Shi, Shan-Rong; Taylor, Clive R; Fowler, Carol B; Mason, Jeffrey T
Tissue-based proteomic approaches (tissue proteomics) are essential for discovering and evaluating biomarkers for personalized medicine. In any proteomics study, the most critical issue is sample extraction and preparation. This problem is especially difficult when recovering proteins from formalin-fixed, paraffin-embedded (FFPE) tissue sections. However, improving and standardizing protein extraction from FFPE tissue is a critical need because of the millions of archival FFPE tissues available in tissue banks worldwide. Recent progress in the application of heat-induced antigen retrieval principles for protein extraction from FFPE tissue has resulted in a number of published FFPE tissue proteomics studies. However, there is currently no consensus on the optimal protocol for protein extraction from FFPE tissue or accepted standards for quantitative evaluation of the extracts. Standardization is critical to ensure the accurate evaluation of FFPE protein extracts by proteomic methods such as reverse phase protein arrays, which is now in clinical use. In our view, complete solubilization of FFPE tissue samples is the best way to achieve the goal of standardizing the recovery of proteins from FFPE tissues. However, further studies are recommended to develop standardized protein extraction methods to ensure quantitative and qualitative reproducibility in the recovery of proteins from FFPE tissues. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Viegas, M S; Martins, T C; Seco, F; do Carmo, A
Interference by autofluorescence is one of the major shortcomes of immunofluorescence analysis by confocal laser scanning microscopy (CLSM). CLSM requires minimal tissue autofluorescence and reduced unspecific fluorescence background, requisites that become more critical when direct immunofluorescence studies are concerned. To control autofluorescence, different reagents and treatments can be used. Until now, the efficacy of the processes described depended on the tissue type and on the processing technique, no general recipe for the control of autofluorescence being available. Using paraffin sections of archival formalin-fixed murine liver, kidney and pancreas, we have found that previously described techniques were not able to reduce autofluorescence to levels that allowed direct immunofluorescence labelling. In this work, we aimed at improving currently described methodologies so that they would allow reduction of the autofluorescent background without affecting tissue integrity or direct immunofluorescence labelling. We have found that the combination of short-duration, high-intensity UV irradiation and Sudan Black B was the best approach to reduce autofluorescence in highly vascularised, high lipofuscins' content tissues, such as murine liver and kidney, and poorly vascularised, low lipofuscins' content tissues such as the pancreas. In addition, we herein show that this methodology is highly effective in reducing autofluorescent background to levels that allow detection of specific signals by direct immunofluorescence.
Lundemose, A G; Banner, Jytte; Birkelund, Svend
A procedure to detect Chlamydia in postmortal formalin-fixed tissue is described. Monoclonal antibodies against a genus specific chlamydia epitope were used in immunofluorescence to detect chlamydia inclusions in formalin-fixed tissue sections. Lung sections from chlamydia-infected mice were exam...
Dowd, S E; Gerba, C P; Enriquez, F J; Pepper, I L
The term microsporidia is used to describe several species of opportunistic protozoan parasites. Encephalitozoon intestinalis and Enterocytozoon bieneusi have been found in stools of more than 40% of AIDS patients with diarrhea. Diagnosis of infection with these small protozoans has been difficult, and until recently their occurrence has not been well documented. Formalin is widely used to preserve clinical specimens, but due to the nature of the fixation process, subsequent analysis, especially analysis by the PCR, is difficult. This study evaluated methods used to prepare formalin-fixed fecal specimens for PCR amplification of microsporidial DNA. Two methods were devised to allow PCR detection and subsequent identification of microsporidia in formalin-fixed fecal specimens to the species level. One method involved immunomagnetic separation to concentrate microsporidial spores from fecal specimens. In the second method Chelex resin (Bio-Rad, Hercules, Calif.) was used to remove inhibitory substances, followed by a DNA concentration step. Both methods resulted in reproducible, confirmed detection of microsporidia in formalinized fecal specimens and subsequent species determination by PCR sequencing. The detection sensitivity was two in vitro culture-derived spores (Encephalitozoon intestinalis) for the direct PCR. The reproducible detection sensitivity for DNA amplification from formalin-fixed fecal samples was 200 spores for either the Chelex method or the immunomagnetic bead separation method. Thus, we developed two methods for rapid, inexpensive detection of microsporidial spores in formalin-fixed fecal specimens.
Hormone Receptor Expression Analyses in Neoplastic and Non-Neoplastic Canine Mammary Tissue by a Bead Based Multiplex Branched DNA Assay: A Gene Expression Study in Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Samples.
Full Text Available Immunohistochemistry (IHC is currently considered the method of choice for steroid hormone receptor status evaluation in human breast cancer and, therefore, it is commonly utilized for assessing canine mammary tumors. In case of low hormone receptor expression, IHC is limited and thus is complemented by molecular analyses. In the present study, a multiplex bDNA assay was evaluated as a method for hormone receptor gene expression detection in canine mammary tissues. Estrogen receptor (ESR1, progesterone receptor (PGR, prolactin receptor (PRLR and growth hormone receptor (GHR gene expressions were evaluated in neoplastic and non-neoplastic canine mammary tissues. A set of 119 fresh frozen and 180 formalin-fixed, paraffin-embedded (FFPE was comparatively analyzed and used for assay evaluation. Furthermore, a possible association between the hormone receptor expression in different histological subtypes of canine malignant mammary tumors and the castration status, breed and invasive growth of the tumor were analyzed. The multiplex bDNA assay proved to be more sensitive for fresh frozen specimens. Hormone receptor expression found was significantly decreased in malignant mammary tumors in comparison to non-neoplastic tissue and benign mammary tumors. Among the histological subtypes the lowest gene expression levels of ESR1, PGR and PRLR were found in solid, anaplastic and ductal carcinomas. In summary, the evaluation showed that the measurement of hormone receptors with the multiplex bDNA assay represents a practicable method for obtaining detailed quantitative information about gene expression in canine mammary tissue for future studies. Still, comparison with IHC or quantitative real-time PCR is needed for further validation of the present method.
Tardío, Juan C; Cambero, Olivia; Sánchez-Estévez, Carolina; Sánchez-García, Ana B; Angulo, Fernando; Moreno, Amalia
Roche cobas 4800 human papillomavirus (HPV) test is an automated real-time polymerase chain reaction-based system that allows the simultaneous detection of 14 human papillomavirus high-risk (HR-HPV) genotypes. This test is Food and Drug Administration approved since 2011 for HPV determination in liquid-based cytologic samples, but a clinically validated technique for formalin-fixed, paraffin-embedded (FFPE) tissue specimens is presently not commercially available. In our laboratory, we have developed an HPV detection procedure in FFPE tissue by cobas 4800 HPV test. In order to validate our method, we retrospectively studied 165 FFPE cervical biopsy and conization specimens with varied diagnoses from our files. In 50 of them, we contrasted the results with those obtained from simultaneous liquid-based cytologies from the same patients. Finally, seeking the possible complementary clinical usefulness of the procedure, we compared the HPV genotypes detected in cervical intraepithelial neoplasia grade 1 (CIN1)-diagnosed biopsies from 20 patients with a subsequent high-grade CIN (CIN2+) diagnosis with those from another group of 20 patients without a posterior CIN2+ diagnosis. Eighty-seven percent of the assays provided informative results. HR-HPV was detected in 28 of 32 (88%) invasive cervical squamous carcinomas. Coincidental HR-HPV genotypes were obtained in 32 of 50 (64%) cases with simultaneous cervical biopsy and liquid-based cytologic samples. A significant higher risk of progression to CIN2+ was found when HPV16 (P=0.022) or any HR-HPV genotype (P=0.037) was detected in CIN1 biopsies. The reported procedure provides an automated, technically time-saving, easy to integrate into laboratory routine, and reliable method of HR-HPV determination in FFPE specimens.
Lundemose, A G; Banner, Jytte; Birkelund, Svend
A procedure to detect Chlamydia in postmortal formalin-fixed tissue is described. Monoclonal antibodies against a genus specific chlamydia epitope were used in immunofluorescence to detect chlamydia inclusions in formalin-fixed tissue sections. Lung sections from chlamydia-infected mice were....... Background and non-specific fluorescence were reduced by treating the tissue sections with trypsin, rabbit serum and Evans blue counterstain. Besides giving an exact diagnosis at autopsy, the method provides the possibility of determining the occurrence of chlamydia infections in various tissues, based...
Davis, A. Sally; Richter, Anke; Becker, Steven; Moyer, Jenna E.; Sandouk, Aline; Skinner, Jeff
Tissue autofluorescence frequently hampers visualization of immunofluorescent markers in formalin-fixed paraffin-embedded respiratory tissues. We assessed nine treatments reported to have efficacy in reducing autofluorescence in other tissue types. The three most efficacious were Eriochrome black T, Sudan black B and sodium borohydride, as measured using white light laser confocal Λ2 (multi-lambda) analysis. We also assessed the impact of steam antigen retrieval and serum application on human tracheal tissue autofluorescence. Functionally fitting this Λ2 data to 2-dimensional Gaussian surfaces revealed that steam antigen retrieval and serum application contribute minimally to autofluorescence and that the three treatments are disparately efficacious. Together, these studies provide a set of guidelines for diminishing autofluorescence in formalin-fixed paraffin-embedded human respiratory tissue. Additionally, these characterization techniques are transferable to similar questions in other tissue types, as demonstrated on frozen human liver tissue and paraffin-embedded mouse lung tissue fixed in different fixatives. PMID:24722432
Dorj, Gantsetseg; Okada, Hiroyuki; Miyazawa, Kohtaro; Masujin, Kentaro; Kimura, Kumiko; Mohri, Shirou; Yokoyama, Takashi
An abnormal isoform of prion protein (PrP(Sc)) was extracted from formalin-fixed paraffin-embedded (FFPE) tissues from sheep and analyzed by western blotting. PrP(Sc) immunoreactivity against anti-PrP monoclonal antibody T2, which recognizes discontinuous PrP sequences, differed amongst individual scrapie sheep cases. This may reflect structural differences in PrP(Sc) that have been formalin-fixed prior to their extraction. This study indicates that western blotting by using FFPE tissues is useful for the retrospective analysis of transmissible spongiform encephalopathies in which only formalin-fixed samples are available and in conducting transmissible spongiform encephalopathies surveillance where freezing system is insufficient.
Sørensen, Margrethe; Redsted Rasmussen, Arne; Simonsen, Kim Pilkjær
Abstract.—A simple enzymatic screening method has been developed to detect whether a tissue sample has been preserved with formalin or with ethanol only because such a method is a useful tool for predicting the quality of genetic test results. The method is based on enzymatic digestion at 55 C...... in museums where DNA analyses of archived specimens are performed. Wasted time and resources can be avoided through the detection of formalin-fixed specimens because these specimens yield low-quality, damaged DNA. In addition to the screening method, it is shown that formalin-preserved specimens can...
Rostami, Sima; Torbaghan, Shams Shariat; Dabiri, Shahriar; Babaei, Zahra; Mohammadi, Mohammad Ali; Sharbatkhori, Mitra; Harandi, Majid Fasihi
Cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus, presents an important medical and veterinary problem globally, including that in Iran. Different genotypes of E. granulosus have been reported from human isolates worldwide. This study identifies the genotype of the parasite responsible for human hydatidosis in three provinces of Iran using formalin-fixed paraffin-embedded tissue samples. In this study, 200 formalin-fixed paraffin-embedded tissue samples from human CE cases were collected from Alborz, Tehran, and Kerman provinces. Polymerase chain reaction amplification and sequencing of the partial mitochondrial cytochrome c oxidase subunit 1 gene were performed for genetic characterization of the samples. Phylogenetic analysis of the isolates from this study and reference sequences of different genotypes was done using a maximum likelihood method. In total, 54.4%, 0.8%, 1%, and 40.8% of the samples were identified as the G1, G2, G3, and G6 genotypes, respectively. The findings of the current study confirm the G1 genotype (sheep strain) to be the most prevalent genotype involved in human CE cases in Iran and indicates the high prevalence of the G6 genotype with a high infectivity for humans. Furthermore, this study illustrates the first documented human CE case in Iran infected with the G2 genotype. PMID:25535316
Full Text Available One of the major breakthroughs in molecular pathology during the last decade was the successful extraction of full-length proteins from formalin-fixed and paraffin-embedded (FFPE clinical tissues. However, only limited data are available for the protein extraction efficiency of over-fixed tissues and FFPE blocks that had been stored for more than 15 years in pathology archives. In this study we evaluated the protein extraction efficiency of FFPE tissues which had been formalin-fixed for up to 144 hours and tissue blocks that were stored for 20 years, comparing an established and a new commercial buffer system. Although there is a decrease in protein yield with increasing fixation time, the new buffer system allows a protein recovery of 66% from 144 hours fixed tissues compared to tissues that were fixed for 6 hours. Using the established extraction procedure, less than 50% protein recovery was seen. Similarly, the protein extraction efficiency decreases with longer storage times of the paraffin blocks. Comparing the two buffer systems, we found that 50% more proteins can be extracted from FFPE blocks that were stored for 20 years when the new buffer system is used. Taken together, our data show that the new buffer system is superior compared to the established one. Because tissue fixation times vary in the routine clinical setting and pathology archives contain billions of FFPE tissues blocks, our data are highly relevant for research, diagnosis, and treatment of disease.
Nicholson Eric M
Full Text Available Abstract Background Formalin-fixed paraffin-embedded tissue is regularly employed in the diagnosis of transmissible spongiform encephalopathies (TSE by immunohistochemistry (IHC, the standard by which all other TSE diagnostic protocols are judged. While IHC affords advantages over diagnostic approaches that typically utilize fresh or frozen tissue, such as Western blot and ELISA, the process of fixing, staining, and analyzing individual sections by hand does not allow for rapid or high throughput screening. However, preservation of tissues in formalin is not dependent upon the availability of refrigeration. Findings Formalin-fixed paraffin-embedded tissues from TSE transmission studies of scrapie in sheep, chronic wasting disease in white-tailed deer or transmissible mink encephalopathy in cattle were cut at 5 μm thickness. Samples containing the tissue equivalent of as little as one 5 μm section can be used to readily discriminate positive from negative samples. Conclusions This approach cannot replace IHC but may be used along with IHC as both a more rapid and readily high throughput screen where fresh or frozen tissues are not available or impractical.
Li, Zhigang; Breitwieser, Florian P; Lu, Jennifer; Jun, Albert S; Asnaghi, Laura; Salzberg, Steven L; Eberhart, Charles G
We test the ability of next-generation sequencing, combined with computational analysis, to identify a range of organisms causing infectious keratitis. This retrospective study evaluated 16 cases of infectious keratitis and four control corneas in formalin-fixed tissues from the pathology laboratory. Infectious cases also were analyzed in the microbiology laboratory using culture, polymerase chain reaction, and direct staining. Classified sequence reads were analyzed with two different metagenomics classification engines, Kraken and Centrifuge, and visualized using the Pavian software tool. Sequencing generated 20 to 46 million reads per sample. On average, 96% of the reads were classified as human, 0.3% corresponded to known vectors or contaminant sequences, 1.7% represented microbial sequences, and 2.4% could not be classified. The two computational strategies successfully identified the fungal, bacterial, and amoebal pathogens in most patients, including all four bacterial and mycobacterial cases, five of six fungal cases, three of three Acanthamoeba cases, and one of three herpetic keratitis cases. In several cases, additional potential pathogens also were identified. In one case with cytomegalovirus identified by Kraken and Centrifuge, the virus was confirmed by direct testing, while two where Staphylococcus aureus or cytomegalovirus were identified by Centrifuge but not Kraken could not be confirmed. Confirmation was not attempted for an additional three potential pathogens identified by Kraken and 11 identified by Centrifuge. Next generation sequencing combined with computational analysis can identify a wide range of pathogens in formalin-fixed corneal specimens, with potential applications in clinical diagnostics and research.
Nikolaou, Panagiota; Papoutsis, Ioannis; Dona, Artemisia; Spiliopoulou, Chara; Athanaselis, Sotiris
During the autopsy of forensic cases, when there is no suspicion of drug use or chemical exposure, biological fluids may not be obtained for toxicological analysis, while specimens of tissues may be collected and preserved in a formalin solution for histological examination. When specific questions arise after the burial, the only possible options are the exhumation of an embalmed body or the toxicological analysis of the formalin-fixed specimens. The drug concentrations in these specimens can be altered due to the extraction efficiency and/or the chemical activity of the formalin solutions used during chemical fixation or embalming process. The aim of this paper is to review the published studies about the determination of specific groups of drugs in formalin-fixed or embalmed specimens and their stability after chemical fixation or embalming process. The analytical aspects of this determination are also discussed. The stability of drugs in formalin environment and the possible reaction of the drugs with formaldehyde, which is a highly reactive chemical substance, should always be considered during post-mortem/post-embalming forensic analysis. The additional analysis of the formalin solution in which the tissue was preserved is considered necessary. The identification and the evaluation of the possible degradation products or chemical derivatives are extremely useful during the interpretation of the results. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.
Laub Petersen, Bodil; Zeuthen, Mette Christa; Pedersen, Sanni
, such as quantitation of signals as in triploidy, it is possible to isolate nuclei from paraffin-embedded tissue. However, using formalin-fixed paraffin-embedded tissue, either in thin sections or as isolated nuclei, one encounters a range of technical problems, paralleling those met in immunohistochemistry. Variations...... nuclei and tissue sections from formalin-fixed, paraffin-embedded tissue....
Warf, M Bryan; Flake, Darl D; Adams, Doug; Gutin, Alexander; Kolquist, Kathryn A; Wenstrup, Richard J; Roa, Benjamin B
These studies were to validate the analytical performance of a gene expression signature that differentiates melanoma and nevi, using RNA expression from 14 signature genes and nine normalization genes that generates a melanoma diagnostic score (MDS). Formalin-fixed paraffin-embedded melanocytic lesions were evaluated in these studies. The overall SD of the assay was determined to be 0.69 MDS units. Individual amplicons within the signature had an average amplification efficiency of 92% and a SD less than 0.5 CT. The MDS was reproducible across a 2000-fold dilution range of input RNA. Melanin, an inhibitor of PCR, does not interfere with the signature. These studies indicate this signature is robust and reproducible and is analytically validated on formalin-fixed paraffin-embedded melanocytic lesions.
Nicholson, Eric M
Diagnosis of prion disease is primarily through immunodetection of the infectious agent. Typically, 2 distinct procedures are recommended for a definitive diagnosis, with immunohistochemistry and Western blot providing the most information as to the specific isolate in question. In the past, these approaches required formalin-fixed, paraffin-embedded tissue and fresh or frozen tissue, respectively; however, methods have been developed that allow for use of fixed tissue for Western blot. The present study describes a method of enriching PrP(Sc) in formalin-fixed, paraffin-embedded tissues prior to Western blot analysis for the detection of PrP(Sc). With this modified procedure, 5 times the previously reported sample size may be used for analysis, greatly enhancing the sensitivity of this procedure.
Fowler, Carol B.; Chesnick, Ingrid E.; Moore, Cedric D.; O'Leary, Timothy J.; Mason, Jeffrey T.
Background Proteomic studies of formalin-fixed paraffin-embedded (FFPE) tissues are frustrated by the inability to extract proteins from archival tissue in a form suitable for analysis by 2-D gel electrophoresis or mass spectrometry. This inability arises from the difficulty of reversing formaldehyde-induced protein adducts and cross-links within FFPE tissues. We previously reported the use of elevated hydrostatic pressure as a method for efficient protein recovery from a hen egg-white lysozy...
Anand, Suresh; Cicchi, Riccardo; Giordano, Flavio; Buccoliero, Anna Maria; Pavone, Francesco Saverio
In the recent years, there has been a considerable surge in the application of spectroscopy for disease diagnosis. Raman and fluorescence spectra provide characteristic spectral profile related to biochemical and morphological changes when tissues progress from normal state towards malignancy. Spectroscopic techniques offer the advantage of being minimally invasive compared to traditional histopathology, real time and quantitative. In biomedical optical diagnostics, freshly excised specimens are preferred for making ex-vivo spectroscopic measurements. With regard to fresh tissues, if the lab is located far away from the clinic it could pose a problem as spectral measurements have to be performed immediately after dissection. Tissue samples are usually placed in a fixative agent such as 4% formaldehyde to preserve the samples before processing them for routine histopathological studies. Fixation prevents the tissues from decomposition by arresting autolysis. In the present study, we intend to investigate the possibility of using formalin fixed samples for discrimination of brain tumours from dysplastic tissue using Raman and fluorescence spectroscopy. Formalin fixed samples were washed with phosphate buffered saline for about 5 minutes in order to remove the effects of formalin during spectroscopic measurements. In case of fluorescence spectroscopy, changes in spectral profile have been observed in the region between 550-670 nm between dysplastic and tumor samples. For Raman measurements, we found significant differences in the spectral profiles between dysplasia and tumor. In conclusion, formalin fixed samples can be potentially used for the spectroscopic discrimination of tumor against dysplastic tissue in brain samples.
Wojakowska, Anna; Marczak, Łukasz; Jelonek, Karol; Polanski, Krzysztof; Widlak, Piotr; Pietrowska, Monika
Formalin-fixed paraffin-embedded (FFPE) tissue specimens constitute a highly valuable source of clinical material for retrospective molecular studies. However, metabolomic assessment of such archival material remains still in its infancy. Hence, there is an urgent need for efficient methods enabling extraction and profiling of metabolites present in FFPE tissue specimens. Here we demonstrate the methodology for isolation of primary metabolites from archival tissues; either fresh-frozen, formalin-fixed or formalin-fixed and paraffin-embedded specimens of mouse kidney were analysed and compared in this work. We used gas chromatography followed by mass spectrometry (GC/MS approach) to identify about 80 metabolites (including amino acids, saccharides, carboxylic acids, fatty acids) present in such archive material. Importantly, about 75% of identified compounds were detected in all three types of specimens. Moreover, we observed that fixation with formalin itself (and their duration) did not affect markedly the presence of particular metabolites in tissue-extracted material, yet fixation for 24h could be recommended as a practical standard. Paraffin embedding influenced efficiency of extraction, which resulted in reduced quantities of several compounds. Nevertheless, we proved applicability of FFPE specimens for non-targeted GS/MS-based profiling of tissue metabolome, which is of great importance for feasibility of metabolomics studies using retrospective clinical material.
P. D. Luka
Full Text Available Aim: Formalin fixing and paraffin embedding of tissue samples is one of the techniques for preserving the structural integrity of cells for a very long time. However, extraction and analysis of genomic material from formalin fixed tissue (FFT remains a challenge despite numerous attempts to develop a more effective method. The success of polymerase chain reaction (PCR depends on the quality of DNA extract. Materials and Methods: Here we assessed the conventional method of DNA extraction from FFT for African swine fever virus (ASFV detection. The modified conventional method gave a higher quality DNA when compared with commercially available DNA extraction kits (QIAamp® DNA Mini Kit, DNeasy® Blood and Tissue Kit, and ZR Genomic DNA™ Tissue MiniPrep. Results: An average A260/A280 DNA purity of 0.86-1.68 and 3.22-5.32 μg DNA/mg for formalin fixed and non-fixed tissues, respectively using a conventional method. In a reproducible and three times repeat PCR, the ASFV DNA expected product size of 278 bp was obtained from the DNA extract of the conventional method but not from the DNA extract of the commercial kits. Conclusion: The present study has demonstrated that the conventional method extracts ASFV genome better than commercial kit. In summary, the commercial kit extraction appeared not suitable to purify ASFV DNA from FFT. We, therefore, recommend that the use of the conventional method be considered for African swine fever DNA extraction from FFT.
Wong, Stephen Q; Li, Jason; Tan, Angela Y-C; Vedururu, Ravikiran; Pang, Jia-Min B; Do, Hongdo; Ellul, Jason; Doig, Ken; Bell, Anthony; MacArthur, Grant A; Fox, Stephen B; Thomas, David M; Fellowes, Andrew; Parisot, John P; Dobrovic, Alexander
Clinical specimens undergoing diagnostic molecular pathology testing are fixed in formalin due to the necessity for detailed morphological assessment. However, formalin fixation can cause major issues with molecular testing, as it causes DNA damage such as fragmentation and non-reproducible sequencing artefacts after PCR amplification. In the context of massively parallel sequencing (MPS), distinguishing true low frequency variants from sequencing artefacts remains challenging. The prevalence of formalin-induced DNA damage and its impact on molecular testing and clinical genomics remains poorly understood. The Cancer 2015 study is a population-based cancer cohort used to assess the feasibility of mutational screening using MPS in cancer patients from Victoria, Australia. While blocks were formalin-fixed and paraffin-embedded in different anatomical pathology laboratories, they were centrally extracted for DNA utilising the same protocol, and run through the same MPS platform (Illumina TruSeq Amplicon Cancer Panel). The sequencing artefacts in the 1-10% and the 10-25% allele frequency ranges were assessed in 488 formalin-fixed tumours from the pilot phase of the Cancer 2015 cohort. All blocks were less than 2.5 years of age (mean 93 days). Consistent with the signature of DNA damage due to formalin fixation, many formalin-fixed samples displayed disproportionate levels of C>T/G>A changes in the 1-10% allele frequency range. Artefacts were less apparent in the 10-25% allele frequency range. Significantly, changes were inversely correlated with coverage indicating high levels of sequencing artefacts were associated with samples with low amounts of available amplifiable template due to fragmentation. The degree of fragmentation and sequencing artefacts differed between blocks sourced from different anatomical pathology laboratories. In a limited validation of potentially actionable low frequency mutations, a NRAS G12D mutation in a melanoma was shown to be a false
Dijkstra, Jeroen R; Opdam, Frank J M; Boonyaratanakornkit, Jerry
. We assessed a novel synthetic control for formalin-fixed, paraffin-embedded (FFPE) tumor samples in a blind study conducted within nine laboratories across Europe. We show that FFPE material can, at least in part, mimic clinical samples and we demonstrate this control to be a valuable tool...
Formalin-fixed paraffin-embedded (FFPE) tissue samples represent a potentially invaluable resource for genomic research into the molecular basis of disease. However, use of FFPE samples in gene expression studies has been limited by technical challenges resulting from degradation...
Hunter, Amanda; Eisma, Roos; Lamb, Clare
By soft fixing cadavers using the Thiel embalming method, our cadavers now exhibit a greater degree of flexibility and color retention compared to that of traditional formalin-fixed cadavers. The aim of this experiment was to discover whether Thiel embalming fluid could be used to revive and soften the muscles of formalin-fixed prosected specimens. Earlier this year, two severely dehydrated formalin-fixed forearm and hand specimens were fully submerged in a tank containing Thiel embalming fluid. After a period of six months the specimens were removed from the tank and noticeable changes were observed in flexibility, quality of the tissue, and color of the specimens. © 2014 Wiley Periodicals, Inc.
Wang, Jian-Hua; Gouda-Vossos, Amany; Dzamko, Nicolas; Halliday, Glenda; Huang, Yue
Both fresh-frozen and formalin-fixed, paraffin-embedded (FFPE) human brain tissues are invaluable resources for molecular genetic studies of central nervous system diseases, especially neurodegenerative disorders. To identify the optimal method for DNA extraction from human brain tissue, we compared methods on differently-processed tissues. Fragments of LRRK2 and MAPT (257 bp and 483 bp/245 bp) were amplified for evaluation. We found that for FFPE samples, the success rate of DNA extraction was greater when using a commercial kit than a laboratory-based method (successful DNA extraction from 76% versus 33% of samples). PCR amplicon size and storage period were key factors influencing the success rate of DNA extraction from FFPE samples. In the fresh-frozen samples, the DNA extraction success rate was 100% using either a commercial kit (QIAamp DNA Micro) or a laboratory-based method (sample boiling in 0.1 mol/L NaOH, followed by proteinase K digestion, and then DNA extraction using Chelex-100) regardless of PCR amplicon length or tissue storage time. Although the present results demonstrate that PCR-amplifiable genomic DNA can be extracted from both fresh-frozen and FFPE samples, fresh brain tissue is recommended for DNA extraction in future neuropathological studies.
Kocjan, Boštjan J; Hošnjak, Lea; Poljak, Mario
Formalin-fixed, paraffin-embedded (FFPE) tissue specimens stored in pathology departments worldwide are an invaluable source for diagnostic purposes when fresh clinical material is unavailable as well as for retrospective molecular and epidemiological studies, especially when dealing with rare clinical conditions for which prospective collection is not feasible. Accurate detection of HPV infection in these specimens is particularly challenging because nucleic acids are often degraded and therefore, not suitable for amplification of larger fragments of the viral genome or viral gene transcripts. This review provides a brief summary of molecular methods for detecting alpha-HPV DNA/RNA in FFPE tissue specimens. We specifically address the key procedural and environmental factors that have the greatest impact on the quality of nucleic acids extracted from FFPE tissue specimens, and describe some solutions that can be used to increase their integrity and/or amplifiability. Moreover, commonly used methods for HPV DNA/RNA detection in FFPE tissue specimens are presented and discussed, focusing on studies using polymerase chain reaction as an HPV detection method and published after 1999. Finally, we briefly summarize our 22 years of experience with HPV detection in FFPE tissue specimens. Copyright © 2015 Elsevier B.V. All rights reserved.
Potluri, Keerti; Mahas, Ahmed; Kent, Michael N; Naik, Sameep; Markey, Michael
As new technologies come within reach for the average cytogenetic laboratory, the study of chromosome structure has become increasingly more sophisticated. Resolution has improved from karyotyping (in which whole chromosomes are discernible) to fluorescence in situ hybridization and comparative genomic hybridization (CGH, with which specific megabase regions are visualized), array-based CGH (aCGH, examining hundreds of base pairs), and next-generation sequencing (providing single base pair resolution). Whole genome next-generation sequencing remains a cost-prohibitive method for many investigators. Meanwhile, the cost of aCGH has been reduced during recent years, even as resolution has increased and protocols have simplified. However, aCGH presents its own set of unique challenges. DNA of sufficient quantity and quality to hybridize to arrays and provide meaningful results is required. This is especially difficult for DNA from formalin-fixed paraffin-embedded (FFPE) tissues. Here, we compare three different methods for acquiring DNA of sufficient length, purity, and "amplifiability" for aCGH and other downstream applications. Phenol-chloroform extraction and column-based commercial kits were compared with adaptive focused acoustics (AFA). Of the three extraction methods, AFA samples showed increased amplicon length and decreased polymerase chain reaction (PCR) failure rate. These findings support AFA as an improvement over previous DNA extraction methods for FFPE tissues. Copyright © 2015 Elsevier Inc. All rights reserved.
Full Text Available Abstract Background Quantification of microRNAs in specific cell populations microdissected from tissues can be used to define their biological roles, and to develop and deploy biomarker assays. In this study, a number of variables were examined for their effect on the yield of microRNAs in samples obtained from formalin-fixed paraffin-embedded tissues by laser microdissection. Results MicroRNA yield was improved by using cresyl violet instead of hematoxylin-eosin to stain tissue sections in preparation for microdissection, silicon carbide instead of glass fiber as matrix in RNA-binding columns, and overnight digestion of dissected samples with proteinase K. Storage of slides carrying stained tissue sections at room temperature for up to a week before microdissection, and storage of the microdissectates at room temperature for up to a day before RNA extraction did not adversely affect microRNA yield. Conclusions These observations should be of value for the efficient isolation of microRNAs from microdissected formalin-fixed tissues with a flexible workflow.
Boisen, Mogens Karsbøl; Dehlendorff, Christian; Linnemann, Dorte
BACKGROUND: Archival formalin-fixed paraffin-embedded (FFPE) cancer tissue samples are a readily available resource for microRNA (miRNA) biomarker identification. No established standard for reference miRNAs in FFPE tissue exists. We sought to identify stable reference miRNAs for normalization...
Archival formalin-fixed paraffin-embedded (FFPE) samples offer a vast, untapped source of genomic data for biomarker discovery. However, the quality of FFPE samples is often highly variable, and conventional methods to assess RNA quality for RNA-sequencing (RNA-seq) are not infor...
Formalin-fixed paraffin-embedded tissue is regularly employed in the diagnosis of transmissible spongiform encephalopathies (TSE) by immunohistochemistry (IHC), the standard by which all other TSE diagnostic protocols are judged. While IHC affords advantages over diagnostic approaches that typically...
Danielle Mercatante Carrick
Full Text Available Next Generation Sequencing (NGS technologies are used to detect somatic mutations in tumors and study germ line variation. Most NGS studies use DNA isolated from whole blood or fresh frozen tissue. However, formalin-fixed paraffin-embedded (FFPE tissues are one of the most widely available clinical specimens. Their potential utility as a source of DNA for NGS would greatly enhance population-based cancer studies. While preliminary studies suggest FFPE tissue may be used for NGS, the feasibility of using archived FFPE specimens in population based studies and the effect of storage time on these specimens needs to be determined. We conducted a study to determine whether DNA in archived FFPE high-grade ovarian serous adenocarcinomas from Surveillance, Epidemiology and End Results (SEER registries Residual Tissue Repositories (RTR was present in sufficient quantity and quality for NGS assays. Fifty-nine FFPE tissues, stored from 3 to 32 years, were obtained from three SEER RTR sites. DNA was extracted, quantified, quality assessed, and subjected to whole exome sequencing (WES. Following DNA extraction, 58 of 59 specimens (98% yielded DNA and moved on to the library generation step followed by WES. Specimens stored for longer periods of time had significantly lower coverage of the target region (6% lower per 10 years, 95% CI: 3-10% and lower average read depth (40x lower per 10 years, 95% CI: 18-60, although sufficient quality and quantity of WES data was obtained for data mining. Overall, 90% (53/59 of specimens provided usable NGS data regardless of storage time. This feasibility study demonstrates FFPE specimens acquired from SEER registries after varying lengths of storage time and under varying storage conditions are a promising source of DNA for NGS.
Koepsell, Scott A.; Hinrichs, Steven H.
A real-time PCR assay to detect Histoplasma capsulatum in formalin-fixed, paraffin-embedded (FFPE) tissue is described. The assay had an analytical sensitivity of 6 pg/μl of fungal DNA, analytical specificity of 100%, and clinical sensitivity of 88.9%. This proof-of-concept study may aid in the diagnosis of histoplasmosis from FFPE tissue. PMID:22855519
Andrea J Adams
Full Text Available Museum collections provide indispensable repositories for obtaining information about the historical presence of disease in wildlife populations. The pathogenic amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd has played a significant role in global amphibian declines, and examining preserved specimens for Bd can improve our understanding of its emergence and spread. Quantitative PCR (qPCR enables Bd detection with minimal disturbance to amphibian skin and is significantly more sensitive to detecting Bd than histology; therefore, developing effective qPCR methodologies for detecting Bd DNA in formalin-fixed specimens can provide an efficient and effective approach to examining historical Bd emergence and prevalence. Techniques for detecting Bd in museum specimens have not been evaluated for their effectiveness in control specimens that mimic the conditions of animals most likely to be encountered in museums, including those with low pathogen loads. We used American bullfrogs (Lithobates catesbeianus of known infection status to evaluate the success of qPCR to detect Bd in formalin-fixed specimens after three years of ethanol storage. Our objectives were to compare the most commonly used DNA extraction method for Bd (PrepMan, PM to Macherey-Nagel DNA FFPE (MN, test optimizations for Bd detection with PM, and provide recommendations for maximizing Bd detection. We found that successful detection is relatively high (80-90% when Bd loads before formalin fixation are high, regardless of the extraction method used; however, at lower infection levels, detection probabilities were significantly reduced. The MN DNA extraction method increased Bd detection by as much as 50% at moderate infection levels. Our results indicate that, for animals characterized by lower pathogen loads (i.e., those most commonly encountered in museum collections, current methods may underestimate the proportion of Bd-infected amphibians. Those extracting DNA from
Adams, Andrea J; LaBonte, John P; Ball, Morgan L; Richards-Hrdlicka, Kathryn L; Toothman, Mary H; Briggs, Cheryl J
Museum collections provide indispensable repositories for obtaining information about the historical presence of disease in wildlife populations. The pathogenic amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd) has played a significant role in global amphibian declines, and examining preserved specimens for Bd can improve our understanding of its emergence and spread. Quantitative PCR (qPCR) enables Bd detection with minimal disturbance to amphibian skin and is significantly more sensitive to detecting Bd than histology; therefore, developing effective qPCR methodologies for detecting Bd DNA in formalin-fixed specimens can provide an efficient and effective approach to examining historical Bd emergence and prevalence. Techniques for detecting Bd in museum specimens have not been evaluated for their effectiveness in control specimens that mimic the conditions of animals most likely to be encountered in museums, including those with low pathogen loads. We used American bullfrogs (Lithobates catesbeianus) of known infection status to evaluate the success of qPCR to detect Bd in formalin-fixed specimens after three years of ethanol storage. Our objectives were to compare the most commonly used DNA extraction method for Bd (PrepMan, PM) to Macherey-Nagel DNA FFPE (MN), test optimizations for Bd detection with PM, and provide recommendations for maximizing Bd detection. We found that successful detection is relatively high (80-90%) when Bd loads before formalin fixation are high, regardless of the extraction method used; however, at lower infection levels, detection probabilities were significantly reduced. The MN DNA extraction method increased Bd detection by as much as 50% at moderate infection levels. Our results indicate that, for animals characterized by lower pathogen loads (i.e., those most commonly encountered in museum collections), current methods may underestimate the proportion of Bd-infected amphibians. Those extracting DNA from archived museum
Guyard, Alice; Boyez, Alice; Pujals, Anaïs; Robe, Cyrielle; Tran Van Nhieu, Jeanne; Allory, Yves; Moroch, Julien; Georges, Odette; Fournet, Jean-Christophe; Zafrani, Elie-Serge; Leroy, Karen
Formalin-fixed paraffin-embedded (FFPE) tissue blocks are widely used to identify clinically actionable molecular alterations or perform retrospective molecular studies. Our goal was to quantify degradation of DNA occurring during mid to long-term storage of samples in usual conditions. We selected 46 FFPE samples of surgically resected carcinomas of lung, colon, and urothelial tract, of which DNA had been previously extracted. We performed a second DNA extraction on the same blocks under identical conditions after a median period of storage of 5.5 years. Quantitation of DNA by fluorimetry showed a 53% decrease in DNA quantity after storage. Quantitative PCR (qPCR) targeting KRAS exon 2 showed delayed amplification of DNA extracted after storage in all samples but one. The qPCR/fluorimetry quantification ratio decreased from 56 to 15% after storage (p DNA analyzable by qPCR represented only 11% of the amount obtained at first extraction. Maximal length of amplifiable DNA fragments assessed with a multiplex PCR was reduced in DNA extracted from stored tissue, indicating that DNA fragmentation had increased in the paraffin blocks during storage. Next-generation sequencing was performed on 12 samples and showed a mean 3.3-fold decrease in library yield and a mean 4.5-fold increase in the number of single-nucleotide variants detected after storage. In conclusion, we observed significant degradation of DNA extracted from the same FFPE block after 4 to 6 years of storage. Better preservation strategies should be considered for storage of FFPE biopsy specimens.
O. M. Barth
Full Text Available The presence of viral antigen in sections from formalin-fixed and paraffin-embedded human tissues was demonstrated by trypsin digestion followed by direct or indirect immunofluorescence. The specimens may be used for retrospective diagnosis. The immunofluorescence technique has to be adapted to the suspected virus infection on the basis of previous histopathology study. Variations of trypsin concentration time and temperature of incubation, expose different viral antigens and have to be previously tested for each unknown system. For measles virus detection in lung a stronger digestion has to be applied as compared to adenovirus or respiratory disease viruses in the same tisue. Flavivirus in liver tissue needs a weaker digestion. The reproducibility of the method makes it useful as a routine technique in diagnosis of virus infection.A presença de antígeno viral em cortes de tecidos humanos fixados em formol e emblocados em parafina foi demonstrada pela digestão com tripsina foi demonstrada pela ingestão com tripsina seguida de imunofluorescência direta ou indireta. Os espécimens podem ser utilizados para diagnoses retrospectivas. A técnica da imunofluorescência deve ser adaptada à infecção viral suspeita segundo diagnosie histopatológica prévia. Os parâmetros para a digestão do tecido pela tripsina, relacionados à concentração, duração de atuação e temperatura, expõem diferentes antígenos virais e devem ser previamente testados para cada sistema a ser estabelecido. Uma digestão mais intensa deve ser aplicada para a detecção do vírus do sarampo em tecido pulmonar do que para adenovírus ou vírus respiratório sincicial no mesmo tecido. Por outro lado, o vírus da febre amarela em tecido de fígado necessita de uma digestão mais fraca.
Hykin, Sarah M; Bi, Ke; McGuire, Jimmy A
For 150 years or more, specimens were routinely collected and deposited in natural history collections without preserving fresh tissue samples for genetic analysis. In the case of most herpetological specimens (i.e. amphibians and reptiles), attempts to extract and sequence DNA from formalin-fixed, ethanol-preserved specimens-particularly for use in phylogenetic analyses-has been laborious and largely ineffective due to the highly fragmented nature of the DNA. As a result, tens of thousands of specimens in herpetological collections have not been available for sequence-based phylogenetic studies. Massively parallel High-Throughput Sequencing methods and the associated bioinformatics, however, are particularly suited to recovering meaningful genetic markers from severely degraded/fragmented DNA sequences such as DNA damaged by formalin-fixation. In this study, we compared previously published DNA extraction methods on three tissue types subsampled from formalin-fixed specimens of Anolis carolinensis, followed by sequencing. Sufficient quality DNA was recovered from liver tissue, making this technique minimally destructive to museum specimens. Sequencing was only successful for the more recently collected specimen (collected ~30 ybp). We suspect this could be due either to the conditions of preservation and/or the amount of tissue used for extraction purposes. For the successfully sequenced sample, we found a high rate of base misincorporation. After rigorous trimming, we successfully mapped 27.93% of the cleaned reads to the reference genome, were able to reconstruct the complete mitochondrial genome, and recovered an accurate phylogenetic placement for our specimen. We conclude that the amount of DNA available, which can vary depending on specimen age and preservation conditions, will determine if sequencing will be successful. The technique described here will greatly improve the value of museum collections by making many formalin-fixed specimens available for
Loiacono, Christina M; Beckwith, Nadine; Kunkle, Robert A; Orcutt, Dennis; Hall, S Mark
Transmissible, spongiform encephalopathies including bovine spongiform encephalopathy (BSE) and scrapie are fatal neurodegenerative disorders associated with the presence of an infectious abnormal isoform of normal mammalian proteins called prions. Identification of the prion protein associated with scrapie (PrP(Sc)) in the central nervous system is typically based upon immunoassays including immunohistochemistry (IHC) using formalin-fixed tissues or Western blot (WB) assays using fresh and/or frozen, non-formalin-fixed tissues. Each assay can discriminate between BSE, classical scrapie, and a previously reported strain of scrapie recently identified in the United States named Nor98 scrapie. Different tissue samples are required from the same animal to run these 2 different immunoassays. This may result in inconsistent test results for the same animal. Sampling problems such as collecting insufficient volumes of fresh tissue or less than optimal anatomic location of brainstem for IHC can affect the ability of the test procedures to offer definitive and discriminatory results. Recently, a WB method using formalin-fixed, paraffin-embedded (FFPE) tissue to identify PrP(Sc) was developed that successfully identified PrP(Sc) in sheep affected by classical scrapie. In the current study, the use of this technique to produce discriminatory results identifying classical BSE in bovine tissue and both classical and Nor98 scrapie in ovine tissue using paraffin-embedded brain samples is described. Protein-banding patterns from WB using FFPE tissue were similar to protein-banding patterns produced by WB assays utilizing fresh tissues from the same animals, and results correlated well with the IHC PrP(Sc)-positive staining present in the cerebellum and obex regions of brain samples from these animals.
Mini P Singh
Full Text Available Detecting high-risk-human papillomavirus (HPV types has become an integral part of the cervical cancer screening programmes. This study aimed to develop a multiplex polymerase chain reaction (PCR for identification of HPV types 16 and 18 along with the beta globin gene in formalin-fixed and paraffin-embedded cervical biopsy specimens. A total of 59 samples from patients with cervical abnormalities were tested. HPV 16 positivity was 50% in cervical cancers and 52.9% in cervical intraepithelial neoplasia. Our multiplex PCR protocol can be used as a simple and cost-effective tool for high-risk-HPV detection in cervical cancer screening programmes.
Vaca-Paniagua, Felipe; Alvarez-Gomez, Rosa María; Maldonado-Martínez, Hector Aquiles; Pérez-Plasencia, Carlos; Fragoso-Ontiveros, Veronica; Lasa-Gonsebatt, Federico; Herrera, Luis Alonso; Cantú, David; Bargallo-Rocha, Enrique; Mohar, Alejandro; Durand, Geoffroy; Forey, Nathalie; Voegele, Catherine; Vallée, Maxime; Le Calvez-Kelm, Florence; McKay, James; Ardin, Maude; Villar, Stéphanie; Zavadil, Jiri; Olivier, Magali
Triple negative breast cancer (TNBC), defined by the lack of expression of the estrogen receptor, progesterone receptor and human epidermal receptor 2, is an aggressive form of breast cancer that is more prevalent in certain populations, in particular in low- and middle-income regions. The detailed molecular features of TNBC in these regions remain unexplored as samples are mostly accessible as formalin-fixed paraffin embedded (FFPE) archived tissues, a challenging material for advanced genomic and transcriptomic studies. Using dedicated reagents and analysis pipelines, we performed whole exome sequencing and miRNA and mRNA profiling of 12 FFPE tumor tissues collected from pathological archives in Mexico. Sequencing analyses of the tumor tissues and their blood pairs identified TP53 and RB1 genes as the most frequently mutated genes, with a somatic mutation load of 1.7 mutations/exome Mb on average. Transcriptional analyses revealed an overexpression of growth-promoting signals (EGFR, PDGFR, VEGF, PIK3CA, FOXM1), a repression of cell cycle control pathways (TP53, RB1), a deregulation of DNA-repair pathways, and alterations in epigenetic modifiers through miRNA:mRNA network de-regulation. The molecular programs identified were typical of those described in basal-like tumors in other populations. This work demonstrates the feasibility of using archived clinical samples for advanced integrated genomics analyses. It thus opens up opportunities for investigating molecular features of tumors from regions where only FFPE tissues are available, allowing retrospective studies on the search for treatment strategies or on the exploration of the geographic diversity of breast cancer.
