecular background of bone tumors. Three groups of bone tumors are recognized, namely, tumors with specific translocations combined with a relatively simple karyotype involving chromosomal translocations (Ewing sarcoma, aneurysmal bone cyst), tumors with specific gene mutations or amplifications (chondrosarcoma, fibrous dysplasia, chordoma), and sarcomas with genetic instability and as a consequence complex karyotypes (osteosarcoma). Technical advancements will rapidly reveal new alterations in the more rare sarcoma subtypes for which the molecular background has remained enigmatic. Opening the archives and using new technologies, as well as refinement of existing technologies for decalcified paraffin-embedded tissue, may bring to light more specific genetic aberrations in bone tumors that can be applied in molecular diagnostics in the near future.

- by Pancras C . W . Hogendoorn and +1
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- Molecular Diagnostics, Molecular Pathology, Cancer Genetics, Genetic Testing

The 2nd Milan Meeting on Adverse Reactions to Bovine Proteins was the venue for the presentation of the first consensus-based approach to the management of cow's milk allergy. It was also the first time that the Grading of Recommendations, Assessments, Development, and Evaluation approach for formulating guidelines and recommendations was applied to the field of food allergy. In this report we present the contributions in allergen science, epidemiology, natural history, evidence-based diagnosis, and therapy synthesized in the World Allergy Organization Diagnosis and Rationale for Action against Cow's Milk Allergy guidelines and presented during the meeting. A consensus emerged between discussants that cow's milk allergy management should reflect not only basic research but also a newer and better appraisal of the literature in the light of the values and preferences shared by patients and their caregivers in partnership. In the field of diagnosis, atopy patch testing and microarray technology have not yet evolved for use outside the research setting. With foreseeable breakthroughs (eg, immunotherapy and molecular diagnosis) in the offing, the step ahead in leadership can only stem from a worldwide organization implementing consensus-based clinical practice guidelines to diffuse and share clinical knowledge. (J Allergy Clin Immunol

- by Gabriel R Bouygue
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- Immunology, Molecular Pathology, Allergy, Immunotherapy

Triple negative breast cancer (TNBC) is a subtype of breast tumor which comprises 24% of newly diagnosed tumors that lacks hormone receptor expression and HER2 gene amplification. This study illustrated the pathological features of triple negative breast cancer with special reference to the landmark research that molecularly characterize this subtype of breast cancer. Additionally, this article discusses functional problems with arisen in clinical routine as a result of advent genetic expression breast cancer profiling and it's novel prognostic and predictive effects on triple-negative breast cancer pathology. Additionally, histopathological features of triple-negative neoplasms are discussed, emphasizing the critical nature of histologic detection in specific cancer subtypes with a significant effect on clinical results. Notably, emphasis is placed on the emerging clinical frontier represented by immunotherapy, with special emphasis on the implementation of immune checkpoint inhibitors in TNBC therapy and their effect on potential treatments.

Molecular organization of a cell is dynamically transformed along the course of cellular physiological processes, pathologic developments or derived from interactions with drugs. The capability to measure and monitor concentrations of macromolecules in a single cell would greatly enhance studies of cellular processes in heterogeneous populations. In this communication, we introduce and experimentally validate a bio-analytical single-cell assay, wherein the overall concentration of macromolecules is estimated in specific subcellular domains, such as structure-function compartments of the cell nucleus as well as in nucleoplasm. We describe quantitative mapping of local biomolecular concentrations, either intrinsic relating to the functional and physiological state of a cell, or altered by a therapeutic drug action, using two-photon excited fluorescence lifetime imaging (FLIM). The proposed assay utilizes a correlation between the fluorescence lifetime of fluorophore and the refractive index of its microenvironment varying due to changes in the concentrations of macromolecules, mainly proteins. Two-photon excitation in Near-Infra Red biological transparency window reduced the photo-toxicity in live cells, as compared with a conventional single-photon approach. Using this new assay, we estimated average concentrations of proteins in the compartments of nuclear speckles and in the nucleoplasm at ~150 mg/ml, and in the nucleolus at ~284 mg/ml. Furthermore, we show a profound influence of pharmaceutical inhibitors of RNA synthesis on intracellular protein density. The approach proposed here will significantly advance theranostics, and studies of drug-cell interactions at the single-cell level, aiding development of personal molecular medicine.

- by Artem Pliss
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- Optical Imaging, Molecular Pathology, Single cell analysis, Theranostics

The past decade has witnessed significant advances in the application of molecular diagnostics for the pre-operative risk-stratification of cytologically indeterminate thyroid nodules. The tests that are currently marketed in the United States for this purpose combine aspects of tumor genotyping with gene and/or microRNA expression profiling. This review compares the general methodology and clinical validation studies for the three tests currently offered in the United States: ThyroSeq v3, Afirma GSC and Xpression Atlas, and ThyGeNEXT/ThyraMIR.