Bjerg Bennike, Tue; Kastaniegaard, Kenneth; Padurariu, Simona
Large repositories of well characterized RNAlater preserved samples and formalin-fixed, paraffin-embedded samples have been generated worldwide. However, the impact on the proteome of the preservation methods remain poorly described. Therefore, we analyzed the impact on the proteome of preserving...... samples in RNAlater, and by formalin-fixation, paraffin-embedding on human soft tissue, using directly frozen samples as a control ("Comparing the proteome of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human tissue samples" ). We here report the data from the analysis...
Thompson, Seonaid M; Craven, Rachel A; Nirmalan, Niroshini J; Harnden, Patricia; Selby, Peter J; Banks, Rosamonde E
Formalin-fixed paraffin-embedded (FFPE) tissue samples represent a tremendous potential resource for biomarker discovery, with large numbers of samples in hospital pathology departments and links to clinical information. However, the cross-linking of proteins and nucleic acids by formalin fixation has hampered analysis and proteomic studies have been restricted to using frozen tissue, which is more limited in availability as it needs to be collected specifically for research. This means that rare disease subtypes cannot be studied easily. Recently, improved extraction techniques have enabled analysis of FFPE tissue by a number of proteomic techniques. As with all clinical samples, pre-analytical factors are likely to impact on the results obtained, although overlooked in many studies. The aim of this review is to discuss the various pre-analytical factors, which include warm and cold ischaemic time, size of sample, fixation duration and temperature, tissue processing conditions, length of storage of archival tissue and storage conditions, and to review the studies that have considered these factors in more detail. In those areas where investigations are few or non-existent, illustrative examples of the possible importance of specific factors have been drawn from studies using frozen tissue or from immunohistochemical studies of FFPE tissue. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Full Text Available Abstract Background The identification of genomic signatures of colorectal cancer for risk stratification requires the study of large series of cancer patients with an extensive clinical follow-up. Multicentric clinical studies represent an ideal source of well documented archived material for this type of analyses. Methods To verify if this material is technically suitable to perform matrix-CGH, we performed a pilot study using macrodissected 29 formalin-fixed, paraffin-embedded tissue samples collected within the framework of the EORTC-GI/PETACC-2 trial for colorectal cancer. The scientific aim was to identify prognostic genomic signatures differentiating locally restricted (UICC stages II-III from systemically advanced (UICC stage IV colorectal tumours. Results The majority of archived tissue samples collected in the different centers was suitable to perform matrix-CGH. 5/7 advanced tumours displayed 13q-gain and 18q-loss. In locally restricted tumours, only 6/12 tumours showed a gain on 13q and 7/12 tumours showed a loss on 18q. Interphase-FISH and high-resolution array-mapping of the gain on 13q confirmed the validity of the array-data and narrowed the chromosomal interval containing potential oncogenes. Conclusion Archival, paraffin-embedded tissue samples collected in multicentric clinical trials are suitable for matrix-CGH analyses and allow the identification of prognostic signatures and aberrations harbouring potential new oncogenes.
Full Text Available The aim of the study was to determine optimal hydrolysis time for the Feulgen DNA staining of archival formalin fixed paraffin‐embedded surgical samples, prepared as single cell suspensions for image cytometric measurements. The nuclear texture features along with the IOD (integrated optical density of the tumor nuclei were analysed by an automated high resolution image cytometer as a function of duration of hydrolysis treatment (in 5 N HCl at room temperature. Tissue blocks of breast carcinoma, ovarian serous carcinoma, ovarian serous tumor of borderline malignancy and leiomyosarcoma were included in the study. IOD hydrolysis profiles showed plateau between 30 and 60 min in the breast carcinoma and leiomyosarcoma, and between 40 and 60 min in the ovarian serous carcinoma and ovarian serous tumor of borderline malignancy. Most of the nuclear texture features remained stable after 20 min of hydrolysis treatment. Our results indicate that the optimal hydrolysis time for IOD and for nuclear texture feature measurements, was between 40 and 60 min in the cell preparations from tissue blocks of three epithelial and one soft tissue tumor.
Kajimura, Junko; Ito, Reiko; Manley, Nancy R; Hale, Laura P
Performance of immunofluorescence staining on archival formalin-fixed paraffin-embedded human tissues is generally not considered to be feasible, primarily due to problems with tissue quality and autofluorescence. We report the development and application of procedures that allowed for the study of a unique archive of thymus tissues derived from autopsies of individuals exposed to atomic bomb radiation in Hiroshima, Japan in 1945. Multiple independent treatments were used to minimize autofluorescence and maximize fluorescent antibody signals. Treatments with NH3/EtOH and Sudan Black B were particularly useful in decreasing autofluorescent moieties present in the tissue. Deconvolution microscopy was used to further enhance the signal-to-noise ratios. Together, these techniques provide high-quality single- and dual-color fluorescent images with low background and high contrast from paraffin blocks of thymus tissue that were prepared up to 60 years ago. The resulting high-quality images allow the application of a variety of image analyses to thymus tissues that previously were not accessible. Whereas the procedures presented remain to be tested for other tissue types and archival conditions, the approach described may facilitate greater utilization of older paraffin block archives for modern immunofluorescence studies. © 2016 The Histochemical Society.
Hizem, A; M'rad, S; Oudni-M'rad, M; Mestiri, S; Hammedi, F; Mezhoud, H; Zakhama, A; Mokni, M; Babba, H
Cystic echinococcosis (CE) caused by Echinococcus granulosus remains a serious problem worldwide for issues relating to public health and the economy. The most predominantly affected sites are the liver and the lungs, but other organs such as the heart, the spleen and the peritoneum can also be infected. Access to cysts from uncommon sites has limited genomic and molecular investigations. In the present study, genotypes of E. granulosus sensu lato were identified from formalin-fixed paraffin-embedded tissues (FF-PETs) implicated in human CE. Tissue samples were obtained from 57 patients with histologically confirmed CE. DNA samples were analysed using Egss 1 polymerase chain reaction (PCR) specific to the mitochondrial 12S rRNA gene of E. granulosus sensu stricto. All cysts were typed as E. granulosus sensu stricto with up to 35% of the liver and 16.6% of lungs being the most frequently infected, and up to 48.4% of samples being from rare sites. No correlation was found between cyst site and either the gender or the age of patients. This study demonstrates the possibility of exploiting atypical cysts using FF-PET samples and highlights the predominance of E. granulosus sensu stricto species in the Tunisian population, even in unusual infection sites.
Luebker, Stephen A; Koepsell, Scott A
Urea based protein extraction of formalin-fixed paraffin-embedded (FFPE) tissue provides the most efficient workflow for proteomics due to its compatibility with liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). This study optimizes the use of urea for proteomic analysis of clinical FFPE tissue. A series of protein extraction conditions manipulating temperature and buffer composition were compared to reduce carbamylation introduced by urea and increase protein detection. Each extraction was performed on a randomized pair of serial sections of homogenous FFPE tissue and analyzed with LC-ESI-MS/MS. Results were compared in terms of yield, missed cleavages, and peptide carbamylation. Lowering extraction temperature to 60°C decreased carbamylation at the cost of decreased protein detection and yield. Protein extraction for at least 20 minutes at 95°C followed by 60°C for 2 hours maximized total protein yield while maintaining protein detection and reducing carbamylation by 7.9%. When accounting for carbamylation during analysis, this modified extraction temperature provides equivalent peptide and protein detection relative to the commercially available Qproteome® FFPE Tissue Kit. No changes to buffer composition containing 7 M urea, 2 M thiourea, and 1 M ammonium bicarbonate resulted in improvements to control conditions. Optimized urea in-solution digestion provides an efficient workflow with maximized yields for proteomic analysis of clinically relevant FFPE tissue.
Moens, Lotte N J; Falk-Sörqvist, Elin; Ljungström, Viktor; Mattsson, Johanna; Sundström, Magnus; La Fleur, Linnéa; Mathot, Lucy; Micke, Patrick; Nilsson, Mats; Botling, Johan
In recent years, the advent of massively parallel next-generation sequencing technologies has enabled substantial advances in the study of human diseases. Combined with targeted DNA enrichment methods, high sequence coverage can be obtained for different genes simultaneously at a reduced cost per sample, creating unique opportunities for clinical cancer diagnostics. However, the formalin-fixed, paraffin-embedded (FFPE) process of tissue samples, routinely used in pathology departments, results in DNA fragmentation and nucleotide modifications that introduce a number of technical challenges for downstream biomolecular analyses. We evaluated the HaloPlex target enrichment system for somatic mutation detection in 80 tissue fractions derived from 20 clinical cancer cases with paired tumor and normal tissue available in both FFPE and fresh-frozen format. Several modifications to the standard method were introduced, including a reduced target fragment length and two strand capturing. We found that FFPE material can be used for HaloPlex-based target enrichment and next-generation sequencing, even when starting from small amounts of DNA. By specifically capturing both strands for each target fragment, we were able to reduce the number of false-positive errors caused by FFPE-induced artifacts and lower the detection limit for somatic mutations. We believe that the HaloPlex method presented here will be broadly applicable as a tool for somatic mutation detection in clinical cancer settings. Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Wiśniewski, Jacek R; Duś, Kamila; Mann, Matthias
Archival formalin-fixed and paraffin-embedded clinical samples represent a very diverse source of material for proteomic investigation of diseases, often with follow-up patient information. Here, we describe an analytical workflow for analysis of laser-capture microdissected formalin-fixed and paraffin-embedded samples that allows studying proteomes to a depth of 10 000 proteins per sample. The workflow involves lysis of tissue in SDS-containing buffer, detergent removal, and consecutive digestion of the proteins with two enzymes by the multienzyme digestion filter-aided sample preparation method. Resulting peptides are fractionated by pipette-tip based strong anion exchange into six fractions and analyzed by LC-MS/MS on a bench top quadrupole Orbitrap mass spectrometer. Analysis of the data using the MaxQuant software resulted in the identification of 9502 ± 28 protein groups per a 110 nL sample of microdissected cells from human colonic adenoma. This depth of proteome analysis enables systemic insights into the organization of the adenoma cells and an estimation of the abundances of known biomarkers. It also allows the identification of proteins expressed from tumor suppressors, oncogenes, and other key players in the development and progression of the colorectal cancer. Our proteomic platform can be used for quantitative comparisons between samples representing different stages of diseases and thus can be applied to the discovery of biomarkers or drug targets. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Kuranishi, Fumito; Imaoka, Yuki; Sumi, Yuusuke; Uemae, Yoji; Yasuda-Kurihara, Hiroko; Ishihara, Takeshi; Miyazaki, Tsubasa; Ohno, Tadao
No effective treatment has been developed for bone-metastatic breast cancer. We found 3 cases with clinical complete response (cCR) of the bone metastasis and longer overall survival of the retrospectively examined cohort treated comprehensively including autologous formalin-fixed tumor vaccine (AFTV). AFTV was prepared individually for each patient from their own formalin-fixed and paraffin-embedded breast cancer tissues. Three patients maintained cCR status of the bone metastasis for 17 months or more. Rate of cCR for 1 year or more appeared to be 15% (3/20) after comprehensive treatments including AFTV. The median overall survival time (60.0 months) and the 3- to 8-year survival rates after diagnosis of bone metastasis were greater than those of historical control cohorts in Japan (1988-2002) and in the nationwide population-based cohort study of Denmark (1999-2007). Bone-metastatic breast cancer may be curable after comprehensive treatments including AFTV, although larger scale clinical trial is required.
Full Text Available Introduction. No effective treatment has been developed for bone-metastatic breast cancer. We found 3 cases with clinical complete response (cCR of the bone metastasis and longer overall survival of the retrospectively examined cohort treated comprehensively including autologous formalin-fixed tumor vaccine (AFTV. Patients and Methods. AFTV was prepared individually for each patient from their own formalin-fixed and paraffin-embedded breast cancer tissues. Results. Three patients maintained cCR status of the bone metastasis for 17 months or more. Rate of cCR for 1 year or more appeared to be 15% (3/20 after comprehensive treatments including AFTV. The median overall survival time (60.0 months and the 3- to 8-year survival rates after diagnosis of bone metastasis were greater than those of historical control cohorts in Japan (1988–2002 and in the nationwide population-based cohort study of Denmark (1999–2007. Conclusion. Bone-metastatic breast cancer may be curable after comprehensive treatments including AFTV, although larger scale clinical trial is required.
Michelsen, Nete Vinstrup; Brusgaard, Klaus; Tan, Qihua
The use of formalin-fixed, paraffin-embedded (FFPE) tissue overcomes the most prominent issues related to research on relatively rare diseases: limited sample size, availability of control tissue, and time frame. The use of FFPE pancreatic tissue in GEM may be especially challenging due to its very...
Bennike, Tue Bjerg; Kastaniegaard, Kenneth; Padurariu, Simona
Large biobanks exist worldwide containing formalin-fixed, paraffin-embedded samples and samples stored in RNAlater. However, the impact of tissue preservation on the result of a quantative proteome analysis remains poorly described.Human colon mucosal biopsies were extracted from the sigmoideum...
Use of archival resources has been limited to date by inconsistent methods for genomic profiling of degraded RNA from formalin-fixed paraffin-embedded (FFPE) samples. RNA-sequencing offers a promising way to address this problem. Here we evaluated transcriptomic dose responses us...
Full Text Available Abstract Background Analysis of key therapeutic targets such as epidermal growth factor receptor (EGFR in clinical tissue samples is typically done by immunohistochemistry (IHC and is only subjectively quantitative through a narrow dynamic range. The development of a standardized, highly-sensitive, linear, and quantitative assay for EGFR for use in patient tumor tissue carries high potential for identifying those patients most likely to benefit from EGFR-targeted therapies. Methods A mass spectrometry-based Selected Reaction Monitoring (SRM assay for the EGFR protein (EGFR-SRM was developed utilizing the Liquid Tissue®-SRM technology platform. Tissue culture cells (n = 4 were analyzed by enzyme-linked immunosorbent assay (ELISA to establish quantitative EGFR levels. Matching formalin fixed cultures were analyzed by the EGFR-SRM assay and benchmarked against immunoassay of the non-fixed cultured cells. Xenograft human tumor tissue (n = 10 of non-small cell lung cancer (NSCLC origin and NSCLC patient tumor tissue samples (n = 23 were microdissected and the EGFR-SRM assay performed on Liquid Tissue lysates prepared from microdissected tissue. Quantitative curves and linear regression curves for correlation between immunoassay and SRM methodology were developed in Excel. Results The assay was developed for quantitation of a single EGFR tryptic peptide for use in FFPE patient tissue with absolute specificity to uniquely distinguish EGFR from all other proteins including the receptor tyrosine kinases, IGF-1R, cMet, Her2, Her3, and Her4. The assay was analytically validated against a collection of tissue culture cell lines where SRM analysis of the formalin fixed cells accurately reflects EGFR protein levels in matching non-formalin fixed cultures as established by ELISA sandwich immunoassay (R2 = 0.9991. The SRM assay was applied to a collection of FFPE NSCLC xenograft tumors where SRM data range from 305amol/μg to 12,860amol/μg and
Nederlof Petra M
Full Text Available Abstract Background Array Comparative Genomic Hybridization (aCGH is a rapidly evolving technology that still lacks complete standardization. Yet, it is of great importance to obtain robust and reproducible data to enable meaningful multiple hybridization comparisons. Special difficulties arise when aCGH is performed on archival formalin-fixed, paraffin-embedded (FFPE tissue due to its variable DNA quality. Recently, we have developed an effective DNA quality test that predicts suitability of archival samples for BAC aCGH. Methods In this report, we first used DNA from a cancer cell-line (SKBR3 to optimize the aCGH protocol for automated hybridization, and subsequently optimized and validated the procedure for FFPE breast cancer samples. We aimed for highest throughput, accuracy, and reproducibility applicable to FFPE samples, which can also be important in future diagnostic use. Results Our protocol of automated array-CGH on archival FFPE ULS-labeled DNA showed very similar results compared with published data and our previous manual hybridization method. Conclusion This report combines automated aCGH on unamplified archival FFPE DNA using non-enzymatic ULS labeling, and describes an optimized protocol for this combination resulting in improved quality and reproducibility.
Fu, Zongming; Yan, Kun; Rosenberg, Avraham; Jin, Zhicheng; Crain, Barbara; Athas, Grace; Heide, Richard S Vander; Howard, Timothy; Everett, Allen D; Herrington, David; Van Eyk, Jennifer E
Evaluate combination of heat and elevated pressure to enhance protein extraction and quality of formalin-fixed (FF), and FF paraffin-embedded (FFPE) aorta for proteomics. Proteins were extracted from fresh frozen aorta at room temperature (RT). FF and FFPE aortas (3 months and 15 years) were extracted at RT, heat alone, or a combination of heat and high pressure. Protein yields were compared, and digested peptides from the extracts were analyzed with MS. Combined heat and elevated pressure increased protein yield from human FF or FFPE aorta compared to matched tissues with heat alone (1.5-fold) or at RT (8.3-fold), resulting in more proteins identified and with more sequence coverage. The length of storage did adversely affect the quality of proteins from FF tissue. For long-term storage, aorta was preserved better with FFPE than FF alone. Periostin and MGF-E8 were demonstrated suitable for MRM assays from FFPE aorta. Combination of heat and high pressure is an effective method to extract proteins from FFPE aorta for downstream proteomics. This method opens the possibility for use of archival and often rare FFPE aortas and possibly other tissues available to proteomics for biomarker discovery and quantification. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Full Text Available Prostate cancer (PCa is the most frequently diagnosed type of cancer in developed countries. The decisive method of diagnosis is based on the results of biopsies, morphologically evaluated to determine the presence or absence of cancer. Although this approach leads to a confident diagnosis in most cases, it can be improved by using the molecular markers present in the tissue. Both miRNAs and proteins are considered excellent candidates for biomarkers in formalin-fixed paraffin-embedded (FFPE tissues, due to their stability over long periods of time. In the last few years, a concerted effort has been made to develop the necessary tools for their reliable measurement in these types of samples. Furthermore, the use of these kinds of markers may also help in establishing tumor grade and aggressiveness, as well as predicting the possible outcomes in each particular case for the different treatments available. This would aid clinicians in the decision-making process. In this review, we attempt to summarize and discuss the potential use of microRNA and protein profiles in FFPE tissue samples as markers to better predict PCa diagnosis, progression, and response to therapy.
Fowler, Carol B; Chesnick, Ingrid E; Moore, Cedric D; O'Leary, Timothy J; Mason, Jeffrey T
Proteomic studies of formalin-fixed paraffin-embedded (FFPE) tissues are frustrated by the inability to extract proteins from archival tissue in a form suitable for analysis by 2-D gel electrophoresis or mass spectrometry. This inability arises from the difficulty of reversing formaldehyde-induced protein adducts and cross-links within FFPE tissues. We previously reported the use of elevated hydrostatic pressure as a method for efficient protein recovery from a hen egg-white lysozyme tissue surrogate, a model system developed to study formalin fixation and histochemical processing. In this study, we demonstrate the utility of elevated hydrostatic pressure as a method for efficient protein recovery from FFPE mouse liver tissue and a complex multi-protein FFPE tissue surrogate comprised of hen egg-white lysozyme, bovine carbonic anhydrase, bovine ribonuclease A, bovine serum albumin, and equine myoglobin (55∶15∶15∶10∶5 wt%). Mass spectrometry of the FFPE tissue surrogates retrieved under elevated pressure showed that both the low and high-abundance proteins were identified with sequence coverage comparable to that of the surrogate mixture prior to formaldehyde treatment. In contrast, non-pressure-extracted tissue surrogate samples yielded few positive and many false peptide identifications. Studies with soluble formalin-treated bovine ribonuclease A demonstrated that pressure modestly inhibited the rate of reversal (hydrolysis) of formaldehyde-induced protein cross-links. Dynamic light scattering studies suggest that elevated hydrostatic pressure and heat facilitate the recovery of proteins free of formaldehyde adducts and cross-links by promoting protein unfolding and hydration with a concomitant reduction in the average size of the protein aggregates. These studies demonstrate that elevated hydrostatic pressure treatment is a promising approach for improving the recovery of proteins from FFPE tissues in a form suitable for proteomic analysis.
Fowler, Carol B.; Chesnick, Ingrid E.; Moore, Cedric D.; O'Leary, Timothy J.; Mason, Jeffrey T.
Background Proteomic studies of formalin-fixed paraffin-embedded (FFPE) tissues are frustrated by the inability to extract proteins from archival tissue in a form suitable for analysis by 2-D gel electrophoresis or mass spectrometry. This inability arises from the difficulty of reversing formaldehyde-induced protein adducts and cross-links within FFPE tissues. We previously reported the use of elevated hydrostatic pressure as a method for efficient protein recovery from a hen egg-white lysozyme tissue surrogate, a model system developed to study formalin fixation and histochemical processing. Principal Findings In this study, we demonstrate the utility of elevated hydrostatic pressure as a method for efficient protein recovery from FFPE mouse liver tissue and a complex multi-protein FFPE tissue surrogate comprised of hen egg-white lysozyme, bovine carbonic anhydrase, bovine ribonuclease A, bovine serum albumin, and equine myoglobin (55∶15∶15∶10∶5 wt%). Mass spectrometry of the FFPE tissue surrogates retrieved under elevated pressure showed that both the low and high-abundance proteins were identified with sequence coverage comparable to that of the surrogate mixture prior to formaldehyde treatment. In contrast, non-pressure-extracted tissue surrogate samples yielded few positive and many false peptide identifications. Studies with soluble formalin-treated bovine ribonuclease A demonstrated that pressure modestly inhibited the rate of reversal (hydrolysis) of formaldehyde-induced protein cross-links. Dynamic light scattering studies suggest that elevated hydrostatic pressure and heat facilitate the recovery of proteins free of formaldehyde adducts and cross-links by promoting protein unfolding and hydration with a concomitant reduction in the average size of the protein aggregates. Conclusions These studies demonstrate that elevated hydrostatic pressure treatment is a promising approach for improving the recovery of proteins from FFPE tissues in a form
Carol B Fowler
Full Text Available Proteomic studies of formalin-fixed paraffin-embedded (FFPE tissues are frustrated by the inability to extract proteins from archival tissue in a form suitable for analysis by 2-D gel electrophoresis or mass spectrometry. This inability arises from the difficulty of reversing formaldehyde-induced protein adducts and cross-links within FFPE tissues. We previously reported the use of elevated hydrostatic pressure as a method for efficient protein recovery from a hen egg-white lysozyme tissue surrogate, a model system developed to study formalin fixation and histochemical processing.In this study, we demonstrate the utility of elevated hydrostatic pressure as a method for efficient protein recovery from FFPE mouse liver tissue and a complex multi-protein FFPE tissue surrogate comprised of hen egg-white lysozyme, bovine carbonic anhydrase, bovine ribonuclease A, bovine serum albumin, and equine myoglobin (55∶15∶15∶10∶5 wt%. Mass spectrometry of the FFPE tissue surrogates retrieved under elevated pressure showed that both the low and high-abundance proteins were identified with sequence coverage comparable to that of the surrogate mixture prior to formaldehyde treatment. In contrast, non-pressure-extracted tissue surrogate samples yielded few positive and many false peptide identifications. Studies with soluble formalin-treated bovine ribonuclease A demonstrated that pressure modestly inhibited the rate of reversal (hydrolysis of formaldehyde-induced protein cross-links. Dynamic light scattering studies suggest that elevated hydrostatic pressure and heat facilitate the recovery of proteins free of formaldehyde adducts and cross-links by promoting protein unfolding and hydration with a concomitant reduction in the average size of the protein aggregates.These studies demonstrate that elevated hydrostatic pressure treatment is a promising approach for improving the recovery of proteins from FFPE tissues in a form suitable for proteomic analysis.
Full Text Available Formalin-fixed and paraffin-embedded (FFPE tissues represent a valuable source for biomarker studies and clinical routine diagnostics. However, they suffer from degradation of nucleic acids due to the fixation process. Since genetic and epigenetic studies usually require PCR amplification, this degradation hampers its use significantly, impairing PCR robustness or necessitating short amplicons. In routine laboratory medicine a highly robust PCR performance is mandatory for the clinical utility of genetic and epigenetic biomarkers. Therefore, methods to improve PCR performance using DNA from FFPE tissue are highly desired and of wider interest. The effect of template DNA derived from FFPE tissues on PCR performance was investigated by means of qPCR and conventional PCR using PCR fragments of different sizes. DNA fragmentation was analyzed via agarose gel electrophoresis. This study showed that poor PCR amplification was partly caused by inhibition of the DNA polymerase by fragmented DNA from FFPE tissue and not only due to the absence of intact template molecules of sufficient integrity. This PCR inhibition was successfully minimized by increasing the polymerase concentration, dNTP concentration and PCR elongation time thereby allowing for the robust amplification of larger amplicons. This was shown for genomic template DNA as well as for bisulfite-converted template DNA required for DNA methylation analyses. In conclusion, PCR using DNA from FFPE tissue suffers from inhibition which can be alleviated by adaptation of the PCR conditions, therefore allowing for a significant improvement of PCR performance with regard to variability and the generation of larger amplicons. The presented solutions to overcome this PCR inhibition are of tremendous value for clinical chemistry and laboratory medicine.
Full Text Available Abstract Introduction Protein extraction from formalin-fixed paraffin-embedded (FFPE tissues is challenging due to extensive molecular crosslinking that occurs upon formalin fixation. Reverse-phase protein array (RPPA is a high-throughput technology, which can detect changes in protein levels and protein functionality in numerous tissue and cell sources. It has been used to evaluate protein expression mainly in frozen preparations or FFPE-based studies of limited scope. Reproducibility and reliability of the technique in FFPE samples has not yet been demonstrated extensively. We developed and optimized an efficient and reproducible procedure for extraction of proteins from FFPE cells and xenografts, and then applied the method to FFPE patient tissues and evaluated its performance on RPPA. Results Fresh frozen and FFPE preparations from cell lines, xenografts and breast cancer and renal tissues were included in the study. Serial FFPE cell or xenograft sections were deparaffinized and extracted by six different protein extraction protocols. The yield and level of protein degradation were evaluated by SDS-PAGE and Western Blots. The most efficient protocol was used to prepare protein lysates from breast cancer and renal tissues, which were subsequently subjected to RPPA. Reproducibility was evaluated and Spearman correlation was calculated between matching fresh frozen and FFPE samples. The most effective approach from six protein extraction protocols tested enabled efficient extraction of immunoreactive protein from cell line, breast cancer and renal tissue sample sets. 85% of the total of 169 markers tested on RPPA demonstrated significant correlation between FFPE and frozen preparations (p Conclusions With optimized protein extraction methods, FFPE tissues can be a valuable source in generating reproducible and biologically relevant proteomic profiles using RPPA, with specific marker performance varying according to tissue type.
Guo, Huifang; Liu, Wenbin; Ju, Zhenlin; Tamboli, Pheroze; Jonasch, Eric; Mills, Gordon B; Lu, Yiling; Hennessy, Bryan T; Tsavachidou, Dimitra
Protein extraction from formalin-fixed paraffin-embedded (FFPE) tissues is challenging due to extensive molecular crosslinking that occurs upon formalin fixation. Reverse-phase protein array (RPPA) is a high-throughput technology, which can detect changes in protein levels and protein functionality in numerous tissue and cell sources. It has been used to evaluate protein expression mainly in frozen preparations or FFPE-based studies of limited scope. Reproducibility and reliability of the technique in FFPE samples has not yet been demonstrated extensively. We developed and optimized an efficient and reproducible procedure for extraction of proteins from FFPE cells and xenografts, and then applied the method to FFPE patient tissues and evaluated its performance on RPPA. Fresh frozen and FFPE preparations from cell lines, xenografts and breast cancer and renal tissues were included in the study. Serial FFPE cell or xenograft sections were deparaffinized and extracted by six different protein extraction protocols. The yield and level of protein degradation were evaluated by SDS-PAGE and Western Blots. The most efficient protocol was used to prepare protein lysates from breast cancer and renal tissues, which were subsequently subjected to RPPA. Reproducibility was evaluated and Spearman correlation was calculated between matching fresh frozen and FFPE samples.The most effective approach from six protein extraction protocols tested enabled efficient extraction of immunoreactive protein from cell line, breast cancer and renal tissue sample sets. 85% of the total of 169 markers tested on RPPA demonstrated significant correlation between FFPE and frozen preparations (p extraction methods, FFPE tissues can be a valuable source in generating reproducible and biologically relevant proteomic profiles using RPPA, with specific marker performance varying according to tissue type.
Li, Man; Zhou, Xingang; Li, Jingjing; Sun, Lei; Chen, Xiangmei; Wang, Peng
It is difficult to diagnose brucellar spondylitis because of its nonspecific clinical, radiological, and histological characteristics. This study aimed to determine whether real-time polymerase chain reaction (PCR) using formalin-fixed paraffin-embedded (FFPE) tissues was superior to conventional serum-based methods for diagnosing brucellar spondylitis.This retrospective study included 31 patients with brucellosis and a control group of 20 people with no history of brucellosis or exposure to Brucella spp. Samples from all patients with brucellar spondylitis were evaluated using Giemsa staining, the standard tube agglutination (STA) test, blood culture, and real-time PCR.The brucellar spondylitis was acute in 7 patients (22.6%), subacute in 15 patients (48.4%), and chronic in 9 patients (29%). Serological assays provided positive results for 25 patients (80.1%), real-time PCR provided positive results for 29 patients (93.5%), and blood cultures provided positive results for 11 patients (35.5%). The real-time PCR provided sensitivity of 93.5%, specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 100%. The corresponding values for the STA test were 80.1%, 100%, 100%, and 76.9%, respectively. Real-time PCR provided better sensitivity than Giemsa staining, the STA test, and blood culture, although the difference between PCR and STA was not statistically significant (P = .22). B melitensis was the only pathogen that was detected in patient with brucellar spondylitis using real-time PCR.These results suggest that real-time PCR provides a high sensitivity for diagnosing brucellar spondylitis. Furthermore, the real-time PCR results indicate that B melitensis was the causative pathogen in these cases.
Yamamoto, Satoko; Ibusuki, Mutsuko; Yamamoto, Yutaka; Fu, Peifen; Fujiwara, Saori; Murakami, Keiichi; Iwase, Hirotaka
Ki67 is a protein associated with cell cycle activity and shows a good correlation with the growth fraction, which has been proposed as a prognostic or predictive marker in breast cancer. In this study, we aimed to analyze the expression levels of Ki67 (MKI67) messenger RNA (mRNA) derived from formalin-fixed paraffin-embedded (FFPE) tissues for comparison with the immunohistochemical Ki67 labeling index, and investigate the correlation coefficients with clinical outcomes. We analyzed the data of Ki67 mRNA from FFPE and matched fresh-frozen (FF) tissues based on a real-time quantitative reverse-transcription polymerase chain reaction (RT-qPCR) assay system in 203 cases of primary invasive breast cancer. The correlation between Ki67 mRNA expression of either FFPE or FF specimens and Ki67 labeling index was positive, as was the correlation between the FFPE and FF results (P Ki67 mRNA expression of FFPE specimens was significantly associated with clinicopathological characteristics: tumor size, lymph node status, nuclear grade, hormone receptors, human epidermal growth factor receptor 2 (Her2) status, and tumor subtype. In prognostic results, Ki67 gene expression in the FFPE specimens revealed almost similar patterns of significance in Kaplan-Meier curves and univariate and multivariate relapse-free survival results as the Ki67 labeling index. Gene expression analysis of Ki67 of FFPE specimens could be successfully performed using RT-qPCR, closely resembling the significant clinical characteristics of Ki67 labeling index. These results confirm that Ki67 gene expression of FFPE specimens has potential for evaluation of cell cycle activity of breast cancer specimens.
Full Text Available Nonepithelial ovarian cancer (NEOC is a rare cancer that is often misdiagnosed as other malignant tumors. Research on this cancer using fresh tissues is nearly impossible because of its limited number of samples within a limited time provided. The study is to identify potential genes and their molecular pathways related to NEOC using formalin-fixed paraffin embedded samples. Total RNA was extracted from eight archived NEOCs and seven normal ovaries. The RNA samples with RNA integrity number >2.0, purity >1.7 and cycle count value <28 cycles were hybridized to the Illumina Whole-Genome DASL assay (cDNA-mediated annealing, selection, extension, and ligation. We analyzed the results using the GeneSpring GX11.0 and FlexArray software to determine the differentially expressed genes. Microarray results were validated using an immunohistochemistry method. Statistical analysis identified 804 differentially expressed genes with 443 and 361 genes as overexpressed and underexpressed in cancer, respectively. Consistent findings were documented for the overexpression of eukaryotic translation elongation factor 1 alpha 1, E2F transcription factor 2, and fibroblast growth factor receptor 3, except for the down-regulated gene, early growth response 1 (EGR1. The immunopositivity staining for EGR1 was found in the majority of cancer tissues. This finding suggested that the mRNA level of a transcript did not always match with the protein expression in tissues. The current gene profile can be the platform for further exploration of the molecular mechanism of NEOC.
Full Text Available Formalin fixing with paraffin embedding (FFPE has been a standard sample preparation method for decades, and archival FFPE samples are still very useful resources. Nonetheless, the use of FFPE samples in cancer genome analysis using next-generation sequencing, which is a powerful technique for the identification of genomic alterations at the nucleotide level, has been challenging due to poor DNA quality and artificial sequence alterations. In this study, we performed whole-exome sequencing of matched frozen samples and FFPE samples of tissues from 4 cancer patients and compared the next-generation sequencing data obtained from these samples. The major differences between data obtained from the 2 types of sample were the shorter insert size and artificial base alterations in the FFPE samples. A high proportion of short inserts in the FFPE samples resulted in overlapping paired reads, which could lead to overestimation of certain variants; >20% of the inserts in the FFPE samples were double sequenced. A large number of soft clipped reads was found in the sequencing data of the FFPE samples, and about 30% of total bases were soft clipped. The artificial base alterations, C>T and G>A, were observed in FFPE samples only, and the alteration rate ranged from 200 to 1,200 per 1M bases when sequencing errors were removed. Although high-confidence mutation calls in the FFPE samples were compatible to that in the frozen samples, caution should be exercised in terms of the artifacts, especially for low-confidence calls. Despite the clearly observed artifacts, archival FFPE samples can be a good resource for discovery or validation of biomarkers in cancer research based on whole-exome sequencing.
Meyer, Julia A; Zhou, Delu; Mason, Clinton C; Downie, Jonathan M; Rodic, Vladimir; Abromowitch, Minnie; Wistinghausen, Birte; Termuhlen, Amanda M; Angiolillo, Anne L; Perkins, Sherrie L; Lones, Mark A; Barnette, Phillip; Schiffman, Joshua D; Miles, Rodney R
Recurrent genomic changes in B-lymphoblastic leukemia (B-ALL) identified by genome-wide single-nucleotide polymorphism (SNP) microarray analysis provide important prognostic information, but gene copy number analysis of its rare lymphoma counterpart, B-lymphoblastic lymphoma (B-LBL), is limited by the low incidence and lack of fresh tissue for genomic testing. We used molecular inversion probe (MIP) technology to analyze and compare copy number alterations (CNAs) in archival formalin-fixed paraffin-embedded pediatric B-LBL (n = 23) and B-ALL (n = 55). Similar to B-ALL, CDKN2A/B deletions were the most common alteration identified in 6/23 (26%) B-LBL cases. Eleven of 23 (48%) B-LBL patients were hyperdiploid, but none showed triple trisomies (chromosomes 4, 10, and 17) characteristic of B-ALL. IKZF1 and PAX5 deletions were observed in 13 and 17% of B-LBL, respectively, which was similar to the reported frequency in B-ALL. Immunoglobulin light chain lambda (IGL) locus deletions consistent with normal light chain rearrangement were observed in 5/23 (22%) B-LBL cases, compared with only 1% in B-ALL samples. None of the B-LBL cases showed abnormal, isolated VPREB1 deletion adjacent to IGL locus, which we identified in 25% of B-ALL. Our study demonstrates that the copy number profile of B-LBL is distinct from B-ALL, suggesting possible differences in pathogenesis between these closely related diseases. © 2016 Wiley Periodicals, Inc.
Vera-Lozada, Gabriela; Scholl, Vanesa; Barros, Mário Henrique M; Sisti, Davide; Guescini, Michele; Stocchi, Vilberto; Stefanoff, Claudio Gustavo; Hassan, Rocio
Formalin-fixed paraffin-embedded (FFPE) tissues are invaluable sources of biological material for research and diagnostic purposes. In this study, we aimed to identify biological and technical variability in RT-qPCR TaqMan® assays performed with FFPE-RNA from lymph nodes of classical Hodgkin lymphoma samples. An ANOVA-nested 6-level design was employed to evaluate BCL2, CASP3, IRF4, LYZ and STAT1 gene expression. The most variable genes were CASP3 (low expression) and LYZ (high expression). Total variability decreased after normalization for all genes, except by LYZ. Genes with moderate and low expression were identified and suffered more the effects of the technical manipulation than high-expression genes. Pre-amplification was shown to introduce significant technical variability, which was partially alleviated by lowering to a half the amount of input RNA. Ct and Cy0 quantification methods, based on cycle-threshold and the kinetic of amplification curves, respectively, were compared. Cy0 method resulted in higher quantification values, leading to the decrease of total variability in CASP3 and LYZ genes. The mean individual noise was 0.45 (0.31 to 0.61 SD), indicating a variation of gene expression over ~1.5 folds from one case to another. We showed that total variability in RT-qPCR from FFPE-RNA is not higher than that reported for fresh complex tissues, and identified gene-, and expression level-sources of biological and technical variability, which can allow better strategies for designing RT-qPCR assays from highly degraded and inhibited samples. Copyright © 2014 Elsevier Inc. All rights reserved.
Regezi, J A; Zarbo, R J; Keren, D F
An immunofluorescent technique was used on formalin-fixed, paraffin-embedded tissue to characterize and diagnose plasma cell lesions of the head and neck. Infiltrates were classified as either monoclonal or polyclonal with rhodamine and fluorescein staining of light chains and heavy chains. In the cases in which histopathologic diagnoses were relatively certain, immunofluorescence provided good correlation. In those cases in which histopathologic diagnoses were equivocal, immunofluorescence distinguished between reactive and neoplastic infiltrates through the determination of clonality of the infiltrates.
Marosy, Beth A.; Craig, Brian D.; Hetrick, Kurt N.; Witmer, P. Dane; Ling, Hua; Griffith, Sean M.; Myers, Ben; Ostrander, Elaine A.; Stanford, Janet L.; Brody, Lawrence C.; Doheny, Kimberly F.
This unit describes a protocol for generating exome enriched sequencing libraries using DNA extracted from Formalin Fixed Paraffin Embedded (FFPE) samples. Utilizing commercially available kits, we present a low input FFPE workflow starting with 50ng of DNA. This procedure includes a repair step to address damage caused by FFPE preservation that improves sequence quality. Subsequently, libraries undergo an in-solution targeted selection for exons, followed by sequencing using the Illumina next generation short read sequencing platform. PMID:28075488
Full Text Available Serologic diagnosis is one of the most widely used diagnostic methods for Q fever, but the window period in antibody response of 2 to 3 weeks after symptom onset results in significant diagnostic delay. We investigated the diagnostic utility of Q fever PCR from formalin-fixed liver tissues in Q fever patients with acute hepatitis.We reviewed the clinical and laboratory data in patients with Q fever hepatitis who underwent liver biopsy during a 17-year period, and whose biopsied tissues were available. We also selected patients who revealed granuloma in liver biopsy and with no Q fever diagnosis within the last 3 years as control. Acute Q fever hepatitis was diagnosed if two or more of the following clinical, serologic, or histopathologic criteria were met: (1 an infectious hepatitis-like clinical feature such as fever (≥ 38°C with elevated hepatic transaminase levels; (2 exhibition of a phase II immunoglobulin G (IgG antibodies titer by IFA of ≥ 1:128 in single determination, or a four-fold or greater rise between two separate samples obtained two or more weeks apart; (3 histologic finding of biopsy tissue showing characteristic fibrin ring granuloma.A total of 11 patients with acute Q fever hepatitis were selected and analyzed. Of the 11 patients, 3 (27% had exposure to zoonotic risk factors and 7 (63% met the serologic criteria. Granulomas with either circumferential or radiating fibrin deposition were observed in 10 cases on liver biopsy and in 1 case on bone marrow biopsy. 8 (73% revealed positive Coxiella burnetii PCR from their formalin-fixed liver tissues. In contrast, none of 10 patients with alternative diagnosis who had hepatic granuloma revealed positive C. burnetii PCR from their formalin-fixed liver tissues.Q fever PCR from formalin-fixed liver tissues appears to be a useful adjunct for diagnosing Q fever hepatitis.
Stefanović, D; Samardžija, G; Redžek, A; Arnaut, M; Nikin, Z; Stefanović, M
Romanowsky-Giemsa (RG) stains were devised during the 19th century for identifying plasmodia parasites in blood smears. Later, RG stains became standard procedures for hematology and cytology. Numerous attempts have been made to apply RG staining to formalin-fixed paraffin-embedded tissue sections, with varied success. Most published work on this topic described RG staining methods in which sections were overstained, then subjected to acid differentiation; unfortunately, the differentiation step often caused inconsistent staining outcomes. If staining is performed under optimal conditions with control of dye concentration, pH, solution temperature and staining time, no differentiation is required. We used RG and 0.002 M buffer, pH 42, for staining and washing sections. All steps were performed at room temperature. After staining and air drying, sections were washed in 96-100% ethanol to remove extraneous stain. Finally, sections were washed in xylene and mounted using DPX. Staining results were similar to routine hemalum and eosin (H & E) staining. Nuclei were blue; intensity depended largely on chromatin density. RNA-rich sites were purple. Collagen fibers, keratin, muscle cells, erythrocytes and white matter of the central nervous system were stained pinkish and reddish hues. Cartilage matrix, mast cell granules and areas of myxomatous degeneration were purple. Sulfate-rich mucins were stained pale blue, while those lacking sulfate groups were unstained. Deposits of hemosiderin, lipofuscin and melanin were greenish, and calcium deposits were blue. Helicobacter pylori bacteria were violet to purple. The advantages of the method are its close similarity to H & E staining and technical simplicity. Hemosiderin, H. pylori, mast cell granules, melanin and specific granules of different hematopoietic cells, which are invisible or barely distinguishable by H & E staining, are visualized. Other advantages over previous RG stains include shorter staining time and avoidance
Matsuda, Y; Nagao, M; Takatori, T; Niijima, H; Nakajima, M; Iwase, H; Kobayashi, M; Iwadate, K
One of the hydrolysis products of sarin (isopropyl methylphosphonofluoridate) was detected in formalin-fixed brain tissues of victims poisoned in the Tokyo subway terrorist attack. Part of this procedure, used for the detection of sarin hydrolysis products in erythrocytes of sarin victims, has been described previously. The test materials were four individual cerebellums, which had been stored in formalin fixative for about 2 years. Sarin-bound acetylcholinesterase (AChE) was solubilized from these cerebellums, purified by immunoaffinity chromatography, and digested with trypsin. Then the sarin hydrolysis products bound to AChE were released by alkaline phosphatase digestion, subjected to trimethylsilyl derivatization (TMS), and detected by gas chromatography-mass spectrometry. Peaks at m/z 225 and m/z 240, which are indicative of TMS-methylphosphonic acid, were observed within the retention time range of authentic methylphosphonic acid. However, no isopropyl methylphosphonic acid was detected in the formalin-fixed cerebellums of these 4 sarin victims, probably because the isopropoxy group of isopropyl methylphosphonic acid underwent chemical hydrolysis during storage. This procedure will be useful for the forensic diagnosis of poisoning by protein-bound, highly toxic agents, such as sarin, which are easily hydrolysed. This appears to be the first time that intoxication by a nerve agent has been demonstrated by analyzing formalin-fixed brains obtained at autopsy.
Johnston, Louise; Power, Michael; Sloan, Philip
system. The Idylla NRAS-BRAF mutation test has been developed for the qualitative detection of mutations in NRAS and BRAF oncogenes, facilitating genetic profiling of patients with cancer. The aim of this study was to carry out a formal clinical performance evaluation. METHODS: Two-hundred and forty......-two formalin-fixed paraffin-embedded (FFPE) human malignant colorectal cancer (CRC) tissue samples were identified in departmental archives and tested with both the Idylla NRAS-BRAF mutation test and the Agena Bioscience MassARRAY test. RESULTS: The overall concordance between the Idylla NRAS-BRAF mutation...... reference test. Reanalysis of this sample by droplet digital PCR confirmed that the mutation was present, but at an allelic frequency below the stated sensitivity level of the MassARRAY system. CONCLUSION: These results confirm that the Idylla NRAS-BRAF mutation test has high concordance with a widely used...
Djidja, Marie-Claude; Claude, Emmanuelle; Scriven, Peter; Allen, David W; Carolan, Vikki A; Clench, Malcolm R
MALDI-mass spectrometry imaging (MALDI-MSI) has been shown to allow the study of protein distribution and identification directly within formalin-fixed paraffin-embedded (FFPE) tissue sections. However, direct protein identification from tissue sections remains challenging due to signal interferences and/or existing post-translational or other chemical modifications. The use of antigen retrieval (AR) has been demonstrated for unlocking proteins prior to in situ enzymatic digestion and MALDI-MSI analysis of FFPE tissue sections. In the work reported here, the identification of proline oxidation, which may occur when performing the AR protocol, is described. This facilitated and considerably increased the number of identified peptides when adding proline oxidation as a variable modification to the MASCOT search criteria. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. Copyright © 2016 Elsevier B.V. All rights reserved.
Wang, Xiaoning; Deng, Zaian; Huang, Chuncui; Zhu, Tong; Lou, Jiatao; Wang, Lin; Li, Yan
N-glycan profiling is a powerful approach for analyzing the functional relationship between N-glycosylation and cancer. Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. Furthermore, fresh tissue samples are difficult to ship and store. Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. We found that our method was highly reproducible. We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MS n -based structure prediction. We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. Copyright © 2017 Elsevier B.V. All rights reserved.