- by Zubair Baloch
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- Medicine, Molecular Pathology

Background: Clinicians have long appreciated the distinct phenotype of systemic juvenile idiopathic arthritis (SJIA) compared to polyarticular juvenile idiopathic arthritis (POLY). We hypothesized that gene expression profiles of peripheral blood mononuclear cells (PBMC) from children with each disease would reveal distinct biological pathways when analyzed for significant associations with elevations in two markers of JIA activity, erythrocyte sedimentation rate (ESR) and number of affected joints (joint count, JC). Methods: PBMC RNA from SJIA and POLY patients was profiled by kinetic PCR to analyze expression of 181 genes, selected for relevance to immune response pathways. Pearson correlation and Student's t-test analyses were performed to identify transcripts significantly associated with clinical parameters (ESR and JC) in SJIA or POLY samples. These transcripts were used to find related biological pathways.

- by Sheng-Yung P Chang
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- Juvenile Idiopathic Arthritis, Molecular Pathology, Child, Joints

To assess the effect of red wine on atherosclerosis, New Zealand rabbits were given 1% cholesterol diet for 12 weeks and compared to animals that received the diet plus either red wine or nonalcoholic wine products (NAWP). Diet induced marked increases in total and LDL cholesterol; yet no significant changes in HDL and triglyceride concentrations occurred. In the control group, plaque area was 69 +-9% of the aortic surface, while in the wine and NAWP groups it was only 38 _+ 9 and 47 -+ 12%, respectively (P < 0.0001). The average intima/media thickness ratio was 0.60 __ 0.2 in control animals, 0.14 ~ 0.09 in the wine group, and 0.39 -+ 0.19 in the NAWP group (P < 0.0001). No significant differences were noted in LDL oxidizability among treatments. Thus, both red wine and NAWP can prevent plaque formation in hypercholesterolemic rabbits despite significant increases in LDL. We speculate that anti-platelet effect, blockade of expression of endothelial cell adhesion molecules, and/or NO stimulation by red wine flavonoids are possible explanations.

- by Carlos Serrano
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- Wine, Molecular Pathology, Lipids, New Zealand

Carcinogenic human papillomaviruses (HPVs) that cause cervical cancer preferentially infect basal, metaplastic squamous cells of the transformation zone. If infection persists, and a vegetative infection ensues, a premalignant lesion may develop with the potential to progress into an invasive squamous cell carcinoma. Papillomavirus prophylactic vaccines target the systemic immune system for induction of neutralizing antibodies that protect the basal cells against infection. Because the carcinogenic HPVs are susceptible to neutralization by antibodies for 9-48 h after reaching the basal cells, both low and high titered HPV type-specific antibodies induced by HPV L1 and L2-based vaccines are highly efficacious. The greatest burden of HPV-associated cancers occurs in poor areas of the world where women do not have access to routine gynecological care. The burden of HIV/AIDS in these same regions of the world has added to the burden of HPV-associated disease. There is an urgent need for a cost-effective, broad-spectrum HPV prophylactic vaccine in developing countries, which necessitates substantial cost subsidization of the virus-like particle (VLP) based vaccines licensed in industrialized countries or an alternative approach with second-generation vaccines that are specifically designed for delivery to women in resource-poor communities.

- by Kenneth Palmer and +1
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- Molecular Pathology, HUMAN PAPILLOMAVIRUS, Clinical Sciences

The renin-angiotensin system (RAS) conceived as a coordinated hormonal cascade plays an important role in controlling multiple functions in many organs and is much more complex than previously thought. The RAS has continued to expand, with the identification of new components, functions and subsystems. Angiotensin-converting enzyme (ACE) and its novel homolog angiotensin converting enzyme 2 (ACE2) are two key enzymes involved in the synthesis of bioactive components of the RAS. The main active peptides of the RAS include angiotensin II (Ang II), Ang III, Ang IV, and angiotensin-(1-7) [Ang-(1-7)] among which Ang II and Ang-(1-7) are much more important in health and disease. The axis formed by ACE2 represents an endogenous counter-regulatory pathway within the RAS, and its actions are opposite to those of the ACE axis. Conventionally the RAS has been considered to be important in the cardiovascular system, metabolism, cell growth and homeostasis. In recent years, a key role of ACE and ACE2 and their peptides has been recognized in the inflammatory process in conditions such as cardiac hypertrophy, pulmonary hypertension, glomerulonephritis, lung injury, sepsis, and acute pancreatitis. Investigations are ongoing to better understand the role of the RAS in inflammation. A comprehensive understanding of the RAS components in inflammation can provide new possibilities for therapeutic approaches against inflammatory diseases.