Zhang, Liangxuan; Chen, Liangjing; Sah, Sachin; Latham, Gary J; Patel, Rajesh; Song, Qinghua; Koeppen, Hartmut; Tam, Rachel; Schleifman, Erica; Mashhedi, Haider; Chalasani, Sreedevi; Fu, Ling; Sumiyoshi, Teiko; Raja, Rajiv; Forrest, William; Hampton, Garret M; Lackner, Mark R; Hegde, Priti; Jia, Shidong
The success of precision oncology relies on accurate and sensitive molecular profiling. The Ion AmpliSeq Cancer Panel, a targeted enrichment method for next-generation sequencing (NGS) using the Ion Torrent platform, provides a fast, easy, and cost-effective sequencing workflow for detecting genomic "hotspot" regions that are frequently mutated in human cancer genes. Most recently, the U.K. has launched the AmpliSeq sequencing test in its National Health Service. This study aimed to evaluate the clinical application of the AmpliSeq methodology. We used 10 ng of genomic DNA from formalin-fixed, paraffin-embedded human colorectal cancer (CRC) tumor specimens to sequence 46 cancer genes using the AmpliSeq platform. In a validation study, we developed an orthogonal NGS-based resequencing approach (SimpliSeq) to assess the AmpliSeq variant calls. Validated mutational analyses revealed that AmpliSeq was effective in profiling gene mutations, and that the method correctly pinpointed "true-positive" gene mutations with variant frequency >5% and demonstrated high-level molecular heterogeneity in CRC. However, AmpliSeq enrichment and NGS also produced several recurrent "false-positive" calls in clinically druggable oncogenes such as PIK3CA. AmpliSeq provided highly sensitive and quantitative mutation detection for most of the genes on its cancer panel using limited DNA quantities from formalin-fixed, paraffin-embedded samples. For those genes with recurrent "false-positive" variant calls, caution should be used in data interpretation, and orthogonal verification of mutations is recommended for clinical decision making.
Full Text Available We have developed a gene expression assay (Whole-Genome DASL, capable of generating whole-genome gene expression profiles from degraded samples such as formalin-fixed, paraffin-embedded (FFPE specimens.We demonstrated a similar level of sensitivity in gene detection between matched fresh-frozen (FF and FFPE samples, with the number and overlap of probes detected in the FFPE samples being approximately 88% and 95% of that in the corresponding FF samples, respectively; 74% of the differentially expressed probes overlapped between the FF and FFPE pairs. The WG-DASL assay is also able to detect 1.3-1.5 and 1.5-2 -fold changes in intact and FFPE samples, respectively. The dynamic range for the assay is approximately 3 logs. Comparing the WG-DASL assay with an in vitro transcription-based labeling method yielded fold-change correlations of R(2 approximately 0.83, while fold-change comparisons with quantitative RT-PCR assays yielded R(2 approximately 0.86 and R(2 approximately 0.55 for intact and FFPE samples, respectively. Additionally, the WG-DASL assay yielded high self-correlations (R(2>0.98 with low intact RNA inputs ranging from 1 ng to 100 ng; reproducible expression profiles were also obtained with 250 pg total RNA (R(2 approximately 0.92, with approximately 71% of the probes detected in 100 ng total RNA also detected at the 250 pg level. When FFPE samples were assayed, 1 ng total RNA yielded self-correlations of R(2 approximately 0.80, while still maintaining a correlation of R(2 approximately 0.75 with standard FFPE inputs (200 ng.Taken together, these results show that WG-DASL assay provides a reliable platform for genome-wide expression profiling in archived materials. It also possesses utility within clinical settings where only limited quantities of samples may be available (e.g. microdissected material or when minimally invasive procedures are performed (e.g. biopsied specimens.
Paulsen, I M S; Dimke, H; Frische, S
Heat-induced epitope retrieval (HIER) is widely used for immunohistochemistry on formalin fixed paraffin-embedded tissue and includes temperatures well above the melting point of paraffin. We therefore tested whether traditional xylene-based removal of paraffin is required on sections from paraffin...... of dewaxing in xylene. In conclusion, the HIER procedure described and tested can be used as a single procedure enabling dewaxing, hydration and epitope retrieval for immunohistochemistry in formalin fixed paraffin-embedded tissue....
Zhu, Yazhen; Lu, Dan; Lira, Maruja E; Xu, Qing; Du, Yunzhi; Xiong, Jianghong; Mao, Mao; Chung, Hyun Cheol; Zheng, Guangjuan
Human epidermal growth factor receptor 2 (HER2) is a key driver of tumorigenesis, and over-expression as a result of HER2 gene amplification has been observed in a number of solid tumors. Recently HER2 has become an important biomarker for the monoclonal antibody treatment of HER2-positive metastatic breast and advanced gastric cancer. The HER2 targeting antibody trastuzumab treatment requires accurate measurement of HER2 levels for proper diagnosis. Droplet digital PCR (ddPCR) with highly direct, precise and absolute nucleic acid quantification could be used to detect HER2 amplification levels. Our objective was to evaluate a robust, accurate and less subjective application of ddPCR for HER2 amplification levels and test the assay performance in clinical formalin-fixed paraffin-embedded (FFPE) breast and gastric carcinoma samples. Genomic DNA from HER2 amplified cell line SK-BR-3 was used to set up the ddPCR assays. The copy number of HER2 was compared to the chromosome 17 centromere reference gene (CEP17), expressed as HER2:CEP17 ratio. Genomic DNAs of FFPE specimens from 145 Asian patients with breast and gastric carcinomas were assayed using both standard methods, immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH), and ddPCR. Based on 145 clinical breast and gastric carcinoma cases, our study demonstrated a high concordance of ddPCR results to FISH and IHC. In breast cancer specimens, the ddPCR results had high concordance with FISH and IHC defined HER2 status with a sensitivity of 90.9% (30/33) and a specificity of 100% (77/77). In gastric cancer specimens that were concordant in both FISH and IHC, our assay was 95.5% concordant with FISH and IHC (21/22). ddPCR has the advantage of automation and also allows levels of HER2 amplification to be easily evaluated in large numbers of samples, and presents a potential option to define HER2 status. Copyright © 2015 Elsevier Inc. All rights reserved.
Marosy, Beth A; Craig, Brian D; Hetrick, Kurt N; Witmer, P Dane; Ling, Hua; Griffith, Sean M; Myers, Benjamin; Ostrander, Elaine A; Stanford, Janet L; Brody, Lawrence C; Doheny, Kimberly F
This unit describes a technique for generating exome-enriched sequencing libraries using DNA extracted from formalin-fixed paraffin-embedded (FFPE) samples. Utilizing commercially available kits, we present a low-input FFPE workflow starting with 50 ng of DNA. This procedure includes a repair step to address damage caused by FFPE preservation that improves sequence quality. Subsequently, libraries undergo an in-solution-targeted selection for exons, followed by sequencing using the Illumina next-generation short-read sequencing platform. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.
Schiefer, Ana-Iris; Parlow, Laura; Gabler, Lisa; Mesteri, Ildiko; Koperek, Oskar; von Deimling, Andreas; Streubel, Berthold; Preusser, Matthias; Lehmann, Annika; Kellner, Udo; Pauwels, Patrick; Lambin, Suzan; Dietel, Manfred; Hummel, Michael; Klauschen, Frederick; Birner, Peter; Möbs, Markus
The mutated BRAF oncogene represents a therapeutic target in malignant melanoma. Because BRAF mutations are also involved in the pathogenesis of other human malignancies, the use of specific BRAF inhibitors might also be extended to other diseases in the future. A prerequisite for the clinical application of BRAF inhibitors is the reliable detection of activating BRAF mutations in routine histopathological samples. In a multicenter approach, we evaluated a novel and fully automated PCR-based system (Idylla) capable of detecting BRAF V600 mutations in formalin-fixed, paraffin-embedded tissue within 90 minutes with high sensitivity. We analyzed a total of 436 samples with the Idylla system. Valid results were obtained in 421 cases (96.56%). Its performance was compared with conventional methods (pyrosequencing or Sanger sequencing). Concordant results were obtained in 406 cases (96.90%). Reanalysis of eight discordant samples by next-generation sequencing and/or pyrosequencing with newly extracted DNA and the BRAF RGQ Kit confirmed the Idylla result in seven cases, resulting in an overall agreement of 98.57%. In conclusion, the Idylla system is a highly reliable and sensitive platform for detection of BRAF V600 mutations in formalin-fixed, paraffin-embedded material, providing an efficient alternative to conventional diagnostic methods, particularly for routine diagnostics laboratories with limited experience in molecular pathology. Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Guil-Luna, S.; Stenvang, Jan; Brünner, Nils
and its isoforms in formalin-fixed, paraffin-embedded tissue samples from canine mammary lesions (4 dysplasias, 10 benign tumors, and 46 carcinomas) using 1-step SYBR Green quantitative real-time polymerase chain reaction (RT-qPCR). Progesterone receptor was expressed in 75% of dysplasias, all benign...... in the expression of isoform A versus B. Analysis of progesterone receptor mRNA isoforms by RT-qPCR was successful in routinely formalin-fixed, paraffin-embedded tissue samples and enabled the distribution of isoforms A and B to be identified for the first time in dysplasias, benign tumors, and malignant tumors...
Droplet digital PCR (ddPCR) vs quantitative real-time PCR (qPCR) approach for detection and quantification of Merkel cell polyomavirus (MCPyV) DNA in formalin fixed paraffin embedded (FFPE) cutaneous biopsies.
Arvia, Rosaria; Sollai, Mauro; Pierucci, Federica; Urso, Carmelo; Massi, Daniela; Zakrzewska, Krystyna
Merkel cell polyomavirus (MCPyV) is associated with Merkel cell carcinoma and high viral load in the skin was proposed as a risk factor for the occurrence of this tumour. MCPyV DNA was detected, with lower frequency, in different skin cancers but since the viral load was usually low, the real prevalence of viral DNA could be underestimated. To evaluate the performance of two assays (qPCR and ddPCR) for MCPyV detection and quantification in formalin fixed paraffin embedded (FFPE) tissue samples. Both assays were designed to simultaneous detection and quantification of both MCPyV as well as house-keeping DNA in clinical samples. The performance of MCPyV quantification was investigated using serial dilutions of cloned target DNA. We also evaluated the applicability of both tests for the analysis of 76 FFPE cutaneous biopsies. The two approaches resulted equivalent with regard to the reproducibility and repeatability and showed a high degree of linearity in the dynamic range tested in the present study. Moreover, qPCR was able to quantify ≥10 5 copies per reaction, while the upper limit of ddPCR was 10 4 copies. There was not significant difference between viral load measured by the two methods The detection limit of both tests was 0,15 copies per reaction, however, the number of positive samples obtained by ddPCR was higher than that obtained by qPCR (45% and 37% respectively). The ddPCR represents a better method for detection of MCPyV in FFPE biopsies, mostly these containing low copies number of viral genome. Copyright © 2017 Elsevier B.V. All rights reserved.
Canteros, Cristina E; Vélez H, Alejandro; Toranzo, Adriana I; Suárez-Alvarez, Roberto; Tobón O, Ángela; Jimenez A, María del Pilar; Restrepo M, Ángela
Coccidioides immitis and C. posadasii are the etiologic agents of coccidioidomycosis, an endemic fungal disease of the Americas. In Colombia, this mycosis is uncommon, and only five cases, two of them imported, have been documented.By means of DNA sequencing, C. immitis was identified in formalin-fixed, paraffin-embedded archival tissues samples from the 5th Colombian patient diagnosed in 1997. The patient was born in Pinto, Department of Magdalena, and had never visited other geographic regions, a reason to consider that the mycosis had been acquired locally.This species is primarily found in California although it has been occasionally reported in other geographic areas such as Mexico and Brazil. This is the first indigenous report of C. immitis-associated coccidioidomycosis in a Colombian patient. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.
Donczo, Boglarka; Guttman, Andras
More than a century ago in 1893, a revolutionary idea about fixing biological tissue specimens was introduced by Ferdinand Blum, a German physician. Since then, a plethora of fixation methods have been investigated and used. Formalin fixation with paraffin embedment became the most widely used types of fixation and preservation method, due to its proper architectural conservation of tissue structures and cellular shape. The huge collection of formalin-fixed, paraffin-embedded (FFPE) sample archives worldwide holds a large amount of unearthed information about diseases that could be the Holy Grail in contemporary biomarker research utilizing analytical omics based molecular diagnostics. The aim of this review is to critically evaluate the omics options for FFPE tissue sample analysis in the molecular diagnostics field. Copyright © 2018. Published by Elsevier B.V.
Reis Patricia P
Full Text Available Abstract Background MicroRNAs (miRs are non-coding RNA molecules involved in post-transcriptional regulation, with diverse functions in tissue development, differentiation, cell proliferation and apoptosis. miRs may be less prone to degradation during formalin fixation, facilitating miR expression studies in formalin-fixed paraffin-embedded (FFPE tissue. Results Our study demonstrates that the TaqMan Human MicroRNA Array v1.0 (Early Access platform is suitable for miR expression analysis in FFPE tissue with a high reproducibility (correlation coefficients of 0.95 between duplicates, p 35, we show that reproducibility between technical replicates, equivalent dilutions, and FFPE vs. frozen samples is best in the high abundance stratum. We also demonstrate that the miR expression profiles of FFPE samples are comparable to those of fresh-frozen samples, with a correlation of up to 0.87 (p Conclusion Our study thus demonstrates the utility, reproducibility, and optimization steps needed in miR expression studies using FFPE samples on a high-throughput quantitative PCR-based miR platform, opening up a realm of research possibilities for retrospective studies.
Chabert, Steren; Villalobos, Manuel; Ulloa, Patricia; Salas, Rodrigo; Tejos, Cristian; San Martin, Sebastian; Pereda, Jaime
Human tissues are usually studied using a series of two-dimensional visualizations of in vivo or cutout specimens. However, there is no precise anatomical description of some of the processes of human fetal development. The purpose of our study is to develop a quantitative description of the normal axial skeleton by means of high-resolution three-dimensional magnetic resonance (MR) images, collected from six normal 20-week-old human fetuses fixed in formaldehyde. Fetuses were collected after spontaneous abortion and subsequently fixed with formalin. They were imaged using a 1.5 T MR scanner with an isotropic spatial resolution of 200 µm. The correct tissue discrimination between ossified and cartilaginous bones was confirmed by comparing the images achieved by MR scans and computerized axial tomographies. The vertebral column was segmented out from each image using a specially developed semi-automatic algorithm. Vertebral body dimensions and inter-vertebral distances were larger in the lumbar region, in agreement with the beginning of the ossification process from the thoracolumbar region toward the sacral and cephalic ends. In this article, we demonstrate the feasibility of using MR images to study the ossification process in formalin-fixed fetal tissues. A quantitative description of the ossification centers of vertebral bodies and arches is presented. © 2012 John Wiley & Sons, Ltd.
Jarošová, Marie; Kučerová, Jana; Flodr, Patrik; Mikešová, Michaela; Procházka, Vít; Papajík, Tomáš
The currently valid molecular genetic subclassification of patients with diffuse large B-cell lymphoma (DLBCL) into three prognostic subgroups based on expression profiling has been the objective of numerous genetic studies. In routine clinical practice, however, expression profiling technology remains unavailable for the most of centers. Apart from the technology, in some cases molecular genetic laboratories have problems obtaining high-quality material, i.e. fresh tissues, for RNA isolation to determine gene expression. One possibility is to determine the gene expression from RNA obtained by isolation from formalin-fixed, paraffin-embedded (FFPE) tissue. This pilot study aimed at isolating RNA from FFPE in patients diagnosed with DLBCL and verifying the potential use of such RNA for the expression analysis of 7 selected genes. Although the study showed that it is possible to isolate RNA and determine the expression of the selected genes from archival material, the values of relative expression of some genes in the set were too variable to be used for unambiguous prognostic classification. It was confirmed that retrospective analyses of selected genes may be performed with sufficient material obtained, and that properly archived blocks may be used for molecular biology analyses even after 8 years.
Tewari, Deepanker; Del Piero, Fabio; Cieply, Stephen; Feria, Willard; Acland, Helen
Equine herpesvirus-1 (EHV-1) strains with a single point mutation at the 2254 nucleotide position with a G2254 constitution within the DNA polymerase gene are associated strongly with equine myeloencephalopathies. Infections with non-neuropathogenic EHV-1 strains without the G2254 nucleotide but with an A2254 nucleotide are associated less frequently with equine neurologic disease. A retrospective study utilizing DNA extracted from formalin fixed paraffin embedded tissues was conducted with real time PCR and pyrosequencing, to determine the infecting EHV-1 strains. Infection with EHV-1 A2254 and or G2254 strain was detected with real time PCR, and was confirmed with a rapid pyrosequencing technique. Pyrosequencing was useful in at least 2 cases where real time PCR was equivocal in determining the infecting EHV-1 strain type. The strain with G2254 mutation was detected in 9.4% of 21 studied abortion cases, and in 86.6% of 15 neurologic cases. Copyright © 2013 Elsevier B.V. All rights reserved.
Ehrlein, J; Wanke, R; Weis, S; Brem, G; Hermanns, W
A sensitive technique of non-isotopic in situ hybridization (NISH) is presented, which permits the detection of human growth hormone (hGH) mRNA in routinely formalin-fixed, paraffin-embedded transgenic mouse tissues and human post mortem pituitaries; the latter were used as positive tissue controls in this study. In addition, a double staining procedure combining NISH and immunohistochemistry for the visualization of both hGH and hGH mRNA in the same paraffin section is described. Digoxigenin-labelled antisense hGH RNA was used for NISH of hGH mRNA. The NISH protocol was based upon an established radioactive method. Alkaline phosphatase and horseradish peroxidase-based immunoenzymatic procedures for the detection of digoxigenin-labelled RNA probes using different chromogens [4-nitro blue tetrazolium chloride (NBT), Fast Blue BB, New Fuchsin, and 3,3'-diaminobenzidine tetrahydrochloride (DAB) with or without intensification of the DAB staining] were compared. The proteolytic tissue pretreatment and the detection procedure were found to be the most critical steps for successful visualization of hGH mRNA. After optimization of the permeabilization conditions, hGH mRNA could be visualized in each case studied when alkaline phosphatase/NBT-based detection was employed. The NISH technique presented here, performed either separately or in combination with immunohistochemistry, permits retrospective analyses, of hGH (trans)gene expression in archival, paraffin-embedded specimens.
Mansour, Anthony; Chatila, Rajaa; Bejjani, Noha; Dagher, Carole; Faour, Wissam H
Protein detection methods in formalin-fixed paraffin embedded (FFPE) tissue blocks are widely used in research and clinical setting in order to diagnose or to confirm a diagnosis of various types of diseases. Therefore, multiple protein extraction methods from FFPE tissue sections have been developed in this regard. However, the yield and the quality of proteins extracted from FFPE tissues are significantly reduced in blocks stored for longer periods of time. Regardless the protein extraction method used, tissue sections must be first deparaffinized with xylene, and then washed in serial dilutions of ethanol in order to remove the toxic organic solvent "xylene" and rehydrate the tissue. The objective of this study was first to develop a method to deparaffinize FFPE blocks that excludes the use of toxic solvent "xylene". Second minimize the time required to perform the extraction. Here we describe a method where:•The entire paraffin embedded blocks are deparaffinized and rehydrated using only hot distilled water as a substitute for both xylene and ethanol•The entire procedure takes about 15 min•Deparaffinized blocks are immediately homogenized in lysis buffer, and the obtained lysate analyzed by Western blot. With this new modified technique, we were able to successfully detect actin and AKT proteins in lysates from blocks embedded in paraffin for up to 9 years.
Donover, P Scott; Wojciechowski, Brian S; Thirumaran, Rajesh; Zemba-Palko, Vlasta; Prendergast, George C; Wallon, U Margaretha
Overexpression of the extracellular metalloproteinase inhibitor TIMP-4 in estrogen receptor-negative breast cancers was found recently to be associated with a poor prognosis for survival. To pursue exploration of the theranostic applications of TIMP-4, specific antibodies with favorable properties for immunohistochemical use and other clinical assays are needed. Here we report the characterization of a monoclonal antibody (clone 9:4-7) specific for full-length human TIMP-4 with suitable qualities. The antibody was determined to be an IgG(2b) immunoglobulin. In enzyme-linked immunosorbent assay (ELISA) and immunoblotting assays, it did not exhibit any detectable crossreactivity with recombinant forms of the other human TIMPs 1, 2, and 3. In contrast, the antibody displayed high specificity and sensitivity for TIMP-4 including in formalin-fixed and paraffin-embedded specimens of human breast specimens. An analysis of tissue microarrays of human cancer and corresponding normal tissues revealed specific staining patterns with excellent signal-to-noise ratios. This study documents TIMP-4 monoclonal antibody clone 9:4-7 as an effective tool for preclinical and clinical investigations. Published 2010 Wiley-Liss, Inc.
Kim, Dowhan; Lee, Da Hye; Choi, Junjeong; Shim, Kyu Won; Kim, Se Hoon
The therapeutic protocols for treatment of germinomas and non-germinomatous germ cell tumors (NGGCTs) are completely different, so it is important to distinguish pure germinomas from NGGCTs. As it can be difficult to diagnose by morphology alone, immunohisto-chemistry (IHC) has been widely used as an ancillary test to improve diagnostic accuracy. However, IHC has limitations due to the misinterpretation of results or the aberrant loss of immunoreactivity. However, real-time RT-PCR has certain advantages over IHC, including its quantitative nature. The aim of our study was to evaluate the usefulness of real-time RT-PCR on formalin-fixed paraffin-embedded (FFPE) tissue blocks for the diagnosis of germ cell tumors of the central nervous system. We selected eight markers of germ cell tumors using a literature search, and validated them using real-time RT-PCR. Among them, POU5F1, NANOG and TGFB2 were statistically significant (P=0.05) in multiple comparisons (MANOVA) of three groups (pure germinomas, mature teratomas and malignant germ cell tumors). Two-group (pure germinomas and NGGCTs) discriminant analysis achieved a 70.0% success rate in cross-validation. We concluded that real-time RT-PCR using FFPE tissue has adequate validating power comparable to IHC in the diagnosis of central nervous system germ cell tumors; therefore, when IHC is not available, not conclusive or not informative, RT-PCR is a potential alternative to a repeat biopsy.
Glud, Martin; Klausen, Mikkel; Gniadecki, Robert; Rossing, Maria; Hastrup, Nina; Nielsen, Finn C; Drzewiecki, Krzysztof T
MicroRNAs (miRNAs) are small, noncoding RNA molecules that regulate cellular differentiation, proliferation, and apoptosis. MiRNAs are expressed in a developmentally regulated and tissue-specific manner. Aberrant expression may contribute to pathological processes such as cancer, and miRNA may therefore serve as biomarkers that may be useful in a clinical environment for diagnosis of various diseases. Most miRNA profiling studies have used fresh tissue samples. However, in some types of cancer, including malignant melanoma, fresh material is difficult to obtain from primary tumors, and most surgical specimens are formalin fixed and paraffin embedded (FFPE). To explore whether FFPE material would be suitable for miRNA profiling in melanocytic lesions, we compared miRNA expression patterns in FFPE versus fresh frozen samples, obtained from 15 human melanocytic nevi. Out of microarray data, we identified 84 miRNAs that were expressed in both types of samples and represented an miRNA profile of melanocytic nevi. Our results showed a high correlation in miRNA expression (Spearman r-value of 0.80) between paired FFPE and fresh frozen material. The data were further validated by quantitative RT-PCR. In conclusion, FFPE specimens of melanocytic lesions are suitable as a source for miRNA microarray profiling.
Mubemba, B; Thompson, P N; Odendaal, L; Coetzee, P; Venter, E H
Rift Valley fever (RVF), caused by an arthropod borne Phlebovirus in the family Bunyaviridae, is a haemorrhagic disease that affects ruminants and humans. Due to the zoonotic nature of the virus, a biosafety level 3 laboratory is required for isolation of the virus. Fresh and frozen samples are the preferred sample type for isolation and acquisition of sequence data. However, these samples are scarce in addition to posing a health risk to laboratory personnel. Archived formalin-fixed, paraffin-embedded (FFPE) tissue samples are safe and readily available, however FFPE derived RNA is in most cases degraded and cross-linked in peptide bonds and it is unknown whether the sample type would be suitable as reference material for retrospective phylogenetic studies. A RT-PCR assay targeting a 490 nt portion of the structural G N glycoprotein encoding gene of the RVFV M-segment was applied to total RNA extracted from archived RVFV positive FFPE samples. Several attempts to obtain target amplicons were unsuccessful. FFPE samples were then analysed using next generation sequencing (NGS), i.e. Truseq ® (Illumina) and sequenced on the Miseq ® genome analyser (Illumina). Using reference mapping, gapped virus sequence data of varying degrees of shallow depth was aligned to a reference sequence. However, the NGS did not yield long enough contigs that consistently covered the same genome regions in all samples to allow phylogenetic analysis. Copyright © 2017 Elsevier B.V. All rights reserved.
Florenza Lüder Ripoli
Full Text Available Mammary neoplasms are the tumors most affecting female dogs and women. Formalin-fixed, paraffin-embedded (FFPE tissues are an invaluable source of archived biological material. Fresh frozen (FF tissue is considered ideal for gene expression analysis. However, strategies based on FFPE material offer several advantages. Branched-DNA assays permit a reliable and fast workflow when analyzing gene expression. The aim of this study was to assess the comparability of the branched-DNA assay when analyzing certain gene expression patterns between FF and FFPE samples in canine mammary tumors. RNA was isolated from 109 FFPE samples and from 93 FF samples of different canine mammary tissues. Sixteen (16 target genes (Tp53; Myc; HMGA1; Pik3ca; Mcl1; MAPK3; FOXO3; PTEN; GATA4; PFDN5; HMGB1; MAPK1; BRCA2; BRCA1; HMGA2; and Her2 were analyzed via branched-DNA assay (b-DNA. ACTB, GAPDH, and HPRT1 were used as data normalizers. Overall, the relative gene expression of the two different origins of samples showed an agreement of 63%. Still, care should be taken, as FFPE specimens showed lower expression of the analyzed targets when compared to FF samples. The fact that the gene expression in FFPE proved to be lower than in FF specimens is likely to have been caused by the effect of storage time. ACTB had the best performance as a data normalizer.
Adam, Benjamin; Afzali, Bahman; Dominy, Katherine M; Chapman, Erin; Gill, Reeda; Hidalgo, Luis G; Roufosse, Candice; Sis, Banu; Mengel, Michael
Histopathologic diagnoses in transplantation can be improved with molecular testing. Preferably, molecular diagnostics should fit into standard-of-care workflows for transplant biopsies, that is, formalin-fixed paraffin-embedded (FFPE) processing. The NanoString(®) gene expression platform has recently been shown to work with FFPE samples. We aimed to evaluate its methodological robustness and feasibility for gene expression studies in human FFPE renal allograft samples. A literature-derived antibody-mediated rejection (ABMR) 34-gene set, comprised of endothelial, NK cell, and inflammation transcripts, was analyzed in different retrospective biopsy cohorts and showed potential to molecularly discriminate ABMR cases, including FFPE samples. NanoString(®) results were reproducible across a range of RNA input quantities (r = 0.998), with different operators (r = 0.998), and between different reagent lots (r = 0.983). There was moderate correlation between NanoString(®) with FFPE tissue and quantitative reverse transcription polymerase chain reaction (qRT-PCR) with corresponding dedicated fresh-stabilized tissue (r = 0.487). Better overall correlation with histology was observed with NanoString(®) (r = 0.354) than with qRT-PCR (r = 0.146). Our results demonstrate the feasibility of multiplexed gene expression quantification from FFPE renal allograft tissue. This represents a method for prospective and retrospective validation of molecular diagnostics and its adoption in clinical transplantation pathology. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Kennedy, Richard D
Current prognostic factors are poor at identifying patients at risk of disease recurrence after surgery for stage II colon cancer. Here we describe a DNA microarray-based prognostic assay using clinically relevant formalin-fixed paraffin-embedded (FFPE) samples.
Glud, M.; Klausen, M.; Gniadecki, R.
surgical specimens are formalin fixed and paraffin embedded (FFPE). To explore whether FFPE material would be suitable for miRNA profiling in melanocytic lesions, we compared miRNA expression patterns in FFPE versus fresh frozen samples, obtained from 15 human melanocytic nevi. Out of microarray data, we...
Boye, Mette; Feenstra, Anne Avlund; Tegtmeier, Conny
and the immunohistochemical methods were used for detection of S. suis in formalin-fixed, paraffin-embedded tissue sections of brain, endocardium, and lung from pigs infected with S. suis. The methods developed were able to detect single cells of S. suis in situ in the respective samples, whereas no signal was observed from...
Christensen, M; Funder, A D; Bendix, K
AIM: To compare clonal T cell receptor gamma (TCRgamma) gene rearrangements in frozen and formalin-fixed paraffin wax-embedded (FFPE) tissue, using capillary electrophoresis for use in diagnostics, as T cell lymphomas may be difficult to diagnose by conventional methods. METHODS: The DNA for PCR...
Archival samples represent a vast resource for identification of chemical and pharmaceutical targets. Previous use of formalin-fixed paraffin-embedded (FFPE) samples has been limited due to changes in RNA introduced by fixation and embedding procedures. Recent advances in RNA-seq...
Tramm, Trine; Hennig, Guido; Kyndi, Marianne
Gene expression analysis on messenger RNA (mRNA) purified from formalin-fixed, paraffin-embedded tissue is increasingly used for research purposes. Tissue heterogeneity may question specificity and interpretation of results from mRNA isolated from a whole slide section, and thresholds for minimal...
Taverna, Domenico; Pollins, Alonda C; Nanney, Lillian B; Sindona, Giovanni; Caprioli, Richard M
Herein we present a simple, reproducible and versatile approach for in situ protein digestion and identification on formalin-fixed and paraffin-embedded (FFPE) tissues. This adaptation is based on the use of an enzyme delivery platform (hydrogel discs) that can be positioned on the surface of a tissue section. By simultaneous deposition of multiple hydrogels over select regions of interest within the same tissue section, multiple peptide extracts can be obtained from discrete histological areas. After enzymatic digestion, the hydrogel extracts are submitted for LC-MS/MS analysis followed by database inquiry for protein identification. Further, imaging mass spectrometry (IMS) is used to reveal the spatial distribution of the identified peptides within a serial tissue section. Optimization was achieved using cutaneous tissue from surgically excised pressure ulcers that were subdivided into two prime regions of interest: the wound bed and the adjacent dermal area. The robust display of tryptic peptides within these spectral analyses of histologically defined tissue regions suggests that LC-MS/MS in combination with IMS can serve as useful exploratory tools. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Boskamp, Tobias; Lachmund, Delf; Oetjen, Janina; Cordero Hernandez, Yovany; Trede, Dennis; Maass, Peter; Casadonte, Rita; Kriegsmann, Jörg; Warth, Arne; Dienemann, Hendrik; Weichert, Wilko; Kriegsmann, Mark
Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) shows a high potential for applications in histopathological diagnosis, and in particular for supporting tumor typing and subtyping. The development of such applications requires the extraction of spectral fingerprints that are relevant for the given tissue and the identification of biomarkers associated with these spectral patterns. We propose a novel data analysis method based on the extraction of characteristic spectral patterns (CSPs) that allow automated generation of classification models for spectral data. Formalin-fixed paraffin embedded (FFPE) tissue samples from N=445 patients assembled on 12 tissue microarrays were analyzed. The method was applied to discriminate primary lung and pancreatic cancer, as well as adenocarcinoma and squamous cell carcinoma of the lung. A classification accuracy of 100% and 82.8%, resp., could be achieved on core level, assessed by cross-validation. The method outperformed the more conventional classification method based on the extraction of individual m/z values in the first application, while achieving a comparable accuracy in the second. LC-MS/MS peptide identification demonstrated that the spectral features present in selected CSPs correspond to peptides relevant for the respective classification. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. Copyright © 2016 Elsevier B.V. All rights reserved.
Wadsworth, Jonathan D F; Dalmau-Mena, Inmaculada; Joiner, Susan; Linehan, Jacqueline M; O'Malley, Catherine; Powell, Caroline; Brandner, Sebastian; Asante, Emmanuel A; Ironside, James W; Hilton, David A; Collinge, John
Anonymous screening of lymphoreticular tissues removed during routine surgery has been applied to estimate the UK population prevalence of asymptomatic vCJD prion infection. The retrospective study of Hilton et al (J Pathol 2004; 203: 733-739) found accumulation of abnormal prion protein in three formalin-fixed appendix specimens. This led to an estimated UK prevalence of vCJD infection of ∼1 in 4000, which remains the key evidence supporting current risk reduction measures to reduce iatrogenic transmission of vCJD prions in the UK. Confirmatory testing of these positives has been hampered by the inability to perform immunoblotting of formalin-fixed tissue. Animal transmission studies offer the potential for 'gold standard' confirmatory testing but are limited by both transmission barrier effects and known effects of fixation on scrapie prion titre in experimental models. Here we report the effects of fixation on brain and lymphoreticular human vCJD prions and comparative bioassay of two of the three prevalence study formalin-fixed, paraffin-embedded (FFPE) appendix specimens using transgenic mice expressing human prion protein (PrP). While transgenic mice expressing human PrP 129M readily reported vCJD prion infection after inoculation with frozen vCJD brain or appendix, and also FFPE vCJD brain, no infectivity was detected in FFPE vCJD spleen. No prion transmission was observed from either of the FFPE appendix specimens. The absence of detectable infectivity in fixed, known positive vCJD lymphoreticular tissue precludes interpreting negative transmissions from vCJD prevalence study appendix specimens. In this context, the Hilton et al study should continue to inform risk assessment pending the outcome of larger-scale studies on discarded surgical tissues and autopsy samples.
Hedegaard, Jakob; Thorsen, Kasper; Lund, Mette Katrine; Hein, Anne-Mette K.; Hamilton-Dutoit, Stephen Jacques; Vang, Søren; Nordentoft, Iver; Birkenkamp-Demtröder, Karin; Kruhøffer, Mogens; Hager, Henrik; Knudsen, Bjarne; Andersen, Claus Lindbjerg; Sørensen, Karina Dalsgaard; Pedersen, Jakob Skou; Ørntoft, Torben Falck; Dyrskjøt, Lars
Formalin-fixed, paraffin-embedded (FFPE) tissues are an invaluable resource for clinical research. However, nucleic acids extracted from FFPE tissues are fragmented and chemically modified making them challenging to use in molecular studies. We analysed 23 fresh-frozen (FF), 35 FFPE and 38 paired FF/FFPE specimens, representing six different human tissue types (bladder, prostate and colon carcinoma; liver and colon normal tissue; reactive tonsil) in order to examine the potential use of FFPE samples in next-generation sequencing (NGS) based retrospective and prospective clinical studies. Two methods for DNA and three methods for RNA extraction from FFPE tissues were compared and were found to affect nucleic acid quantity and quality. DNA and RNA from selected FFPE and paired FF/FFPE specimens were used for exome and transcriptome analysis. Preparations of DNA Exome-Seq libraries was more challenging (29.5% success) than that of RNA-Seq libraries, presumably because of modifications to FFPE tissue-derived DNA. Libraries could still be prepared from RNA isolated from two-decade old FFPE tissues. Data were analysed using the CLC Bio Genomics Workbench and revealed systematic differences between FF and FFPE tissue-derived nucleic acid libraries. In spite of this, pairwise analysis of DNA Exome-Seq data showed concordance for 70–80% of variants in FF and FFPE samples stored for fewer than three years. RNA-Seq data showed high correlation of expression profiles in FF/FFPE pairs (Pearson Correlations of 0.90 +/- 0.05), irrespective of storage time (up to 244 months) and tissue type. A common set of 1,494 genes was identified with expression profiles that were significantly different between paired FF and FFPE samples irrespective of tissue type. Our results are promising and suggest that NGS can be used to study FFPE specimens in both prospective and retrospective archive-based studies in which FF specimens are not available. PMID:24878701
Al-Attas, Asmaa; Assidi, Mourad; Al-Maghrabi, Jaudah; Dallol, Ashraf; Schulten, Hans-Juergen; Abu-Elmagd, Muhammad; Chaudhary, Adeel; Abuzenadah, Adel; Budowle, Bruce; Buhmeida, Abdelbaset; Al-Qahtani, Mohammed
To date, the conventional formalin-fixed, paraffin-embedded (FFPE) technique is the gold-standard for preserving histomorphology. Once FFPE tissues are stained, slides are routinely archived along with their blocks at biobanks/hospitals. However, the reuse of fixed and stained biospecimens as DNA source is not a common routine practice worldwide and, thus, indicates the need of studies to investigate the feasibility of extracting DNA from already immunohistochemistry (IHC) FFPE-stained slides and its possible reuse in subsequent downstream molecular analyses. FFPE IHC slides from colorectal cancer (CRC) patients were prepared and stored in the CEGMR Biobank. The workflow consists of digitalization of IHC stained slide's image, removing the slide cover-slip, crude dissection and DNA extraction. Following DNA quality assessment, mutation analysis of CTNNB1 and methylation profile of CDH1 were performed. High-quality DNA was obtained allowing 60% concordance between CDH1 methylation and membranous E-cadherin expression pattern. Clean CTNNB1 DNA chromatograms with evenly-spaced peaks were observed. This study is a proof of concept to recycle and reuse DNA from IHC stained slides with suitable concentration and integrity for further downstream molecular applications. These findings will enhance the pathologists' knowledge, attitudes and practices (KAP) towards the use of these biospecimens and support the implementation of this approach in clinical pathology practice. Therefore, the scientific community will benefit from the largest comprehensive database of human fully annotated FFPE biospecimens already available at their disposal in order to demystify the complexity and the heterogeneity of many challenging diseases and foster the transition towards precision medicine. Copyright© 2016, International Institute of Anticancer Research (Dr. John G. Delinasios), All rights reserved.
Full Text Available Bladder carcinogenesis is believed to follow two alternative pathways driven by the loss of chromosome 9 and the gain of chromosome 7, albeit other nonrandom copy number alterations (CNAs were identified. However, confirmation studies are needed since many aspects of this model remain unclear and considerable heterogeneity among cases has emerged. One of the purposes of this study was to evaluate the performance of a targeted test (UroVysion assay widely used for the detection of Transitional Cell Carcinoma (TCC of the bladder, in two different types of material derived from the same tumor. We compared the results of UroVysion test performed on Freshly Isolated interphasic Nuclei (FIN and on Formalin Fixed Paraffin Embedded (FFPE tissues from 22 TCCs and we didn't find substantial differences. A second goal was to assess the concordance between array-CGH profiles and the targeted chromosomal profiles of UroVysion assay on an additional set of 10 TCCs, in order to evaluate whether UroVysion is an adequately sensitive method for the identification of selected aneuploidies and nonrandom CNAs in TCCs. Our results confirmed the importance of global genomic screening methods, that is array based CGH, to comprehensively determine the genomic profiles of large series of TCCs tumors. However, this technique has yet some limitations, such as not being able to detect low level mosaicism, or not detecting any change in the number of copies for a kind of compensatory effect due to the presence of high cellular heterogeneity. Thus, it is still advisable to use complementary techniques such as array-CGH and FISH, as the former is able to detect alterations at the genome level not excluding any chromosome, but the latter is able to maintain the individual data at the level of single cells, even if it focuses on few genomic regions.
Zhang, Yuying; Wang, Xin-Ping; Perner, Sven; Bankfalvi, Agnes; Schlücker, Sebastian
Immunohistochemical analysis of formalin-fixed paraffin-embedded (FFPE) tissues provides important diagnostic and prognostic information in pathology. Metal nanoparticles (NPs) and, in particular, surface-enhanced Raman scattering (SERS) nanotags as a new class of labeling reagents are promising to be used for multiplexed protein profiling on tissue sections. However, nonspecific binding of NPs onto the tissue specimens greatly hampers their clinical applications. In this study, we found that the antigen retrieval method strongly influences the extent of nonspecific binding of the antibody-SERS NP conjugates to the tissue. Our SERS labels comprised ca. 70 nm Au nanostars coated with ethylene glycol-modified Raman reporter molecules for hydrophilic stabilization and subsequent covalent bioconjugation to antibodies. We systematically investigated the influence of heat- and protease-induced epitope retrieval (HIER and PIER, respectively) on the immunostaining quality of prostate-specific antigen (PSA) on human prostate tissue sections. The best staining results were obtained with PIER. Pretreatment of the tissue sections by HIER led to selective but nonspecific adsorption of the antibody-Au nanostar conjugates onto epithelial cells, while enzymatic treatment within PIER did not. In addition to gold nanostars, also other types of metal NPs with different shapes and sizes (including ca. 20 nm quasi-spherical Au NPs and ca. 60 nm quasi-spherical Au/Ag nanoshells) as well as tissue sections from different organs (including prostate and breast) were tested; in each case the same tendency was observed, i.e., PIER yielded better results than HIER. Therefore, we recommend PIER for future NP-based tissue immunostaining such as immuno-SERS microscopy. Alternatively, for antigens that can only be unmasked by heating, PEGylation of the NPs is recommended to avoid nonspecific binding.
Finne, Kenneth; Vethe, Heidrun; Skogstrand, Trude; Leh, Sabine; Dahl, Tone D; Tenstad, Olav; Berven, Frode S; Reed, Rolf K; Vikse, Bjørn Egil
It is well known that hypertension may cause glomerular damage, but the molecular mechanisms involved are still incompletely understood. In the present study, we used formalin-fixed paraffin-embedded (FFPE) tissue to investigate changes in the glomerular proteome in the non-clipped kidney of two-kidney one-clip (2K1C) hypertensive rats, with special emphasis on the glomerular filtration barrier. 2K1C hypertension was induced in 6-week-old Wistar Hannover rats (n = 6) that were sacrificed 23 weeks later and compared with age-matched sham-operated controls (n = 6). Tissue was stored in FFPE tissue blocks and later prepared on tissue slides for laser microdissection. Glomeruli without severe morphological damage were isolated, and the proteomes were analysed using liquid chromatography-tandem mass spectrometry. 2K1C glomeruli showed reduced abundance of proteins important for slit diaphragm complex, such as nephrin, podocin and neph1. The podocyte foot process had a pattern of reduced abundance of transmembrane proteins but unchanged abundances of the podocyte cytoskeletal proteins synaptopodin and α-actinin-4. Lower abundance of important glomerular basement membrane proteins was seen. Possible glomerular markers of damage with increased abundance in 2K1C were transgelin, desmin and acyl-coenzyme A thioesterase 1. Microdissection and tandem mass spectrometry could be used to investigate the proteome of isolated glomeruli from FFPE tissue. Glomerular filtration barrier proteins had reduced abundance in the non-clipped kidney of 2K1C hypertensive rats. © The Author 2014. Published by Oxford University Press on behalf of ERA-EDTA.
Gade, Lalitha; Hurst, Steven; Balajee, S Arunmozhi; Lockhart, Shawn R; Litvintseva, Anastasia P
Molecular methods of detection based on DNA-sequencing of the internal transcribed spacer 1 and 2 (ITS1 and ITS2) or 5΄ end region of 28S (D1-D2 region) of ribosomal RNA gene (rDNA) have been used extensively for molecular identification and detection of fungal infections. However, these regions are not always informative for identification of mucormycetes and other rare fungal pathogens as they often contain large introns, heterogenic regions, and/or cannot be PCR-amplified using broad range fungal PCR primers. In addition, because of the difficulties of recovering intact fungal DNA from human specimens, smaller regions of DNA are more useful for the direct detection of fungal DNA in tissues and fluids. In this study, we investigated the utility of 12F/13R PCR primers targeting a 200-230 bp region of the extended 28S region of rDNA for molecular identification of fungal DNA in formalin fixed paraffin embedded tissues and other clinical specimens. We demonstrated that this region can be successfully used for identification of all genera and some species of clinically relevant mucormycetes, as well as other medically important fungi, such as Aspergillus, Fusarium, Coccidioides, and Cryptococcus. We also demonstrated that PCR amplification and direct sequencing of the extended 28S region of rDNA was more sensitive compared to targeting the ITS2 region, as we were able to detect and identify mucormycetes and other fungal pathogens in tissues from patients with histopathological and/or culture evidence of fungal infections that were negative with PCR using ITS-specific primers. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.
Full Text Available The demand for nucleic acid and protein derivatives from formalin-fixed paraffin-embedded (FFPE tissue has greatly increased due to advances in extraction and purification methods, making these derivatives available for numerous genomic and proteomic platforms. Previously, DNA, RNA, microRNA (miRNA, or protein derived from FFPE tissue blocks were considered "unfit" for such platforms, as the process of tissue immobilization by FFPE resulted in cross-linked, fragmented, and chemically modified macromolecules. We conducted a systematic examination of nucleic acids and proteins co-extracted from 118 FFPE blocks sampled from the AIDS and Cancer Specimen Resource (ACSR at The George Washington University after stratification by storage duration and the three most common tumor tissue types at the ACSR (adenocarcinoma, squamous cell carcinoma, and papillary carcinoma. DNA, RNA, miRNA, and protein could be co-extracted from 98% of the FFPE blocks sampled, with DNA and miRNA "fit" for diverse genomic purposes including sequencing. While RNA was the most labile of the FFPE derivatives, especially when assessed by RNA integrity number (RIN, it was still "fit" for genomic methods that use smaller sequence lengths, e.g., quantitative PCR. While more than half of the protein derivatives were fit for proteomic purposes, our analyses indicated a significant interaction effect on the absorbance values for proteins derived from FFPE, implying that storage duration may affect protein derivatives differently by tumor tissue type. The mean absorbance value for proteins derived from more recently stored FFPE was greater than protein derived from older FFPE, with the exception of adenocarcinoma tissue. Finally, the fitness of one type of derivative was weakly associated with the fitness of derivatives co-extracted from the same FFPE block. The current study used several novel quality assurance approaches and metrics to show that archival FFPE tissue blocks are a
Van Allen, Eliezer M; Wagle, Nikhil; Stojanov, Petar; Perrin, Danielle L; Cibulskis, Kristian; Marlow, Sara; Jane-Valbuena, Judit; Friedrich, Dennis C; Kryukov, Gregory; Carter, Scott L; McKenna, Aaron; Sivachenko, Andrey; Rosenberg, Mara; Kiezun, Adam; Voet, Douglas; Lawrence, Michael; Lichtenstein, Lee T; Gentry, Jeff G; Huang, Franklin W; Fostel, Jennifer; Farlow, Deborah; Barbie, David; Gandhi, Leena; Lander, Eric S; Gray, Stacy W; Joffe, Steven; Janne, Pasi; Garber, Judy; MacConaill, Laura; Lindeman, Neal; Rollins, Barrett; Kantoff, Philip; Fisher, Sheila A; Gabriel, Stacey; Getz, Gad; Garraway, Levi A
Translating whole-exome sequencing (WES) for prospective clinical use may have an impact on the care of patients with cancer; however, multiple innovations are necessary for clinical implementation. These include rapid and robust WES of DNA derived from formalin-fixed, paraffin-embedded tumor tissue, analytical output similar to data from frozen samples and clinical interpretation of WES data for prospective use. Here, we describe a prospective clinical WES platform for archival formalin-fixed, paraffin-embedded tumor samples. The platform employs computational methods for effective clinical analysis and interpretation of WES data. When applied retrospectively to 511 exomes, the interpretative framework revealed a 'long tail' of somatic alterations in clinically important genes. Prospective application of this approach identified clinically relevant alterations in 15 out of 16 patients. In one patient, previously undetected findings guided clinical trial enrollment, leading to an objective clinical response. Overall, this methodology may inform the widespread implementation of precision cancer medicine.
Full Text Available Hereditary breast cancer constitutes only 5–10% of all breast cancer cases and is characterized by strong family history of breast and/or other associated cancer types. Only ~25% of hereditary breast cancer cases carry a mutation in BRCA1 or BRCA2 gene, while mutations in other rare high and moderate-risk genes and common low penetrance variants may account for additional 20% of the cases. Thus the majority of cases are still unaccounted for and designated as BRCAX tumors. MicroRNAs are small non-coding RNAs that play important roles as regulators of gene expression and are deregulated in cancer. To characterize hereditary breast tumors based on their miRNA expression profiles we performed global microarray miRNA expression profiling on a retrospective cohort of 80 FFPE breast tissues, including 66 hereditary breast tumors (13 BRCA1, 10 BRCA2 and 43 BRCAX, 10 sporadic breast carcinomas and 4 normal breast tissues, using Exiqon miRCURY LNA™ microRNA Array v.11.0. Here we describe in detail the miRNA microarray expression data and tumor samples used for the study of BRCAX tumor heterogeneity (Tanic et al., 2013 and biomarkers associated with positive BRCA1/2 mutation status (Tanic et al., 2014. Additionally, we provide the R code for data preprocessing and quality control.
Johansen, Isik Somuncu; Thomsen, Vibeke Østergaard; Forsgren, Arne
Rapid, reliable diagnosis of tuberculosis is essential to initiate correct treatment, avoid severe complications, and prevent transmission. Conventional microbiological methods may not be an option if samples are formalin-fixed and paraffin-embedded (FFPE) for histopathological examination....... With the demonstration of necrotizing granulomatous inflammation, tuberculosis becomes an important differential diagnosis, although it was not initially suspected. Following paraffin extraction, BDProbeTec ET strand displacement amplification for detection of Mycobacterium tuberculosis complex (MTC) was applied to 47...
Wakabayashi, Koichi; Mori, Fumiaki; Kakita, Akiyoshi; Takahashi, Hitoshi; Tanaka, Shinya; Utsumi, Jun; Sasaki, Hidenao
MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression. Recently, we have shown that informative miRNA data can be derived from archived formalin-fixed paraffin-embedded (FFPE) samples from postmortem cases of amyotrophic lateral sclerosis and normal controls. miRNA analysis has now been performed on FFPE samples from affected brain regions in patients with multiple system atrophy (MSA) and the same areas in neurologically normal controls. We evaluated 50 samples from patients with MSA (n=13) and controls (n=13). Twenty-six samples were selected for miRNA analysis on the basis of the criteria reported previously: (i) a formalin fixation time of less than 4 weeks, (ii) a total RNA yield per sample of more than 500ng, and (iii) sufficient quality of the RNA electrophoresis pattern. These included 11 cases of MSA and 5 controls. Thus, the success rate for analysis of RNA from FFPE samples was 52% (26 of 50). For MSA, a total of 395 and 383 miRNAs were identified in the pons and cerebellum, respectively; 5 were up-regulated and 33 were down-regulated in the pons and 5 were up-regulated and 18 were down-regulated in the cerebellum. Several miRNAs down-regulated in the pons (miR-129-2-3p and miR-129-5p) and cerebellum (miR-129-2-3p, miR-129-5p and miR-132-3p) had already been identified in frozen cerebellum from MSA patients. These findings suggest that archived FFPE postmortem samples can be a valuable source for miRNA profiling in MSA. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Bak, Stine T; Staunstrup, Nicklas H; Starnawska, Anna; Daugaard, Tina F; Nyengaard, Jens R; Nyegaard, Mette; Børglum, Anders; Mors, Ole; Dorph-Petersen, Karl-Anton; Nielsen, Anders L
We here characterize the usability of archival formalin-fixed paraffin-embedded (FFPE) brain tissue as a resource for genetic and DNA methylation analyses with potential relevance for brain-manifested diseases. We analyzed FFPE samples from The Brain Collection, Aarhus University Hospital Risskov, Denmark (AUBC), constituting 9479 formalin-fixated brains making it one of the largest collections worldwide. DNA extracted from brain FFPE tissue blocks was interrogated for quality and usability in genetic and DNA methylation analyses by different molecular techniques. Overall, we found that DNA quality was inversely correlated with storage time and DNA quality was insufficient for Illumina methylation arrays; data from methylated DNA immunoprecipitation, clonal bisulfite sequencing, and pyrosequencing of BDNF and ST6GALNAC1 suggested that the original methylation pattern is indeed preserved. Proof-of-principle experiments predicting sex based on the methylation status of the X-inactivated SLC9A7 gene, or genotype differences of the Y and X chromosomes, showed consistency between predicted and actual sex for a subset of FFPE samples. In conclusion, even though DNA from FFPE samples is of low quality and technically challenging, it is likely that a subset of samples can provide reliable data given that the methodology used is designed for small DNA fragments. We propose that simple PCR-based quality control experiments at the genetic and DNA methylation level, carried out at the beginning of any given project, can be used to enrich for the best-performing FFPE samples. The apparent preservation of genetic and DNA methylation patterns in archival FFPE samples may bring along new perspectives for the identification of genetic and epigenetic changes associated with brain-manifested diseases.
Kalmár, Alexandra; Péterfia, Bálint; Hollósi, Péter; Wichmann, Barnabás; Bodor, András; Patai, Árpád V; Schöller, Andrea; Krenács, Tibor; Tulassay, Zsolt; Molnár, Béla
We aimed to test the applicability of formalin-fixed and paraffin-embedded (FFPE) tissue samples for gene specific DNA methylation analysis after using two commercially available DNA isolation kits. Genomic DNA was isolated from 5 colorectal adenocarcinomas and 5 normal adjacent tissues from "recent", collected within 6 months, and "archived", collected more than 5 years ago, FFPE tissues using either High Pure FFPET DNA Isolation kit or QIAamp DNA FFPE Tissue kit. DNA methylation analysis of MAL, SFRP1 and SFRP2 genes, known to be hypermethylated in CRC, was performed using methylation-sensitive high resolution melting (MS-HRM) analysis and sequencing. QIAamp (Q) method resulted in slightly higher recovery in archived (HP: 1.22 ± 3.18 μg DNA; Q: 3.00 ± 4.04 μg DNA) and significantly (p < 0.05) higher recovery in recent samples compared to High Pure method (HP) (HP: 4.10 ± 2.91 μg DNA; Q: 11.51 ± 7.50 μg DNA). Both OD260/280 and OD260/230 ratios were lower, but still high in the High Pure isolated archived and recent samples compared to those isolated with QIAamp. Identical DNA methylation patterns were detected for all 3 genes tested by MS-HRM with both isolation kits in the recent group. However, despite of higher DNA recovery in QIAamp slightly more reproducible methylation results were obtained from High Pure isolated archived samples. Sequencing confirmed DNA hypermethylation in CRCs. In conclusion, reproducible DNA methylation patterns were obtained from recent samples using both isolation kits. However, long term storage may affect the reliability of the results leading to moderate differences between the efficiency of isolation kits.
Nitta, Hiroaki; Kishimoto, Jiro; Grogan, Thomas M
Recently, an automated mRNA in situ hybridization application was introduced for the Ventana Discovery instrument. The application was designed so that all necessary steps from baking through signal detection were completed within 1 day on the instrument. We applied this technology for visualizing the expression site of versican in formalin-fixed mouse skin paraffin tissue sections. Our focus of this study was to demonstrate the effects of protease digestion or heating pretreatment, termed cell conditioning, on the hybridization signal using a well characterized versican antisense riboprobe. Paraffin sections were automatically deparaffinized, fixed, and acid-treated. Then, the tissue sections were subjected to protease digestion alone (3 strengths), cell conditioning alone, or the combination of cell conditioning and protease digestion. Hybridization was performed with digoxigenin-labeled versican antisense probe (20 ng/slide) for 6 hours, and the signal was detected using a Nitro blue Tetrazolium chloride 5-Bromo-4-cloro-3-indolyl phosphate toluidine salt (NBT/BCLIP) substrate solution for 3 hours on the instrument. Cell conditioning alone did not produce any signal, whereas the highest strength of protease digestion produced noticeable background staining. However, when cell conditioning and mild protease digestion were combined, the signal for versican mRNA was clearly demonstrated in the hair papilla region. Thus, we demonstrated the effects of the cell conditioning step followed by mild protease digestion for enhancing the mRNA target staining compared with protease digestion or the cell conditioning step alone. We verified that the automated in situ hybridization process was applicable for formalin-fixed mouse skin paraffin sections and that the automated 1-day protocol is simple and reproducible. The precise control of automation allows fine tuning of temperature and enzyme dose to find the optimized assay condition for the signal to noise ratio and
Full Text Available The requirement for large amounts of good quality DNA for whole-genome applications prohibits their use for small, laser capture micro-dissected (LCM, and/or rare clinical samples, which are also often formalin-fixed and paraffin-embedded (FFPE. Whole-genome amplification of DNA from these samples could, potentially, overcome these limitations. However, little is known about the artefacts introduced by amplification of FFPE-derived DNA with regard to genotyping, and subsequent copy number and loss of heterozygosity (LOH analyses. Using a ligation adaptor amplification method, we present data from a total of 22 Affymetrix SNP 6.0 experiments, using matched paired amplified and non-amplified DNA from 10 LCM FFPE normal and dysplastic oral epithelial tissues, and an internal method control. An average of 76.5% of SNPs were called in both matched amplified and non-amplified DNA samples, and concordance was a promising 82.4%. Paired analysis for copy number, LOH, and both combined, showed that copy number changes were reduced in amplified DNA, but were 99.5% concordant when detected, amplifications were the changes most likely to be 'missed', only 30% of non-amplified LOH changes were identified in amplified pairs, and when copy number and LOH are combined ∼50% of gene changes detected in the unamplified DNA were also detected in the amplified DNA and within these changes, 86.5% were concordant for both copy number and LOH status. However, there are also changes introduced as ∼20% of changes in the amplified DNA are not detected in the non-amplified DNA. An integrative network biology approach revealed that changes in amplified DNA of dysplastic oral epithelium localize to topologically critical regions of the human protein-protein interaction network, suggesting their functional implication in the pathobiology of this disease. Taken together, our results support the use of amplification of FFPE-derived DNA, provided sufficient samples are used
Hunt, Nicholas J; Phillips, Leo; Waters, Karen A; Machaalani, Rita
Matrix assisted laser desorption/ionisation imaging mass spectrometry (MALDI-IMS) has not previously been utilised to examine sudden infant death syndrome (SIDS). This study aimed to optimise MALDI IMS for use on archived formalin-fixed-paraffin-embedded human infant medulla tissue (n=6, controls; n=6, SIDS) to evaluate differences between multiple nuclei of the medulla by using high resolution IMS. Profiles were compared between SIDS and age/sex matched controls. LC-MALDI identified 55 proteins based on 321 peptides across all samples; 286 peaks were found using IMS, corresponding to these 55 proteins that were directly compared between controls and SIDS. Control samples were used to identify common peptides for neuronal/non-neuronal structures allowing identification of medullary regions. In SIDS, abnormal expression patterns of 41 peptides (p≤0.05) corresponding to 9 proteins were observed; these changes were confirmed with immunohistochemistry. The protein abnormalities varied amongst nuclei, with the majority of variations in the raphe nuclei, hypoglossal and pyramids. The abnormal proteins are not related to a previously identified neurological disease pathway but consist of developmental neuronal/glial/axonal growth, cell metabolism, cyto-architecture and apoptosis components. This suggests that SIDS infants have abnormal neurological development in the raphe nuclei, hypoglossal and pyramids of the brainstem, which may contribute to the pathogenesis of SIDS. This study is the first to perform an imaging mass spectrometry investigation in the human brainstem and also within sudden infant death syndrome (SIDS). LC MALDI and MALDI IMS identified 55 proteins based on 285 peptides in both control and SIDS tissue; with abnormal expression patterns present for 41/285 and 9/55 proteins in SIDS using IMS. The abnormal proteins are critical for neurological development; with the impairment supporting the hypothesis that SIDS may be due to delayed neurological
Full Text Available Abstract Background Formalin-fixed, paraffin-embedded (FFPE samples are a highly desirable resource for epigenetic studies, but there is no suitable platform to assay genome-wide methylation in these widely available resources. Recently, Thirlwell et al. (2010 have reported a modified ligation-based DNA repair protocol to prepare FFPE DNA for the Infinium methylation assay. In this study, we have tested the accuracy of methylation data obtained with this modification by comparing paired fresh-frozen (FF and FFPE colon tissue (normal and tumor from colorectal cancer patients. We report locus-specific correlation and concordance of tumor-specific differentially methylated loci (DML, both of which were not previously assessed. Methods We used Illumina's Infinium Methylation 27K chip for 12 pairs of FF and 12 pairs of FFPE tissue from tumor and surrounding healthy tissue from the resected colon of the same individual, after repairing the FFPE DNA using Thirlwell's modified protocol. Results For both tumor and normal tissue, overall correlation of β values between all loci in paired FF and FFPE was comparable to previous studies. Tissue storage type (FF or FFPE was found to be the most significant source of variation rather than tissue type (normal or tumor. We found a large number of DML between FF and FFPE DNA. Using ANOVA, we also identified DML in tumor compared to normal tissue in both FF and FFPE samples, and out of the top 50 loci in both groups only 7 were common, indicating poor concordance. Likewise, while looking at the correlation of individual loci between FFPE and FF across the patients, less than 10% of loci showed strong correlation (r ≥ 0.6. Finally, we checked the effect of the ligation-based modification on the Infinium chemistry for SNP genotyping on an independent set of samples, which also showed poor performance. Conclusion Ligation of FFPE DNA prior to the Infinium genome-wide methylation assay may detect a reasonable
Movassaghi, Masoud; Shabihkhani, Maryam; Hojat, Seyed A; Williams, Ryan R; Chung, Lawrance K; Im, Kyuseok; Lucey, Gregory M; Wei, Bowen; Mareninov, Sergey; Wang, Michael W; Ng, Denise W; Tashjian, Randy S; Magaki, Shino; Perez-Rosendahl, Mari; Yang, Isaac; Khanlou, Negar; Vinters, Harry V; Liau, Linda M; Nghiemphu, Phioanh L; Lai, Albert; Cloughesy, Timothy F; Yong, William H
Commercial targeted genomic profiling with next generation sequencing using formalin-fixed paraffin embedded (FFPE) tissue has recently entered into clinical use for diagnosis and for the guiding of therapy. However, there is limited independent data regarding the accuracy or robustness of commercial genomic profiling in gliomas. As part of patient care, FFPE samples of gliomas from 71 patients were submitted for targeted genomic profiling to one commonly used commercial vendor, Foundation Medicine. Genomic alterations were determined for the following grades or groups of gliomas; Grade I/II, Grade III, primary glioblastomas (GBMs), recurrent primary GBMs, and secondary GBMs. In addition, FFPE samples from the same patients were independently assessed with conventional methods such as immunohistochemistry (IHC), Quantitative real-time PCR (qRT-PCR), or Fluorescence in situ hybridization (FISH) for three genetic alterations: IDH1 mutations, EGFR amplification, and EGFRvIII expression. A total of 100 altered genes were detected by the aforementioned targeted genomic profiling assay. The number of different genomic alterations was significantly different between the five groups of gliomas and consistent with the literature. CDKN2A/B, TP53, and TERT were the most common genomic alterations seen in primary GBMs, whereas IDH1, TP53, and PIK3CA were the most common in secondary GBMs. Targeted genomic profiling demonstrated 92.3%-100% concordance with conventional methods. The targeted genomic profiling report provided an average of 5.5 drugs, and listed an average of 8.4 clinical trials for the 71 glioma patients studied but only a third of the trials were appropriate for glioma patients. In this limited comparison study, this commercial next generation sequencing based-targeted genomic profiling showed a high concordance rate with conventional methods for the 3 genetic alterations and identified mutations expected for the type of glioma. While it may not be feasible to
Full Text Available BACKGROUND: Retrospective studies of archived human specimens, with known clinical follow-up, are used to identify predictive and prognostic molecular markers of disease. Due to biochemical differences, however, formalin-fixed paraffin-embedded (FFPE DNA and RNA have generally been extracted separately from either different tissue sections or from the same section by dividing the digested tissue. The former limits accurate correlation whilst the latter is impractical when utilizing rare or limited archived specimens. PRINCIPAL FINDINGS: For effective recovery of genomic DNA and total RNA from a single FFPE specimen, without splitting the proteinase-K digested tissue solution, we optimized a co-extraction method by using TRIzol and purifying DNA from the lower aqueous and RNA from the upper organic phases. Using a series of seven different archived specimens, we evaluated the total amounts of genomic DNA and total RNA recovered by our TRIzol-based co-extraction method and compared our results with those from two commercial kits, the Qiagen AllPrep DNA/RNA FFPE kit, for co-extraction, and the Ambion RecoverAll™ Total Nucleic Acid Isolation kit, for separate extraction of FFPE-DNA and -RNA. Then, to accurately assess the quality of DNA and RNA co-extracted from a single FFPE specimen, we used qRT-PCR, gene expression profiling and methylation assays to analyze microRNAs, mRNAs, and genomic DNA recovered from matched fresh and FFPE MCF10A cells. These experiments show that the TRIzol-based co-extraction method provides larger amounts of FFPE-DNA and -RNA than the two other methods, and particularly provides higher quality microRNAs and genomic DNA for subsequent molecular analyses. SIGNIFICANCE: We determined that co-extraction of genomic DNA and total RNA from a single FFPE specimen is an effective recovery approach to obtain high-quality material for parallel molecular and high-throughput analyses. Our optimized approach provides the option of
Liu, Christina; Lin, Juan; Ye, Kenny; Kim, Ryung; Hazan, Rachel; Rohan, Thomas; Fineberg, Susan; Loudig, Olivier
Background Retrospective studies of archived human specimens, with known clinical follow-up, are used to identify predictive and prognostic molecular markers of disease. Due to biochemical differences, however, formalin-fixed paraffin-embedded (FFPE) DNA and RNA have generally been extracted separately from either different tissue sections or from the same section by dividing the digested tissue. The former limits accurate correlation whilst the latter is impractical when utilizing rare or limited archived specimens. Principal Findings For effective recovery of genomic DNA and total RNA from a single FFPE specimen, without splitting the proteinase-K digested tissue solution, we optimized a co-extraction method by using TRIzol and purifying DNA from the lower aqueous and RNA from the upper organic phases. Using a series of seven different archived specimens, we evaluated the total amounts of genomic DNA and total RNA recovered by our TRIzol-based co-extraction method and compared our results with those from two commercial kits, the Qiagen AllPrep DNA/RNA FFPE kit, for co-extraction, and the Ambion RecoverAll™ Total Nucleic Acid Isolation kit, for separate extraction of FFPE-DNA and -RNA. Then, to accurately assess the quality of DNA and RNA co-extracted from a single FFPE specimen, we used qRT-PCR, gene expression profiling and methylation assays to analyze microRNAs, mRNAs, and genomic DNA recovered from matched fresh and FFPE MCF10A cells. These experiments show that the TRIzol-based co-extraction method provides larger amounts of FFPE-DNA and –RNA than the two other methods, and particularly provides higher quality microRNAs and genomic DNA for subsequent molecular analyses. Significance We determined that co-extraction of genomic DNA and total RNA from a single FFPE specimen is an effective recovery approach to obtain high-quality material for parallel molecular and high-throughput analyses. Our optimized approach provides the option of collecting DNA, which
Watanabe, Mototsugu; Hashida, Shinsuke; Yamamoto, Hiromasa; Matsubara, Takehiro; Ohtsuka, Tomoaki; Suzawa, Ken; Maki, Yuho; Soh, Junichi; Asano, Hiroaki; Tsukuda, Kazunori; Toyooka, Shinichi; Miyoshi, Shinichiro
Techniques for the extraction and use of nucleic acids from formalin-fixed and paraffin-embedded (FFPE) tissues, preserved over long time periods in libraries, have been developed. However, DNA extracted from FFPE tissues is generally damaged, and long-term storage may affect DNA quality. Therefore, it is important to elucidate the effect of long-term storage on FFPE tissues and evaluate the techniques used to extract DNA from them. In the present study, the yield, purity, and integrity of DNA in FFPE tissue samples was evaluated. Two DNA extraction techniques were used: A silica-binding DNA collection method using QIAamp DNA FFPE Tissue kit (QIA) and a total tissue DNA collection method using a WaxFree DNA extraction kit (WAX). A total of 25 FFPE tissues from lung adenocarcinomas were studied, which had been surgically resected and fixed at Okayama University Hospital prior to examination and subsequent storage at room temperature for 0.5, 3, 6, 9 and 12 years. Extracted DNA was quantified using ultraviolet absorbance, fluorescent dye, and quantitative polymerase chain reaction (qPCR). The quality of the DNA was defined by the absorbance ratio of 260 to 280 nm (A260/280) and Q-score, which is the quantitative value of qPCR product size ratio. The results demonstrated that the yield of total DNA extracted using WAX was significantly greater than when QIA was used (PDNA extracted using WAX included more contaminants and was significantly more fragmented compared with DNA extracted using QIA (PDNA yield or DNA purity, although it did significantly contribute to increased DNA degradation for both QIA and WAX extraction (QIA P=0.02, WAX P=0.03; 0.5 years vs. 3 years, QIA Pextraction methods are viable depending on whether high yield or high quality of extracted DNA is required. However, due to the increased degradation with age, storage time limits the available DNA in FFPE tissues regardless of the extraction method.
Wen, Xianyu; Jeong, Seorin; Kim, Younghoon; Bae, Jeong Mo; Cho, Nam Yun; Kim, Jung Ho; Kang, Gyeong Hoon
Formalin-fixed, paraffin-embedded (FFPE) tissues are important resources for profiling DNA methylation changes and for studying a variety of diseases. However, formalin fixation introduces inter-strand crosslinking, which might cause incomplete bisulfite conversion of unmethylated cytosines, which might lead to falsely elevated measurements of methylation levels in pyrosequencing assays. Long interspersed nucleotide element-1 (LINE-1) is a major constituent of repetitive transposable DNA elements, and its methylation is referred to correlates with global DNA methylation. To identify whether formalin fixation might impact the measured values of methylation in LINE-1 repetitive elements and whether prolonged heat-induced denaturation of DNA might reduce the artificial increases in measured values caused by formalin fixation, we analyzed paired fresh-frozen (FF) and FFPE xenograft tissue samples for their methylation levels in LINE-1 using a pyrosequencing assay. To further confirm the effect of a heating step in the measurement of LINE-1 or single gene methylation levels, we analyzed FFPE tissue samples of gastric cancer and colorectal cancer for their methylation status in LINE-1 and eight single genes, respectively. Formalin fixation led to an increase in the measured values of LINE-1 methylation regardless of the duration of fixation. Prolonged heating of the DNA at 95 °C for 30 min before bisulfite conversion was found (1) to decrease the discrepancy in the measured values between the paired FF and FFPE tissue samples, (2) to decrease the standard deviation of the measured value of LINE-1 methylation levels in FFPE tissue samples of gastric cancer, and (3) to improve the performance in the measurement of single gene methylation levels in FFPE tissue samples of colorectal cancer. Formalin fixation leads to artificial increases in the measured values of LINE-1 methylation, and the application of prolonged heating of DNA samples decreases the discrepancy in the
Frickmann, Hagen; Tenner-Racz, Klara; Eggert, Petra; Schwarz, Norbert G; Poppert, Sven; Tannich, Egbert; Hagen, Ralf M
We report on the reliability of polymerase chain reaction (PCR) for the detection of Entamoeba histolytica from formalin-fixed, paraffin-embedded tissue in comparison with microscopy and have determined predictors that may influence PCR results. E. histolytica-specific and Entamoeba dispar-specific real-time PCR and microscopy from adjacent histologic sections were performed using a collection of formalin-fixed, paraffin-embedded tissue specimens obtained from patients with invasive amebiasis. Specimens had been collected during the previous 4 decades. Association of sample age, parasite density, and reliability of PCR was analyzed. E. histolytica PCR was positive in 20 of 34 biopsies (58.8%); 2 of these 20 were microscopically negative for amebae in neighboring tissue sections. PCR was negative in 9 samples with visible amebae in neighboring sections and in 5 samples without visible parasites in neighboring sections. PCR was negative in all specimens that were older than 3 decades. Low parasite counts and sample ages older than 20 years were predictors for false-negative PCR results. All samples were negative for E. dispar DNA. PCR is suitable for the detection of E. histolytica in formalin-fixed, paraffin-embedded tissue samples that are younger than 2 decades and that contain intermediate to high parasite numbers. Negative results in older samples were due to progressive degradation of DNA over time as indicated by control PCRs targeting the human 18S rRNA gene. Moreover, our findings support previous suggestions that only E. histolytica but not E. dispar is responsible for invasive amebiasis.
Tewari, Deepanker; Kim, Hyun; Feria, Willard; Russo, Brigite; Acland, Helen
West Nile virus (WNV) RNA was quantified in WNV infected crows and horses with the help of a real-time reverse transcriptase-PCR assay. A 5' nuclease assay, based on NS5 gene detection with a fluorescent probe was used for quantifying WNV RNA using formalin fixed paraffin embedded tissue specimens. Quantitative detection of WNV RNA showed the presence of a higher amount of the viral RNA in crow tissues compared to equine tissues and these results correlated well with the detection of WNV antigen by immunostaining. In crows, the highest amount of virus was seen in the intestine and in horses in the brain.
Full Text Available Abstract We describe a novel antigen-retrieval method using a micro-sized chamber for mass spectrometry (MS analysis to identify proteins that are preferentially eluted from formalin-fixed paraffin-embedded (FFPE samples. This approach revealed that heat-induced antigen retrieval (HIAR from an FFPE sample fixed on a glass slide not only improves protein identification, but also facilitates preferential elution of protein subsets corresponding to the properties of antigen-retrieval buffers. Our approach may contribute to an understanding of the mechanism of HIAR.
Thomas W Powers
Full Text Available A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE tissues. Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers.
Powers, Thomas W; Neely, Benjamin A; Shao, Yuan; Tang, Huiyuan; Troyer, Dean A; Mehta, Anand S; Haab, Brian B; Drake, Richard R
A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers.
Del Chierico, F; Onori, M; Di Bella, S; Bordi, E; Petrosillo, N; Menichella, D; Cacciò, S M; Callea, F; Putignani, L
Nine cases of cryptosporidiosis co-infections in AIDS patients were clinically categorised into severe (patients 1, 3, 8 and 9), moderate (patients 4 and 5) and mild (patients 2, 6 and 7). Formalin-fixed faecal specimens from these patients were treated to obtain high quality DNA competent for amplification and sequencing of the 60-kDa glycoprotein (GP60) gene. Sequence analysis revealed that one patient was infected with Cryptosporidium hominis whereas the remaining eight patients were infected with C. parvum. Interestingly, the patients showing severe cryptosporidiosis harboured two subtypes within the C. parvum allelic family IIc (IIcA5G3 and IIcA5G3R2), whereas patients with moderate or mild infections showed various subtypes of the C. parvum allelic family IIa (IIaA14G2R1, IIaA15G2R1, IIaA17G3R1 and IIaA18G3R1). DNA extraction and genotyping of Cryptosporidium spp. is a challenging task on formalin-fixed stool samples, whose diagnostic outcome is age-dependent. The method herein reported represents a step forward routine diagnosis and improves epidemiology of HIV-related clinical cases. Due to the need to elucidate genetic richness of Cryptosporidium human isolates, this approach represents a useful tool to correlate individual differences in symptoms to subgenotyping lineages.
Glud, Martin; Klausen, Mikkel; Gniadecki, Robert
surgical specimens are formalin fixed and paraffin embedded (FFPE). To explore whether FFPE material would be suitable for miRNA profiling in melanocytic lesions, we compared miRNA expression patterns in FFPE versus fresh frozen samples, obtained from 15 human melanocytic nevi. Out of microarray data, we...... identified 84 miRNAs that were expressed in both types of samples and represented an miRNA profile of melanocytic nevi. Our results showed a high correlation in miRNA expression (Spearman r-value of 0.80) between paired FFPE and fresh frozen material. The data were further validated by quantitative RT......-PCR. In conclusion, FFPE specimens of melanocytic lesions are suitable as a source for miRNA microarray profiling....
Tramm, Trine; Hennig, Guido; Kyndi, Marianne
Gene expression analysis on messenger RNA (mRNA) purified from formalin-fixed, paraffin-embedded tissue is increasingly used for research purposes. Tissue heterogeneity may question specificity and interpretation of results from mRNA isolated from a whole slide section, and thresholds for minimal...... tumor content in the paraffin block or macrodissection are used to avoid contamination from non-neoplastic tissue. The aim was to test if mRNA from tissue surrounding breast cancer affected quantification of estrogen receptor α (ESR1), progesterone receptor (PGR) and human epidermal growth factor...... receptor 2 (ERBB2), by comparing gene expression from whole slide and tumor-enriched sections, and correlating gene expression from whole slide sections with corresponding immunohistochemistry. Gene expression, based on mRNA extracted from a training set (36 paraffin blocks) and two validation sets (133...
Christensen, M; Funder, A D; Bendix, K
AIM: To compare clonal T cell receptor gamma (TCRgamma) gene rearrangements in frozen and formalin-fixed paraffin wax-embedded (FFPE) tissue, using capillary electrophoresis for use in diagnostics, as T cell lymphomas may be difficult to diagnose by conventional methods.METHODS: The DNA for PCR...... was extracted from frozen and FFPE tissue, cell lines and blood. PCR primers Vgamma1-8, Vgamma9, Vgamma10 or Vgamma11 (5' end labelled) combined with a mixture of JgammaP1/JgammaP/JgammaP2/Jgamma2 (unlabelled) were used. Monoclonal cases were sequenced and clonality, reproducibility, sensitivity and specificity......% for patient specimens and the specificity 100%. The junctional region between the Vgamma and Jgamma segments was specific for each patient.CONCLUSIONS: Capillary electrophoresis of PCR products from frozen and FFPE tissue is suitable for detecting clonal TCRgamma gene rearrangements. It is important, however...
Kuranishi, Fumito; Ohno, Tadao
Skeletal metastasis of breast carcinoma is refractory to intensive chemo-radiation therapy and therefore is assumed impossible to cure. Here, we report an advanced case of breast cancer with vertebra-Th7 metastasis that showed complete response to combined treatments with formalin-fixed autologous tumor vaccine (AFTV), palliative radiation therapy with 36 Gy, and adjuvant chemotherapy with standardized CEF (cyclophosphamide, epirubicin, and 5FU), zoledronic acid, and aromatase inhibitors following mastectomy for the breast tumor. The patient has been disease-free for more than 4 years after the mammary surgery and remains well with no evidence of metastasis or local recurrence. Thus, a combination of AFTV, palliative radiation therapy, and adjuvant chemotherapy may be an effective treatment for this devastating disease.
Ohizumi, Takuya; Nakamura, Kikuyasu; Yamamoto, Yu; Mase, Masaji; Yamada, Manabu
Detection of fowl adenovirus (FAV) DNA from formalin-fixed and paraffin-embedded (FFPE) sections was attempted by PCR. Serotypes of FAV were classified by sequencing the PCR products. In trials of PCR using a positive control infected with serotype 2 FAV, the best primer set was 57F forward primer (5'-CAARTTCAGRCAGACGGT-3') and 26R reverse primer (5'-GGCTTGACGTACGCTCCGTA-3'). A second PCR with the same primer set revealed a clearer band in the electrophoresis of generated PCR products. Generated PCR products were confirmed to be derived from infected FAV. In addition, PCR and sequencing of PCR products of the liver FFPE sections, from two natural inclusion body hepatitis cases that were not examined for virologic isolation, suggested that the detected FAV was serotype 8a. The PCR of FFPE sections, and serotyping by the sequencing of PCR products, are useful for diagnosis and epidemiologic analysis of FAV infections.
Tekin, Nilgun; Conget, Paulette; Bruna, Flavia; Timar, Botond; Gagyi, Eva; Basak, Ranjan; Naik, Omkar; Auewarakul, Chirayu; Sritana, Narongrit; Levy, Debora; Cerci, Juliano Julio; Bydlowski, Sergio Paulo; Pereira, Juliana; Dimamay, Mark Pierre; Natividad, Filipinas; Chung, June-Key; Belder, Nevin; Kuzu, Isinsu; Paez, Diana; Dondi, Maurizio; Carr, Robert
As a part of an international study on the molecular analysis of Diffuse Large B-cell Lymphoma (DLBCL), a robust protocol for gene expression analysis from RNA extraction to qRT-PCR using Formalin Fixed Paraffin Embedded tissues was developed. Here a study was conducted to define a strategy to validate the previously reported 6-gene (LMO2, BCL6, FN1, CCND2, SCYA3 and BCL2) model as predictor of prognosis in DLBCL. To avoid variation, all samples were tested in a single centre and single platform. This study comprised 8 countries (Brazil, Chile, Hungary, India, Philippines, S. Korea, Thailand and Turkey). Using the Kaplan-Meier and log rank test on patients (n=162) and two mortality risk groups (with those above and below the mean representing high and low risk groups) confirmed that the 6-gene predictor score correlates significantly with overall survival (OS, p<0.01) but not with event free survival (EFS, p=0.18). Adding the International Prognostic Index (IPI) shows that the 6-gene predictor score correlates significantly with high IPI scores for OS (p<0.05), whereas those with low IPI scores show a trend not reaching significance (p=0.08). This study defined an effective and economical qRT-PCR strategy and validated the 6-gene score as a predictor of OS in an international setting. PMID:27825111
Hu, Yu-Chang; Zhang, Qian; Huang, Yan-Hua; Liu, Yu-Fei; Chen, Hong-Lei
Molecular pathology tests are often carried for clinicopathological diagnosis and pathologists have established large collections of formalin-fixed, paraffin-embedded tissue (FFPE) banks. However, extraction of DNA from FFPE is a laborious and challenging for researchers in clinical laboratories. The aim of this study was to compare two widely used DNA extraction methods: using a QIAamp DNA FFPE kit from Qiagen and a Cobas Sample Preparation Kit from Roche, and evaluated the effect of the DNA quality on molecular diagnostics. DNA from FFPE non-small cell lung carcinoma tissues including biopsy and surgical specimens was extracted with both QIAamp DNA FFPE and Cobas Sample Preparation Kits and EGFR mutations of non-small cell lung carcinomas were detected by real-time quantitative PCR using the extracted DNA. Our results showed that DNA extracted by QIAamp and Cobas methods were both suitable to detect downstream EGFR mutation in surgical specimens. Howover, Cobas method could yield more DNA from biopsy specimens, and gain much better EGFR mutation results.
Einaga, Naoki; Yoshida, Akio; Noda, Hiroko; Suemitsu, Masaaki; Nakayama, Yuki; Sakurada, Akihisa; Kawaji, Yoshiko; Yamaguchi, Hiromi; Sasaki, Yasushi; Tokino, Takashi; Esumi, Mariko
Formalin-fixed, paraffin-embedded (FFPE) tissues used for pathological diagnosis are valuable for studying cancer genomics. In particular, laser-capture microdissection of target cells determined by histopathology combined with FFPE tissue section immunohistochemistry (IHC) enables precise analysis by next-generation sequencing (NGS) of the genetic events occurring in cancer. The result is a new strategy for a pathological tool for cancer diagnosis: 'microgenomics'. To more conveniently and precisely perform microgenomics, we revealed by systematic analysis the following three details regarding FFPE DNA compared with paired frozen tissue DNA. 1) The best quality of FFPE DNA is obtained by tissue fixation with 10% neutral buffered formalin for 1 day and heat treatment of tissue lysates at 95°C for 30 minutes. 2) IHC staining of FFPE tissues decreases the quantity and quality of FFPE DNA to one-fourth, and antigen retrieval (at 120°C for 15 minutes, pH 6.0) is the major reason for this decrease. 3) FFPE DNA prepared as described herein is sufficient for NGS. For non-mutated tissue specimens, no artifactual mutation occurs during FFPE preparation, as shown by precise comparison of NGS of FFPE DNA and paired frozen tissue DNA followed by validation. These results demonstrate that even FFPE tissues used for routine clinical diagnosis can be utilized to obtain reliable NGS data if appropriate conditions of fixation and validation are applied.
Solassol, Jérôme; Vendrell, Julie; Märkl, Bruno
, was assessed on archived formalin-fixed paraffin-embedded (FFPE) tissue sections by comparing its results with the results previously obtained by routine reference approaches for KRAS genotyping. In case of discordance, samples were assessed further by additional methods. Among the 374 colorectal cancer FFPE...
Hillig, Thore; Thode, Jørgen; Breinholt, Ellen Marie
We compare HER2 receptor amplification analysis by immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and real-time polymerase chain reaction (real-time PCR) DNA copy-number assay following laser capture microdissection (LCM) in formalin-fixed paraffin embedded tissue from 40...
Kuramoto, Junko; Kawamura, Akio; Dembo, Tomohisa; Kimura, Tokuhiro; Fukuda, Keiichi; Okada, Yasunori
Patent foramen ovale (PFO) can cause ischemic stroke because of paradoxical embolism. Autopsy studies have shown that the prevalence of PFO is 25% in whites or blacks. However, there is a paucity of data on the prevalence of PFO in Asians. The aim of this study was to clarify the prevalence of PFO in the Japanese population. We reviewed 52,717 autopsy reports, which were collected and edited by the Japanese Society of Pathology from 2009 to 2012. Next, we inspected consecutive 103 formalin-fixed specimens that had already been examined by certified pathologists from 2009 to 2013 to find PFO and atrial septal aneurysm (ASA). ASA was defined as ≥10 mm protrusion of the septum into the left or the right atrium. In the database of the Japanese Society of Pathology, the incidence of PFO was 0.08% (43/52,717). Inspection of heart specimens disclosed that the prevalence of PFO was 13.6% (14/103). None of the PFO cases was reported at the original autopsy. PFO was more frequently found in the subjects with ASA (50%) than in those without ASA (9.7%) (P=0.004). PFO is under-reported in autopsy reports. Re-evaluation of heart specimens disclosed that the prevalence of PFO was 13.6%. The prevalence was lower than reported in the past.
Detection of Lawsonia intracellularis in formalin-fixed porcine intestinal tissue samples: comparison of immunofluorescence and in-situ hybridization, and evaluation of the effects of controlled autolysis.
Jensen, T K; Boesen, H T; Vigre, H; Boye, M
Two methods, an immunofluorescence assay (IFA; with a Lawsonia intracellularis-specific monoclonal antibody) and fluorescent in-situ hybridization (FISH; with a specific oligonucleotide probe targeting 16S ribosomal RNA of the bacterium), were compared for their ability to detect L. intracellularis (the cause of porcine proliferative enteritis [PE]) in formalin-fixed samples of intestinal tissue. Of 69 intestinal samples with gross lesions of PE, 63 were positive by both FISH and IFA, but six were positive only by IFA. This indicated that the sensitivity of FISH was 91% that of IFA. However, both methods had a specificity of 100%. Fifty normal porcine intestines were negative by both tests. IFA was much less susceptible than FISH to the effects of autolysis. Thus, three of nine samples from pigs with PE were FISH-negative after being kept at 20 degrees C for 4 days, and seven were FISH negative after 2 weeks; after 4 weeks at this temperature, however, six of the nine samples were still IFA positive. After being kept at 4 degrees C for 12 weeks, the majority of samples (> or = 66%) were positive by both methods.
In situ hybridization (ISH) allows evaluation of genetic abnormalities, such as changes in chromosome number, chromosome translocations or gene amplifications, by hybridization of tagged DNA (or RNA) probes with complementary DNA (or RNA) sequences in interphase nuclei of target tissue. However, chromogenic in situ hybridization (CISH) is also applicable to formalin-fixed, paraffin-embedded (FFPE) tissues, besides metaphase chromosome spreads. CISH is similar to fluorescent in situ hybridization (FISH) regarding pretreatments and hybridization protocols but differs in the way of visualization. Indeed, CISH signal detection is similar to that used in immunohistochemistry, making use of a peroxidase-based chromogenic reaction instead of fluorescent dyes. In particular, tagged DNA probes are indirectly detected using an enzyme-conjugated antibody targeting the tags. The enzymatic reaction of the chromogenic substrate leads to the formation of strong permanent brown signals that can be visualized by bright-field microscopy at 40 × magnification. The advantage of CISH is that it allows the simultaneous observation of gene amplification and tissue morphology and the slides can be stored for a long time.
Erich, Katrin; Sammour, Denis A; Marx, Alexander; Hopf, Carsten
On-slide digestion of formalin-fixed and paraffin-embedded human biopsy tissue followed by mass spectrometry imaging of resulting peptides may have the potential to become an additional analytical modality in future ePathology. Multiple workflows have been described for dewaxing, antigen retrieval, digestion and imaging in the past decade. However, little is known about suitable statistical scores for method comparison and systematic workflow standardization required for development of processes that would be robust enough to be compatible with clinical routine. To define scores for homogeneity of tissue processing and imaging as well as inter-day repeatability for five different processing methods, we used human liver and gastrointestinal stromal tumor tissue, both judged by an expert pathologist to be >98% histologically homogeneous. For mean spectra-based as well as pixel-wise data analysis, we propose the coefficient of determination R 2 , the natural fold-change (natFC) value and the digest efficiency DE% as readily accessible scores. Moreover, we introduce two scores derived from principal component analysis, the variance of the mean absolute deviation, MAD, and the interclass overlap, J overlap , as computational scores that may help to avoid user bias during future workflow development. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. Copyright © 2016 Elsevier B.V. All rights reserved.
Lai, Xianyin; Schneider, Bryan P
Because fresh-frozen tissue samples associated with long-term clinical data and of rare diseases are often unobtainable at the present time, formalin-fixed paraffin-embedded (FFPE) tissue samples are considered a highly valuable resource for researchers. However, protein extraction from FFPE tissues faces challenges of deparaffinization and cross-link reversion. Current procedures for protein extraction from FFPE tissue require separate steps and toxic solvents, resulting in inconvenience in protein extraction. To overcome these limitations, an integrated method was developed using nontoxic solvents in four types of FFPE tissues. The average amount of proteins from three replicates of bladder, kidney, liver, and lung FFPE tissues were 442.6, 728.9, 736.4, and 694.7 μg with CVs of 7.5, 5.8, 2.4, and 4.5%, respectively. Proteomic analysis showed that 348, 417, 607, and 304 unique proteins were identified and quantified without specification of isoform by a least two peptides from bladder, kidney, liver, and lung FFPE tissue samples, respectively. The analysis of individual protein CV demonstrated that 97-99% of the proteins were quantified with a CV ≤ 30%, verifying the reproducibility of the integrated protein extraction method. In summary, the developed method is high-yield, reproducible, convenient, simple, low cost, nonvolatile, nonflammable, and nontoxic. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Full Text Available In cancer drug discovery, it is important to investigate the genetic determinants of response or resistance to cancer therapy as well as factors that contribute to adverse events in the course of clinical trials. Despite the emergence of new technologies and the ability to measure more diverse analytes (e.g., circulating tumor cell (CTC, circulating tumor DNA (ctDNA, etc., tumor tissue is still the most common and reliable source for biomarker investigation. Because of its worldwide use and ability to preserve samples for many decades at ambient temperature, formalin-fixed, paraffin-embedded tumor tissue (FFPE is likely to be the preferred choice for tissue preservation in clinical practice for the foreseeable future. Multiple analyses are routinely performed on the same FFPE samples (such as Immunohistochemistry (IHC, in situ hybridization, RNAseq, DNAseq, TILseq, Methyl-Seq, etc.. Thus, specimen prioritization and optimization of the isolation of analytes is critical to ensure successful completion of each assay. FFPE is notorious for producing suboptimal DNA quality and low DNA yield. However, commercial vendors tend to request higher DNA sample mass than what is actually required for downstream assays, which restricts the breadth of biomarker work that can be performed. We evaluated multiple genomics service laboratories to assess the current state of NGS pre-analytical processing of FFPE. Significant differences in pre-analytical capabilities were observed. Key aspects are highlighted and recommendations are made to improve the current practice in translational research.
Mansour, Anthony G; Khalil, Pamela Abou; Bejjani, Noha; Chatila, Rajaa; Dagher-Hamalian, Carole; Faour, Wissam H
Deparaffinization of formalin-fixed paraffin embedded (FFPE) tissues with xylene currently remains a major challenge to the biomedical community. We developed an efficient xylene-free protocol to isolate proteins from archived FFPE human tissue sections. A total of 79 different types of FFPE tissue sections of 8 µm thickness were obtained from various archived FFPE specimens. Deparaffinization was conducted by gently washing each section with around 1 ml of hot distilled water (≈80°C). The deparaffinized tissues were homogenized in lysis buffer, and the isolated proteins were quantified and efficiently resolved using western blot analysis for the presence of Protein kinase B (PKB/AKT) and β-actin. Moreover, a significant amount of proteins was successfully isolated with an average of 2.31 µg/µl. The migration pattern of AKT and β-actin obtained from the specimens was similar to the positive control obtained from protein lysates prepared from in vitro cultured MDA231 cancer cell lines. AKT was successfully identified in all specimens, and β-actin protein was resolved with an efficiency higher than 80%. The entire extraction procedure requires only 20 minutes. This newly developed technique is an efficient, safe, cost-effective, and rapid method to isolate proteins from FFPE tissue sections adequate for molecular analysis.
Technical Advances for the Clinical Genomic Evaluation of Sudden Cardiac Death: Verification of Next-Generation Sequencing Panels for Hereditary Cardiovascular Conditions Using Formalin-Fixed Paraffin-Embedded Tissues and Dried Blood Spots.