- by David Buttle
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- Chemistry, Medicine, Molecular Pathology, Clinical Sciences

Approximately 30 % of colorectal carcinomas develop via the serrated neoplasia pathway characterized by widespread DNA methylation and frequent BRAF mutation. Serrated polyps represent a heterogeneous group of polyps which are the precursor lesions to serrated pathway colorectal carcinomas. The histological classification of serrated polyps has evolved over the last two decades to distinguish three separate entities: hyperplastic polyp, sessile serrated adenoma (SSA), and traditional serrated adenoma (TSA). The malignant potential of SSAs and TSAs has been clearly demonstrated. SSAs are more challenging to detect by colonoscopy and are likely to account for some interval carcinomas of the proximal colon. Serrated polyposis syndrome is now widely recognized as conferring a high risk of colorectal carcinoma although its cause remains elusive. The current understanding of the actual malignant potential of each serrated polyp subtype is still limited due to the lack of large-scale prospective studies. Patient management guidelines have been recently updated although high-level evidence to support them is still required.

- by Vicki Whitehall
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- Gastroenterology, Histology, Molecular Pathology, Risk factors

Widespread use of DNA restriction fragment length polymorphism (RFLP) to differentiate strains of Mycobacterium tuberculosis to monitor the transmission of tuberculosis has been hampered by the need to culture this slow-growing organism and by the level of technical sophistication needed for RFLP typing. We have developed a simple method which allows simultaneous detection and typing of M. tuberculosis in clinical specimens and reduces the time between suspicion of the disease and typing from 1 or several months to 1 or 2 days. The method is based on polymorphism of the chromosomal DR locus, which contains a variable number of short direct repeats interspersed with nonrepetitive spacers. The method is referred to as spacer oligotyping or "spoligotyping" because it is based on strain-dependent hybridization patterns of in vitro-amplified DNA with multiple spacer oligonucleotides. Most of the clinical isolates tested showed unique hybridization patterns, whereas outbreak strains shared the same spoligotype. The types obtained from direct examination of clinical samples were identical to those obtained by using DNA from cultured M. tuberculosis. This novel preliminary study shows that the novel method may be a useful tool for rapid disclosure of linked outbreak cases in a community, in hospitals, or in other institutions and for monitoring of transmission of multidrugresistant M. tuberculosis. Unexpectedly, spoligotyping was found to differentiate M. bovis from M. tuberculosis, a distinction which is often difficult to make by traditional methods.

- by Leo Miranda
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- Molecular Pathology, Clinical Microbiology, Biological Sciences, Molecular

The molecular pathways which promote lung cancer cell features have been broadly explored, leading to significant improvement in prognostic and diagnostic strategies. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have dramatically altered the treatment approach for patients with metastatic non-small cell lung cancer (NSCLC). Latest investigations by using next-generation sequencing (NGS) have shown that other oncogenic driver mutations, believed mutually exclusive for decades, could coexist in EGFR-mutated NSCLC patients. However, the exact clinical and pathological role of concomitant genomic aberrations needs to be investigated. In this systematic review, we aimed to summarize the recent data on the oncogenic role of concurrent genomic alterations, by specifically evaluating the characteristics, the pathological significance, and their potential impact on the treatment approach.

- by Valentina Calò
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- Medicine, Molecular Pathology

Belina, D., Demissie, T., Ashenafi, H., Tadesse, A. (2015). Comparative pathological study of liver fluke infection in ruminants. Indian J. Vet. Pathol., 39(2): 113-120 A comparative study was conducted from October 2013 to June 2014 on 135 cattle, 175 sheep and 190 goats with a total of 500 ruminants to assess pathological changes on liver infected with fluke at ELFORA export abattoir in Bishoftu, Ethiopia. The animals were included in the study using systematic random sampling. The gross lesions included firm and enlarged livers with tense capsule, haemorrhagic spots, multi focal nodules and enlarged hepatic lymph nodes. The bile ducts were thickened and distended with adult fluke especially in chronic cases. Frequently observed histologic lesions were, hepatic portal fibrosis with large amount of fibrin in the portal area, hepatocytes degeneration, fatty changes and periportal necrosis. Fibrous connective tissue of various amount with fibroblasts and infiltration of mononuclear cells, in particular lymphocytes were common lesions in tracts migrated by parasites. Biliary cirrhosis with epithelial hyperplasia were also observed in cattle and sheep. Eosinophilic hepatitis, talengechtasis and hepatocytes degeneration and necrosis were evident in the parenchyma. Chronic lesions were less prominent in goats' and even no cirrhotic case examined in goat. Cytological examination showed large number of lymphocytes and few plasma cells and eosinophils. The findings of the present study indicated that gross, histopathological and cytological examination supports one another in characterization of liver lesions hence could be used together in diagnosis of ruminant fasciolosis. However, additional diagnostic tests would be added for better confirmation.