Baudhuin, Linnea M; Leduc, Charles; Train, Laura J; Avula, Rajeswari; Kluge, Michelle L; Kotzer, Katrina E; Lin, Peter T; Ackerman, Michael J; Maleszewski, Joseph J
Postmortem genetic testing for heritable cardiovascular (CV) disorders is often lacking because ideal specimens (ie, whole blood) are not retained routinely at autopsy. Formalin-fixed paraffin-embedded tissue (FFPET) is ubiquitously collected at autopsy, but DNA quality hampers its use with traditional sequencing methods. Targeted next-generation sequencing may offer the ability to circumvent such limitations, but a method has not been previously described. The primary aim of this study was to develop and evaluate the use of FFPET for heritable CV disorders via next-generation sequencing. Nineteen FFPET (heart) and blood (whole blood or dried blood spot) specimens underwent targeted next-generation sequencing using a custom panel of 101 CV-associated genes. Nucleic acid yield and quality metrics were evaluated in relation to FFPET specimen age (6 months to 15 years; n=14) and specimen type (FFPET versus whole blood and dried blood spot; n=12). Four FFPET cases with a clinical phenotype of heritable CV disorder were analyzed. Accuracy and precision were 100% concordant between all sample types, with read depths >100× for most regions tested. Lower read depth, as low as 40×, was occasionally observed with FFPET and dried blood spot. High-quality DNA was obtained from FFPET samples as old as 15 years. Genomic analysis of FFPET from the 4 phenotype-positive/genotype unknown cases all revealed putative disease-causing variants. Similar performance characteristics were observed for next-generation sequencing of FFPET, whole blood, and dried blood spot in the evaluation of inherited CV disorders. Although blood is preferable for genetic analyses, this study offers an alternative when only FFPET is available. © 2017 American Heart Association, Inc.
Nechifor-Boilă, Adela Corina; Loghin, Andrada; Vacariu, Victor; Halaţiu, Vasile Bogdan; Borda, Angela
Optimal recovery of nucleic acids from formalin-fixed paraffin-embedded (FFPE) tissues is highly dependent on a series of pre-extraction steps, mainly related (but not limited) to fixation. The aim of our study was to investigate if the storage period of the FFPE blocks had a significant effect on the isolated DNA. We examined the quantity and purity of the isolated DNA from 83 FFPE blocks, corresponding to malignant thyroid (n=28) and renal (n=55) carcinomas that had been stored in our department for up to eight years. The DNA extraction protocol was based on a precipitation method (MasterPure™ DNA Purification Kit, Epicentre), in accordance to the manufacturer instructions, optimized in our laboratory. A spectrophotometer was used to determine the yield (A260) and purity (A260/A280 ratio) of the isolated DNA. We successfully isolated good DNA quantity and purity from all our study cases (mean concentration: 223.4 ± 104.16 ng/μL; mean A260/A280 ratio: 1.68 ± 0.09). Moreover, no statistically significant differences were observed between tumor blocks stored for 2-3 years and 7-8 years, respectively, both in terms of DNA quantity (p=0.196) and purity (p=0.663). In conclusion, we successfully validated an efficient, reproducible DNA extraction technique that provided a good range of DNA concentrations and purity, regardless the type of tissue (thyroid or kidney). Moreover, we demonstrated that the storage period of the FFPE blocks does not have a significant influence on the DNA quantity and purity.
Improving the Prediction of Prostate Cancer Overall Survival by Supplementing Readily Available Clinical Data with Gene Expression Levels of IGFBP3 and F3 in Formalin-Fixed Paraffin Embedded Core Needle Biopsy Material.
Full Text Available A previously reported expression signature of three genes (IGFBP3, F3 and VGLL3 was shown to have potential prognostic value in estimating overall and cancer-specific survivals at diagnosis of prostate cancer in a pilot cohort study using freshly frozen Fine Needle Aspiration (FNA samples.We carried out a new cohort study with 241 prostate cancer patients diagnosed from 2004-2007 with a follow-up exceeding 6 years in order to verify the prognostic value of gene expression signature in formalin fixed paraffin embedded (FFPE prostate core needle biopsy tissue samples. The cohort consisted of four patient groups with different survival times and death causes. A four multiplex one-step RT-qPCR test kit, designed and optimized for measuring the expression signature in FFPE core needle biopsy samples, was used. In archive FFPE biopsy samples the expression differences of two genes (IGFBP3 and F3 were measured. The survival time predictions using the current clinical parameters only, such as age at diagnosis, Gleason score, PSA value and tumor stage, and clinical parameters supplemented with the expression levels of IGFBP3 and F3, were compared.When combined with currently used clinical parameters, the gene expression levels of IGFBP3 and F3 are improving the prediction of survival time as compared to using clinical parameters alone.The assessment of IGFBP3 and F3 gene expression levels in FFPE prostate cancer tissue would provide an improved survival prediction for prostate cancer patients at the time of diagnosis.
Lass, Anna; Karanis, Panagiotis; Korzeniewski, Krzysztof
It is estimated that faecal-orally transmitted diseases are common in Afghanistan, as a consequence of poor hygienic standards of life and widespread contamination of water and food with both human and animal faeces. However, there is little information in the literature concerning infections caused by intestinal parasites in the Afghan population. In this study, we report the occurrence of Giardia intestinalis assemblages (A and B) in formalin-fixed stool samples collected from 245 Afghan schoolchildren living in Ghazni Province in eastern Afghanistan. Detection of the parasite's DNA and genotyping was performed using real-time PCR, specific to the β-giardin gene of G. intestinalis. Positive results were recorded in 52 (21.2%) samples. Genotyping was successful in 39 faecal samples and showed the predominance of assemblage B of G. intestinalis in this population (15 assemblage A and 24 assemblage B). Co-infection with both genotypes A and B was detected in four samples. Additionally, we evaluated the effect of 10% buffered formalin fixative on the detection of G. intestinalis DNA using real-time PCR and nested PCR characterised by different lengths of PCR products (74 and 479 bp, respectively). The human faeces containing the Giardia cysts were tested for 16 weeks. Amplification of G. intestinalis DNA with real-time PCR was possible up to 6 weeks of preservation of stool sample in formalin, compared to only 2 weeks with nested PCR. This suggests that real-time PCR is a more suitable tool in cases where stool samples have to be kept in formalin for longer periods of time.
Chromogenic In Situ Hybridization and p16/Ki67 Dual Staining on Formalin-Fixed Paraffin-Embedded Cervical Specimens: Correlation with HPV-DNA Test, E6/E7 mRNA Test, and Potential Clinical Applications
Full Text Available Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness of p16INK4a/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+. p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH and p16INK4a/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, while p16INK4a/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure.
Chromogenic In Situ Hybridization and p16/Ki67 Dual Staining on Formalin-Fixed Paraffin-Embedded Cervical Specimens: Correlation with HPV-DNA Test, E6/E7 mRNA Test, and Potential Clinical Applications
Zappacosta, Roberta; Colasante, Antonella; Viola, Patrizia; D'Antuono, Tommaso; Lattanzio, Giuseppe; Capanna, Serena; Gatta, Daniela Maria Pia; Rosini, Sandra
Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV) infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH) technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness of p16INK4a/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+). p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH and p16INK4a/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, while p16INK4a/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure. PMID:24369532
HPV16 detection by qPCR method in relation to quantity and quality of DNA extracted from archival formalin fixed and paraffin embedded head and neck cancer tissues by three commercially available kits.
Biesaga, Beata; Janecka, Anna; Mucha-Małecka, Anna; Adamczyk, Agnieszka; Szostek, Sława; Słonina, Dorota; Halaszka, Krzysztof; Przewoźnik, Marcin
The aim of the present study was to compare HPV16 detection by quantitative polymerase chain reaction (qPCR) in relation to the quantity and quality of DNA isolated from 21 formalin fixed and paraffin embedded (FFPE) head and neck cancer tissues by three commercially available kits: EX-WAX™ DNA Extraction Kit (M) (Merck Millipore, Darmstadt, Germany), QIAamp(®) DNA FFPE Tissue (Q) (Qiagen, Hilden, Germany) and ReliaPrep™ FFPE gDNA Miniprep System (P) (Promega, Madison, USA). Quantity of extracted DNA was assessed spectrophometrically and fluorometrically. Its quality was analyzed using A260/280 and A260/230 ratios and the β-actin fragment amplifiability in qPCR. HPV16 presence was detected by qPCR, using specific primers and TaqMan probe. HPV infection was found in 8 DNA samples extracted with M kit (38.1%) and in 7 (33.3%) isolated with Q and P kits. Three samples from M and Q kits were characterized by HPV16 positivity and lack of β-actin amplifiability. They had significantly lower A260/280 ratio (M: 1.6±0.0, p=0.044 and Q: 1.7±0.0, p=0.016) compared to samples with both fragments amplification (M: 1.7±0.0 and Q: 1.9±0.0). Therefore, for HPV detection by qPCR in FFPE tissues we recommend ReliaPrep™ FFPE gDNA Miniprep System. Copyright © 2016 Elsevier B.V. All rights reserved.
Nm23/nucleoside diphosphate kinase-A as a potent prognostic marker in invasive pancreatic ductal carcinoma identified by proteomic analysis of laser micro-dissected formalin-fixed paraffin-embedded tissue
Full Text Available Abstract Background Pancreatic cancer is among the most lethal malignancies worldwide. This study aimed to identify a novel prognostic biomarker, facilitating treatment selection, using mass spectrometry (MS-based proteomic analysis with formalin-fixed paraffin-embedded (FFPE tissue. Results The two groups with poor prognosis (n = 4 and with better prognosis (n = 4 had been carefully chosen among 96 resected cases of pancreatic cancer during 1998 to 2007 in Tohoku University Hospital. Although those 2 groups had adjusted background (UICC-Stage IIB, Grade2, R0, gemcitabine adjuvant, there was a significant difference in postoperative mean survival time (poor 21.0 months, better 58.1 months, P = 0.0067. Cancerous epithelial cells collected from FFPE tissue sections by laser micro-dissection (LMD were processed for liquid chromatography-tandem mass spectrometry (LC-MS/MS. In total, 1099 unique proteins were identified and 6 proteins showed different expressions in the 2 groups by semi-quantitative comparison. Among these 6 proteins, we focused on Nm23/Nucleoside Diphosphate Kinase A (NDPK-A and immunohistochemically confirmed its expression in the cohort of 96 cases. Kaplan-Meier analysis showed high Nm23/NDPK-A expression to correlate with significantly worse overall survival (P = 0.0103. Moreover, in the multivariate Cox regression model, Nm23/NDPK-A over-expression remained an independent predictor of poor survival with a hazard ratio of 1.97 (95% CI 1.16-3.56, P = 0.0110. Conclusions We identified 6 candidate prognostic markers for postoperative pancreatic cancer using FFPE tissues and immunohistochemically demonstrated high Nm23/NDPK-A expression to be a useful prognostic marker for pancreatic cancer.
Cederbye, Camilla Natasha; Palshof, Jesper Andreas; Hansen, Tine Plato
Overexpression of the ATP-dependent drug efflux pump ABCG2 is a major molecular mechanism of multidrug resistance in cancer and might be a predictive biomarker for drug response. Contradictory results have been reported for immunohistochemical studies of ABCG2 protein expression in colorectal...... cancer (CRC), probably because of the use of different antibodies and scoring approaches. In this study, we systematically studied six commercially available anti-ABCG2 antibodies, using cell lines with up-regulation of ABCG2, and selected one antibody for validation in CRC tissue. Furthermore, we...... established scoring guidelines for ABCG2 expression based on the clinically used guidelines for HER2 immunohistochemistry assessment in gastric cancer. The guidelines provide a semi-quantitative measure of the basolateral membrane staining of ABCG2 and disregard the apical membrane staining...
Gilbert, Marcus T P; Sanchez, Juan J; Haselkorn, Tamara
, multiplex PCR with minisequencing (MPMS), on 92 DNA extractions performed on six archival FFPE samples of variable DNA quality, which date between 9 and 25 years old. On the three extracts with highest quality, we found the assay efficiency to be near 100%. However, the efficiency of the lowest quality...... extracts varied significantly. In this study, we demonstrate that although direct measures of DNA concentration in the extracts provide no useful information with regard to subsequent MPMS success, the success of the assay can be determined to some degree a priori, through initial screening of the DNA...... quality using a simple quantitative real-time PCR (qPCR) assay for nuclear DNA, and/or an assay of the maximum PCR amplifiable size of nuclear DNA. MPMS promises to be of significant use in future genetic studies on FFPE material. It provides a streamlined approach for retrieving a large amount of genetic...
Davies, G N; Bevan, I S; Banner, Jytte
Fresh tissue from cases of sudden infant death syndrome is becoming increasingly scarce and therefore researchers interesting in studying the aetiology of this syndrome have had to resort to archival tissue, usually in the form of paraffin wax sections. A simple method for isolating mRNA from for....... Transcripts of a housekeeping gene were detected in all tissues. This method should be useful for examining gene expression in archival material....
Full Text Available Background. Proper rRNA depletion is crucial for the successful utilization of FFPE specimens when studying gene expression. We performed a study to evaluate two major rRNA depletion methods: Ribo-Zero and RNase H. RNAs extracted from 4 samples were treated with the two rRNA depletion methods in duplicate and sequenced (N=16. We evaluated their reducibility, ability to detect RNA, and ability to molecularly subtype these triple negative breast cancer specimens. Results. Both rRNA depletion methods produced consistent data between the technical replicates. We found that the RNase H method produced higher quality RNAseq data as compared to the Ribo-Zero method. In addition, we evaluated the RNAseq data generated from the FFPE tissue samples for noncoding RNA, including lncRNA, enhancer/super enhancer RNA, and single nucleotide variation (SNV. We found that the RNase H is more suitable for detecting high-quality, noncoding RNAs as compared to the Ribo-Zero and provided more consistent molecular subtype identification between replicates. Unfortunately, neither method produced reliable SNV data. Conclusions. In conclusion, for FFPE specimens, the RNase H rRNA depletion method performed better than the Ribo-Zero. Neither method generates data sufficient for SNV detection.
Bustamante, Javier Andres; Astudillo, Miryam; Pazos, Alvaro Jairo; Bravo, Luis Eduardo
Paraffin wax embedded tissues are an invaluable material for retrospective studies requiring the application of molecular analysis. Multiple methods are available to extract DNA from these kinds of samples. However, the most common methods are slow and the reagents often contribute to the fragmentation of genetic material. In order to optimize the procedure, two methods for DNA extraction from paraffin embedded tissue non-optimal conditions were used. 47 blocks containing paraffin-embedded biopsies of pleura, lung and pericardium from 24 patients (66.6% males) older than 18 years, with biopsy proven chronic granulomatous inflammation referred to the department of pathology at University Hospital of Valle between 2002 and 2007 were selected. Each sample was subjected to 10 cuts and was to two methods of DNA extraction: 1. conventional and 2. QIAamp - DNA mini kit. The efficiency of the extracted DNA was assessed by spectrophotometry and PCR amplification of a fragment of the housekeeping gene GAPDH. The concentration of DNA samples extracted by the conventional method was of 65.52 ng/Mu l ± 11.47 (mean ± SE) and the 260/280 absorbance ratio ranged between 0.52 and 2.30 the average concentration of DNA of the samples extracted by the commercial method was 60.89 ng/Mu l ± 6.02 (mean ± SE), with an absorbance that fluctuated between 0 and 2.64. The DNA obtained was amplified by PCR, of 47 samples extracted by methods, 25 and 23 respectively the GAPDH gene amplified successfully. The methods used to obtain DNA showed similar performance, highlighting the potential utility of both extraction methods for the retrospective studies from paraffin embedded tissues in unsuitable conditions.
Kerr, Darcy A; Sweeney, Brenda; Arpin, Ronald N; Ring, Melissa; Pitman, Martha B; Wilbur, David C; Faquin, William C
-Testing for high-risk human papillomavirus (HR-HPV) in head and neck squamous cell carcinomas (HNSCCs) is important for both prognostication and clinical management. Several testing platforms are available for HR-HPV; however, effective alternative automated approaches are needed. -To assess the performance of the automated Roche cobas 4800 HPV real-time polymerase chain reaction-based system on formalin-fixed, paraffin-embedded HNSCC specimens and compare results with standard methods of in situ hybridization (ISH) and p16 immunohistochemistry. -Formalin-fixed, paraffin-embedded samples of HNSCC were collected from archival specimens in the Department of Pathology, Massachusetts General Hospital (Boston), and prepared using the automated system by deparaffinization and dehydration followed by tissue lysis. Samples were integrated into routine cervical cytology testing runs by cobas. Corresponding formalin-fixed, paraffin-embedded samples were evaluated for HR-HPV by ISH and p16 by immunohistochemistry. Discrepant cases were adjudicated by polymerase chain reaction. -Sixty-two HNSCC samples were analyzed using the automated cobas system, ISH, and immunohistochemistry. Fifty-two percent (n = 32 of 62) of formalin-fixed, paraffin-embedded tumors were positive for HR-HPV by cobas. Eighty-eight percent (n = 28 of 32) of cases were the HPV 16 subtype and 12% (n = 4 of 32) were other HR-HPV subtypes. Corresponding testing with ISH was concordant in 92% (n = 57 of 62) of cases. Compared with the adjudication polymerase chain reaction standard, there were 3 false-positive cases by cobas. -Concordance in HNSCC HR-HPV status between cobas and ISH was more than 90%. The cobas demonstrated a sensitivity of 100% and a specificity of 91% for detection of HR-HPV. Advantages favoring cobas include its automation, cost efficiency, objective results, and ease of performance.
Detection of a putative novel adenovirus by PCR amplification, sequencing and phylogenetic characterisation of two gene fragments from formalin-fixed paraffin-embedded tissues of a cat diagnosed with disseminated adenovirus disease.
Lakatos, Béla; Hornyák, Ákos; Demeter, Zoltán; Forgách, Petra; Kennedy, Frances; Rusvai, Miklós
Adenoviral nucleic acid was detected by polymerase chain reaction (PCR) in formalin-fixed paraffin-embedded tissue samples of a cat that had suffered from disseminated adenovirus infection. The identity of the amplified products from the hexon and DNA-dependent DNA polymerase genes was confirmed by DNA sequencing. The sequences were clearly distinguishable from corresponding hexon and polymerase sequences of other mastadenoviruses, including human adenoviruses. These results suggest the possible existence of a distinct feline adenovirus.
Feng, Zipei; Puri, Sachin; Moudgil, Tarsem; Wood, William; Hoyt, Clifford C; Wang, Chichung; Urba, Walter J; Curti, Brendan D; Bifulco, Carlo B; Fox, Bernard A
Adoptive T cell therapy (ACT) has shown great promise in melanoma, with over 50 % response rate in patients where autologous tumor-reactive tumor-infiltrating lymphocytes (TIL) can be cultured and expanded. A major limitation of ACT is the inability to generate or expand autologous tumor-reactive TIL in 25-45 % of patients tested. Methods that successfully identify tumors that are not suitable for TIL generation by standard methods would eliminate the costs of fruitless expansion and enable these patients to receive alternate therapy immediately. Multispectral fluorescent immunohistochemistry with a panel including CD3, CD8, FoxP3, CD163, PD-L1 was used to analyze the tumor microenvironment in 17 patients with melanoma among our 36-patient cohort to predict successful TIL generation. Additionally, we compared tumor fragments and enzymatic digestion of tumor samples for efficiency in generating tumor-reactive TIL. Tumor-reactive TIL were generated from 21/36 (58 %) of melanomas and for 12/13 (92 %) tumors where both enzymatic and fragment methods were compared. TIL generation was successful in 10/13 enzymatic preparations and in 10/13 fragment cultures; combination of both methods resulted in successful generation of autologous tumor-reactive TIL in 12/13 patients. In 17 patients for whom tissue blocks were available, IHC analysis identified that while the presence of CD8(+) T cells alone was insufficient to predict successful TIL generation, the CD8(+) to FoxP3(+) ratio was predictive with a positive-predictive value (PPV) of 91 % and negative-predictive value (NPV) of 86 %. Incorporation of CD163+ macrophage numbers and CD8:PD-L1 ratio did not improve the PPV. However, the NPV could be improved to 100 % by including the ratio of CD8(+):PD-L1(+) expressing cells. This is the first study to apply 7-color multispectral immunohistochemistry to analyze the immune environment of tumors from patients with melanoma. Assessment of the data using unsupervised
Seiler, Caroline; Sharpe, Alan; Barrett, J Carl; Harrington, Elizabeth A; Jones, Emma V; Marshall, Gayle B
Advanced genomic techniques such as Next-Generation-Sequencing (NGS) and gene expression profiling, including NanoString, are vital for the development of personalised medicines, as they enable molecular disease classification. This has become increasingly important in the treatment of cancer, aiding patient selection. However, it requires efficient nucleic acid extraction often from formalin-fixed paraffin-embedded tissue (FFPE). Here we provide a comparison of several commercially available manual and automated methods for DNA and/or RNA extraction from FFPE cancer cell line samples from Qiagen, life Technologies and Promega. Differing extraction geometric mean yields were evaluated across each of the kits tested, assessing dual DNA/RNA extraction vs. specialised single extraction, manual silica column based extraction techniques vs. automated magnetic bead based methods along with a comparison of subsequent nucleic acid purity methods, providing a full evaluation of nucleic acids isolated. Out of the four RNA extraction kits evaluated the RNeasy FFPE kit, from Qiagen, gave superior geometric mean yields, whilst the Maxwell 16 automated method, from Promega, yielded the highest quality RNA by quantitative real time RT-PCR. Of the DNA extraction kits evaluated the PicoPure DNA kit, from Life Technologies, isolated 2-14× more DNA. A miniaturised qPCR assay was developed for DNA quantification and quality assessment. Careful consideration of an extraction kit is necessary dependent on quality or quantity of material required. Here we provide a flow diagram on the factors to consider when choosing an extraction kit as well as how to accurately quantify and QC the extracted material.
Snow, Anthony N; Stence, Aaron A; Pruessner, Jonathan A; Bossler, Aaron D; Ma, Deqin
Extraction of DNA from formalin-fixed, paraffin-embedded (FFPE) tissue is a critical step in molecular oncologic testing. As molecular oncology testing becomes more important for prognostic and therapeutic decision making and tissue specimens become smaller due to earlier detection of suspicious lesions and the use of fine needle aspiration methods for tissue collection, it becomes more challenging for the typical molecular pathology laboratory to obtain reliable test results. We developed a DNA extraction method to obtain sufficient quantity and high quality genomic DNA from limited FFPE tissue for molecular oncology testing using a combination of H&E stained slides, a matrix capture method and the Qiagen DNA column. THREE DNA EXTRACTION METHODS WERE COMPARED: our standard procedure of manually scraping tissue from unstained slides followed by DNA extraction using the QIAamp FFPE column (Qiagen, Valencia, CA), a glue capture method (Pinpoint Solution, Zymo Research Corp, Inc) on H&E stained slides followed by DNA extraction using either the QIAamp column or the column included with the Pinpoint kit (Zymo Research). The DNA extraction protocol was optimized. Statistical analysis was performed using the paired two-sample student's t-test. The combination of the matrix capture method with the QIAamp column gave an equivalent amount of DNA as our standard extraction method using the unstained slides and a 4.6-fold higher DNA yield than using the Zymo column included in the Pinpoint Slide Solution kit. Several molecular tests were performed and DNA purified using the new method gave the same results as for the previous methods. Using H&E stained slides allows visual confirmation of tumor cells during microdissection. The Pinpoint solution made removal of specific tissue from the slides easier and reduced the risk of contamination and tissue loss. This DNA extraction method is simple, cost-effective, and blends with our current workflow requiring no additional equipment.
Salawu, Abdulazeez; Ul-Hassan, Aliya; Hammond, David; Fernando, Malee; Reed, Malcolm; Sisley, Karen
Most soft tissue sarcomas are characterized by genetic instability and frequent genomic copy number aberrations that are not subtype-specific. Oligonucleotide microarray-based Comparative Genomic Hybridisation (array CGH) is an important technique used to map genome-wide copy number aberrations, but the traditional requirement for high-quality DNA typically obtained from fresh tissue has limited its use in sarcomas. Although large archives of Formalin-fixed Paraffin-embedded (FFPE) tumour samples are available for research, the degradative effects of formalin on DNA from these tissues has made labelling and analysis by array CGH technically challenging. The Universal Linkage System (ULS) may be used for a one-step chemical labelling of such degraded DNA. We have optimised the ULS labelling protocol to perform aCGH on archived FFPE leiomyosarcoma tissues using the 180k Agilent platform. Preservation age of samples ranged from a few months to seventeen years and the DNA showed a wide range of degradation (when visualised on agarose gels). Consistently high DNA labelling efficiency and low microarray probe-to-probe variation (as measured by the derivative log ratio spread) was seen. Comparison of paired fresh and FFPE samples from identical tumours showed good correlation of CNAs detected. Furthermore, the ability to macro-dissect FFPE samples permitted the detection of CNAs that were masked in fresh tissue. Aberrations were visually confirmed using Fluorescence in situ Hybridisation. These results suggest that archival FFPE tissue, with its relative abundance and attendant clinical data may be used for effective mapping for genomic copy number aberrations in such rare tumours as leiomyosarcoma and potentially unravel clues to tumour origins, progression and ultimately, targeted treatment. PMID:23209738
Nuovo, Allison J; Garofalo, Michela; Mikhail, Alexandria; Nicol, Alcina F; Vianna-Andrade, Cecilia; Nuovo, Gerard J
Formalin-fixed, paraffin-embedded tissues are widely used in biomedical research but little is known about the effect of the age of the block or unstained slides on the in situ hybridization or immunohistochemistry signal. We compared the in situ-based and immunohistochemistry-based signals for cervical intraepithelial neoplasia samples that ranged from 0 to 15 years of age. There was a progressive and statistically significant decrease in the strength of the p16 signal when comparing tissues prepared from recent unstained slides (0 to 1 y old, mean score of 92%) to those of intermediate age (5 to 7 y old, mean score of 49%) to old unstained slides (cut 13 to 15 y ago, mean score of 10%). Equivalent, progressive, and significant decreases in the intensity of the signals for microRNAs, CD45, and human papillomavirus DNA were seen in tissues stored on slides from 5 to 7 years and 13 to 15 years, respectively. However, the diminution of signal was much less, although still statistically significant, if the sections from the 13- to 15-year-old paraffin blocks were prepared in 2012. The data likely does not represent degradation of the targets as extraction of several microRNA from the old blocks showed no detectable degradation, despite the markedly weakened in situ hybridization signal. It is concluded that in situ-based signal for DNA, microRNAs, and proteins in paraffin-embedded tissues are significantly reduced over time, especially when stored long term on glass slides which, in turn, can lead to a significant underestimation of the amount and presence of the nucleic acid or protein target.
Zhang, Shenli; Tan, Iain B; Sapari, Nur S; Grabsch, Heike I; Okines, Alicia; Smyth, Elizabeth C; Aoyama, Toru; Hewitt, Lindsay C; Inam, Imran; Bottomley, Dan; Nankivell, Matthew; Stenning, Sally P; Cunningham, David; Wotherspoon, Andrew; Tsuburaya, Akira; Yoshikawa, Takaki; Soong, Richie; Tan, Patrick
DNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissues has been used in the past to analyze genetic polymorphisms. We evaluated the technical reproducibility of different types of assays for gene polymorphisms using DNA extracted from FFPE material. By using the MassARRAY iPLEX system, we investigated polymorphisms in DPYD (rs1801159 and rs3918290), UMPS (rs1801019), ERCC1 (rs11615), ERCC1 (rs3212986), and ERCC2 (rs13181) in 56 FFPE DNA samples. By using PCR, followed by size-based gel electrophoresis, we also examined TYMS 5' untranslated region 2R/3R repeats and GSTT1 deletions in 50 FFPE DNA samples and 34 DNAs extracted from fresh-frozen tissues and cell lines. Each polymorphism was analyzed by two independent runs. We found that iPLEX biomarker assays measuring single-nucleotide polymorphisms provided consistent concordant results. However, by using FFPE DNA, size-based PCR biomarkers (GSTT1 and TYMS 5' untranslated region) were discrepant in 32.7% (16/49, with exact 95% CI, 19.9%-47.5%; exact binomial confidence limit test) and 4.2% (2/48, with exact 95% CI, 0.5%-14.3%) of cases, respectively, whereas no discrepancies were observed using intact genomic DNA. Our findings suggest that DNA from FFPE material can be used to reliably test single-nucleotide polymorphisms. However, results based on size-based PCR biomarkers, and particularly GSTT1 deletions, using FFPE DNA need to be interpreted with caution. Independent repeated assays should be performed on all cases to assess potential discrepancies. Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Dana A M Mustafa
Full Text Available Gene expression profiling of tumors is a successful tool for the discovery of new cancer biomarkers and potential targets for the development of new therapeutic strategies. Reliable profiling is preferably performed on fresh frozen (FF tissues in which the quality of nucleic acids is better preserved than in formalin-fixed paraffin-embedded (FFPE material. However, since snap-freezing of biopsy materials is often not part of daily routine in pathology laboratories, one may have to rely on archival FFPE material. Procedures to retrieve the RNAs from FFPE materials have been developed and therefore, datasets obtained from FFPE and FF materials need to be made compatible to ensure reliable comparisons are possible.To develop an efficient method to compare gene expression profiles obtained from FFPE and FF samples using the same platform.Twenty-six FFPE-FF sample pairs of the same tumors representing various cancer types, and two FFPE-FF sample pairs of breast cancer cell lines, were included. Total RNA was extracted and gene expression profiling was carried out using Illumina's Whole-Genome cDNA-mediated Annealing, Selection, extension and Ligation (WG-DASL V3 arrays, enabling the simultaneous detection of 24,526 mRNA transcripts. A sample exclusion criterion was created based on the expression of 11 stably expressed reference genes. Pearson correlation at the probe level was calculated for paired FFPE-FF, and three cut-off values were chosen. Spearman correlation coefficients between the matched FFPE and FF samples were calculated for three probe lists with varying levels of significance and compared to the correlation based on all measured probes. Unsupervised hierarchical cluster analysis was performed to verify performance of the included probe lists to compare matched FPPE-FF samples.Twenty-seven FFPE-FF pairs passed the sample exclusion criterion. From the profiles of 27 FFPE and FF matched samples, the best correlating probes were identified
Vukmirovic, Milica; Herazo-Maya, Jose D; Blackmon, John; Skodric-Trifunovic, Vesna; Jovanovic, Dragana; Pavlovic, Sonja; Stojsic, Jelena; Zeljkovic, Vesna; Yan, Xiting; Homer, Robert; Stefanovic, Branko; Kaminski, Naftali
Idiopathic Pulmonary Fibrosis (IPF) is a lethal lung disease of unknown etiology. A major limitation in transcriptomic profiling of lung tissue in IPF has been a dependence on snap-frozen fresh tissues (FF). In this project we sought to determine whether genome scale transcript profiling using RNA Sequencing (RNA-Seq) could be applied to archived Formalin-Fixed Paraffin-Embedded (FFPE) IPF tissues. We isolated total RNA from 7 IPF and 5 control FFPE lung tissues and performed 50 base pair paired-end sequencing on Illumina 2000 HiSeq. TopHat2 was used to map sequencing reads to the human genome. On average ~62 million reads (53.4% of ~116 million reads) were mapped per sample. 4,131 genes were differentially expressed between IPF and controls (1,920 increased and 2,211 decreased (FDR < 0.05). We compared our results to differentially expressed genes calculated from a previously published dataset generated from FF tissues analyzed on Agilent microarrays (GSE47460). The overlap of differentially expressed genes was very high (760 increased and 1,413 decreased, FDR < 0.05). Only 92 differentially expressed genes changed in opposite directions. Pathway enrichment analysis performed using MetaCore confirmed numerous IPF relevant genes and pathways including extracellular remodeling, TGF-beta, and WNT. Gene network analysis of MMP7, a highly differentially expressed gene in both datasets, revealed the same canonical pathways and gene network candidates in RNA-Seq and microarray data. For validation by NanoString nCounter® we selected 35 genes that had a fold change of 2 in at least one dataset (10 discordant, 10 significantly differentially expressed in one dataset only and 15 concordant genes). High concordance of fold change and FDR was observed for each type of the samples (FF vs FFPE) with both microarrays (r = 0.92) and RNA-Seq (r = 0.90) and the number of discordant genes was reduced to four. Our results demonstrate that RNA sequencing of RNA
Immunohistochemistry of sarcolemmal membrane-associated proteins in formalin-fixed and paraffin-embedded skeletal muscle tissue: a promising tool for the diagnostic evaluation of common muscular dystrophies.
Suriyonplengsaeng, Chinnawut; Dejthevaporn, Charungthai; Khongkhatithum, Chaiyos; Sanpapant, Suda; Tubthong, Nattha; Pinpradap, Koset; Srinark, Nippa; Waisayarat, Jariya
The analysis of fresh frozen muscle specimens is standard following routine muscle biopsy, but this service is not widely available in countries with limited medical facilities, such as Thailand. Nevertheless, immunohistochemistry (IHC) analysis is essential for the diagnosis of patients with a strong clinical suspicion of muscular dystrophy, in the absence of mutations detected by molecular genetics. As the successful labelling of sarcolemmal membrane-associated proteins in formalin-fixed and paraffin-embedded (FFPE) muscle sections using IHC staining has rarely been described, this study aimed to develop a reproducible IHC method for such an analysis. Thirteen cases were studied from the files of the Department of Pathology, Mahidol University. Diagnoses included three Duchenne muscular dystrophy (DMD), one Becker muscular dystrophy (BMD), one dysferlinopathy, and several not-specified muscular dystrophies. IHC was performed on FFPE sections at different thicknesses (3 μm, 5 μm, and 8 μm) using the heat-mediated antigen retrieval method with citrate/EDTA buffer, followed by an overnight incubation with primary antibodies at room temperature. Antibodies against spectrin, dystrophin (rod domain, C-terminus, and N-terminus), dysferlin, sarcoglycans (α, β, and γ), and β-dystroglycan were used. Frozen sections were tested in parallel for comparative analysis. Antibodies labelling spectrin, dystrophin (rod domain and C-terminus), dysferlin, sarcoglycans (α, β, and γ), and β-dystroglycan clearly exhibited sarcolemmal staining in FFPE sections. However, staining of FFPE sections using the antibody directed against the N-terminus of dystrophin was unsuccessful. The absence of labeling for dystrophins and dysferlin in FFPE sections was documented in all three DMD patients and the dysferlinopathy patient. The BMD diagnosis could not be made using IHC in FFPE sections alone because of a lack of staining for the dystrophin N-terminus, indicating a limitation of
TruSeq-Based Gene Expression Analysis of Formalin-Fixed Paraffin-Embedded (FFPE Cutaneous T-Cell Lymphoma Samples: Subgroup Analysis Results and Elucidation of Biases from FFPE Sample Processing on the TruSeq Platform
Full Text Available Cutaneous T-cell lymphomas (CTCLs are a heterogeneous group of malignancies with courses ranging from indolent to potentially lethal. We recently studied in a 157 patient cohort gene expression profiles generated by the TruSeq targeted RNA gene expression sequencing. We observed that the sequencing library quality and depth from formalin-fixed paraffin-embedded (FFPE skin samples were significantly lower when biopsies were obtained prior to 2009. We also observed that the fresh CTCL samples clustered together, even though they included stage I–IV disease. In this study, we compared TruSeq gene expression patterns in older (≤2008 vs. more recent (≥2009 FFPE samples to determine whether these clustering analyses and earlier described differentially expressed gene findings are robust when analyzed based on the year of biopsy. We also explored biases found in FFPE samples when subjected to the TruSeq analysis of gene expression. Our results showed that ≤2008 and ≥2009 samples clustered equally well to the full data set and, importantly, both analyses produced nearly identical trends and findings. Specifically, both analyses enriched nearly identical DEGs when comparing benign vs. (1 stage I–IV and (2 stage IV (alone CTCL samples. Results obtained using either ≤2008 or ≥2009 samples were strongly correlated. Furthermore, by using subgroup analyses, we were able to identify additional novel differentially expressed genes (DEGs, which did not reach statistical significance in the prior full data set analysis. Those included CTCL-upregulated BCL11A, SELL, IRF1, SMAD1, CASP1, BIRC5, and MAX and CTCL-downregulated MDM4, SERPINB3, and THBS4 genes. With respect to sample biases, no matter if we performed subgroup analyses or full data set analysis, fresh samples tightly clustered together. While principal component analysis revealed that fresh samples were spatially closer together, indicating some preprocessing batch effect, they remained
Utzinger, J; Botero-Kleiven, S; Castelli, F; Chiodini, P L; Edwards, H; Köhler, N; Gulletta, M; Lebbad, M; Manser, M; Matthys, B; N'Goran, E K; Tannich, E; Vounatsou, P; Marti, H
The present study aimed to compare the diagnostic performance of different European reference laboratories in diagnosing helminths and intestinal protozoa, using an ether-concentration method applied to sodium acetate-acetic acid-formalin (SAF)-preserved faecal samples. In total, 102 stool specimens were analysed during a cross-sectional parasitological survey in urban farming communities in Côte d'Ivoire. Five SAF-preserved faecal samples were prepared from each specimen and forwarded to the participating reference laboratories, processed and examined under a microscope adhering to a standard operating procedure (SOP). Schistosoma mansoni (cumulative prevalence: 51.0%) and hookworm (cumulative prevalence: 39.2%) were the predominant helminths. There was excellent agreement (kappa > 0.8; p protozoa were Entamoeba coli (median prevalence: 67.6%), Blastocystis hominis (median prevalence: 55.9%) and Entamoeba histolytica/Entamoeba dispar (median prevalence: 47.1%). Substantial agreement among reference laboratories was found for E. coli (kappa = 0.69), but only fair or moderate agreement was found for other Entamoeba species, Giardia intestinalis and Chilomastix mesnili. There was only poor agreement for B. hominis, Isospora belli and Trichomonas intestinalis. In conclusion, although common helminths were reliably diagnosed by European reference laboratories, there was only moderate agreement between centres for pathogenic intestinal protozoa. Continued external quality assessment and the establishment of a formal network of reference laboratories is necessary to further enhance both accuracy and uniformity in parasite diagnosis.
Kunder, Ratika; Jalali, Rakesh; Sridhar, Epari; Moiyadi, Aliasgar; Goel, Naina; Goel, Atul; Gupta, Tejpal; Krishnatry, Rahul; Kannan, Sadhana; Kurkure, Purna; Deopujari, Chandrashekhar; Shetty, Prakash; Biyani, Naresh; Korshunov, Andrey; Pfister, Stefan M; Northcott, Paul A; Shirsat, Neelam Vishwanath
Medulloblastoma has recently been found to consist of 4 molecularly and clinically distinct subgroups: WNT, Sonce hedgehog (SHH), Group 3, and Group 4. Deregulated microRNA expression is known to contribute to pathogenesis and has been shown to have diagnostic and prognostic potential in the classification of various cancers. Molecular subgrouping and microRNA expression analysis of 44 frozen and 59 formalin-fixed paraffin embedded medulloblastomas from an Indian cohort were carried out by real-time RT-PCR assay. The differential expression of 9 microRNAs in the 4 molecular subgroups was validated in a set of 101 medulloblastomas. The tumors in the WNT subgroup showed significant (P classification of medulloblastomas into the 4 molecular subgroups was obtained using a set of 12 protein-coding genes and 9 microRNAs as markers in a real-time RT-PCR assay with an accuracy of 97% as judged by the Prediction Analysis of Microarrays. Age at diagnosis, histology, gender-related incidence, and the relative survival rates of the 4 molecular subgroups in the present Indian cohort were found to be similar to those reported for medulloblastomas from the American and European subcontinent. Non-WNT, non-SHH medulloblastomas underexpressing miR-592 or overexpressing miR-182 were found to have significantly inferior survival rates, indicating utility of these miRNAs as markers for risk stratification. The microRNA based real-time PCR assay is rapid, simple, inexpensive, and useful for molecular classification and risk stratification of medulloblastomas, in particular formalin-fixed paraffin embedded tissues, wherein the expression profile of protein-coding genes is often less reliable due to RNA fragmentation.
Reference size-matching, whole-genome amplification, and fluorescent labeling as a method for chromosomal microarray analysis of clinically actionable copy number alterations in formalin-fixed, paraffin-embedded tumor tissue.
Gunn, Shelly R; Govender, Shailin; Sims, Cynthe L; Khurana, Aditi; Koo, Samuel; Scoggin, Jayne; Moore, Mathew W; Cotter, Philip D
Cancer genome copy number alterations (CNAs) assist clinicians in selecting targeted therapeutics. Solid tumor CNAs are most commonly evaluated in formalin-fixed, paraffin-embedded (FFPE) tissue by fluorescence in situ hybridization. Although fluorescence in situ hybridization is a sensitive and specific assay for interrogating pre-selected genomic regions, it provides no information about co-existing clinically significant copy number changes. Chromosomal microarray analysis is an alternative DNA-based method for interrogating genome-wide CNAs in solid tumors. However, DNA extracted from FFPE tumor tissue produces an essential, yet problematic, sample type. The College of American Pathologists/American Society of Clinical Oncology guidelines for optimal tumor tissue handling published in 2007 for breast cancer, and in 2016 for gastroesophageal adenocarcinomas are lacking for other solid tumors. Thus, cold ischemia times are seldom monitored in non-breast cancer, non-gastroesophageal adenocarcinomas, and all tumor biospecimens are affected by chemical fixation. Although intended to preserve specimens for long-term storage, formalin fixation causes loss of genetic information through DNA damage. Here, we describe a reference size matching, whole-genome amplification, and fluorescent labeling method for FFPE-derived DNA designed to improve chromosomal microarray results from sub-optimal nucleic acids and salvage highly degraded samples. With this technological advance, whole-genome copy number analysis of tumor DNA can be reliably performed in the clinical laboratory for a wide variety of tissue conditions and tumor types. Copyright © 2018. Published by Elsevier Inc.
Evaluation of Mutational Testing of Preneoplastic Barrett's Mucosa by Next-Generation Sequencing of Formalin-Fixed, Paraffin-Embedded Endoscopic Samples for Detection of Concurrent Dysplasia and Adenocarcinoma in Barrett's Esophagus
Del Portillo, Armando; Lagana, Stephen M.; Yao, Yuan; Uehara, Takeshi; Jhala, Nirag; Ganguly, Tapan; Nagy, Peter; Gutierrez, Jorge; Luna, Aesis; Abrams, Julian; Liu, Yang; Brand, Randall; Sepulveda, Jorge L.; Falk, Gary W.; Sepulveda, Antonia R.
Barrett's intestinal metaplasia (BIM) may harbor genomic mutations before the histologic appearance of dysplasia and cancer and requires frequent surveillance. We explored next-generation sequencing to detect mutations with the analytical sensitivity required to predict concurrent high-grade dysplasia (HGD) and esophageal adenocarcinoma (EAC) in patients with Barrett's esophagus by testing nonneoplastic BIM. Formalin-fixed, paraffin-embedded (FFPE) routine biopsy or endoscopic mucosal resection samples from 32 patients were tested: nonprogressors to HGD or EAC (BIM-NP) with BIM, who never had a diagnosis of dysplasia or EAC (N = 13); progressors to HGD or EAC (BIM-P) with BIM and a worse diagnosis of HGD or EAC (N = 15); and four BIM-negative samples. No mutations were detected in the BIM-NP (0 of 13) or BIM-negative samples, whereas the BIM-P samples had mutations in 6 (75%) of 8 cases in TP53, APC, and CDKN2A (P = 0.0005), detected in samples with as low as 20% BIM. We found that next-generation sequencing from routine FFPE nonneoplastic Barrett's esophagus samples can detect multiple mutations in minute areas of BIM with high analytical sensitivity. Next-generation sequencing panels for detection of TP53 and possibly combined mutations in other genes, such as APC and CDKN2A, may be useful in the clinical setting to improve dysplasia and cancer surveillance in patients with Barrett's esophagus. PMID:26068095
Nested-PCR for the detection of Mycoplasma hyopneumoniae in bronchial alveolar swabs, frozen tissues and formalin-fixed paraffin-embedded swine lung samples: comparative evaluation with immunohistochemical findings and histological features
Paula R. Almeida
Full Text Available The diagnosis of Mycoplasma hyopneumoniae infection is often performed through histopathology, immunohistochemistry (IHC and polymerase chain reaction (PCR or a combination of these techniques. PCR can be performed on samples using several conservation methods, including swabs, frozen tissue or formalin-fixed and paraffin-embedded (FFPE tissue. However, the formalin fixation process often inhibits DNA amplification. To evaluate whether M. hyopneumoniae DNA could be recovered from FFPE tissues, 15 lungs with cranioventral consolidation lesions were collected in a slaughterhouse from swine bred in herds with respiratory disease. Bronchial swabs and fresh lung tissue were collected, and a fragment of the corresponding lung section was placed in neutral buffered formalin for 48 hours. A PCR assay was performed to compare FFPE tissue samples with samples that were only refrigerated (bronchial swabs or frozen (tissue pieces. M. hyopneumoniae was detected by PCR in all 15 samples of the swab and frozen tissue, while it was detected in only 11 of the 15 FFPE samples. Histological features of M. hyopneumoniae infection were presented in 11 cases and 7 of these samples stained positive in IHC. Concordance between the histological features and detection results was observed in 13 of the FFPE tissue samples. PCR was the most sensitive technique. Comparison of different sample conservation methods indicated that it is possible to detect M. hyopneumoniae from FFPE tissue. It is important to conduct further research using archived material because the efficiency of PCR could be compromised under these conditions.
Imaoka, Yuki; Kuranishi, Fumito; Miyazaki, Tsubasa; Yasuda, Hiroko; Ohno, Tadao
The prognosis of advanced (stage IV) cancer of the digestive organs is very poor. We have previously reported a case of advanced breast cancer with bone metastasis that was successfully treated with combined treatments including autologous formalin-fixed tumor vaccine (AFTV). Herein, we report the success of this approach in advanced stage IV (heavily metastasized) cases of gall bladder cancer and colon cancer. Case 1: A 61-year-old woman with stage IV gall bladder cancer (liver metastasis and lymph node metastasis) underwent surgery in May 2011, including partial resection of the liver. She was treated with AFTV as the first-line adjuvant therapy, followed by conventional chemotherapy. This patient is still alive without any recurrence, as confirmed with computed tomography, for more than 5 years. Case 2: A 64-year-old man with stage IV colon cancer (multiple para-aortic lymph node metastases and direct abdominal wall invasion) underwent non-curative surgery in May 2006. Following conventional chemotherapy, two courses of AFTV and radiation therapy were administered sequentially. This patient has had no recurrence for more than 5 years. We report the success of combination therapy including AFTV in cases of liver-metastasized gall bladder cancer and abdominal wall-metastasized colon cancer. Both patients experienced long-lasting, complete remission. Therefore, combination therapies including AFTV should be considered in patients with advanced cancer of the digestive organs.
Mahsa Akbari Oryani
Full Text Available Introduction: Several methods are available for the diagnosis of autoimmune bullous disease. Since the immunohistochemistry of complement component is easy and more accessible compared to other methods, it is thought that this technique as an efficient method can replace other difficult, and time-consuming procedures. Therefore, in this study we aimed to systematically review the literatures in which the diagnostic value of complement component 3d (C3d and C4d had been investigated in bullous pemphigoid. Methods: A systematic search was conducted in the PubMed, Google scholar, and Scopus using following search method (((C3d OR C4d OR complement component 3d OR complement component 4d immunohistochemistry OR (C3d OR C4d marker OR complement component 3d OR complement component 4d marker AND (bullous pemphigoid OR cutaneous pemphigoid to evaluate the diagnostic value of C3d and/or C4d for early and accurate detection of bullous pemphigoid on November 2015. Subsequently, the extracted data were described.Result: Total of 28 documents were collected and reviewed based on the purpose of this study. Of the collected articles, 21 documents were excluded in several steps of article selection process and only 7 relevant articles were included for data assessment. The results showed that the deposits of C3d and/or C4d in skin biopsies were found in 125 of 134 patients, indicating that immunohistochemistry is a reliable technique for the diagnosis of inflammatory skin diseases.Conclusion: The results of this review showed that C3d and/or C4d immunohistochemistry in skin biopsies is a reliable technique for the diagnosis of inflammatory skin diseases, particularly bullous pemphigoid.
Bastos da Silva, Inara; Batista, Tatiana Pimental de Andrade; Martines, Roosecelis Brasil; Kanamura, Cristina Takami; Ferreira, Isabelle Martins Ribeiro; Vidal, Jose Ernesto; Pereira-Chioccola, Vera Lucia
This study investigated the genetic features of Toxoplasma gondii isolated directly in autopsies of HIV-infected patients who died with severe disseminated toxoplasmosis. This retrospective analysis was conducted in a cohort of 15 HIV-infected patients with clinical and laboratory data. They had previous cerebral toxoplasmosis at least 6 months before the disseminated toxoplasmosis episode. The hypothesis was that they were infected with highly virulent parasites due to the condition in which they died. T. gondii genotyping was done directly in DNA extracted from 30 autopsy brain and lung samples (2 per patient) and mutilocus PCR-RFLP genotyping was done using 12 molecular markers. The 30 clinical samples were genotyped successfully in 8 or more loci and six suggestive genotypes were identified. One of them was Toxo DB #11, previously identified in different domestic animals and virulent in experimental animals. The other five suggestive genotypes identified in 14 patients were not described. TgHuDis1 was the most frequent and was determined in 8 patients. TgHuDis3 and TgHuDis5 were identified in two patients each. TgHuDis2 and TgHuDis4 have been identified in one patient each. These suggestive genotypes could be considered as virulent, since they caused severe tissue damage and had similar characteristics as Toxo # DB 11. Copyright © 2016 Elsevier Inc. All rights reserved.
Huang, Meggie Mo Chao; Leong, Sai Mun; Chua, Hui Wen; Tucker, Steven; Cheong, Wai Chye; Chiu, Lily; Li, Mo-Huang; Koay, Evelyn Siew-Chuan
Among patients with colorectal cancer (CRC), KRAS mutations were reported to occur in 30-51 % of all cases. CRC patients with KRAS mutations were reported to be non-responsive to anti-epidermal growth factor receptor (EGFR) monoclonal antibody (MoAb) treatment in many clinical trials. Hence, accurate detection of KRAS mutations would be critical in guiding the use of anti-EGFR MoAb therapies in CRC. In this study, we carried out a detailed investigation of the efficacy of a wild-type (WT) blocking real-time polymerase chain reaction (PCR), employing WT KRAS locked nucleic acid blockers, and Sanger sequencing, for KRAS mutation detection in rare cells. Analyses were first conducted on cell lines to optimize the assay protocol which was subsequently applied to peripheral blood and tissue samples from patients with CRC. The optimized assay provided a superior sensitivity enabling detection of as little as two cells with mutated KRAS in the background of 10(4) WT cells (0.02 %). The feasibility of this assay was further investigated to assess the KRAS status of 45 colorectal tissue samples, which had been tested previously, using a conventional PCR sequencing approach. The analysis showed a mutational discordance between these two methods in 4 of 18 WT cases. Our results present a simple, effective, and robust method for KRAS mutation detection in both paraffin embedded tissues and circulating tumour cells, at single-cell level. The method greatly enhances the detection sensitivity and alleviates the need of exhaustively removing co-enriched contaminating lymphocytes.
Kimura, Yurika; Kubo, Sachiho; Koda, Hiroko; Shigemoto, Kazuhiro; Sawabe, Motoji; Kitamura, Ken
Molecular analysis using archival human inner ear specimens is challenging because of the anatomical complexity, long-term fixation, and decalcification. However, this method may provide great benefit for elucidation of otological diseases. Here, we extracted mRNA for RT-PCR from tissues dissected from archival FFPE human inner ears by laser microdissection. Three human temporal bones obtained at autopsy were fixed in formalin, decalcified by EDTA, and embedded in paraffin. The samples were isolated into spiral ligaments, outer hair cells, spiral ganglion cells, and stria vascularis by laser microdissection. RNA was extracted and heat-treated in 10 mM citrate buffer to remove the formalin-derived modification. To identify the sites where COCH and SLC26A5 mRNA were expressed, semi-nested RT-PCR was performed. We also examined how long COCH mRNA could be amplified by semi-nested RT-PCR in archival temporal bone. COCH was expressed in the spiral ligament and stria vascularis. However, SLC26A5 was expressed only in outer hair cells. The maximum base length of COCH mRNA amplified by RT-PCR was 98 bp in 1 case and 123 bp in 2 cases. We detected COCH and SLC26A5 mRNA in specific structures and cells of the inner ear from archival human temporal bone. Our innovative method using laser microdissection and semi-nested RT-PCR should advance future RNA study of human inner ear diseases. Copyright © 2013 Elsevier B.V. All rights reserved.
Hieu, Nguyen Minh; Nam, Nguyen Hoang; Huyen, Nguyen Thi; Van Anh, Nguyen Thi; Nghia, Phan Tuan; Khoa, Nguyen Ba; Toan, Nguyen Linh; Luong, Nguyen Hoang
SiO2-coated Fe3O4 nanoparticles (Fe3O4@SiO2 NPs) were successfully synthesized using ultrasound in order to extract DNA from cancer tissues for application in diagnostics. The core 10.7-nm-diameter Fe3O4 nanoparticles were synthesized by co-precipitation of Fe3+ and Fe2+ as reaction substrates and NH4OH as precipitant, then coated with a thin layer of amorphous silica by a modified Stober method. Further SiO2 coating using alkaline hydrolysis of tetraethyl orthosilicate in ethanol and water mixture was accelerated in the presence of a 37-kHz ultrasound, resulting in the NPs having different sizes of 14.5 nm (version M1), 24.4 nm (version M2), and 34.9 nm (version M3) with saturation magnetization values of 50.2 emu/g, 18.6 emu/g, 10.3 emu/g, respectively. Among the three Fe3O4@SiO2 NPs versions, the M1 NPs allowed extraction of DNAs from 10 mg formalin-fixed and paraffin-embedded (FFPE) tissues of nasopharyngeal carcinoma patients with the highest recovery of about 100-500 ng/ μl and good purity (A260/A280: 1.8-1.9). The extracted DNAs could be used as templates for downstream amplification of 252-bp sequencing specifically for the Braf cancer biomarker gene using polymerase chain reaction (PCR), as well as detection of the pathogenic Epstein-Barr virus (EBV) and the human papilloma-virus (HPV) using real-time PCR. DNA extraction recoveries of both EBV and HPV using Fe3O4@SiO2 NPs M1 were significantly better that those using commercialized Fe3O4@SiO2 microbeads, as indicated by lower threshold cycles of all fluorescent signals including fluorescein amidite (FAM) dye representative for EBV infection, hexachlorofluorescein (HEX) dye representative for β-globin (internal control), and SYBR Green dye representative for HPV infection in tested clinical samples from patients with nasopharyngeal carcinoma (NPC).
Technical validation of an RT-qPCR in vitro diagnostic test system for the determination of breast cancer molecular subtypes by quantification of ERBB2, ESR1, PGR and MKI67 mRNA levels from formalin-fixed paraffin-embedded breast tumor specimens.
Laible, Mark; Schlombs, Kornelia; Kaiser, Katharina; Veltrup, Elke; Herlein, Stefanie; Lakis, Sotiris; Stöhr, Robert; Eidt, Sebastian; Hartmann, Arndt; Wirtz, Ralph M; Sahin, Ugur
MammaTyper is a novel CE-marked in vitro diagnostic RT-qPCR assay which assigns routinely processed breast cancer specimens into the molecular subtypes Luminal A-like, Luminal B-like (HER2 positive or negative), HER2 positive (non-luminal) and Triple negative (ductal) according to the mRNA expression of ERBB2, ESR1, PGR and MKI67 and the St Gallen consensus surrogate clinical definition. Until now and regarding formalin-fixed, paraffin-embedded material (FFPE), this has been a task mostly accomplished by immunohistochemistry (IHC). However the discrepancy rates of IHC for the four breast cancer biomarkers are frequently under debate, especially for Ki-67 which carries the highest degree of inter- and even intra-observer variability. Herein we describe a series of studies in FFPE specimens which aim to fully validate the analytical performance of the MammaTyper assay, including the site to site reproducibility of the individual marker measurements. Tumor RNA was extracted with the novel RNXtract RNA extraction kit. Synthetic RNA was used to assess the sensitivity of the RNXtract kit. DNA and RNA specific qPCR assays were used so as to determine analyte specificity of RNXtract. For the assessment of limit of blank, limit of detection, analytical measurement range and PCR efficiency of the MammaTyper kit serial dilutions of samples were used. Analytical precision studies of MammaTyper were built around two different real time PCR platforms and involved breast tumor samples belonging to different subtypes analyzed across multiple sites and under various stipulated conditions. The MammaTyper assay robustness was tested against RNA input variations, alternative extraction methods and tumor cell content. Individual assays were linear up to at least 32.33 and 33.56 Cqs (quantification cycles) for the two qPCR platforms tested. PCR efficiency ranged from 99 to 109 %. In qPCR platform 1, estimates for assay specific inter-site standard deviations (SD) were between 0.14 and
Technical validation of an RT-qPCR in vitro diagnostic test system for the determination of breast cancer molecular subtypes by quantification of ERBB2, ESR1, PGR and MKI67 mRNA levels from formalin-fixed paraffin-embedded breast tumor specimens
Laible, Mark; Schlombs, Kornelia; Kaiser, Katharina; Veltrup, Elke; Herlein, Stefanie; Lakis, Sotiris; Stöhr, Robert; Eidt, Sebastian; Hartmann, Arndt; Wirtz, Ralph M.; Sahin, Ugur
MammaTyper is a novel CE-marked in vitro diagnostic RT-qPCR assay which assigns routinely processed breast cancer specimens into the molecular subtypes Luminal A-like, Luminal B-like (HER2 positive or negative), HER2 positive (non-luminal) and Triple negative (ductal) according to the mRNA expression of ERBB2, ESR1, PGR and MKI67 and the St Gallen consensus surrogate clinical definition. Until now and regarding formalin-fixed, paraffin-embedded material (FFPE), this has been a task mostly accomplished by immunohistochemistry (IHC). However the discrepancy rates of IHC for the four breast cancer biomarkers are frequently under debate, especially for Ki-67 which carries the highest degree of inter- and even intra-observer variability. Herein we describe a series of studies in FFPE specimens which aim to fully validate the analytical performance of the MammaTyper assay, including the site to site reproducibility of the individual marker measurements. Tumor RNA was extracted with the novel RNXtract RNA extraction kit. Synthetic RNA was used to assess the sensitivity of the RNXtract kit. DNA and RNA specific qPCR assays were used so as to determine analyte specificity of RNXtract. For the assessment of limit of blank, limit of detection, analytical measurement range and PCR efficiency of the MammaTyper kit serial dilutions of samples were used. Analytical precision studies of MammaTyper were built around two different real time PCR platforms and involved breast tumor samples belonging to different subtypes analyzed across multiple sites and under various stipulated conditions. The MammaTyper assay robustness was tested against RNA input variations, alternative extraction methods and tumor cell content. Individual assays were linear up to at least 32.33 and 33.56 Cqs (quantification cycles) for the two qPCR platforms tested. PCR efficiency ranged from 99 to 109 %. In qPCR platform 1, estimates for assay specific inter-site standard deviations (SD) were between 0.14 and 0
Risk of radiation-induced subcutaneous fibrosis in relation to single nucleotide polymorphisms in TGFB1, SOD2, XRCC1, XRCC3, APEX and ATM--a study based on DNA from formalin fixed paraffin embedded tissue samples
Andreassen, C N; Alsner, J; Overgaard, M
) codon 148 polymorphisms were assessed based on archival histological material. Differences in fibrosis risk were quantified from dose-response assessments. RESULTS: For none of the investigated polymorphisms, significant associations with risk of subcutaneous fibrosis were observed. A detailed analysis...
Prasetyo, Rizki Eko; Mastutik, Gondo; Mustokoweni, Sjahjenny
HPV infection is known to cause cervical cancer. This study aimed to identify the variant of HPV genotypes of cervical precancerous lesions from low grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesion (HSIL). This was an explorative study using formalin fix paraffin embedded (FFPE) from cervical precancerous lesions at Dr. Soetomo Hospital, Surabaya. DNA was extracted from FFPE and hybridized for HPV genotyping using Ampliquality HPV Type Express kit (...
Sabol, Ivan; Milutin Gašperov, Nina; Matovina, Mihaela; Božinović, Ksenija; Grubišić, Goran; Fistonić, Ivan; Belci, Dragan; Alemany, Laia; Džebro, Sonja; Dominis, Mara; Šekerija, Mario; Tous, Sara; de Sanjosé, Silvia; Grce, Magdalena
The main etiological factor of precancerous lesion and invasive cervical cancer are oncogenic human papillomaviruses types (HPVs). The objective of this study was to establish the distribution of the most common HPVs in different cervical lesions and cancer prior to the implementation of organized population-based cervical screening and HPV vaccination in Croatia. In this study, 4,432 cervical specimens, collected through a 16-year period, were tested for the presence of HPV-DNA by polymerase chain reaction (PCR) with three sets of broad-spectrum primers and type-specific primers for most common low-risk (LR) types (HPV-6, 11) and the most common high-risk (HR) types (HPV-16, 18, 31, 33, 45, 52, 58). Additional 35 archival formalin-fixed, paraffin embedded tissue of cervical cancer specimens were analyzed using LiPA25 assay. The highest age-specific HPV-prevalence was in the group 18-24 years, which decreased continuously with age (Ptypes significantly increased (Ptype found with a prevalence (with or without another HPV-type) of 6.9% in normal cytology, 15.5% in atypical squamous cells of undetermined significance, 14.4% in low-grade squamous intraepithelial lesions, 33.3% in high-grade squamous intraepithelial lesions, and 60.9% in cervical cancer specimens (Ptypes among Croatian women, which will enable to predict and to monitor the impact of HPV-vaccination and to design effective screening strategies in Croatia.
Background NanoString’s Prosigna™ Breast Cancer Prognostic Gene Signature Assay is based on the PAM50 gene expression signature. The test outputs a risk of recurrence (ROR) score, risk category, and intrinsic subtype (Luminal A/B, HER2-enriched, Basal-like). The studies described here were designed to validate the analytical performance of the test on the nCounter Analysis System across multiple laboratories. Methods Analytical precision was measured by testing five breast tumor RNA samples across 3 sites. Reproducibility was measured by testing replicate tissue sections from 43 FFPE breast tumor blocks across 3 sites following independent pathology review at each site. The RNA input range was validated by comparing assay results at the extremes of the specified range to the nominal RNA input level. Interference was evaluated by including non-tumor tissue into the test. Results The measured standard deviation (SD) was less than 1 ROR unit within the analytical precision study and the measured total SD was 2.9 ROR units within the reproducibility study. The ROR scores for RNA inputs at the extremes of the range were the same as those at the nominal input level. Assay results were stable in the presence of moderate amounts of surrounding non-tumor tissue (<70% by area). Conclusions The analytical performance of NanoString’s Prosigna assay has been validated using FFPE breast tumor specimens across multiple clinical testing laboratories. PMID:24625003
Nielsen, Torsten; Storhoff, James; Wallden, Brett; Schaper, Carl; Ferree, Sean; Liu, Shuzhen; Gao, Dongxia; Barry, Garrett; Dowidar, Naeem; Maysuria, Malini
NanoString’s Prosigna™ Breast Cancer Prognostic Gene Signature Assay is based on the PAM50 gene expression signature. The test outputs a risk of recurrence (ROR) score, risk category, and intrinsic subtype (Luminal A/B, HER2-enriched, Basal-like). The studies described here were designed to validate the analytical performance of the test on the nCounter Analysis System across multiple laboratories. Analytical precision was measured by testing five breast tumor RNA samples across 3 sites. Reproducibility was measured by testing replicate tissue sections from 43 FFPE breast tumor blocks across 3 sites following independent pathology review at each site. The RNA input range was validated by comparing assay results at the extremes of the specified range to the nominal RNA input level. Interference was evaluated by including non-tumor tissue into the test. The measured standard deviation (SD) was less than 1 ROR unit within the analytical precision study and the measured total SD was 2.9 ROR units within the reproducibility study. The ROR scores for RNA inputs at the extremes of the range were the same as those at the nominal input level. Assay results were stable in the presence of moderate amounts of surrounding non-tumor tissue (<70% by area). The analytical performance of NanoString’s Prosigna assay has been validated using FFPE breast tumor specimens across multiple clinical testing laboratories
Evensen, O.; Lorenzen, Ellen
An immunohistochemical method is described for the detection of Flexibacter psychrophilus in formalin-fixed, parafiin-wax-embedded fry of rainbow trout. Rabbit antiserum as well as rainbow trout hyperimmune serum were used in the study. The distribution and tissue localization of the bacterium...... are typically found during the chronic stage of the disease....
In situ hybridization technique for the detection of swine enteric and respiratory coronaviruses, transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), in formalin-fixed paraffin-embedded tissues.
Sirinarumitr, T; Paul, P S; Kluge, J P; Halbur, P G
The in situ hybridization (ISH) technique was developed to detect the swine coronaviruses, transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), in cell culture and tissue sections from TGEV-or PRCV-infected pigs. The 35S-labeled RNA probes were generated from two plasmids pPSP.FP1 and pPSP.FP2 containing part of the S gene of TGEV. The procedure was first standardized in cell cultures. The radiolabeled pPSP.FP2 probe detected both TGEV and PRCV in virus-inoculated cell cultures, whereas pPSP.FP1 probe detected TGEV but not PRCV. The probe was then used to detect TGEV or PRCV in tissues of pigs experimentally infected with TGEV or PRCV or naturally infected with TGEV. Again, the probes detected TGEV in intestines of experimentally and naturally infected pigs and PRCV in the lungs of experimentally infected pigs. TGEV RNA was detected mainly within the enterocytes at the tips of villi and, less often, within some crypt epithelial cells. PRCV was shown to replicate mainly in the bronchiolar epithelial cells and in lesser amount in type II pneumocytes, type I pneumocytes, alveolar macrophages and bronchial epithelial cells, respectively. ISH has potential applications as a diagnostic test for the detection and differentiation of TGEV and PRCV in tissues and in studies to gain a better understanding of the mechanism of pathogenesis of enteric and respiratory coronavirus infections.
Identificação imuno-histoquímica de Listeria monocytogenes em placentas fixadas em formol e embebidas em parafina Immunohistochemical identification of Listeria monocytogenes in formalin-fixed and paraffin-embedded placentas
Jussara Pires Schwab
pregnancy trimester, age of pregnant women, cases of abortion and premature delivery, and to the occurrence of habitual abortion. METHODS: a retrospective study was carried out at the pathology service of a teaching hospital in the city of Porto Alegre in 2000. The paraffin blocks of 254 placentas, obtained from abortion, premature delivery and full-term birth, were analyzed by conventional histology using hematoxylin and eosin (HE staining. The IHC assay consisted of a rabbit anti-listeria polyclonal antibody B65420R (Biodesign® diluted 1:1000, in addition to the avidin-biotin-streptavidin complex; 148 placentas revealed inflammatory disorders, hemorrhage, necrosis and thrombosis. The c² test was used for statistical analysis. RESULTS: Listeria monocytogenes was detected in 33.78% of the placentas analyzed by IHC. Chorioamnionitis and villitis showed significant statistical difference in the positive placentas. Lm occurred in the 1st, 2nd and 3rd trimester of pregnancy. The age of pregnant women, the cases of abortion and/or premature births were not statistically different as to the presence or absence of Lm in the placentas. Habitual abortions occurred in patients with or without Lm in the placental tissue. CONCLUSION: Immunohistochemistry may be used to confirm the histopathological diagnosis of listeriosis in all trimesters of pregnancy.
Lundemose, AG; Lundemose, JB; Birkelund, Svend
examined and the effect of autolysis and tetracycline treatment was evaluated. Furthermore, lung tissue from two patients who died of ornithosis was examined. Inclusions detected in lung sections showed a bright apple-green fluorescence, and had a characteristic and easily recognizable morphology...
Full Text Available Background: Carcinomas of esophagus, mostly squamous cell carcinomas, occur throughout the world. There are a number of suspected genetic or environmental etiologies. Human papilloma virus (HPV is said to be a major etiology in areas with high incidence of esophageal carcinoma, while it is hardly detectable in low incidence regions. This study was designed to evaluate the prevalence of HPV in esophageal squamous cell carcinoma (ESCC cases diagnosed in Pathology Department, Medical School, Shiraz University of Medical Sciences.Methods: DNA material for PCR amplification of HPV genome was extracted from formalin-fixed paraffin-embedded tissue blocks of 92 cases of ESCC, diagnosed during 20 years from 1982 to 2002. Polymerase chain reaction was performed for amplification and detection of common HPV and type specific HPV-16 and HPV-18 genomic sequences in the presence of positive control (HPV-18 and HPV positive biopsies of uterine exocervix and additional internal controls i.e. beta-globin and cytotoxic T lymphocyte antigen 4 (CTLA4.Result: Good amplification of positive control and internal controls was observed. However, no amplification of HPV genome was observed.Conclusion: There is no association between HPV infection and the development of esophageal squamous cell carcinoma in the cases evaluated.
Otimização da imunoistoquímica para detecção de herpesvírus bovino tipo 5 (BHV-5 em tecidos do sistema nervoso central fixados com formaldeído Optimization of immunohistochemistry for Bovine Herpesvirus 5 (BHV-5 detection on formalin-fixed tissues of the central nervous system
Full Text Available Com o objetivo de otimizar a técnica de imunoistoquímica para detecção de herpesvírus bovino tipo 5 (BHV-5 em tecidos do sistema nervoso central fixado em formaldeído, foram avaliados diferentes métodos de digestão enzimática, diferentes anticorpos e reagentes para bloqueio de reações inespecíficas. As reações apresentaram a máxima intensidade de coloração específica e a quantidade mínima de coloração de fundo quando foram usadas protease de Streptomyces griseus (0,1% ou proteinase K de Tritirachium album limber (0,05%, mediante incubação durante 15 minutos a 37°C. Entre os anticorpos monoclonais analisados, dois foram capazes de detectar BHV-5. As reações inespecíficas foram bloqueadas mais efetivamente pela incubação do tecido com caseína (0,5%, durante cinco minutos, ou leite em pó (2,5%, durante 60 minutos, ou soro eqüino (2,5%, durante 60 minutos. Com a técnica otimizada foi possível a detecção de BHV-5 em material de arquivo.In order to optimize immunohistochemical technique (IHC for detection of Bovine herpesvirus type 5 (BHV-5 on formalin-fixed sections of central nervous system, different methods of enzymatic digestion, use of different antibodies and products for blocking of nonspecific reactivity were evaluated. The reactions showed the highest intensity of specific coloration and the minimum amounts of background when protease from Streptomyces griseus (0.1% or proteinase K from Tritirachium album limber (0.05% were used, incubating for 15 minutes at 37°C. Only two of the tested monoclonal antibodies specifically labelled BHV-5 antigen. The nonspecific reactions were blocked through incubation of tissues with casein (0.5% for five minutes or powdered milk (2.5% for 60 minutes or equine serum (2.5% for 60 minutes. The optimized immunohistochemical method allowed the detection of BHV-5 antigen in histopathological archives.
Ahmed E. Mansour
Conclusion: The prevalence of spontaneous bacterial pleuritis in the studied group of patients with hepatic hydrothorax was 14.3%. Patients with advanced liver disease, low pleural fluid protein, or SBP are at risk for spontaneous bacterial pleuritis.
The roles of causative factors responsible for prevalence of malaria in the village of Solana, India, were studied. Mosquitoes and larvae density in and around the area were measured by process of random sampling and counting their numbers under microscopy. Malaria in population of the village was diagnosed by ...
Conclusion: Results indicated that dental caries prevalence among school children in Qatar has reached critical levels, and is influenced by socio-demographic factors. The mean decayed, missing, and filled teeth values obtained in this study were the second highest detected in the Eastern Mediterranean region.
Full Text Available Methicillin-resistant Staphylococcus aureus (MRSA is a major nosocomial pathogen worldwide. Malta is one of the countries with the highest MRSA prevalence in Europe, as identified from hospital blood cultures . However, community prevalence of MRSA has never previously been investigated. This study aimed at establishing the prevalence of community MRSA nasal colonization in Maltese individuals and identifying the clonal characteristics of the detected isolates. Nasal swabs were collected from 329 healthy individuals who were also asked to complete a brief questionnaire about risk factors commonly associated with MRSA carriage and infection. The swabs were transported and enriched in a nutrient broth supplemented with NaCl. The presence of MRSA was then determined by culturing on MRSA Select chromogenic agar and then confirming by several assays, including catalase, coagulase and PBP2a agglutination tests. The isolates were assayed for antibiotic susceptibilities and typed by microarray analysis to determine the clonal characteristics of each strain. The prevalence of MRSA nasal colonization in the healthy Maltese population was found to be 8.81% (95% confidence interval [CI], 5.75–11.87%, much higher than that found in other studies carried out in several countries. No statistical association was found between MRSA carriage and demographics or risk factors; however, this was hindered by the small sample size. Almost all the isolates were fusidic-acid resistant. The majority were found to belong to a local endemic clone (CC5 which seems to be replacing the previously prevalent European clone UK-EMRSA-15 in the country. A new clone (CC50-MRSA-V was also characterized. The presence of such a significant community reservoir of MRSA increases the burdens already faced by the local healthcare system to control the MRSA epidemic. Colonization of MRSA in otherwise healthy individuals may represent a risk for endogenous infection and transmission to
Shirol, Pallavi D; Naik, Veena; Kale, Alka
Lichen planus is a premalignant condition with minimal diagnostic aids. This study is an attempt to use paraffin embedded sections of lichen planus with immunofluorescein stain and to evaluate the immunofluorescent sections to establish pattern of fibrinogen deposition. Thirty-five paraffin embedded sections of old and new cases of oral lichen planus (study group) and five normal oral mucosa (control group) were chosen. Two sections of each (H & E) case were taken, one was stained with hematoxylin and eosin and another with fluorescein isothiocynate conjugate (FITC) polyclonal rabbit antibody against fibrinogen. Fluorescent findings were examined with a fluorescent microscope. A high statistical significant correlation was found in respect to fluorescence positivity, intensity of fluorescence and distribution of fluorescence each with p direct immunofluorescence staining in routine set up where only formalin fixed tissues are received. Paraffin embedded sections can be successfully used in direct immunofluorescence staining when only formalin fixed tissues are received.
Monaco, A; Cattaneo, R; Spadaro, A; Marchetti, E; Barone, A
The aim of this study was to investigate the prevalence of kinesiographic coincidence between the most cranial position during deglutition of mandible and habitual occlusal position and to evaluate the distribution of clinical diagnosis according to the kinesiographic pattern of deglutition. 201 random patients in waiting list for dental treatment and classified as orthodontic patients, prosthetic patients, TMD patients and control patients, were evaluated. Kinesiographic records were acquired using K7I and positioning a magnetic sensor frame integral with the head and with the sensory field balanced on an artificial magnet adhering to the mucosa covering the roots of the lower mandibular incisors. The kinesiographic occlusal position was compared to the kinesiographic most cranial position of the mandible during swallowing. 99 patients displayed a discrepancy between the most cranial position during swallowing and the occlusal position. 102 patients did not show any discrepancy. In this group the kinesiographic most cranial position during swallowing coincided with the occlusal position. The finding suggests that computerised kinesiography could be useful to study deglutition, detecting in a reliable way the movement pattern. Atypical deglutition seems to be less atypical than previously though in dental patient population and, despite these data confirm its correlation with malocclusion, we noted an inverse correlation with necessity of prosthetic treatment and no higher prevalence in TMD patients.
Full Text Available Aim of the study was to clarify the prevalence and status of smoking among today’s teenagers according to the survey. Materials and methods. There was carried out the study of the prevalence of smoking among adolescents in the Dnipro (n = 2299 individuals. Results. According to the results of the survey, there were 20.8 % active smokers among adolescents, among them 89.1 % adolescents smoked every day, including 24.9 % boys and 14.6 % girls. On average, adolescents begin smoking regularly at the age of 14.4 ± 0.1 year. The average length of smoking in adolescents was to 3.4 ± 0.1. The average number of cigarettes adolescents smoked per day made up of 9.1 ± 0.3 units. Studying the type of tobacco products 48.6 % of adolescents were found to smoke light cigarettes. The average index of smoker was 109.1 ± 3.3 and was significantly higher in boys (125.5 ± 4.9 than in girls (91.9 ± 4.0 (p < 0.001. Study of the motivation to quit smoking found that only 20.3 % of teenagers are highly motivated to quit, whereas about 31.7 % of adolescents have no motivation to quit smoking. Conclusions. The study showed that the problem of tobacco use remains highly relevant: in the city every fifth teenager smokes. The findings necessitate widespread introduction of antismoking programs among adolescents, which should be directed first to reduce the intensity of smoking and increase motivation to quit smoking, promote healthy lifestyles, as well as thorough clinical examination of adolescent smokers with the aim of identifying risk groups for the development of chronic obstructive pulmonary disease.
. While there is continued efforts to determine factors associated, there would be need to examine if birth order influences its prevalence. Aim: The present study was aimed to determine the prevalence of psychoactive substance use among ...
Chika, Noriyasu; Eguchi, Hidetaka; Kumamoto, Kensuke; Suzuki, Okihide; Ishibashi, Keiichiro; Tachikawa, Tetsuhiko; Akagi, Kiwamu; Tamaru, Jun-Ichi; Okazaki, Yasushi; Ishida, Hideyuki
We investigated the prevalence of Lynch syndrome and Lynch-like syndrome among Japanese colorectal cancer patients, as there have been no credible data from Japan. Immunohistochemical analyses for mismatch repair proteins (MLH1, MSH2, MSH6 and PMS2) were carried out in surgically resected, formalin-fixed paraffin-embedded specimens obtained from 1,234 newly diagnosed colorectal cancer patients between March 2005 and April 2014. The presence/absence of the BRAF V600E mutation and hypermethylation of the MLH1 promoter was analyzed where necessary. Genetic testing was finally undertaken in patients suspected as having Lynch syndrome. By the universal screening approach with immunohistochemical analysis for mismatch repair proteins followed by analyses for the BRAF V600E mutation and MLH1 promoter methylation status, 11 (0.9%) of the 1,234 patients were identified as candidates for genetic testing. Out of the 11 patients, 9 (0.7%) were finally diagnosed as having Lynch syndrome; the responsible genes included MLH1 (n = 1), MSH2 (n = 4), EPCAM (n = 1) and MSH6 (n = 3). The remaining two patients (0.2%) were regarded as having Lynch-like syndrome, since biallelic somatic deletion of the relevant mismatch repair genes was detected in the absence of germline mismatch repair alterations. None of the cases was identified as having germline MLH1 epimutation. The prevalence of Lynch syndrome among all newly diagnosed cases of colorectal cancer in Japan is in the same range as that recently reported by studies in Western population. The prevalence of Lynch-like syndrome seems to be extremely low. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: email@example.com
Gracia, Elías; Froesch, Patrizia; Mazzucchelli, Luca; Martin, Vittoria; Rodríguez-Abreu, Delvys; Jiménez, Julio; Melgares, María; Santos, Dania; Capó, Virginia; Cavalli, Franco; Zucca, Emanuele; Bertoni, Francesco
Most ocular adnexal lymphomas (OAL) are extranodal marginal zone B-cell lymphomas (EMZL) of mucosa-associated lymphoid tissue (MALT)-type. Chronic antigen stimulation has been suggested to have a pathogenetic role in EMZL and Chlamydia psittaci chronic infection has been recently associated with the development of OAL in a series of patients from Italy. To assess this association, an evaluation of the presence of C. psittaci was made in a different OAL population. DNA samples were obtained from formalin-fixed, paraffin-embedded sections samples of 26 patients with OAL, 20 non-OAL and 20 benign ocular lesions, diagnosed and treated between 1998 and 2003 at National Institute of Oncology in Havana, Cuba. All samples were histologically reviewed by an expert pathologist. Fluorescence in situ hybrization (FISH) analysis of translocations involving MALT1 was performed. The presence of bacterial DNA was assessed with a multiplex touchdown enzyme time release polymerase chain reaction. DNA sequencing was performed to confirm suspicious bands. Seventy-three percent of the OAL cases were EMZL and 81% were in stage IE. FISH analysis was performed in 13 OAL cases and none of them evidenced MALT1 translocations. DNA of C. psittaci was detected in 11% of the 46 lymphomas: two orbital EMZL and three non-OAL. All 20 benign ocular lesions were negative for C. psittaci. The low prevalence of C. psittaci in OAL suggests geographical differences in the etiology of this entity. International studies are needed to clarify the role of C. psittaci in OALs.
Quigley, Patricia A; Palacios, Polly; Spehar, Andrea M
The Veterans Health Administration (VHA) serves the health care needs of an adult, predominantly male, and aging population. The aging profile of VHA patients is 25% greater than the civilian sector (DVA 2001). Aged patients are at higher risk for falls. In February 2002, 6 VHA medical centers profiled their inpatients' fall risk profile as one aspect of program initiatives targeted at reducing veterans' fall risk and fall-related injuries, participating in a one-day collection of fall risk measurement using the Morse Fall Scale (MFS) for all inpatients (n = 1819), acute and long-term care units. Data results are reported for age, MFS score, and the relationship between age and score, and by type of ward/unit, ie, predominately acute and critical care or long-term care. The results of this prevalence study documented that the veteran inpatient population are at high-risk for anticipated physiological falls. This Veteran Integrated Services Network-wide Deployment of an Evidence-based Program to Prevent Patient Falls study was completed as part of a nationally funded clinical initiative, National Program Initiative 20-006-1.
Full Text Available Objectives: Goals of epidemiological studies are the description of the measures of frequency of diseases, the attempt to clarify possible etiopathogenic mechanisms, and the provision of data to support health policy decisions. To increase the familiarity of rheumatologists toward epidemiology, we describe the methodology used in a prevalence study of musculoskeletal complaints performed in Chiavari, Italy. Methods: A questionnaire, originally developed by the Epidemiology Unit of the Arthitis Research Council in Manchester, UK, to investigate the prevalence of rheumatoid arthritis, was used after translation and validation. 4456 subjects aged 16 years or more listed in four general practices were invited to participate in the study and to fill the ARC questionnaire. The 3294 responders reported a any past occurrence of joint swelling lasting more than four weeks and the distribution of the swollen joints on a mannequin; b any joint pain lasting more than four weeks; c current joint pain or swelling; d morning stiffness; e whether they had been previously told by a doctor they had arthritis. Results: Four steps were necessary to obtain a 74% response, i.e. direct contact, two mailings and a phone interview. The performance of the different questions was good. The prevalence of the most common conditions among patients answering positively to the questions regarding morning stiffness and symmetrical swelling of joints was as follows: osteoarthritis 2.60%, fibromyalgia 1.30%, carpal tunnel syndrome 1.14%, rheumatoid arthritis 0.31%, and psoriatic arthritis 0.10%. Conclusions: Methodological issues regarding the selection of the population and sample to study, the development of a questionnaire, and the problems in obtaining valid informations are discussed.
This could be attributed to management and differences in the epidemiology of sampling sites. The prevalence of Amblyomma and. Boophilus in the cattle were 28.33% and 11.25% respectively. The higher prevalence of tick in cattle in this study was due to managemental and epidemiological factors. Ticks require moisture ...
Zhao, Dongdong; Dong, Bao; Yu, Dandan; Ren, Qiongqiong; Sun, Yehuan
A growing number of studies have investigated the prevalence of Molar Incisor Hypomineralization (MIH) around the world. The aim of this study was to systematically estimate the pooled prevalence of MIH. A comprehensive literature research was completed in English and Chinese databases. Random effect models were used to calculate the pooled prevalence. To address the heterogeneity, meta-regression, and sensitivity analyzes were conducted. Publication bias was estimated by trim and fill method. Seventy eligible studies were included. The pooled prevalence of MIH was 14.2% globally. In subgroup analysis, South America (18.0%, 95% CI: 13.8-22.2) and Spain (21.1%, 95% CI: 17.7-24.6) had the highest prevalence. There was no significant difference between males (14.3%, 95% CI: 12.0-16.6) and females (14.4%, 95% CI: 12.8-15.9). The prevalence of MIH among children 10 years of age or younger (15.1%, 95% CI: 12.1-18.2) was much higher than the prevalence of MIH among older children (12.1%, 95% CI: 8.0-16.3). Sample size explained 15.7% heterogeneity. MIH has a high incidence globally, especially among children <10 years old. It is, therefore, imperative to develop more appropriate dental healthcare strategies to care for these children and to identify the etiology of MIH to prevent it occurring. © 2017 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Pilot study for assessment of prevalence of intrafamilial hepatitis C transmission in relation to salivary viral load among infected patients with and without chronic renal failure. H El Tayeb, NA El Nakeeb, MM Sayed, WA Yousry, SHA Agwa ...
Traube, Chani; Silver, Gabrielle; Reeder, Ron W; Doyle, Hannah; Hegel, Emily; Wolfe, Heather A; Schneller, Christopher; Chung, Melissa G; Dervan, Leslie A; DiGennaro, Jane L; Buttram, Sandra D W; Kudchadkar, Sapna R; Madden, Kate; Hartman, Mary E; deAlmeida, Mary L; Walson, Karen; Ista, Erwin; Baarslag, Manuel A; Salonia, Rosanne; Beca, John; Long, Debbie; Kawai, Yu; Cheifetz, Ira M; Gelvez, Javier; Truemper, Edward J; Smith, Rebecca L; Peters, Megan E; O'Meara, A M Iqbal; Murphy, Sarah; Bokhary, Abdulmohsen; Greenwald, Bruce M; Bell, Michael J
To determine prevalence of delirium in critically ill children and explore associated risk factors. Multi-institutional point prevalence study. Twenty-five pediatric critical care units in the United States, the Netherlands, New Zealand, Australia, and Saudi Arabia. All children admitted to the pediatric critical care units on designated study days (n = 994). Children were screened for delirium using the Cornell Assessment of Pediatric Delirium by the bedside nurse. Demographic and treatment-related variables were collected. Primary study outcome measure was prevalence of delirium. In 159 children, a final determination of mental status could not be ascertained. Of the 835 remaining subjects, 25% screened positive for delirium, 13% were classified as comatose, and 62% were delirium-free and coma-free. Delirium prevalence rates varied significantly with reason for ICU admission, with highest delirium rates found in children admitted with an infectious or inflammatory disorder. For children who were in the PICU for 6 or more days, delirium prevalence rate was 38%. In a multivariate model, risk factors independently associated with development of delirium included age less than 2 years, mechanical ventilation, benzodiazepines, narcotics, use of physical restraints, and exposure to vasopressors and antiepileptics. Delirium is a prevalent complication of critical illness in children, with identifiable risk factors. Further multi-institutional, longitudinal studies are required to investigate effect of delirium on long-term outcomes and possible preventive and treatment measures. Universal delirium screening is practical and can be implemented in pediatric critical care units.
Best, Kate E; Addor, Marie-Claude; Arriola, Larraitz
BACKGROUND: Hirschsprung's disease is a congenital gut motility disorder, characterised by the absence of the enteric ganglion cells along the distal gut. The aim of this study was to describe the epidemiology of Hirschsprung's disease, including additional congenital anomalies, total prevalence......, trends, and association with maternal age. METHODS: Cases of Hirschsprung's disease delivered during 1980 to 2009 notified to 31 European Surveillance of Congenital Anomaly registers formed the population-based case-series. Prevalence rates and 95% confidence intervals were calculated as the number...... interval, 1.03-1.15) per 10,000 births and there was a small but significant increase in prevalence over time (relative risk = 1.01; 95% credible interval, 1.00-1.02; p = 0.004). There was evidence of geographical heterogeneity in prevalence (p chromosomal...
Jauhri, Mayank; Bhatnagar, Akanksha; Gupta, Satish; Bp, Manasa; Minhas, Sachin; Shokeen, Yogender; Aggarwal, Shyam
Colorectal cancer incidences are on a rise in India. In this study, we have analyzed the mutation frequencies of six potential biomarkers, their coexistence, association with clinicopathological characteristics, and tumor location in Indian colorectal cancer patients. Next-generation sequencing was performed to identify mutations in the six potential biomarker genes using formalin-fixed paraffin-embedded tissue blocks of 112 colorectal cancer patients. The mutation frequency observed in KRAS, BRAF, PIK3CA, NRAS, TP53, and APC was 35.7%, 7.1%, 16.1%, 6.3%, 39.3%, and 29.5%, respectively. The significant associations of mutations were KRAS with age less than 60 years (p = 0.041), PIK3CA with males (p = 0.032), tumor stage I-II (p = 0.013), lack of metastasis in lymph nodes (p = 0.040), NRAS with rectum (p = 0.002), and APC with T2 stage of tumor growth (p = 0.013). No single patient harbored mutations in these six genes or any five genes simultaneously. Significance was noted in coexistence of KRAS with APC (p = 0.024) and mutual exclusion of KRAS with BRAF (p = 0.029). PIK3CA exon 9 was observed to be more frequently associated with KRAS mutations than PIK3CA exon 20 (p = 0.072). NRAS mutations were mutually exclusive with BRAF and PIK3CA mutations. As per our knowledge, this is the first next-generation sequencing-based biomarker study in Indian colorectal cancer patients. Frequent coexistence of gene mutations in pairs and triplets suggests that synergistic effect of overlapping mutations might further trigger the disease. In addition, infrequent coexistence of multiple gene mutations hints toward different signaling pathways for colorectal cancer tumorigenesis.
Full Text Available Stroke is a leading cause of morbidity and mortality worldwide and its late burden has mainly been attributable to developing countries. Lebanon is one of these countries where epidemiological studies on stroke burden are scarce but necessary. Thus, the present study was conducted to assess the prevalence of stroke survivors among Lebanese inhabitants. A cross-sectional survey was carried out using randomly selected landline phone numbers on all governorates to retrieve data on stroke survivors and their sociodemographic characteristics. Results were then standardized over the Lebanese and the World Health Organization (WHO world populations. A total of 6963 Lebanese inhabitants were included in the study; among these were 56 stroke survivors. This led to an adjusted stroke prevalence of 0.50% [95% confidence interval (CI = 0.33–0.66%] and a world-standardized prevalence of 0.60% (95% CI = 0.42–0.78%. A significantly higher stroke prevalence was found among older age groups and more socioeconomically privileged areas. Overall, the study showed a relatively higher prevalence of stroke in this sample of Lebanese inhabitants when compared to other developing countries. However, larger community-based studies with a clinical assessment of stroke cases are needed to confirm our findings.
Full Text Available BACKGROUND: Human Papilloma virus (HPV is recognized as the etiological agent of benign and malignant ano-genital lesions. The most prevalent genotypes associated with cervical carcinoma are HPV-16 and -18 worldwide. However, recent studies have emphasized the role of other genotypes, such as HPV-51, in the pathogenesis of cervical dysplasia. The aim of the study was to estimate the burden of HPV-51 infection in invasive cervical malignant lesions in Northern Sardinia, Italy. METHODS/PRINCIPAL FINDINGS: An observational, retrospective, prevalence, mono-center study was carried out to evaluate the presence of HPV genotypes in tissues biopsies of cervical lesions (CIN-1, CIN-2, CIN-3 and invasive carcinoma gathered from 1996 to 2009. Biological samples were collected from women admitted consecutively to a tertiary university hospital situated in Sassari, Italy. Molecular methods were used to identify 28 oncogenic HPV types. A total of 155 formalin-fixed and paraffin-embedded cervical tissue samples were analyzed. Approximately half of the cervical lesions were classified as invasive carcinoma. HPV-DNA was detected in 71% of the samples, with a higher frequency (100% in those categorized as invasive neoplasia. Mono- or co-infections were demonstrated in 45.8% and 25.8% of the cervical samples, respectively. Overall, the most prevalent HPV types were -16 (49% and -51 (19.4%, with an increased frequency of detection associated with the severity of the cervical lesions. CONCLUSIONS/SIGNIFICANCE: This survey highlights for the first time the relevant role of HPV-51 infection in the pathogenesis of invasive cervical cancer prior to the introduction of a vaccination program. Although a selection bias could have influenced the results, other recent studies have described the impact of HPV-51. This remarkable epidemiological element should be carefully evaluated, particularly in the view of opting for preventive vaccines, whose cross-protection patterns
Full Text Available Irene Petersen,1,2 Shuk-Li Collings,1,3 Rachel L McCrea,1 Irwin Nazareth,1 David P Osborn,4 Phil J Cowen,5 Cormac J Sammon1 1Department of Primary Care and Population Health, University College London, London, UK; 2Department of Clinical Epidemiology, Aarhus University, Aarhus N, Denmark; 3OXON Epidemiology, London, UK; 4Division of Psychiatry, University College London, London, UK; 5University Department of Psychiatry, Warneford Hospital, Oxford, UK Objective: The aim of this study was to examine the prevalence of major congenital malformations associated with antiepileptic drug (AED treatment in pregnancy.Patients and methods: Using data from The Health Improvement Network, we identified women who have given live birth and their offspring. Four subgroups were selected based on the AED treatment in early pregnancy, valproate, carbamazepine, lamotrigine and women not receiving AED treatment. We compared the prevalence of major congenital malformations within children of these four groups and estimated prevalence ratios (PRs using Poisson regression adjusted for maternal age, sex of child, quintiles of Townsend deprivation score and indication for treatment.Results: In total, 240,071 women were included in the study. A total of 229 women were prescribed valproate in pregnancy, 357 were prescribed lamotrigine and 334 were prescribed carbamazepine and 239,151 women were not prescribed AEDs. Fifteen out of 229 (6.6% women prescribed valproate gave birth to a child with a major congenital malformation. The figures for lamotrigine, carbamazepine and women not prescribed AEDs were 2.7%, 3.3% and 2.2%, respectively. The prevalence of major congenital malformation was similar for women prescribed lamotrigine or carbamazepine compared to women with no AED treatment in pregnancy. For women prescribed valproate in polytherapy, the prevalence was fourfold higher. After adjustments, the effect of estimates attenuated, but the prevalence remained two- to
The main source of animal protein for humans is livestock and their products. Parasitism is one of the main constraints limiting livestock production. A study was conducted to determine the prevalence of helminths in slaughtered cattle, sheep and goats in Mwanza City, Tanzania. The period of study was from 2008 to 2011 ...