- by dinaol Belina and +1
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- Molecular Pathology

Prostatik adenokarsinomlar batı ülkelerinde son derece yüksek sıklıkta
görülmekte olup, özellikle ilk tanı anında prostat bezine sınırlı,
düşük dereceli prostat tümörlü hastaların aktif izleme alınması veya
radikal tedaviye gidilmesi arasında karar verme sürecinde klinisyene
yardımcı olabilecek standardize testlerin bulunmaması, hastalarda
radikal tedaviye daha sıklıkla başvurulmasına sebep olmakta,
bu da beraberinde ciddi morbidite ve maliyet yükü getirmektedir.
Prostat kanseri patogenezi alanında giderek daha da gelişmiş moleküler
tekniklerin uygulamaya geçmesi, prostat kanserinin tanısı,
tedavisi ve takibinde kullanılmaya aday bir çok yeni hedefin ortaya
çıkarılmasını sağlamıştır. Ancak bu gelişmelerin büyük kısmı
henüz klinik pratiğe girecek olgunluğa ulaşmamıştır. Ortaya çıkarılan
moleküler hedeflerin özellikle yeni prostat kanseri tanısı alan
hastalarda aktif izleme karar verme aşamasında etkili olabilecek
testlere uyarlanmasına ihtiyaç duyulmaktadır.
Bu yazıda, prostat kanser tümörigenezinde belirgin rol oynayan
birkaç temel genetik değişikliğin tanıtılması ve yakın zamanda ortaya
çıkarılan yeni moleküler hedeflerin, genetik değişikliklerin ve
yeni test tekniklerinin prostat kanserinin tedavisi ve tedavi sonrası
izleminde alabileceği rollerin klinikte çalışan ürolog ve patologlara
tanıtılması amaçlanmıştır.

- by Bora Gurel
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- Urology, Prostate Cancer, Molecular Pathology, Cancer Diagnosis

Greater numbers of individuals are living to older ages. A major concern at both individual and population levels is how to live these years at a high functional level. If we had physiological markers to identify those at risk for progressive functional decline and impeding death, therapies could be targeted towards these individuals to prevent adverse outcomes. Senescence is presently considered as the consequence of lifelong antigenic stress impinging upon the individual genetic background. We might consider inflammation markers as synthetic measures of lifelong attrition combined with genetic tendency to develop an inflammatory phenotype. Such biomarkers are the most powerful predictors of frailty and mortality in the elderly available today. The aim of this review is to translate results from the research on ageing into a practical view, suggesting new tools for the clinical approach to older people.

- by Daniela Monti
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- Genetics, Aging, Inflammation, Molecular Pathology

“Why do parasites harm their host?” is a recurrent question in evolutionary biology and ecology, and has several implications for the biomedical sciences, particularly public health and epidemiology. Contrasting the meaning(s) of the concept of “virulence” in molecular pathology and evolutionary ecology, we review different explanations proposed as to why, and under what conditions, parasites cause harm to their host: whereas the former uses molecular techniques and concepts to explain changes and the nature of virulence seen as a categorical trait, the latter conceptualizes virulence as a phenotypic quantitative trait (usually related to a reduction in the host’s fitness). After describing the biology of emerging influenza viruses we illustrate how the ecological and the molecular approaches provide distinct
(but incomplete) explanations of the 1918–19 influenza pandemic. We suggest that an evolutionary approach is necessary to understand the dynamics of disease transmission but that a broader understanding of virulence will ultimately benefit from articulating and integrating the ecological dynamics with cellular mechanisms
of virulence. Both ecological and functional perspectives on host-pathogens’ interactions are required to answer the opening question but also to devise appropriate health-care measures in order to prevent (and predict?) future influenza pandemics and other emerging threats. Finally, the difficult co-existence of distinct explanatory
frameworks reflects the fact that scientists can work on a same problem using various methodologies but it also highlights the enduring tension between two scientific styles of practice in biomedicine.

Triclabendazole (TCBZ) has been the drug of choice to treat liver fluke infections in livestock for > 20 years, due to its high activity against both adult and juvenile flukes. More recently, it has been used successfully to treat human cases of fascioliasis. Resistance to TCBZ first appeared in the field in Australia in the mid-1990s. Since then, resistance has been reported from a number of countries throughout Europe: Ireland, Scotland, Wales, Spain and The Netherlands. The heavy reliance on a single drug puts treatment strategies for fascioliasis at risk. Should resistance develop further, the prospect is an alarming one. This review will present an overview of progress in understanding the mechanism of resistance to TCBZ, examining possible changes in the target molecule, in drug influx/efflux mechanisms and in the metabolism of TCBZ by the fluke. The review will also consider ways to deal with resistance, covering drug-oriented options such as: the use of alternative drugs, drug combinations and the search for new compounds.