Studies on Prevalence and Risk Factors for Hepatitis B Surface Antigen among Secondary School Students in North-central, Nigeria. ... high among the studied population. This suggests that public awareness on the virus be accorded urgent attention, while vaccination programme should be improved in the community.
Shah, Sandeep H; Rajani, Kajal; Kataria, Lakhan; Trivedi, Ashish; Patel, Sangita; Mehta, Kedar
Domestic violence is a major contributor to physical and mental ill health of the victim, and it is evident to some degree, in every society of the world. 1) To study perception about domestic violence in the study population. 2) To compare prevalence of domestic violence within the three subgroups of the study population (i.e. spouses of psychotic patients, spouses of non-psychiatric patients and hospital staff). A cross-sectional study was conducted among married men and women coming to Dhiraj General Hospital. Interviews were conducted using a semi-structured questionnaire. Inquiry was done about their perception regarding domestic violence, own experience any time in their life, and about the form of violence. Data was entered and analyzed using SPSS. 42.7% of study participants had never heard the words domestic violence. The overall prevalence of any form of violence in the study population as a whole was 32.3%. There was no significant difference found in the proportion of domestic violence among the three groups. The prevalence of physical, emotional, sexual and economic domestic violence was 16.3%, 25.3%, 2% and 11.3% respectively. Younger age group and female sex were significantly associated with the occurrence of domestic violence. Apart from the high prevalence of domestic violence in the present era, it is evident from the study that the participants' perception about domestic violence was low. Efforts should be made to raise public consciousness and reporting of domestic violence and its attendant consequences.
Brochado, Sandra; Soares, Sara; Fraga, Sílvia
This descriptive scoping aims to understand how the prevalence of cyberbullying has been estimated across studies. A systematic scoping review of cyberbullying empirical studies was conducted by using three bibliographic databases to search for papers published between January 2004 and August 2014. A protocol was defined to identify the relevant papers. Papers selected were included in a data sheet developed by the authors to record specific findings. In total, 159 studies were included in the scoping review. Most of the prevalence studies were conducted in the last 4 years, mainly in North America ( n = 77) and in Europe ( n = 65). High methodological heterogeneity was found among the studies, which may contribute to explain variability in prevalence estimates. Cyberbullying experiences were assessed through several different perspectives: focused only on victims, focused only on perpetrators, or focused on both victims and perpetrators (without differentiating between if they are victims or perpetrators). Most of the studies tend to assess cybervictimization experiences. However, even considering the same perspective, the same country, and the same recall period, a high variability in the estimates was observed. As a main conclusion, the way in which the prevalence of cyberbullying is estimated is influenced by methodological research options.
A prospective longitudinal study to estimate the prevalence of obesity in Egyptian children with nocturnal enuresis and the association between body mass index and ... Egyptian Journal of Medical Human Genetics ... Response to the treatment was evaluated statistically and correlated with body mass index percentile.
Objectives: This was a cross-sectional study to determine the sero prevalence of serum antibodies to three parasitic infections namely Entamoeba histolytica, Schistosoma sp. and Toxoplasma gondii, which are opportunistic infections among HIV/AIDS patients. Methods: One thousand and eighty patients that attended three ...
Comparative study on prevalence of gastrointestinal parasites, ticks and subclinical mastitis as well as milk yield was conducted on local zebu lactating dairy cows exposed to different feeding regimes in dry seasons at Sululta and Mukaturi districts of North Shewa zone, central Ethiopia. A total of 15 households at each ...
Dec 20, 2011 ... Exposure/ sharing of sharp objects had a prevalence of 64.0% in males compared to. 36.0% in females. DISCUSSION. In this study, 35 (18.4%) of the participants were positive for HBsAg. This figure is alarming considering the future role of these youths in the economy of a developing country like Nigeria.
Full Text Available INTRODUCTION: Hepatitis B is a global health problem. Among the South East Asian countries India is in intermediate zone of prevalence I.e. 2 to 5 %. 1 In the state of Tripura the prevalence of Hepatitis B among hemodialysis patients is 7.3%2 and among voluntary blood donors it is 1.2%. 3 The present study is aimed to assess the prevalence of HBV infection in Tripura at community level in particular among the different ethnic communities prevailing in this region. METHODS: A cross sectional study at community level has been done. Total 6202 samples have been collected from healthy individual above 10 years of age and a 1:4 sampling of household done. Serological marker of HBs Ag was done by ELISA.RESULTS: Prevalence of HbsAg sero-positivity was 3.6% (95%CI 3.14 - 4.06 at community level of Tripura. A higher prevalence was found in males than females which was 4.5% (95%CI 3.77-5.23 & 2.65% (95%CI 2.08 – 3.22 respectively. The prevalence of Hepatitis B was found higher among tribal community than non-tribal community which was 5.3% (95%CI 4.49 – 6.10 and 1.97%(95%CI 1.49 – 2.45 respectively. Among tribal population highest prevalence was observed among Chakma community (11.41% which was followed by Reang (7.69%, Noatia (6.09%, Jamatia (5.7%, Murasing (5.15%, Tripuri (4.95%, Halam(4.21%, and Lusai (2.7% respectively. The study shows that HBsAgseropositivity increases with age in community level reaching peak between 61 to 80 years age group I.e. 4.8% which is statistically significant. CONCLUSION: Higher prevalence of Hepatitis B among tribal population is of paramount importance from public health point of view and early intervention by Hepatitis B vaccination will reduce the disease burden among Tribal population in Tripura.
Pavare, Jana; Grope, Ilze; Gardovska, Dace
In accordance with the 1st International pediatric sepsis consensus conference, where sepsis was defined as SIRS associated with suspected or proven infection, we have identified the need to assess the prevalence of SIRS and sepsis in children with abnormal temperatures hospitalized in The Children's Clinical University Hospital in Latvia. A descriptive prospective point prevalence study (using two time periods, each 24 h, randomly chosen) was conducted on all children (n = 943) treated in the hospital. All children with abnormal temperatures - fever or hypothermia (n = 92) - were included in the study. Questionnaires evaluating age-specific SIRS criteria were completed. The prevalence of SIRS was detected with 95% CI. Out of a total of 943 patients treated in the hospital, 10% (n = 92) had abnormal temperatures. In all these cases the abnormal temperature was a fever; hypothermia was not established in any patient. Of the children with fever, 72% (n = 66) had SIRS. Of the SIRS patients, 8% (n = 5) developed sepsis, 5% (n = 3) severe sepsis and 2% (n = 1) septic shock. Seventy-six percent (n = 50) of the SIRS patients had fever in combination with respiratory rate >2 SD above normal for age; 50% (n = 33) had fever with abnormal leukocyte count; 15% (n = 10) had fever with tachycardia >2 SD above normal for age. Most of the SIRS patients (39%, n = 25) were aged 2-5 years. Twenty-one percent (n = 14) of the children with SIRS and 50% (n = 2) of those with severe sepsis and septic shock had an underlying disease. In no case was SIRS and sepsis recognized by doctors and the diagnoses were not recorded on the patients' cards. Our results would indicate a high risk for sepsis development in children with SIRS. Early SIRS diagnosis and awareness of risk of developing sepsis could change the medical approach to the patient in everyday clinical practice, eventually leading to early, goal-directed therapy for sepsis.
Full Text Available Objectives: The aim of the study was the assessment of SLE prevalence in an adult population. The study was perfomed on a sample of more than 30.000 patients over 18 resident in the Florence sub-area of Scandicci- Le Signe (Italy on the basis of the register of general practitioners (GPs. Methods: Twenty GPs gave the Lupus Screening Questionnaire (LQS to their patients. The LQS has been completed by 32.521 patients resident in Scandicci, Lastra a Signa and Signa area. On the basis of the LQS analysis an SLE diagnosis was suspected in 30 patients. These 30 patients have been investigated by routine exams, antinuclear antibodies and have been refered to rheumatologist to assess the SLE diagnosis and disease activity (ECLAM Score. Results: The overall population aged >18 years was 71.204 ( 42.474 living in Scandicci, 15.368 in Lastra a Signa and 13.362 in Signa. The diagnosis was confirmed in 23 out of the 30 suspected SLE patients. Eleven out of the 23 SLE patients were from Scandicci, 6 from Lastra a Signa and the other 6 from Signa. The overall rate of SLE prevalence is 71/100.000 (1 case/1408 with a confidence interval of 95%, (confidence limits: 49-92. The rate of SLE prevalence in Scandicci is 81/100.000 (confidence limits: 42-121, in Lastra a Signa 62/100.000 (confidence limits: 32-92 and in Signa 65/100.000 (confidence limits: 36-94. Conclusion: This is the first epidemiological study on SLE prevalence in Italy. The rate of SLE prevalence is similar to that of other European studies. LQS is confirmed as an easy and reliable tool to assess SLE diagnosis also in the Italian population too.
Naif Abdullah Al-Dhabi
Full Text Available Breast cancer is the most frequently occurring disease among women worldwide. The early stage of breast cancer identification is the key challenge in cancer control and prevention procedures. Although gene expression profiling helps to understand the molecular mechanism of diseases or disorder in the living system, gene expression pattern alone is not sufficient to predict the exact mechanisms. Current proteomics tools hold great application for analysis of cancerous conditions. Hence, the generation of differential protein expression profiles has been optimized for breast cancer and normal tissue samples in our organization. Normal and tumor tissues were collected from 20 people from a local hospital. Proteins from the diseased and normal tissues have been investigated by 2D gel electrophoresis and MALDI-TOF-MS. The peptide mass fingerprint data were fed into various public domains like Mascot, MS-Fit, and Pept-ident against Swiss-Prot protein database and the proteins of interest were identified. Some of the differentially expressed proteins identified were human annexin, glutathione S-transferase, vimentin, enolase-1, dihydrolipoamide dehydrogenase, glutamate dehydrogenase, Cyclin A1, hormone sensitive lipase, beta catenin, and so forth. Many types of proteins were identified as fundamental steps for developing molecular markers for diagnosis of human breast cancer as well as making a new proteomic database for future research.
Liepe, K; Blättner, B
Violence during pregnancy can be associated with health consequences for both the pregnant woman und the unborn child. The available data on prevalence are insufficient in Germany, an underestimation of the problem is probable. Therefore data from other OECD countries are considered. Searching in the databases EMBASE (incl. Medline), CINAHL, PsycINFO and SSCI in English or German language, primary and secondary studies from OECD countries published from 2004 to 2011 were identified. Studies without a valid instrument were excluded. 20 studies using 6 different, partially modified instruments were identified. In OECD countries the prevalence data vary, depending on the instrument applied and the design of the studies. The majority of the studies underestimate the prevalence of violence during pregnancy. Study results from OECD countries are in the analysed range of the reviews. The data collected in Germany are at the bottom of the spectrum. This suggests an underestimation of the problem in Germany. Further research is needed using more valid instruments and better study designs. © Georg Thieme Verlag KG Stuttgart · New York.
Sandeep H Shah
Full Text Available Background: Domestic violence is a major contributor to physical and mental ill health of the victim, and it is evident to some degree, in every society of the world. Objectives: 1 To study perception about domestic violence in the study population. 2 To compare prevalence of domestic violence within the three subgroups of the study population (i.e. spouses of psychotic patients, spouses of non-psychiatric patients and hospital staff. Materials and Methods: A cross-sectional study was conducted among married men and women coming to Dhiraj General Hospital. Interviews were conducted using a semi-structured questionnaire. Inquiry was done about their perception regarding domestic violence, own experience any time in their life, and about the form of violence. Data was entered and analyzed using SPSS. Results: 42.7% of study participants had never heard the words domestic violence. The overall prevalence of any form of violence in the study population as a whole was 32.3%. There was no significant difference found in the proportion of domestic violence among the three groups. The prevalence of physical, emotional, sexual and economic domestic violence was 16.3%, 25.3%, 2% and 11.3% respectively. Younger age group and female sex were significantly associated with the occurrence of domestic violence. Conclusion: Apart from the high prevalence of domestic violence in the present era, it is evident from the study that the participants′ perception about domestic violence was low. Efforts should be made to raise public consciousness and reporting of domestic violence and its attendant consequences.
Tangjitgamol, Siriwan; Kittisiam, Thannaporn; Tanvanich, Sujitra
The study was to evaluate the prevalence of mismatch repair gene defect among Thai patients with endometrial cancer and its association with clinico-pathological features and survivals. The formalin fixed paraffin-embedded blocks of EMC tissue from hysterectomy specimens of patients having surgery in our institution between 1 Jan 1995 and 31 December 2016 were assessed for the immunohistochemical expression of 4 mismatch repair proteins (MLH1, PMS, MSH2, MSH 6). Mismatch repair gene defect was determined by a negative expression of at least 1 protein. Among 385 EMC patients included in the study, mean age was 57.3 ± 10.8 years with 62.3% aged ⩽ 60 years. The most frequent mismatch repair gene defect was MSH6 (38.7%), followed by PMS2 (34.3%), MLH1 (33.2%), and MSH2 (16.4%). Overall, 55.1% showed negative expression of at least one protein. We found significantly higher mismatch repair gene defect in patients aged ⩽ 60 years, with early stage disease, and negative lymph node status than the other comparative groups: 59.2% vs 48.3% for age (p = 0.037), 58.2% vs 45.2% (p = 0.027) for stage, and 58.1% vs 44.6% (p = 0.048) for nodal status. The 5-year progression-free survival, overall survival, and endometrial cancer-specific survival of patients with mismatch repair gene defect was higher than those without gene defect. The differences were statistically significant for only progression-free survival and endometrial cancer-specific survival: 87.7% (95% confidence interval = 83.0%-92.4%) vs 81.5% (95% confidence interval = 75.4%-87.6%) (p = 0.049) for progression-free survival and 91.0% (95% confidence interval = 86.9%-95.1%) vs 85.5% (95% confidence interval = 80.0%-91.0%) (p = 0.044) for endometrial cancer-specific survival, respectively. In conclusion, more than half of Thai endometrial cancer patients had mismatch repair gene defect. The patients with mismatch repair gene defect had significantly younger age (⩽ 60 years) and better prognosis in terms of
Objective: To describe the pattern and prevalence of variations that occur in the supraclavicular part of the brachial plexus in a. Kenyan population. Study design: Descriptive cross-sectional study. Materials and methods: Ninety-four brachial plexuses from forty-seven formalin fixed cadavers were displayed by gross
Horga, Alejandro; Raja Rayan, Dipa L; Matthews, Emma; Sud, Richa; Fialho, Doreen; Durran, Siobhan C M; Burge, James A; Portaro, Simona; Davis, Mary B; Haworth, Andrea; Hanna, Michael G
To obtain minimum point prevalence rates for the skeletal muscle channelopathies and to evaluate the frequency distribution of mutations associated with these disorders. Analysis of demographic, clinical, electrophysiologic, and genetic data of all patients assessed at our national specialist channelopathy service. Only patients living in the United Kingdom with a genetically defined diagnosis of nondystrophic myotonia or periodic paralysis were eligible for the study. Prevalence rates were estimated for England, December 2011. A total of 665 patients fulfilled the inclusion criteria, of which 593 were living in England, giving a minimum point prevalence of 1.12/100,000 (95% confidence interval [CI] 1.03-1.21). Disease-specific prevalence figures were as follows: myotonia congenita 0.52/100,000 (95% CI 0.46-0.59), paramyotonia congenita 0.17/100,000 (95% CI 0.13-0.20), sodium channel myotonias 0.06/100,000 (95% CI 0.04-0.08), hyperkalemic periodic paralysis 0.17/100,000 (95% CI 0.13-0.20), hypokalemic periodic paralysis 0.13/100,000 (95% CI 0.10-0.17), and Andersen-Tawil syndrome (ATS) 0.08/100,000 (95% CI 0.05-0.10). In the whole sample (665 patients), 15 out of 104 different CLCN1 mutations accounted for 60% of all patients with myotonia congenita, 11 out of 22 SCN4A mutations for 86% of paramyotonia congenita/sodium channel myotonia pedigrees, and 3 out of 17 KCNJ2 mutations for 42% of ATS pedigrees. We describe for the first time the overall prevalence of genetically defined skeletal muscle channelopathies in England. Despite the large variety of mutations observed in patients with nondystrophic myotonia and ATS, a limited number accounted for a large proportion of cases.
Bozga, A; Stanciu, R P; Mănuc, D
Tooth agenesis is a phenomenon that occurs relatively commonly. The incidence of the missing teeth presented in the previous reports varies according to the studied population. The aim of this study was to find the prevalence of tooth agenesis in a population group in Bucharest. The prevalence and distribution of dental agenesis was determined in a sample of 518 patients, 285 females and 233 males, aged 6 to 41 years, who had been treated in the Clinic of Orthodontics and Dentofacial Orthopedics in Bucharest. The tooth agenesis was diagnosed by using the orthodontic records and study casts for each patient. 35 of the patients, 17 males and 18 females, were diagnosed with at least one absent permanent tooth and 47 missing permanent teeth were reported. A prevalence of 6.757% was observed for tooth agenesis. The mandibular second premolar was found to be the most affected tooth, followed by the maxillary lateral incisor, maxillary second premolar, mandibular central incisors, mandibular second molar and mandibular lateral incisor. The incidence of dental agenesis, its pattern and distribution per tooth type are in accordance with the previous published studies.
Mahesh, P. A.; Wong, Gary W. K.; Ogorodova, L.; Potts, J.; Leung, T. F.; Fedorova, O.; Holla, Amrutha D.; Fernandez-Rivas, M.; Clare Mills, E. N.; Kummeling, I.; Versteeg, S. A.; van Ree, R.; Yazdanbakhsh, M.; Burney, P.
Data are lacking regarding the prevalence of food sensitization and probable food allergy among general population in India. We report the prevalence of sensitization and probable food allergy to 24 common foods among adults from general population in Karnataka, South India. The study was conducted
Huang, Ke Qiang; Li, Chuan Silvia; Lin, Zhong Qiu; Feng, Guo Fei; Wang, Xiao Hui; Fu, Wen Zhe; Xie, Zhi Quan
This study aimed to determine the prevalence rate of knee osteoarthritis (OA) and the risk factors for OA in hospitalized elderly patients. We conducted this retrospective study in elderly patients (aged 65 years and older) who were hospitalized in the Geriatric Ward of General Hospital of Guangzhou Military Command of the People's Liberation Army between January 2011 and June 2013, including general condition, present history, past history, physical examination, X-ray results, and disease diagnosis. The prevalence, awareness, and treatment rates of knee OA in hospitalized elderly patients were calculated. Risk factors were computed using multiple logistic regression analysis. Of a total of 267 (17.4%) hospitalized elderly patients diagnosed with knee OA, the prevalence rate of OA was 9.95% in males and 37.76% in females. The rate of awareness among those with OA was 51.68%; the rate of treatment was 83.33%; and the rate of control was 77.39%. The medical expenses for both females (1143±315 yuan month-1) and males (1192±357 yuan month-1) in knee OA patients are higher than that of the non-knee OA group (989±274 yuan month-1, 1038±295 yuan month-1). The risk factors for knee OA include gender (OR=2.448), age (OR=1.124), transportation mode (OR= 8.972), exercise (OR=7.374), bowel evacuation position (OR=5.767), family history of knee OA (OR=2.195), and body mass index (OR=2.469). The prevalence of knee OA is unexpectedly high in hospitalized elderly patients, and the rates of awareness and treatment are less than desirable. Prevention and control measures should be taken in patients with concomitant risk factors.
Bani-Hani, Kamal E.; Hammouri, Shadi M.
Helicobacter pylori infection is considered the most common infection worldwide and is associated with many other disorders. The aim of this study is to determine the prevalence of this infection among patients undergoing endoscopy in Northern Jordan. Between November 1998 and September 2000, all patients referred from the Gastro-esophageal Clinic to the Endoscopy Unit at Princess Basma Teaching Hospital, Irbid, Northern Jordan were enrolled in this prospective study. For each patient clinical and epidemiological data was collected and endoscopy was performed. At least 3 antral biopsies were obtained from each patient, and these were examined histologically for the presence of gastritis and stained for Helicobacter pylori using modified Giemsa stain. A total of 197 consecutive patients (113 females) with a mean age of 40.2 years (range 15-91 years) were studied. Abdominal pain was the highest presenting symptom. Gastritis 91% and esophagitis 42% were the most frequent endoscopic findings. Gastritis was documented histologically in 183 (93%) of patients. Helicobacter pylori was found in 161 patients (82%), with all of these having histological gastritis. The 11 patients with gastric ulcer, compared to the 51 out of the 59 (86%) patients with duodenal ulcer, showed Helicobacter pylori in their biopsies. The prevalence of Helicobacter pylori infection in patients subjected to an upper gastrointestinal endoscopy in Jordan is high. This study confirms that Helicobacter pylori is significantly associated with gastritis and peptic ulcer. Further studies are needed to determine the types of Helicobacter pylori strains present in Jordan. (author)
Tic disorders are neurodevelopmental disorders of childhood associated with psychiatric comorbidity and academic problems. Estimating the prevalence and understanding the epidemiology of tic disorders is more complex than was once thought. Until fairly recently, tic disorders were thought to be rare, but today tics are believed to be the most common movement disorder, with 0.2-46.3% of schoolchildren experiencing tics during their lifetime. Tentative explanations for differing prevalence estimates include the multidimensional nature of tics with a varied and heterogeneous presentation, and the use of different epidemiological methods and study designs. Literature review and analysis of methodological issues pertinent to epidemiological studies of tic disorders. Epidemiological studies of tic disorders were reviewed, and the main elements of epidemiological studies, including sample selection, case ascertainment strategy, definition of tic disorders, and the degree of coverage of the eligible population (i.e., the response rate) were examined. In order to improve the quality of epidemiological studies of tic disorders, a number of recommendations were made, including but not limited to a review of the diagnostic criteria for tic disorders, and inclusion of new tic disorder categories for those with tics of secondary etiology.
Full Text Available Introduction: Tic disorders are neurodevelopmental disorders of childhood associated with psychiatric comorbidity and academic problems. Estimating the prevalence and understanding the epidemiology of tic disorders is more complex than was once thought. Until fairly recently, tic disorders were thought to be rare, but today tics are believed to be the most common movement disorder, with 0.2–46.3% of schoolchildren experiencing tics during their lifetime. Tentative explanations for differing prevalence estimates include the multidimensional nature of tics with a varied and heterogeneous presentation, and the use of different epidemiological methods and study designs. Methods: Literature review and analysis of methodological issues pertinent to epidemiological studies of tic disorders. Results: Epidemiological studies of tic disorders were reviewed, and the main elements of epidemiological studies, including sample selection, case ascertainment strategy, definition of tic disorders, and the degree of coverage of the eligible population (i.e., the response rate were examined. Discussion: In order to improve the quality of epidemiological studies of tic disorders, a number of recommendations were made, including but not limited to a review of the diagnostic criteria for tic disorders, and inclusion of new tic disorder categories for those with tics of secondary etiology.
Félix-Redondo, Francisco Javier; Grau, María; Baena-Díez, José Miguel; Dégano, Irene R; de León, Antonio Cabrera; Guembe, Maria Jesús; Alzamora, María Teresa; Vega-Alonso, Tomás; Robles, Nicolás R; Ortiz, Honorato; Rigo, Fernando; Mayoral-Sanchez, Eduardo; Tormo, Maria José; Segura-Fragoso, Antonio; Fernández-Bergés, Daniel
To estimate the prevalence of overweight and obesity in the Spanish population as measured with body mass index (BMI), waist circumference (WC) and waist to height ratio (WHtR) and to determine the associated cardiovascular risk factors. Pooled analysis with individual data from 11 studies conducted in the first decade of the 21st century. Participants aged 35-74 years were asked about the history of cardiovascular diseases, hypertension, diabetes and hypercholesterolemia. Height, weight, WC, blood pressure, glycaemia, total cholesterol, low-density and high-density lipoprotein cholesterol and coronary risk were measured. The prevalence of overweight (BMI 25-29.9 kg/m(2)), general obesity (BMI ≥ 30 kg/m(2)), suboptimal WC (≥ 80 cm and population. We included 28,743 individuals. The prevalence of overweight and suboptimal WC was 51% and 30% in men and 36% and 22% in women, respectively; general obesity was 28% in both sexes and abdominal obesity 36% in men and 55% in women. The prevalence of WHtR ≥0.5 was 89% and 77% in men and women, respectively. All cardiovascular risk factors were significantly associated with abnormal increased values of BMI, WC and WHtR. Hypertension showed the strongest association with overweight [OR = 1.99 (95% confidence interval 1.81-2.21) and OR = 2.10 (1.91-2.31)]; suboptimal WC [OR = 1.78 (1.60-1.97) and OR = 1.45 (1.26-1.66)], with general obesity [OR = 4.50 (4.02-5.04), and OR = 5.20 (4.70-5.75)] and with WHtR ≥0.5 [OR = 2.94 (2.52-3.43), and OR = 3.02 (2.66-3.42)] in men and women respectively, besides abdominal obesity in men only [OR = 3.51 (3.18-3.88)]. Diabetes showed the strongest association with abdominal obesity in women [OR = 3,86 (3,09-4,89). The prevalence of obesity in Spain was high. Overweight, suboptimal WC, general, abdominal obesity and WHtR ≥0.5 was significantly associated with diabetes, hypertension, hypercholesterolemia and coronary risk. The use of lower cut-off points for both BMI and particularly WC
Grønseth, Rune; Vollmer, William M.; Hardie, Jon A.; Ólafsdóttir, Inga Sif; Lamprecht, Bernd; Buist, A. Sonia; Gnatiuc, Louisa; Gulsvik, Amund; Johannessen, Ane; Enright, Paul
Dyspnea is a cardinal symptom for cardiorespiratory diseases. No study has assessed worldwide variation in dyspnea prevalence or predictors of dyspnea. We used cross-sectional data from population-based samples in 15 countries of the BOLD study to estimate prevalence of dyspnea in the full sample as well as in an a priori defined low-risk group (few risk factors or dyspnea-associated diseases). Dyspnea was defined by the modified Medical Research Council questions. We used ordered logistic regression analysis to study the association of dyspnea with site, sex, age, education, smoking habits, low/high BMI, self-reported disease, and spirometry results. Of the 9,484 participants, 27% reported any dyspnea. In the low-risk subsample (N=4,329), 16% reported some dyspnea. In multivariate analyses, all covariates were correlated to dyspnea, but only 13% of dyspnea variation was explained. Women reported more dyspnea than men (odds ratio ≈ 2.1). When forced vital capacity (FVC) fell below 60% of predicted, dyspnea was much more likely. There was considerable geographical variation in dyspnea, even when we adjusted for known risk factors and spirometry results. We were only able to explain 13% of dyspnea variation. PMID:24176991
Full Text Available BACKGROUND: Allergic rhinitis is a common condition, though not life threatening, causes significant morbidity in terms of quality of life. Confirmation of allergen as etiological agent is cumbersome. Hence need for a simple test is vital and eosinophil parameters were looked at to answer the quest. AIM: To find out the prevalence of e osinophilia in Allergic rhinitis . To assess the value of nasal cytogram as an alternative investigation in diagnosing allergic rhinitis . MATERIALS & METHODS: Prospective study of 200 cases divided into two groups of 100 each was done. One group clinically with allergic rhinitis and other without. All cases had clinical examination after history was taken, Blood Absolute eosinophil count, Nasal smear for eosinophils done and assessed. RESULTS: Of the 200 patients examined in two groups of 100 each, mean age of allergic rhinitis patients was 26.22 years . Allergic rhinitis was more common in males than females. Prevalence of nasal eosinophilia was 61%.and blood eosinophilia was 57% in allergic rhinitis patients. Nasal smear sensitivity was 61% and specificity w as 87% . CONCLUSION: Nasal smear eosinophilia is a valid test, can be quickly and easily performed and read. Being an in - expensive test can be used to screen the patients of allergic rhinitis
Full Text Available White lesions of the oral mucosa are of concern to the dental surgeon in view of the fact that some of these may be potentially malignant. Oral lichen plane: (OLP and oral lichenoid reactions (OLR share similar clinical appearances but need to be carefully distinguished because of their different etiologies and clinical behaviour. This study screened 5.017 population, in a house-to-house field survey, for tobacco use and investigated the prevalence of oral lichenoid reactions in the 98 quid users. Six subjects with clinical/clinical and histopathological criteria compatible with the diagnosis of OLR were identified. All these subjects were users of ′Gutka′, a unique chewable variant of tobacco quid containing areca nut and catechu. Statistical analysis revealed a significant association between quid habit and lesion occurrence (p < 0.005.
Ditisheim, Agnès; Wuerzner, Grégoire; Ponte, Belen; Vial, Yvan; Irion, Olivier; Burnier, Michel; Boulvain, Michel; Pechère-Bertschi, Antoinette
Preeclampsia is associated with increased cardiovascular and renal risk. The aim of this prospective cohort study was to characterize the early postpartum blood pressure (BP) profile after preeclampsia. We enrolled 115 women with preeclampsia and 41 women with a normal pregnancy in a prospective cohort study. At 6 to 12 week postpartum, we assessed the prevalence of different hypertensive phenotypes using 24-hour ambulatory BP monitoring (ABPM), as well as the risk of salt sensitivity and the variability of BP derived from ABPM parameters. Among patients with preeclampsia, 57.4% were still hypertensive at the office. Daytime ABP was significantly higher in the preeclampsia group (118.9±15.0/83.2±10.4 mm Hg) than in controls (104.8±7.9/71.6±5.3 mm Hg; P preeclampsia women remained hypertensive on ABPM in the postpartum, of whom 24.3% were still under antihypertensive treatment; 17.9% displayed a white-coat hypertension and 11.6% had masked hypertension. In controls, 2.8% had white-coat hypertension; none had masked hypertension or needed hypertensive treatment. The prevalence of nondippers was similar 59.8% in the preeclampsia group versus 51.4% in controls. High-risk class of salt sensitivity of BP was increased in preeclampsia women (48.6%) compared with controls (17.1%); P preeclampsia. This finding may help identify women who should be included in a postpartum cardiovascular risk management program. URL: https://www.clinicaltrials.gov. Unique identifier: NCT01095939. © 2017 American Heart Association, Inc.
Hooff, M.V.; McFarlane, A.C.; Davies, C.E.; Searle, A.K.; Fairweather-Schmidt, A.K.; Verhagen, A.F.; Benassi, H.; Hodson, S.E.
BACKGROUND: The Australian Defence Force (ADF) Mental Health Prevalence and Wellbeing Study (MHPWS) is the first study of mental disorder prevalence in an entire military population. OBJECTIVE: The MHPWS aims to establish mental disorder prevalence, refine current ADF mental health screening
protozoa in Toledo district, southern Beli ze . . . 18 Results of logistic regression analyses of Ascaris Lumbricoides prevalence in 5 villages of Toledo...selected by the forward stepwise methods. . . 20 Results of logistic regression analyses of Ascaris lumbricoides prevalence in 5 villages of Toledo...are listed) .................•................. 47 Summary of risk factors for being positive for Ascaris lumbricoides infection. Data from contingency
Samples were smeared on glass slides, stained using Ziehl Neelsen Stain and later observed under light (oil immersion) microscopy. The results showed that were positive for tuberculosis had prevalence of 541 (14.7%) out of 3679 subjects. The age group 30-43 years had the highest prevalence of 145 (17.0%) out of 858 ...
Worms or eggs recovered from these necropsied animals were used to determine the prevalence of gastrointestinal helminths. The prevalences of different helminths identified are as follows: Spirocerca lupi, 25 (4.64%), Haemonchus contortus, 223 (41.37%), Toxocara canis, 17 (3.13%), Ancylostoma caninum, 24 (4.45%), ...
A prevalence survey of malaria was carried out in Azia, a rural community in Ihiala Local Government Area of Anambra State, southeast Nigeria. Thick blood films of 1000 individuals attending local hospitals in the community were used to determine the prevalence. Also, structured questionnaire was administered to 1000 ...
Wise, Adina H; Yang, Amy; Naik, Hetanshi; Stauffer, Chanan; Zeid, Natasha; Liong, Christopher; Balwani, Manisha; Desnick, Robert J; Alcalay, Roy N
Recent research has suggested a possible link between Parkinson's disease (PD) and Fabry disease. To test this relationship, we administered a self-report and family history questionnaire to determine the prevalence of PD in Fabry disease patients and family members with likely pathogenic alpha-galactosidase A (GLA) mutations. A total of 90 Fabry patients (77 from the online survey and 13 from the Icahn School of Medicine at Mount Sinai (ISMMS)) were included in the analysis. Two of the Fabry disease patients who completed the online survey were diagnosed with PD (2/90, 2.2%). Among probands older than 60, 8.3% (2/24) were diagnosed with PD. Using Kaplan Meier survival analysis, the age-specific risk of PD by age 70 was 11.1%. Family history was available on 72 Fabry families from the online study and 9 Fabry families from ISMMS. Among these 81 families, 6 (7.4%) had one first degree relative who fit the criteria for a conservative diagnosis of PD. The results of this study suggest that there may be an increased risk of developing PD in individuals with GLA mutations, but these findings should be interpreted with caution given the limitations of the study design.
Cañero-Velasco, M C; Mutti, J E; Gonzalez, J E; Alonso, A; Otegui, L; Adragna, M; Antonuccio, M; Laso, M; Montenegro, M; Repetto, L; Brandi, M; Canepa, J; Baimberg, E
Hemodialized pediatric patients are a risk population for the hepatitis B and C virus infection. The aim of this paper was to study the serum prevalence of HBV and HCV infection in hemodialized children. We study 61 pediatric patients at hemodialisis, 12 on renal transplant, range between 2 and 20 years old (mean: 12.9 years), 23 male and 38 female. The specific anti-HCV IgC were measured by enzyme immunoassay (ELISA Abbott) and confirmed by LIA-TEK (Organon). The anti-HBV were measured by ELISA Abbott and transaminases by cinetic method (ASAT: 29 UI/L and ALT: 33 UI/L). The 19.7% of studied children were HCV (+) and 29.5% were HBV (+), 38.9% of them were HbsAg (+) and 50% anti-HBs (+). The HCV and HBV infection was more elevated in relation to the transfusion number and the hemodilisis time. The elevation of ALT/ASAT activity isn't a right infection index for HCV and HBV in this children.
Baş, Yalçın; Seçkin, Havva Yıldız; Kalkan, Göknur; Takci, Zennure; Çitil, Rıza; Önder, Yalçın; Şahin, Şafak; Demir, Ayşe Kevser
We aimed to determine the prevalence of psoriasis (PS) and seborrheic dermatitis (SD) (erythematous-squamous diseases) in our region and reveal the frequently encountered associated factors to aid in planning appropriate healthcare. A community-based study was conducted with 85 sample groups that reflected the population rate and demography of Tokat Province in northern Anatolia. In this community, the prevalence of PS in people older than 20 years of age was 1.2% and the prevalence of SD was 5.2%. SD rates were higher in patients who used tobacco and especially alcohol. SD prevalence was also higher in patients treated for depression and epilepsy. Furthermore, as education levels increased, SD prevalence increased proportionally. In general, PS prevalence in this region was higher than in Asia and Africa but lower than in Europe and the United States. Whereas SD prevalence varies between 2% and 12% throughout the world, the average SD prevalence was 5.2% in this study.
Vazirianzadeh B.* PhD,
Full Text Available Aims Cutaneous leishmaniasis is a prevalent parasitological disease with diverse clinical manifestations in Iran. Therefore, the present retrospective study carried out to describe the demographic features of cutaneous leishmaniasis in Ramshir, Iran. Materials & Methods This descriptive study was performed on 136 cutaneous leishmaniasis patients whose data were recorded in the Ramshir health center during 2006-9. Demographic information of patients including age, sex, habitat and sites of lesions, month and years of incidence were recorded. The data were analyzed by SPSS 16 software. Findings Totally 79 patients (58.1% resided in urban areas and the born to 9 years (49.3% was recognized as the most infected age group. Hands (41.2% had the highest rates of cutaneous leishmaniasis lesions followed by face (36.0% and foot (22.8%. The maximum number of cutaneous leishmaniasis lesions was reported in March. Conclusion As cutaneous leishmaniasis in Ramshir seemed to be an endemic rural type, the appropriate preventing measures regarding to the rural cutaneous leishmaniasis should be considered to decrease incidence of the disease in the region.
A biomedical survey of stool specimens from 82% of the population (n=672) of S villages in Toledo District, Belize were examined by the formalin-ethyl acetate concentration technique for the prevalence of intestinal parasitic infections...
van de Glind, Geurt; van Emmerik-van Oortmerssen, Katelijne; Carpentier, Pieter Jan; Levin, Frances R.; Koeter, Maarten W. J.; Barta, Csaba; Kaye, Sharlene; Skutle, Arvid; Franck, Johan; Konstenius, Maija; Bu, Eli-Torild; Moggi, Franz; Dom, Geert; Demetrovics, Zolt; Fatséas, Mélina; Schillinger, Arild; Kapitány-Fövény, Máté; Verspreet, Sofie; Seitz, Andrea; Johnson, Brian; Faraone, Stephen V.; Ramos-Quiroga, J. Antoni; Allsop, Steve; Carruthers, Susan; Schoevers, Robert A.; van den Brink, Wim
Attention deficit/hyperactivity disorder (ADHD) is an increasingly recognized comorbid condition in subjects with substance use disorders (SUDs). This paper describes the methods and study population of the International ADHD in Substance Use Disorders Prevalence (IASP) study. Objectives of the IASP
De Glind, Geurt Van; Oortmerssen, Katelijne Van Emmerik-Van; Carpentier, Pieter Jan; Levin, Frances R.; Koeter, Maarten W. J.; Barta, Csaba; Kaye, Sharlene; Skutle, Arvid; Franck, Johan; Konstenius, Maija; Bu, Eli-Torild; Moggi, Franz; Dom, Geert; Demetrovics, Zolt; Fatseas, Melina; Schillinger, Arild; Kapitany-Foeveny, Mate; Verspreet, Sofie; Seitz, Andrea; Johnson, Brian; Faraone, Stephen V.; Ramos-Quiroga, J. Antoni; Allsop, Steve; Carruthers, Susan; Schoevers, Robert A.; Grp, Iasp Res; Van den Brink, Wim
Attention deficit/hyperactivity disorder (ADHD) is an increasingly recognized comorbid condition in subjects with substance use disorders (SUDs). This paper describes the methods and study population of the International ADHD in Substance Use Disorders Prevalence (IASP) study. Objectives of the IASP
EGHBERT ELVAN AMPOU
Full Text Available Monitoring nilai prevalence karang yang mengalarni pemutihan clan penyakit sangat perlu dilakukan secara intensiv di Indonesia yang juga masuk dalam kawasan CTI, informasi tentang hal ini boleh dikatakan relatif belum banyak dilakukan orang. Metode yang dipakai selama survey adalah time swim dimana dibagi pada 2 kedalaman ( 5 dan 10 meter selama 30 menit, untuk analisis data digunakan rum us prevalence. Nilai Prevalence karang yang mengalarni pemutihan dan penyakit di Raja Ampat pada kedalaman 5m=30,67%; 10m=23,50% di bulan November 2009. Taman Nasional Bunaken kedalaman 5m=55,47%; 10m=83,73% di bulan Agustus 2009 clan di Pulau Runduma-Taman Nasional kedalaman 5m=23,55%; 10m=50,94% di bulan Oktober 2009.Jenis karang yang dominan mengalami pemutihan clan penyakit adalah genus Porites clan Acropora, sedangkan tidak dominan adalah genus Pocillopora dan Montipora.
Bussu, Francesco, E-mail: firstname.lastname@example.org [Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Sali, Michela [Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Gallus, Roberto [Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Petrone, Gianluigi; Zannoni, Gian Franco [Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Autorino, Rosa; Dinapoli, Nicola [Institute of Radiotherapy, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Santangelo, Rosaria [Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Vellone, Valerio Gaetano [Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Graziani, Cristina [Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Miccichè, Francesco [Institute of Radiotherapy, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Almadori, Giovanni; Galli, Jacopo [Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Delogu, Giovanni; Sanguinetti, Maurizio [Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); Rindi, Guido [Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma (Italy); and others
Purpose: Human papillomavirus (HPV) 16 infection is associated with oropharyngeal carcinogenesis and is likely the cause of the reported increase in disease incidence. We evaluated the prevalence of HPV infection and the reliability of different diagnostic tools using primary tumor samples from a cohort of 50 patients. Methods and Materials: Formalin-fixed paraffin-embedded (FFPE) tumor samples were collected from all 50 consecutive primary oropharyngeal SCC patients who were enrolled in the study; fresh tumor samples were available in 22 cases. NucliSENS EasyQ HPVv1 was used for RNA, and Digene Hybrid Capture-2(HC2) was used for DNA detection. p16 Expression was evaluated by immunohistochemistry in FPPE specimens. Results: Based on the DNA detection assay on FFPE samples, the frequency of high-risk HPV infection was 32%. The agreement rate between HPV RNA and HPV DNA detection in fresh samples was 100%. The agreement rate between p16 immunohistochemistry (IHC) and the detection of HPV DNA in the FFPE samples was fair but not excellent (κ = 0.618). HPV DNA detection was highly significant, as measured by disease-specific survival and determined using a Wilcoxon test (P=.001). p16 IHC also exhibited a prognostic value but with a lower statistical significance (P=.0475). The detection of HPV DNA, but not p16 IHC, was also significantly correlated with locoregional control (P=.0461). Conclusion: Diagnostic methods based on the detection of HPV nucleic acids appear to be more reliable and objective because they do not require reading by a trained histopathologist. Furthermore, the detection of HPV DNA exhibits an improved correlation with survival, and therefore appears definitely more reliable than p16 IHC for routine use in clinical practice.
Bussu, Francesco; Sali, Michela; Gallus, Roberto; Petrone, Gianluigi; Zannoni, Gian Franco; Autorino, Rosa; Dinapoli, Nicola; Santangelo, Rosaria; Vellone, Valerio Gaetano; Graziani, Cristina; Miccichè, Francesco; Almadori, Giovanni; Galli, Jacopo; Delogu, Giovanni; Sanguinetti, Maurizio; Rindi, Guido
Purpose: Human papillomavirus (HPV) 16 infection is associated with oropharyngeal carcinogenesis and is likely the cause of the reported increase in disease incidence. We evaluated the prevalence of HPV infection and the reliability of different diagnostic tools using primary tumor samples from a cohort of 50 patients. Methods and Materials: Formalin-fixed paraffin-embedded (FFPE) tumor samples were collected from all 50 consecutive primary oropharyngeal SCC patients who were enrolled in the study; fresh tumor samples were available in 22 cases. NucliSENS EasyQ HPVv1 was used for RNA, and Digene Hybrid Capture-2(HC2) was used for DNA detection. p16 Expression was evaluated by immunohistochemistry in FPPE specimens. Results: Based on the DNA detection assay on FFPE samples, the frequency of high-risk HPV infection was 32%. The agreement rate between HPV RNA and HPV DNA detection in fresh samples was 100%. The agreement rate between p16 immunohistochemistry (IHC) and the detection of HPV DNA in the FFPE samples was fair but not excellent (κ = 0.618). HPV DNA detection was highly significant, as measured by disease-specific survival and determined using a Wilcoxon test (P=.001). p16 IHC also exhibited a prognostic value but with a lower statistical significance (P=.0475). The detection of HPV DNA, but not p16 IHC, was also significantly correlated with locoregional control (P=.0461). Conclusion: Diagnostic methods based on the detection of HPV nucleic acids appear to be more reliable and objective because they do not require reading by a trained histopathologist. Furthermore, the detection of HPV DNA exhibits an improved correlation with survival, and therefore appears definitely more reliable than p16 IHC for routine use in clinical practice
BACKGROUND: The assessment of the extent and distribution of Schistosoma infection in every region in Nigeria is important and required for formulating intervention strategies suitable for each endemic area. OBJECTIVE: To assess the prevalence of urinary schistosomiasis in Ogun State, South-West Nigeria. METHODS: ...
Objective: To assess the prevalence of gingival bleeding following gentle periodontal probing during pregnancy as compared to after child birth in the same set of women. Method: Three hundred and eighty-four consecutive pregnant women in third trimester were selected at the antenatal clinics of Adeoyo Maternity ...
emergence of drug resistant pathogens now. Objective: To determine the prevalence and drug susceptibility patterns of bacterial pathogens involved in urinary tract infections within the last three years, January 1st 2008 to December 30th 2010 in Tikur Anbesa Specialized. Teaching Hospital of Addis Ababa, Ethiopia.
Under occupation, business women recorded highest, 19(6.3%)prevalence rate, students rated 4(1.3%) among others. Vaginal discharge rated highest, (81%) among other observable clinical symptoms associated with the disease as recorded in the survey. The findings regarding the disease deserve some quick response ...
Ephraim-Emmanuel, Benson Chukwunweike; Dotimi, Doris Atibinye; Kei, Bodisere Opuowei; Joshua, Respect Isaiah
Background: The prevalence of dental fluorosis in certain parts of the globe has been found to be increasing. In contrast to these prevalence reports in these areas, there is little or no systematic epidemiological study evaluating dental fluorosis in Bayelsa state, Nigeria. We thus studied the prevalence of dental fluorosis among school children in Ogbia town, Ogbia local government of Bayelsa state, Nigeria and evaluated the role played by potential risk factors. Method: A cross-section...