- by Alan Trudgett
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- Proteomics, Drug metabolism, Molecular modeling, Molecular Pathology

Myeloproliferative neoplasms (MPNs; formerly chronic myeloproliferative disorders) are a class of myeloid hematologic malignancies that represent a stem cell-derived expansion of 1 or more hematopoietic cell lineages. The current 2008 World Health Organization system recognizes 8 types of MPN: chronic myelogenous leukemia, chronic neutrophilic leukemia, polycythemia vera, primary myelofibrosis, essential thrombocythemia, chronic eosinophilic leukemia, mastocytosis, and myeloproliferative neoplasm, unclassifiable. This review summarizes the salient characteristics of the MPNs, with emphasis on recent developments in the molecular pathophysiology and therapeutic monitoring of these disorders. by guest on January 7, 2017 http://ajcp.oxfordjournals.org/ Downloaded from Am J Clin Pathol 2010;133:602-615 603 A B ❚Image 1❚ Chronic myelogenous leukemia, chronic phase. A, Bone marrow aspirate smear showing granulocytes in all stages of maturation, eosinophils, basophils, and a small megakaryocyte with decreased nuclear lobation (Wright-Giemsa, ×1,000). B, Bone marrow core biopsy sample showing hypercellular bone marrow for age with expanded myelopoiesis and small megakaryocytes with decreased nuclear lobation (H&E, ×400). ❚Table 1❚

- by ahmed Hassan
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- Genetics, Cancer, Signal Transduction, Molecular Pathology

A model to study microvascular proliferation, the Disc Angiogenesis System (DAS), consists of a synthetic foam disc implanted subcutaneously in experimental animals. After a period of growth, usually 7 to 21 days, the disc is removed. Planar sections are used to measure and characterize the growth. Microvessels grow centripetally into the disc, together with fibroblasts. Concentric growth zones have been defined by light and electron microscopy. Moderate growth occurs spontaneously and is accelerated by angiogenic stimulants placed in the center of the disc. Morphometric analyses have shown that vessel growth is directly proportional to total fibrovascular growth, so the former can be quantified by procedures measuring the latter. These include manual projection of sections and computer-assisted digital image analysis, which is recommended for routine use. The proliieration of endothelial and other cells is determined by incorporation of tritiated thymidine, using scintillation counting and autoradiography. Using the DAS, well-established angiogenie agents such as basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and prostaglandin E, were found to increase proliferation of endothelial cells (EC) and microvessels. Heparin augmented the effect of bFGF. When used by itself heparin increased angiogenesis but not EC proliferation, in keeping with in vitro observations indicating that it stimulates migration but not proliferation of EC. Locally applied hyperthermia and ionizing radiation decreased angiogenesis, even when applied after the angiogenic stimulus. Systemic prostaglandin synthetase inhibitors antagonized the angiogenic effects of bFGF and EGF, in accordance with a postulated role of prostaglandins in the transduction of proliferative signals in microvascular EC. The DAS is easy to assemble and implant in small animals, including mice, which tolerate it well. Hence multiple discs can be used for each time or dose point, which allows reproducible measurements of vascular growth and increases statistical accuracy. Another advantage of the system is the capability of discriminating between proliferation and migration of EC and tibroblasts. The DAS can be used to test putative agonists or antagonists of angiogenesis. More generally, the DAS provides a model of wound healing, either uncomplicated or complicated by intlammation, and of angiogenic responses to solid tumors. 8 wz Academic press, h.

- by LUIS FAJARDO
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- Wound Healing, Molecular Pathology, Mice, Clinical Sciences

Cardiac myxomas are rare benign and slowly proliferating neoplasms of uncertain histogenesis with heterogeneous histomorphology and variable and sometimes clinically quite malignant pathological manifestations. Majority of cardiac myxoma occur sporadically while a relatively small proportion of diagnosed cases develop as a part of Carney complex syndrome with established familial pattern of inheritance. Although histologically indistinguishable these two forms of cardiac myxoma exhibit distinct cytogenetic make-up and apparent pathological differences important for their clinical presentation and prognosis. Additional problem is presented with secondary lesions with more aggressive histology and significantly faster cell proliferation suggesting their successive malignant alteration. Surgical resection of cardiac myxoma is currently the only treatment of choice. However, to avoid potentially hazardous operating procedures and possible postoperative complications and to prevent recurrence of the neoplastic lesions it is necessary to develop alternative approaches and identify a possible drug targets for their successful pharmacological treatment. Due to the rarity of the disease, a small number of cases in one institution and lack of comprehensive experimental data particularly concerning the cases of metastatic dissemination and secondary lesions with malignant nature, a comprehensive multi-institutional approach is required for better understanding of their molecular pathology and illumination of key molecular, genetic as well as epigenetic markers and regulatory pathways responsible for their development. In this article we provide comprehensive pathohistological, molecular and cytogenetic overview of sporadic cardiac myxoma cases restating the major hypothesis concerning their histogenesis and emphasizing potential approaches for their further reexamination.

- by S. Ivčević and +1
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- Cardiology, Molecular Pathology, Differential Diagnosis, Epigenetic

The causes of non-trauma-mediated rhabdomyolysis are not well understood. It has been speculated that ethanol-associated rhabdomyolysis may be attributed to ethanol induction of skeletal muscle cytochrome P450(s), causing drugs such as acetaminophen or cocaine to be metabolized to myotoxic compounds. To examine this possibility, the hypothesis that feeding ethanol induces cytochrome P450 in skeletal muscle was tested. To this end, rats were fed an ethanol-containing diet and skeletal muscle tissue was assessed for induction of CYP2E1 and CYP1A1/2 by immunohistochemical procedures; liver was examined as a positive control tissue. Enzymatic assays and Western blot analyses were also performed on these tissues. In one feeding system, ethanol-containing diets induced CYP1A1/2 in soleus, plantaris, and diaphragm muscles, with immunohistochemical staining predominantly localized to capillaries surrounding myofibers. Antibodies to CYP2E1 did not react with skeletal muscle tissue from animals receiving a control or ethanol-containing diet. However, neither skeletal muscle CYP1A1/2 nor CYP2E1 was induced when ethanol diets were administered by a different feeding system. Ethanol consumption can induce some cytochrome P450 isoforms in skeletal muscle tissue; however, the mechanism of CYP induction is apparently complex and appears to involve factors in addition to ethanol, per se.

- by Gerry Hobbs and +1
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- Skeletal muscle biology, Diet, Immunohistochemistry, Molecular Pathology

Both Fig. 1b and the legend of Fig. 1 were incorrect.

- by R. Schafer
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- Information Systems, Technology, Microscopy, Fuzzy Logic

Friedreich ataxia (FRDA) is a devastating neurodegenerative disease caused by mutations in the frataxin gene (FXN). Frataxin is an essential protein which localizes to the mitochondria and is required for the synthesis of iron-sulfur clusters and heme. Most individuals with FRDA are homozygous for trinucleotide GAA.TTC repeat expansions in intron 1 of FXN. The instability of these GAA.TTC repeats, the formation of non-B DNA GAA.TTC structures, and accompanying epigenetic changes lead to reduced FXN transcript and frataxin protein. This 'loss of frataxin' is considered the main driver of disease pathology with mitochondria-rich tissues such as the heart and the brain most affected. While our understanding of FRDA etiology has advanced in recent years, exactly how reduced frataxin leads to disease remains largely unknown. Most therapeutic strategies aim to increase frataxin, yet there are other underlying aspects of the molecular pathology that could impact disease progression and severity. These include RNA toxicity due to antisense RNAs, dysregulated splicing and microRNAs, and repeat-associated protein toxicity via RAN translation. Here we review the diverse array of molecular events that have been shown to influence clinical outcome in FRDA. We also examine additional pathogenic factors from other trinu-cleotide repeat diseases which could be potentially important in FRDA. [Discovery Medicine 17(91):25-35,

Myeloproliferative neoplasms (MPNs; formerly chronic myeloproliferative disorders) are a class of myeloid hematologic malignancies that represent a stem cell-derived expansion of 1 or more hematopoietic cell lineages. The current 2008 World Health Organization system recognizes 8 types of MPN: chronic myelogenous leukemia, chronic neutrophilic leukemia, polycythemia vera, primary myelofibrosis, essential thrombocythemia, chronic eosinophilic leukemia, mastocytosis, and myeloproliferative neoplasm, unclassifiable. This review summarizes the salient characteristics of the MPNs, with emphasis on recent developments in the molecular pathophysiology and therapeutic monitoring of these disorders. by guest on September 10, 2016 http://ajcp.oxfordjournals.org/ Downloaded from Am J Clin Pathol 2010;133:602-615 603 A B ❚Image 1❚ Chronic myelogenous leukemia, chronic phase. A, Bone marrow aspirate smear showing granulocytes in all stages of maturation, eosinophils, basophils, and a small megakaryocyte with decreased nuclear lobation (Wright-Giemsa, ×1,000). B, Bone marrow core biopsy sample showing hypercellular bone marrow for age with expanded myelopoiesis and small megakaryocytes with decreased nuclear lobation (H&E, ×400). ❚Table 1❚

- by Ravi Vij
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- Genetics, Cancer, Signal Transduction, Molecular Pathology

Renal neoplasms exhibit a wide spectrum of molecular characteristics that are closely associated with their diverse morphologic manifestations and clinical behaviors. A wealth of information has been garnered via methodology ranging from classical cytogenetics to FISH and gene expression profiling; however, the exact mechanisms by which each type of renal tumor develops remain incompletely understood. Oddly, tumors with distinctly different morphology and prognosis sometimes show some overlap in the observed genetic abnormalities; by contrast, morphologically well characterized benign and malignant tumors that seem intuitively related cannot necessarily be shown to exhibit a step-wise progression from benign to malignant biologic behavior. Nevertheless, modern methodologies have been highly successful, not only in subclassifying renal tumors, but also in defining previously unrecognized entities. Further avenues of utility for these methods probably include the resolution of challenging differential diagnoses, as well as application in targeted therapy and prediction of outcome. We review the molecular and genetic characteristics of renal neoplasms, with emphasis on markers that demonstrate utility in differentiating morphologically similar neoplasms and in predicting clinical outcome.

- by Antonio Lopez-beltran
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- Molecular Genetics, Molecular Pathology, Renal cell Carcinoma, Expert

Chondroid tumours comprise a heterogeneous group of lesions with diverse morphological features and clinical behaviour. Over the past decade, a substantial number of chondroid tumours have been shown to harbour specific genetic abnormalities. These genetic alterations can be divided into two groups: tumours with specific translocations, and tumours with specific gene mutations. These genetic events give not only important insight in the tumourigenesis of chondroid tumours but offers new possibilities for molecular diagnosis. Some of these specific genetic changes have been implemented in routine pathology. This review aims to highlight the molecular genetics of chondroid tumours and their diagnostic applications.

- by Pancras C . W . Hogendoorn
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- Immunohistochemistry, Molecular Genetics, Molecular Pathology, PCR

Age-related macular degeneration (AMD) is a leading cause of irreversible blindness in the world. Although the etiology and pathogenesis of AMD remain largely unclear, a complex interaction of genetic and environmental factors is thought to exist. AMD pathology is characterized by degeneration involving the retinal photoreceptors, retinal pigment epithelium, and Bruch's membrane, as well as, in some cases, alterations in choroidal capillaries. Recent research on the genetic and molecular underpinnings of AMD brings to light several basic molecular pathways and pathophysiological processes that might mediate AMD risk, progression, and/or response to therapy. This review summarizes, in detail, the molecular pathological findings in both humans and animal models, including genetic variations in CFH, CX3CR1, and ARMS2/HtrA1, as well as the role of numerous molecules implicated in inflammation, apoptosis, cholesterol trafficking, angiogenesis, and oxidative stress.

- by Chi-Chao Chan
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- Genetics, Retinal Pigment Epithelium, Macular Degeneration, Molecular Pathology

The authors made a preliminary assessment of possible correlations between the intratumoral vascular density (IVD) and the architectural tumoral patterns described by Gleason. The studied material consisted of samples obtained by transurethral resection from 34 patients diagnosed with prostatic adenocarcinoma. Ten fields, five for dominant and five for secondary identified patterns of each case, with no necrosis were selected randomly from CD34 immunomarked sections using ×20 objective. IVD increased with Gleason pattern both for the entire group, but also for "solid" phenotype group of subtypes up to pattern 4, respectively subtype 4B. In "necrotizing" phenotype group of subtypes, IVD had a decreasing trend from the better-differentiated subtypes to the poorest one. These preliminary data showed that the intratumoral vascular network reacts differently to the loss of tumoral differentiation in the two groups of Gleason subtypes suggesting the existence of two different populations of malignant cells.

- by Iancu Emil Plesea
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- Pathology, Breast Cancer, Prostate Cancer, Cancer Research

Analysis of atypical meningococci from invasive disease that either (i) did not produce acid from maltose and glucose or (ii) were gamma-glutamyl peptidase test negative for porA and porB DNA variable region (VR) type, multilocus sequence type, and for presence of capsule transport gene ctrA, conclusively demonstrated that these are Neisseria meningitidis.

- by David Tribe
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- Genetics, Pathology, Biology, Medicine
- by F. Kalinec
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- Electron Microscopy, Biology, Transmission Electron Microscopy, Medicine

Nanoparticle-mediated drug delivery represents the future in terms of treating inner ear diseases. Lipid core nanocapsules (LNCs), 50 nm in size, were shown to pass though the round window membrane (RWM) and reached the spiral ganglion cells and nerve fibers, among other cell types in the inner ear. The present study aimed to evaluate the toxicity of the LNCs in vitro and in vivo, utilizing intact round window membrane delivery in rats. The primary cochlear cells and mouse fibroblast cells treated with LNCs displayed dosage dependant toxicity. In vivo study showed that administration of LNCs did not cause hearing loss, nanoparticle application-related cell death, or morphological changes in the inner ear, at up to 28 days of observation. The cochlear neural elements, such as synaptophysin, ribbon synapses, and S-100, were not affected by the administration of LNCs. However, expression of neurofilament-200 decreased in SGCs and in cochlear nerve in osseous spiral lamina canal after L...

- by Marian Löbler
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- Nanoparticle, Nanoparticles, Nanotechnology, Molecular Pathology

- by Pekka Saukko
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- Genetics, Algorithms, Forensic Science, Death
- by Samata Kakkad
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- Molecular Pathology, Diagnostic Imaging, Clinical Sciences, Neoplasms

A dysregulation in the 2V,5V-oligoadenylate (2-5A)-dependent RNase L antiviral pathway has been detected in peripheral blood mononuclear cells (PBMC) of chronic fatigue syndrome (CFS) patients, which is characterized by upregulated 2-5A synthetase and RNase L activities, as well as by the presence of a low molecular weight (LMW) 2-5A-binding protein of 37-kDa related to RNase L. This truncated protein has been shown to originate from proteolytic cleavage of the native 83-kDa RNase L by m-calpain and human leukocyte elastase (HLE). We investigated the possible role of 2-5A oligomers in the proteolytic action toward the endonuclease and show that incubation of CFS PBMC extracts with 2-5A trimer and tetramer, but not with the dimer, results in a significant protection of the native 83-kDa RNase L against cleavage by endogenous and purified proteases. Similar results are obtained with a purified recombinant RNase L. An analysis of the size of 2-5A oligomers produced by the catalytic activity of the 2-5A synthetase present in PBMC extracts further shows that samples containing the 37-kDa RNase L preferentially produce 2-5A dimers instead of higher oligomers. Taken together, our results indicate that homodimerization of RNase L by 2-5A oligomers higher than the dimer prevents its cleavage by proteolytic enzymes. The presence of the truncated 37-kDa RNase L in PBMC extracts is therefore likely to result, not only from the abnormal activation of inflammatory proteases, but also from a dysregulation in 2-5A synthetase induction or activation towards the preferential production of 2-5A dimers. D

Molecular tests that detect and/or quantify HCV RNA are important in the diagnosis and management of patients with chronic hepatitis C (CHC) undergoing anti-viral therapy. The primary goal of anti-HCV therapy is to achieve a sustained virologic response (SVR) defined as "undetectable" Hepatitis C Virus (HCV) RNA in the serum or plasma at 12 to 24 weeks following the end of treatment.

- by Regis Vilchez
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- Microbiology, Medical Microbiology, Treatment Outcome, Molecular Pathology

Alpha-fetoprotein (AFP)-producing gastric cancer (AFP-GC) is a highly malignant variant of adenocarcinoma with aberrant hepatocellular phenotype. A detailed understanding of the regulation of its liver phenotype is lacking. Liver-enriched nuclear factors (LENFs) are implicated in the transcriptional regulation of AFP in the fetal liver. To investigate the regulatory role of LENFs in AFP-GCs, the expression of LENFs including CCAAT/enhancer binding protein (C/EBP)-β, C/EBP-α, hepatocyte nuclear factor (HNF)-1α, HNF-1β and HNF-4α was investigated in 3 cell lines of AFP-GC and 7 cell lines of control GC. The liver activating protein (LAP), an activating isoform of C/EBP-β, was predominantly expressed in AFP-GCs, whereas the liver inhibitory protein (LIP), an inhibitory isoform of C/EBP-β, predominated in the control GCs. HNF-1α was relatively suppressed in AFP-GCs. HNF-4α was expressed in one of three AFP-GC cell lines. C/EBP-α and HNF-1β were expressed at the same levels in both cell types of GC. AFP-GCs expressed a set of hepatocyte-related proteins (e.g., transferrin and albumin) while they still retained the several glandular cell-related proteins (e.g., MUC2). The induction of LIP reduced transferrin expression and induced CEA expression in an AFP-GC line. Collecting these results, it was suggested that the contribution of LENFs, especially isoforms of C/EBP-β, is possibly important in phenotypic regulation of AFP-GCs.

- by yana supriatna
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- Transcription Regulation, Western blotting, Molecular Pathology, Cell line