Introduction: Tic disorders are neurodevelopmental disorders of childhood associated with psychiatric comorbidity and academic problems. Estimating the prevalence and understanding the epidemiology of tic disorders is more complex than was once thought. Until fairly recently, tic disorders were thought to be rare, but today tics are believed to be the most common movement disorder, with 0.2–46.3% of schoolchildren experiencing tics during their lifetime. Tentative explanations fo...
Fiest, Kirsten M; Sauro, Khara M; Wiebe, Samuel; Patten, Scott B; Kwon, Churl-Su; Dykeman, Jonathan; Pringsheim, Tamara; Lorenzetti, Diane L; Jetté, Nathalie
To review population-based studies of the prevalence and incidence of epilepsy worldwide and use meta-analytic techniques to explore factors that may explain heterogeneity between estimates. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses standards were followed. We searched MEDLINE and EMBASE for articles published on the prevalence or incidence of epilepsy since 1985. Abstract, full-text review, and data abstraction were conducted in duplicate. Meta-analyses and meta-regressions were used to explore the association between prevalence or incidence, age group, sex, country level income, and study quality. A total of 222 studies were included (197 on prevalence, 48 on incidence). The point prevalence of active epilepsy was 6.38 per 1,000 persons (95% confidence interval [95% CI] 5.57-7.30), while the lifetime prevalence was 7.60 per 1,000 persons (95% CI 6.17-9.38). The annual cumulative incidence of epilepsy was 67.77 per 100,000 persons (95% CI 56.69-81.03) while the incidence rate was 61.44 per 100,000 person-years (95% CI 50.75-74.38). The prevalence of epilepsy did not differ by age group, sex, or study quality. The active annual period prevalence, lifetime prevalence, and incidence rate of epilepsy were higher in low to middle income countries. Epilepsies of unknown etiology and those with generalized seizures had the highest prevalence. This study provides a comprehensive synthesis of the prevalence and incidence of epilepsy from published international studies and offers insight into factors that contribute to heterogeneity between estimates. Significant gaps (e.g., lack of incidence studies, stratification by age groups) were identified. Standardized reporting of future epidemiologic studies of epilepsy is needed. © 2016 American Academy of Neurology.
Purpose: Although snake bites occur frequently in Benin City, the prevalence has not been documented. This study was therefore done to determine the prevalence, morbidity, mortality, and the orthodox treatment of victims. Methods: The study was retrospective and data on victims of snake bite covering a period of twenty ...
Schiøtz, Michaela L; Stockmarr, Anders; Høst, Dorte
Prevalences of multimorbidity vary between European studies and several methods and definitions are used. In this study we examine the prevalence of multimorbidity in relation to age, gender and educational attainment and the association between physical and mental health conditions and educational...
Nielsen, Cathrine Wildenschild; Kjøller, Mette; Sabroe, Svend
The purpose of this study was to examine the influence of the increasing prevalence of obesity on the development of health care utilization in Denmark in the period 1987-2005.......The purpose of this study was to examine the influence of the increasing prevalence of obesity on the development of health care utilization in Denmark in the period 1987-2005....
Pascoal, Maria I. N.; Rapoport, Abrão; Chagas, José F. S.; Pascoal, Maria B.N.; Costa, Claudiney C.; Magna, Luis Antonio
Introdução: A presença de sintomas otológicos associados à desordem temporomandibular (DTM) é discutida há seis décadas; entretanto, sua etiologia ainda permanece obscura. Forma de estudo: Prospectivo clínico randomizado. Objetivo: Neste estudo foram avaliadas a prevalência de sintomas otológicos na DTM, sua correlação com a dor muscular e a ausência de dentes posteriores. Material e Método: Foram avaliados 126 pacientes portadores de DTM, através de questionário subjetivo dos sintomas, palpa...
Background: Acute mountain sickness is a major public health problem in high altitudes. Similarly, anecdotal evidence suggests that there is high prevalence of tobacco smoking among this group though prevalence of obstructive airflow limitation is not known. Objectives: The main aims of the study were to measure the awareness of AMS and report the prevalence of AMS and obstructive lung diseases in high altitude Nepalese porters. Setting: This study was done with bases in Namche Bazaar (...
KELLY, GREGORY M.; KONG, YINK HEAY; DOBI, ALBERT; SRIVASTAVA, SHIV; SESTERHENN, ISABELL A.; PATHMANATHAN, RAJADURAI; TAN, HUI MENG; TAN, SHYH-HAN; CHEONG, SOK CHING
Overexpression of the erythroblast transformation-specific-related gene (ERG) oncoprotein due to transmembrane protease, serine 2 (TMPRSS2)-ERG fusion, the most prevalent genomic alteration in prostate cancer (CaP), is more frequently observed among Caucasian patients compared to patients of African or Asian descent. To the best of our knowledge, this is the first study to investigate the prevalence of ERG alterations in a multiethnic cohort of CaP patients. A total of 191 formalin-fixed para...
Full Text Available Objectives: To evaluate the prevalence and clinical significance of cathepsin G antibodies in patients affected with systemic sclerosis (SSc, scleroderma. Methods: 115 patients affected by SSc, 55 (47,8% with diffuse scleroderma (dSSc and 60 (52,2% with limited scleroderma (lSSc, were tested for cathepsin G antibodies by ELISA method. Moreover these sera were evaluated by indirect immunofluorescence (IIF on ethanol and formalin fixed human neutrophils. Results: By means of the ELISA method 16 (13,9% patients were found to be sera positive for anti-cathepsin G, 2 (12.5% of which showed a perinuclear fluorescence pattern (P-ANCA and 4 (25% an atypical ANCA staining, while 10 (62,5% were negative on IIF. The IIF on scleroderma sera revealed 5 (4,3% P-ANCA and 18 (15,7% atypical ANCA patterns. The anti-cathepsin G antibodies significantly prevailed in scleroderma sera (p=0.02 when their frequency was compared with that of healthy controls; while they were not significantly associated to any clinical or serological features of SSc patients. Conclusions: The anti-cathepsin G antibodies were significantly frequent in scleroderma sera; however, no clinical correlations were found. Thus, the significance of their presence in SSc still needs to be clarified.
Full Text Available BACKGROUND: Atopic dermatitis (AD is a common inflammatory and chronically relapsing disorder with increasing prevalence. However, little is known about its prevalence in Shanghai, the top metropolitan of China. This study will estimate and compare the prevalence of AD in urban and rural areas in representative samples of 3 to 6-year-old children in Shanghai. METHODOLOGY/PRINCIPAL FINDINGS: A descriptive cross-sectional study was performed. Pre-school children were obtained by cluster sampling from 8 communities in different districts in Shanghai. The main instrument was the core questionnaire module for AD used in the U.K. Working Party's study. All the data were statistically analyzed by EpiData 3.1 and SPSS16.0. A total of 10,436 children completed the study satisfactorily, with a response rate of 95.8%. The prevalence of AD in 3 to 6-year-old children was 8.3% (Male: 8.5%, Female: 8.2%. The prevalence in urban areas of Shanghai was gradiently and significantly higher than that in rural areas. The highest prevalence was in the core urban area (10.2% in Xuhui Tianping vs. the lowest far from the urban areas (4.6% in Chongming Baozhen. CONCLUSIONS/SIGNIFICANCE: The prevalence of AD was 8.3% (95%CI: 7.6%-9.1% in children aged 3 to 6 in Shanghai. The prevalence of AD decreased from the center to the rural areas in Shanghai.
Chaaya, Monique; Saab, Dahlia; Maalouf, Fadi T.; Boustany, Rose-Mary
In Lebanon, no estimate for autism prevalence exists. This cross-sectional study examines the prevalence of Autism spectrum disorder (ASD) in toddlers in nurseries in Beirut and Mount-Lebanon. The final sample included 998 toddlers (16-48 months) from 177 nurseries. We sent parents the Modified Checklist for Autism in Toddlers (M-CHAT) for…
Aim: To deterime the prevalence of anaemia in patients with clinical AIDS. Methodology: A retrospective small observational study of the prevalence of anaemia in 114 patients with confirmed HIV/AIDS seen at the department of medicine of the university of Maiduguri Teaching Hospital between January, 2002 and June, ...
Introduction: We determined the sero-prevalence of HIV among female sex workers (FSWs) in Togo identified their sexual risk behaviors. Methods: We conducted a cross-sectional study from 17 to 27 December, 2011 on 1106 FSWs in Togo. Venous sample were collected to estimate HIV prevalence as per national ...
Dekkers, Laura M.; Groot, Norbert A.; Díaz Mosquera, Elena N.; Andrade Zúñiga, Ivonne P.; Delfos, Martine F.
This research presents the results of the first phase of the study on the prevalence of pupils with Autism Spectrum Disorder (ASD) in regular education in Quito, Ecuador. One-hundred-and-sixty-one regular schools in Quito were selected with a total of 51,453 pupils. Prevalence of ASD was assessed by an interview with the rector of the school or…
Pedro J. Serrano-Castro
Full Text Available Background. This study assesses the lifetime and active prevalence of epilepsy in Spain in people older than 18 years. Methods. EPIBERIA is a population-based epidemiological study of epilepsy prevalence using data from three representative Spanish regions (health districts in Zaragoza, Almería, and Seville between 2012 and 2013. The study consisted of two phases: screening and confirmation. Participants completed a previously validated questionnaire (EPIBERIA questionnaire over the telephone. Results. A total of 1741 valid questionnaires were obtained, including 261 (14.99% raising a suspicion of epilepsy. Of these suspected cases, 216 (82.75% agreed to participate in phase 2. Of the phase 2 participants, 22 met the International League Against Epilepsy’s diagnostic criteria for epilepsy. The estimated lifetime prevalence, adjusted by age and sex per 1,000 people, was 14.87 (95% CI: 9.8–21.9. Active prevalence was 5.79 (95% CI: 2.8–10.6. No significant age, sex, or regional differences in prevalence were detected. Conclusions. EPIBERIA provides the most accurate estimate of epilepsy prevalence in the Mediterranean region based on its original methodology and its adherence to ILAE recommendations. We highlight that the lifetime prevalence and inactive epilepsy prevalence figures observed here were compared to other epidemiological studies.
Schiøtz, Michaela L; Stockmarr, Anders; Høst, Dorte
Prevalences of multimorbidity vary between European studies and several methods and definitions are used. In this study we examine the prevalence of multimorbidity in relation to age, gender and educational attainment and the association between physical and mental health conditions and educational.......3-3.4) and approximately 50 times more prevalent among older persons than younger ones. Physical and mental health comorbidity was also twice as prevalent among people with lower secondary education than among those with postgraduate education. The presence of a mental health condition was strongly associated...... attainment in a Danish population. A cross-sectional design was used to study the prevalence of multimorbidity, defined as two or more chronic conditions, and of comorbid physical and mental health conditions across age groups and educational attainment levels among 1,397,173 individuals aged 16 years...
Rust, Kristina Yasmin; Baumgartner, Markus R; Dally, Annika M; Kraemer, Thomas
New psychoactive substances are conquering the drug scene. Police seize different colourful packages with exceptional names. They are declared as 'bath salts', 'plant food', or 'research chemical powders'. Little is known about the actual prevalence of these drugs. Reanalysis of hair samples from routine cases concerning the presence of new psychoactive substances or 'smart drugs' should provide insight into changing patterns of designer drugs. All hair samples from 2009 and 2010 that originally tested positive for amphetamines or MDMA (N = 325) were reanalyzed for new or smart drugs such as 4-fluoroamphetamine, piperazines (BZP, mCPP and TFMPP), cathinones (4-MMC (mephedrone), methylone, butylone, ethylone, MDPV, methcathinone and cathinone), methylphenidate and ketamine. Hair snippets were extracted using a two-step extraction procedure. The analytes were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) (electrospray ionization; multiple-reaction-monitoring mode - information dependent acquisition - enhanced product ion scan). New psychoactive substances were found in 120 cases (37%). Concerning the piperazine drugs, mCPP was positive in 34 (10.5%) cases and TFMPP in one case. Five mCPP cases were also positive for trazodone, an antidepressant which is metabolized to mCPP. In 11 (3%) cases, 4-MMC was detected. Concerning the smart drugs, methylphenidate was found in 16 (5%). Ketamine was found in 45 (14%) cases. 4-Fluoroamphetamine was identified in 12 (4%) cases and methylone in one case.In conclusion, there is a high prevalence of these drugs. Consequently, at least the most common ones (e.g. mCPP, KET, 4-MMC and 4-FA) should be included in screening procedures in clinical and forensic toxicology. Copyright © 2012 John Wiley & Sons, Ltd.
Elza C de Bruin
Full Text Available Personalized healthcare relies on accurate companion diagnostic assays that enable the most appropriate treatment decision for cancer patients. Extensive assay validation prior to use in a clinical setting is essential for providing a reliable test result. This poses a challenge for low prevalence mutations with limited availability of appropriate clinical samples harboring the mutation. To enable prospective screening for the low prevalence AKT1 E17K mutation, we have developed and validated a competitive allele-specific TaqMan® PCR (castPCR™ assay for mutation detection in formalin-fixed paraffin-embedded (FFPE tumor tissue. Analysis parameters of the castPCR™ assay were established using an FFPE DNA reference standard and its analytical performance was assessed using 338 breast cancer and gynecological cancer FFPE samples. With recent technical advances for minimally invasive mutation detection in circulating tumor DNA (ctDNA, we subsequently also evaluated the OncoBEAM™ assay to enable plasma specimens as additional diagnostic opportunity for AKT1 E17K mutation testing. The analysis performance of the OncoBEAM™ test was evaluated using a novel AKT1 E17K ctDNA reference standard consisting of sheared genomic DNA spiked into human plasma. Both assays are employed at centralized testing laboratories operating according to quality standards for prospective identification of the AKT1 E17K mutation in ER+ breast cancer patients in the context of a clinical trial evaluating the AKT inhibitor AZD5363 in combination with endocrine (fulvestrant therapy.
Proverbio, D; Perego, R; Spada, E; Ferro, E
To determine the prevalence of adverse food reactions (AFRs) in dogs with dermatological signs presented to the referral dermatological clinic of the University of Milan. The medical records of dogs with dermatological signs were reviewed. Prevalence of AFRs was calculated. Owner and clinician pruritus scores were compared. Breed, sex and age predisposition were statistically tested, as was the association between AFR and selected clinical features. The prevalence of AFRs in dogs with dermatological signs was 12% (16 of 130). AFR was diagnosed in 26% of dogs with allergic disease and 48% of those subjected to a dietary trial. There was a significant association between AFRs and early onset of clinical signs (dogs with perianal fistulas were German shepherd dogs. The prevalence of AFRs in the study population was higher than most reported values. Further studies are warranted to investigate the true prevalence of AFR and its possible association with perianal fistula and other potential markers.
Full Text Available OBJECTIVE: To study the variations in the origin of subscapular artery and its branches in the dissecting room. METHOD: The morphological study of 40 formalin fixed adult human cadavers of both sexes was carried out following a standard dissecting manual. OBSERVATION: Variations in origin of Subscapular artery was observed in 15% of the cadavers, origin from 2 nd part of axillary artery in 15%, as a common branch with Lateral thoracic artery in 10%, associated variations of posterior circumflex humeral artery, thoraco dorsal artery and circumflex scapular artery in 10%. CONCLUSION: In comparison with other similar studies, the present one also shows a near to equal scenario which helps us to strengthen the already existing knowledge. Appreciation of these variations is important to vascular surgeons for vascular graft surgeries, general and on co - surgeons while exposure of axilla and axillary artery so also to radiologists and anatomists
Arnedo-Pena, Alberto; García-Marcos, Luis; Bercedo-Sanz, Alberto; Aguinaga-Ontoso, Inés; González-Díaz, Carlos; García-Merino, Águeda; Busquets-Monge, Rosa; Suárez-Varela, Maria Morales; Batlles-Garrido, Juan; Blanco-Quirós, Alfredo A.; López-Silvarrey, Angel; García-Hernández, Gloria; Fuertes, Jorge
The aim of the present study was to estimate the associations between the prevalence of asthma symptoms in schoolchildren and meteorological variables in west European countries that participated in the International Study of Asthma and Allergies in Children (ISAAC), Phase III 1997-2003. An ecologic study was carried out. The prevalence of asthma was obtained from this study from 48 centers in 14 countries, and meteorological variables from those stations closest to ISAAC centers, together with other socioeconomic and health care variables. Multilevel mixed-effects linear regression models were used. For schoolchildren aged 6-7 years, the prevalence rate of asthma decreased with an increase in mean annual sunshine hours, showed a positive association with rainy weather, and warm temperature, and a negative one with relative humidity and physician density (PD). Current wheeze prevalence was stronger in autumn/winter seasons and decreased with increasing PD. Severe current wheeze decreased with PD. For schoolchildren aged 13-14 years, the prevalence rates of asthma and current wheeze increased with rainy weather, and these rates decreased with increased PD. Current wheeze, as measured by a video questionnaire, was inversely associated with sunny weather, and nurse density. Severe current wheeze prevalence was stronger during autumn/winter seasons, decreased with PD, and indoor chlorinated public swimming pool density, and increased with rainy weather. Meteorological factors, including sunny and rainy weather, and PD may have some effect on the prevalence rates of asthma symptoms in children from west European countries.
Fernanda Mara de Paiva Bertoli
Full Text Available The prevalence of signs and symptoms of temporomandibular disorders (TMD increases during adolescence and adulthood. Few studies have examined TMD prevalence in Brazilian adolescents.To investigate the prevalence of TMD in Brazilian adolescents.A representative population-based sample of 934 adolescents (10-14-years-old was examined. TMD screening was performed using a questionnaire by the American Academy of Orofacial Pain. TMD diagnoses used research diagnostic criteria for temporomandibular disorders (RDC/TMD-Axis-I. Examinations were performed by a single calibrated examiner (kappa > 0.80.The prevalence of TMD symptoms was 34.9%; the most frequently reported symptoms were headache and neck ache (20.9%, followed by joint sounds (18.5%. Myofascial pain was the most prevalent type (10.3%, followed by disc displacement with reduction (8.0% and arthralgia (3.5%. There was a significant association between sex and TMD symptoms; prevalence was significantly higher in girls (RP = 1.37; 95% CI = 1.14-1.65; p = 0.001. Myofascial pain of TMD and displacement with reduction were more prevalent in girls (RP = 1.76; p = 0.007 and RP = 2.06; p = 0.004, respectively.TMD symptoms were present in 34.9% of adolescents, with myofascial pain being the most prevalent type (10.3%. TMD was significantly more common in girls. Routine pediatric dental care should include a TMD screening.
Speich, Benjamin; Marti, Hanspeter; Ame, Shaali M; Ali, Said M; Bogoch, Isaac I; Utzinger, Jürg; Albonico, Marco; Keiser, Jennifer
Pathogenic intestinal protozoa infections are common in school-aged children in the developing world and they are frequently associated with malabsorption syndromes and gastrointestinal morbidity. Since diagnosis of these parasites is difficult, prevalence data on intestinal protozoa is scarce. We collected two stool samples from school-aged children on Pemba Island, Tanzania, as part of a randomized controlled trial before and 3 weeks after treatment with (i) single-dose albendazole (400 mg); (ii) single-dose nitazoxanide (1,000 mg); (iii) nitazoxanide-albendazole combination (1,000 mg-400 mg), with each drug given separately on two consecutive days; and (iv) placebo. Formalin-fixed stool samples were examined for the presence of intestinal protozoa using an ether-concentration method to determine the prevalence and estimate cure rates (CRs). Almost half (48.7%) of the children were diagnosed with at least one of the (potentially) pathogenic protozoa Giardia intestinalis, Entamoeba histolytica/E. dispar and Blastocystis hominis. Observed CRs were high for all treatment arms, including placebo. Nitazoxanide showed a significant effect compared to placebo against the non-pathogenic protozoon Entamoeba coli. Intestinal protozoa infections might be of substantial health relevance even in settings where they are not considered as a health problem. Examination of a single stool sample with the ether-concentration method lacks sensitivity for the diagnosis of intestinal protozoa, and hence, care is indicated when interpreting prevalence estimates and treatment effects.
Smith, W C; Kenicer, M B; Tunstall-Pedoe, H; Clark, E C; Crombie, I K
Data from 10,359 men and women aged 40-59 years from 22 districts in the Scottish Heart Health Study were used to describe the prevalence rates of coronary heart disease in Scotland in 1984-1986 and their relation to the geographical variation in mortality in these districts. Prevalence was measured by previous history, Rose chest pain questionnaire, and the Minnesota code of a 12 lead resting electrocardiogram. The prevalence of coronary heart disease in Scotland was high compared with studi...
Full Text Available Objectives:(1 To find the contraceptive prevalence in different reproductive age group females.(2 What is the correlation between fertility pattern and contraceptive prevalence.Study Design : Cross sectional descriptive study.Setting : Community Development block Sarojini Nagar, Lucknow district.Study universe : Married females in reproductive age group (15-49 Years.Study variables : Age, fertility, contraceptive prevalence, education and social class.Statistical analysis : Chi Square test.Result : Contraceptive prevalence was 29.7%. It was 6.5% for women aged 15-19 year to 45.2% for women aged 3539 years. The acceptors were analyzed based on the number of living children. 74% of the acceptors had 3 or more children as against 26% with 0 to 2 living children.
Nuamah, Mercy A; Browne, Joyce L; Öry, Alexander V; Damale, Nelson K R; Klipstein-Grobusch, Kerstin; Rijken, Marcus J
BACKGROUND: The global increase in Cesarean section rate is associated with short- and long-term complications, including adhesions with potential serious maternal and fetal consequences. This study investigated the prevalence of adhesions and association between adhesions and postoperative
Adank, M. A.; Brogi, E.; Bogomolniy, F.; Wadsworth, E. A.; Lafaro, K. J.; Yee, C. J.; Kirchhoff, T.; Meijers-Heijboer, E. J.; Kauff, N. D.; Boyd, J.; Offit, K.
A major limitation in counseling unaffected women from families with inherited breast and ovarian cancer is that a "true-negative" interpretation of wild type BRCA analysis of the proband cannot be inferred in the absence of demonstration of a BRCA mutation segregating in the kindred. Documentation
Diagnosis of prion disease is primarily through immunodetection of the infectious agent. Typically, 2 distinct procedures are recommended for a definitive diagnosis with immunohistochemistry and Western blot providing the most information as to the specific isolate in question. In the past these app...
van de Glind, Geurt; Van Emmerik-van Oortmerssen, Katelijne; Carpentier, Pieter Jan; Levin, Frances R; Koeter, Maarten W J; Barta, Csaba; Kaye, Sharlene; Skutle, Arvid; Franck, Johan; Konstenius, Maija; Bu, Eli-Torild; Moggi, Franz; Dom, Geert; Demetrovics, Zolt; Fatséas, Mélina; Schillinger, Arild; Kapitány-Fövény, Máté; Verspreet, Sofie; Seitz, Andrea; Johnson, Brian; Faraone, Stephen V; Ramos-Quiroga, J Antoni; Allsop, Steve; Carruthers, Susan; Schoevers, Robert A; van den Brink, Wim
Attention deficit/hyperactivity disorder (ADHD) is an increasingly recognized comorbid condition in subjects with substance use disorders (SUDs). This paper describes the methods and study population of the International ADHD in Substance Use Disorders Prevalence (IASP) study. Objectives of the IASP are to determine the prevalence of ADHD in adult treatment seeking patients with SUD in different countries and SUD populations, determine the reliability and validity of the Adult ADHD Self-report Scale V 1.1 (ASRS) as ADHD screening instrument in SUD populations, investigate the comorbidity profile of SUD patients with and without ADHD, compare risk factors and protective factors in SUD patients with and without a comorbid diagnosis of ADHD, and increase our knowledge about the relationship between ADHD and the onset and course of SUD. In this cross-sectional, multi-centre two stage study, subjects were screened for ADHD with the ASRS, diagnosed with the Conner's Adult ADHD Diagnostic Interview for DSM-IV (CAADID), and evaluated for SUD, major depression, bipolar disorder, anti social personality disorder and borderline personality disorder. Three thousand five hundred and fifty-eight subjects from 10 countries were included. Of these 40.9% screened positive for ADHD. This is the largest international study on this population evaluating ADHD and comorbid disorders. Copyright © 2013 John Wiley & Sons, Ltd.
Kummeling, I.; Mills, E. N. C.; Clausen, M.; Dubakiene, R.; Pérez, C. Farnãndez; Fernández-Rivas, M.; Knulst, A. C.; Kowalski, M. L.; Lidholm, J.; Le, T.-M.; Metzler, C.; Mustakov, T.; Popov, T.; Potts, J.; van Ree, R.; Sakellariou, A.; Töndury, B.; Tzannis, K.; Burney, P.
Background: The epidemiological surveys in children and adults of the EU-funded multidisciplinary Integrated Project EuroPrevall, launched in June 2005, were designed to estimate the currently unknown prevalence of food allergy and exposure to known or suspected risk factors for food allergy across
Full Text Available Objective: To investigate and provide data on the prevalence, epidemiological pattern zoonotic impact of helminth parasites in cattle slaughtered at abattoir in Khorramabad, Lorestan province, southwestern Iran from April 2010 to April 2013. Methods: A total of 1 50 869 livers of cattle were examined. The total prevalence rate of distomatosis and hydaditidosis in different seasons were calculated. Results: The overall prevalence rate of infection was 23.3%. The prevalence rate of hydatid cyst (9.4% infection was significantly higher than the prevalence of fasciolosis (7.6% and dicrocoeliosis (6.3% (P<0.001. Data showed significant seasonal pattern for three parasitic infections (P<0.001. The highest prevalence rate of Fasciola spp. and D. dendriticum infection were seen in spring, while the highest rate of hydatidosis was seen in winter. The highest and lowest of overall infection were recorded during winter 2012 and autumn 2011, respectively. Conclusions: According to this study, it can be concluded that Khorramabad as an endemic region for distomatosis and hydatidosis infection. More surveys are suggested to be carried out to collect more data about the internal organs infection prevalence and risk factors for developing a prediction model in ruminants in southwestern Iran.
Andrés Duque DUQUE
Full Text Available The objective of this study was to assess the prevalence of mucositis and peri-implantitis associated with the use of two types of implants—conventional versus platform switching after one year of loading. A longitudinal study of 64 implants in 25 patients was performed. Clinical variables, such as clinical pocket depth and bleeding upon probing, plaque, mobility, gingival recession, clinical attachment loss, and radiographic bone loss, were analyzed. The case definition for peri-implantitis was established as pockets of ≥ 5 mm with bleeding and bone loss ≥ 2 mm. One year after implant loading, the prevalence of mucositis and peri-implantitis with conventional implants (CIs was 81.2% and 15.6%, respectively. For platform switching implants (PSIs the prevalence was 90% and 6.6%, respectively. These differences were not statistically significant (p = 0.5375. However, there was a trend towards a lower prevalence of peri-implantitis with platform switching Implants.
Capurso, Gabriele; Archibugi, Livia; Pasquali, Piera; Aceti, Alessandro; Balducci, Paolo; Bianchi, Patrizia; Buono, Francesco; Camerucci, Stefano; Cantarini, Rosanna; Centofanti, Sergio; Colantonio, Patrizia; Cremaschi, Riccarda; Crescenzi, Sergio; Di Mauro, Caterina; Di Renzi, Davide; Filabozzi, Andrea; Fiorillo, Alfonso; Giancaspro, Giuseppe; Giovannetti, Paola; Lanna, Giuseppe; Medori, Claudio; Merletti, Emilio; Nunnari, Enzo; Paris, Francesca; Pavone, Marco; Piacenti, Angela; Rossi, Almerindo; Scamuffa, Maria Cristina; Spinelli, Giovanni; Taborchi, Marco; Valente, Biagio; Villanova, Antonella; Chiriatti, Alberto; Delle Fave, Gianfranco
Data on chronic pancreatitis prevalence are scanty and usually limited to hospital-based studies. Investigating chronic pancreatitis prevalence in primary care. Participating primary care physicians reported the prevalence of chronic pancreatitis among their registered patients, environmental factors and disease characteristics. The data were centrally reviewed and chronic pancreatitis cases defined according to M-ANNHEIM criteria for diagnosis and severity and TIGAR-O classification for etiology. Twenty-three primary care physicians participated in the study. According to their judgment, 51 of 36.401 patients had chronic pancreatitis. After reviewing each patient data, 11 turned out to have definite, 5 probable, 19 borderline and 16 uncertain disease. Prevalence was 30.2/100.000 for definite cases and 44.0/100.000 for definite plus probable cases. Of the 16 patients with definite/probable diagnosis, 8 were male, with mean age of 55.6 (±16.7). Four patients had alcoholic etiology, 5 post-acute/recurrent pancreatitis, 6 were deemed to be idiopathic. Four had pancreatic exocrine insufficiency, 10 were receiving pancreatic enzymes, and six had pain. Most patients had initial stage and non-severe disease. This is the first study investigating the prevalence of chronic pancreatitis in primary care. Results suggest that the prevalence in this context is higher than in hospital-based studies, with specific features, possibly representing an earlier disease stage. Copyright © 2016 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.
Dreyfus, Isabelle; Chouquet, Cécile; Ezzedine, Khaled; Henner, Sophie; Chiavérini, Christine; Maza, Aude; Pascal, Sandrine; Rodriguez, Lauriane; Vabres, Pierre; Martin, Ludovic; Mallet, Stéphanie; Barbarot, Sébastien; Dupuis, Jérôme; Mazereeuw-Hautier, Juliette
Inherited ichthyoses represent a group of rare skin disorders characterized by scaling, hyperkeratosis and inconstant erythema, involving most of the tegument. Epidemiology remains poorly described. This study aims to evaluate the prevalence of inherited ichthyosis (excluding very mild forms) and its different clinical forms in France. Capture - recapture method was used for this study. According to statistical requirements, 3 different lists (reference/competence centres, French association of patients with ichthyosis and internet network) were used to record such patients. The study was conducted in 5 areas during a closed period. The prevalence was estimated at 13.3 per million people (/M) (CI95%, [10.9 - 17.6]). With regard to autosomal recessive congenital ichthyosis, the prevalence was estimated at 7/M (CI 95% [5.7 - 9.2]), with a prevalence of lamellar ichthyosis and congenital ichthyosiform erythroderma of 4.5/M (CI 95% [3.7 - 5.9]) and 1.9/M (CI 95% [1.6 - 2.6]), respectively. Prevalence of keratinopathic forms was estimated at 1.1/M (CI 95% [0.9 - 1.5]). Prevalence of syndromic forms (all clinical forms together) was estimated at 1.9/M (CI 95% [1.6 - 2.6]). Our results constitute a crucial basis to properly size the necessary health measures that are required to improve patient care and design further clinical studies.
Full Text Available It is known that health problem represents an object of permanent study. The study to prevent chronic diseases, factors involved to keep the health status, means to improve the effect of various diseases/affections have been and are appreciated.
Filipowicz, Rebecca; Greene, Tom H.; Wei, Guo; Beddhu, Srinivasan
Both higher body size (as indicated by body mass index- BMI) and higher muscle mass (as indicated by serum creatinine- (SCr)) are associated with better survival in HD patients (pts) but the relative importance of muscle vs. body size is not established. In the current study, the associations of SCr, BMI and the ratio of Scr to BMI with time to death were examined in Cox proportional hazards models using HEMO study data. Details of HEMO Study have been published elsewhere. In the current stud...
Chowdhury, Mohammad Ziaul Islam; Anik, Ataul Mustufa; Farhana, Zaki; Bristi, Piali Dey; Abu Al Mamun, B M; Uddin, Mohammad Jasim; Fatema, Jain; Akter, Tanjila; Tani, Tania Akhter; Rahman, Meshbahur; Turin, Tanvir C
Metabolic syndrome (MS) is a cluster of health problems that set the stage for serious health conditions and places individuals at higher risk of cardiovascular disease, diabetes and stroke. The worldwide prevalence of MS in the adult population is on the rise and Bangladesh is no exception. According to some epidemiological study, MS is highly prevalent in Bangladesh and has increased dramatically in last few decades. To provide a clear picture of the current situation, we conducted a systematic review and meta-analysis with an objective to assess the prevalence of metabolic syndrome among the Bangladeshi population using data already published in the scientific literature. We searched MEDLINE, EMBASE and PubMed and manually checked references of all identified relevant publications that described the prevalence of MS in Bangladesh. Random effects meta-analysis was used to pool the prevalence. Heterogeneity was explored using formal tests and subgroup analyses. Study quality and publication bias was also explored. Electronic and grey literature search retrieved 491 potentially relevant papers. After removing duplicates, reviewing titles and abstracts and screening full texts, 10 studies were finally selected. Most of the studies were conducted in rural populations and study participants were mostly females. The weighted pooled prevalence of metabolic syndrome regardless of gender and criteria used to define metabolic syndrome, was 30.0% with high heterogeneity observed. Weighted pooled prevalence of metabolic syndrome is higher in females (32%) compared to males (25%) though not statistically significant (p = 0.434). Prevalence was highest (37%) when Modified NCEP ATP III criteria was used to define MS, while it was lowest (20%) when WHO criteria was used. In most cases, geographical area (urban/rural) was identified as a source of heterogeneity between the studies. Most of the studies met study quality assessment criteria's except adequate sample size
An epidemiologic study was carried out in the dry season on 250 male commercial motor bike riders from five different parks. 69% of the bike riders were in the (year) 31– 40 and 41–50 age range while 31% were in 21–30, 51– 60 and 61 – 70 age range. Half of the population studied were normotensive. Arterial ...
Seyyed Hadi Mo'tamedi
Full Text Available Objective: The need for immigration usually depends on complicated relations between economical, social, familial and political factors. Unavailability to education, occupation, services and respecting to primary human rights are the most important factors in immigration. Materials & Methods: This study designed and performed for detection of frequency of depression in Afghan refugees in Bardsir’s camp. In this survey 300 persons (162 female, 138 male of that camp were selected. They filled out Beck's depression inventory and demographic questionnaire. The method of sampling was cluster sampling and the study was cross-sectional. Results: Total prevalence of depression in these refugees was 53%. The most severity of depression was in age group 20-29 years. Statistically there was no significant difference between depression and age. The relation between the prevalence of depression and sex was studied (57% in female and 47.8% in male. Statistically there was a significant between the prevalence of depression and sex (P<0.04. Depression rate among single people was more than married people, but the relation between the prevalence of depression and marital status was not significant. The most severity of depression in relation with refuges duration was found in the people with refuges period of 141-150 months. Conclusion: Generally the prevalence of depression among refugees except sex doesn't relate with demographic factors and mainly the factors after migration affected the prevalence of depression.
S A Rizwan
Full Text Available In India there is an increasing trend in hypertension prevalence among the general population. Studies have shown that tribal populations in India are also experiencing this burden.The aim was to estimate the pooled prevalence of primary hypertension among adult tribal populations of India.A systematic search was conducted in MEDLINE, IndMed, Web of Science, Google Scholar and major journals for studies published between 1981 and 2011. Two authors independently reviewed the studies, did quality assessment and extracted data in pre-coded spread-sheets. Pooled estimates of prevalence of hypertension were calculated using DerSimonian-Laird random effects model. Subgroup and sensitivity analyses and meta-regression were performed.Twenty studies or 53 subpopulations with 64 674 subjects were included in final review. The pooled estimate of hypertension prevalence was 16.1% (95% CI: 13.5, 19.2. There was significant heterogeneity among the studies (I2 = 99% and Q = 4624.0, df = 53, p<0.001. Subgroup analyses showed that year of study, acculturation status, special features, and BP measurement techniques significantly influenced prevalence, but after meta-regression analyses, 'decade of study' remained the only covariate that significantly and independently influenced prevalence (R2 = 0.57, Q = 119.2, df = 49, p value <0.001.An increasing trend was found in the prevalence of hypertension in adult tribal populations across three decades. Although acculturation was probably the underlying agent that caused this increase, other unmeasured factors that need further research were also important. Concerned policy makers should focus on the changing health needs of tribal communities.
Sally S. Zahra
May 13, 2016 ... Abstract Purpose: Nocturnal enuresis is defined as the involuntary voiding of urine in bed beyond the age at which bladder control is normally obtained. Previous studies have suggested a high rate of obesity in children with nocturnal enuresis. We evaluated this hypothesis and investi- gated the correlation ...
PROF. OLIVER OSUAGWA
Jun 1, 2015 ... used in the study. Malaria parasitemia was determined using rapid diagnostic test kit ((ICT. COMBO, Core Diagnostic, U.K). Other data was collected using .... Vomiting Loss of Diarrhea. Pale eyes Seizure/ Dizziness. Headache Joint/ Don't know. House- cold weakness appetite convulsion body pains holds ...
), and particularly multidrug-resistant TB (MDR-TB), has become a significant public health problem in a number of countries and an obstacle to effective global TB control. Method: This is a prospective randomized cross sectional study to ...
±8.7(5-11) by 4.4±8.8(3-6) while those from the ... they ingest Taenia solium eggs through the contamination of their food or water with human ... is of free range with poor environmental and personal sanitation. This study was conducted to.
Hoda El Tayeb
Mar 2, 2012 ... Duration of hemodialysis, past history of blood transfu- sions, and alanine ... nal failure and its effect on intrafamilial transmission of the virus. 3. Subjects and methods. Twenty patients with HCV were enrolled in this study. They ... Analysis of data was done by IBM computer using SPSS. (statistical program ...
Wu, Yu-Tzu; Grant, William B; Prina, A Matthew; Lee, Hsin-yi; Brayne, Carol
Western diets are associated with obesity, vascular diseases, and metabolic syndrome and might increase dementia risk in later life. If these associations are causal, those low- and middle-income countries experiencing major changes in diet might also see an increasing prevalence of dementia. To investigate the relationship of dietary supply and the prevalence of dementia in mainland China, Hong Kong, and Taiwan over time using existing data and taking diagnostic criteria into account. Estimated total energy supply and animal fat from the United Nations was linked to the 70 prevalence studies in mainland China, Hong Kong, and Taiwan from 1980 to 2012 according to the current, 10 years, and 20 years before starting year of investigation. Studies using newer and older diagnostic criteria were separated into two groups. Spearman's rank correlation was calculated to investigate whether trends in total energy, animal fat supply, and prevalence of dementia were monotonically related. The supply of total energy and animal fat per capita per day in China increased considerably over the last 50 years. The original positive relationship of dietary supply and dementia prevalence disappeared after stratifying by newer and older diagnostic criteria and there was no clear time lag effect. Taking diagnostic criteria into account, there is no cross-sectional or time lag relationship between the dietary trends and changes in dementia prevalence. It may be too early to detect any such changes because current cohorts of older people did not experience these dietary changes in their early to mid-life.
Kakoei, Shahla; Parirokh, Masoud; Nakhaee, Nouzar; Jamshidshirazi, Forogh; Rad, Maryam; Kakooei, Sina
This study was carried out to estimate toothache prevalence among adult residents in Kerman. This cross-sectional, population-based study was conducted among individuals aged over 18 years (n=1800). The relevant data on the prevalence of toothache and associated factors were collected by interviewing the individuals in their homes and filling out a questionnaire designed by the examiners. Prevalence of toothache and associated factors that patients recalled previous to their interview were analyzed by chi-square test and multivariate logistic regression analysis. Nine hundred ninety-one individuals (55.1%) reported toothache during the 6 months before the interview. The participants who flossed daily, had regular dental visits, and had higher education showed a significantly lower prevalence of toothache (Pdental floss (OR=1.5), or having a mental or psychological illness (OR=1.5) were more likely to have a history of toothache. High prevalence of toothache (more than half) among residents of Kerman shows a serious and major public health problem. Toothache prevalence in middle aged adults, lower education, bigger family size, lower dental hygiene habit and/or those having mental or psychological illness were more common in the city of Kerman.
Lee, Seung Youp; Park, In Woo; Jang, In San; Choi, Dong Soon; Cha, Bong Kuen
This retrospective study was performed to investigate the prevalence of the idiopathic osteosclerosis (IO) in Korean malocclusion patients according to age, sex, and the Angle's classification of malocclusion. This study consisted of 2,001 randomly selected patients from the Department of Orthodontics at the Gangneung-Wonju National University Dental Hospital, Korea. The prevalence of IO in Korean malocclusion patients was recorded using their panoramic radiographs, and the following parameters were surveyed; age, sex, and the Angle's classification of malocclusion. The chi-square test was analyzed to determine the statistical significance of differences in the prevalence of IO between age, sex, and the Angle's classification of malocclusion. The prevalence of IO in the jaws was 6.7% in a total of 2,001 examined orthodontic patients. The majority of IO was found in the mandible (96.58%). The 30-39 age group showed the highest prevalence of IO (9.60%). There was a higher prevalence in females (6.89%) than in males (6.45%). The prevalence of IO in Angle Class I group (7.07%) was the most frequent, followed by Angle Class II group (6.72%), and Angle Class III group (6.40%). However, there was no statistical significance in sex and Angle's classification of malocclusion. The prevalence of IO in malocclusion patients showed the differences between various age groups and most of them were found in the mandibular posterior area. However, sex and the type of malocclusion are not to be considered as a contributing factor of IO.
Tan, Victoria Ping-Yi; Wong, Benjamin ChunYu; Wong, Wai Man; Leung, Wai Keung; Tong, Daniel; Yuen, Man Fung; Fass, Ronnie
The prevalence of gastroesophageal reflux disease (GERD) is consistently lower in the Chinese than in white populations. Population-based data tracking the time trend of GERD prevalence in Chinese subjects is conflicting. This study examines the population prevalence, risk factors, and time trend associated with GERD in a Chinese population. A population-based cross-sectional study utilizing a validated GERD questionnaire administered by a telephone survey was performed on 3360 Chinese subjects from Hong Kong. GERD prevalence rates in 2011 were compared with prevalence rates in 2002 and 2006. Multiple logistic regressions were performed to determine the risk factors associated with weekly GERD. A total of 2074 subjects (mean age, 48.1±18.2 y; range 18 to 94; 63.1% female) completed the survey (response rate 61.7%). The prevalence of GERD as defined by the Montreal definition was 3.8%. The prevalence of weekly GERD had increased by 1.3% between 2002 and 2011, which represents an at least 50% relative increase (PGERD was associated with noncardiac chest pain [odds ratio (OR), 1.7; 95% confidence interval (CI), 1.034-2.9; P=0.037], dyspepsia (OR, 5.1; 95% CI, 3.0-8.8; PGERD rates in the ethnic Chinese have risen over the last decade. Despite this, variables associated with a survey diagnosis of GERD remain ostensibly unchanged. GERD research in East Asia should focus on the factors driving the rapid rise in prevalence rates and the association with more atypical symptoms of GERD.
Mota, Maria; Popa, Simona Georgiana; Mota, Eugen; Mitrea, Adina; Catrinoiu, Doina; Cheta, Dan Mircea; Guja, Cristian; Hancu, Nicolae; Ionescu-Tirgoviste, Constantin; Lichiardopol, Radu; Mihai, Bogdan Mircea; Popa, Amorin Remus; Zetu, Cornelia; Bala, Cornelia Gabriela; Roman, Gabriela; Serafinceanu, Cristian; Serban, Viorel; Timar, Romulus; Veresiu, Ioan Andrei; Vlad, Adrian Radu
The PREDATORR (PREvalence of DiAbeTes mellitus, prediabetes, overweight, Obesity, dyslipidemia, hyperuricemia and chronic kidney disease in Romania) study is the first national study analyzing the prevalence of diabetes mellitus (DM) and prediabetes, and their association with cardiometabolic, sociodemographic, and lifestyle risk factors in the Romanian population aged 20-79 years. This was an epidemiological study with a stratified, cross-sectional, cluster random sampling design. Sociodemographic, lifestyle, and anamnestic data were collected through self- and interviewer-administered questionnaires, and biochemical assays and oral glucose tolerance tests were performed. In all, 2728 participants from 101 clinics of general practitioners were randomly selected, with a probability proportional to population size according to the 2002 Romanian Census. The participation rate was 99.6%. Impaired glucose regulation (prediabetes, known and unknown DM) was found in 28.1% of the study population. The overall age- and sex-adjusted prevalence of DM was 11.6% (95% CI 9.6%-13.6%), of which 2.4% (95% CI 1.7%-3.1%) had unknown DM. The prevalence of DM increased with age and was higher in men than in women. The age- and sex-adjusted prevalence of prediabetes was 16.5% (95%CI 14.8%-18.2%), with the highest percentage in the 60-79 year age group and in women. Obesity, abdominal obesity, dyslipidemia, low education level, and a family history of diabetes were associated with glucose metabolism disorders. The PREDATORR study shows a high prevalence of impaired glucose regulation in the adult Romanian population, providing data on the prevalence of DM and prediabetes and their association with several risk factors. © 2015 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and John Wiley Sons & Australia, Ltd.
Yang, Jaeun; Hirsch, Lauren; Martino, Davide; Jette, Nathalie; Roberts, Jodie; Pringsheim, Tamara
The objective of this study was to examine: (1) the prevalence of diagnosed Tourette syndrome in Canada by sex in youth (aged 12-17) and adults and (2) socioeconomic factors in this population. The majority of epidemiological studies of tics have focused on children and youth, with few studies describing the prevalence of tics in adult populations. Canadian data on Tourette syndrome prevalence were derived from the Canadian Community Health Survey 2010 and 2011 cycles, a Statistics Canada population-based cross-sectional survey that collects information related to health status. We determined the prevalence of diagnosed Tourette syndrome and examined sociodemographic factors, including age, sex, education, income, employment, and birthplace. Overall, 122,884 Canadians participated in the surveys, with 122 participants diagnosed with Tourette syndrome. The prevalence of Tourette syndrome was higher in males in youth: 6.03 per 1000 (95% confidence interval: 3.24-8.81) in males versus 0.48 per 1,000 (95% confidence interval: 0.05-0.91) in females, with a prevalence risk ratio of 5.31 (95% confidence interval: 2.38-11.81). In adults, the prevalence of Tourette syndrome was 0.89 per 1,000 (95% confidence interval: 0.48-1.29) in males versus 0.44 (95% confidence interval: 0.16.0-0.71) in females, with a prevalence risk ratio of 1.93 (95% confidence interval: 1.21-3.08). After adjusting for age and sex, adults with Tourette syndrome had lower odds of receiving postsecondary education or being employed and higher odds of having income lower than the median and receiving governmental support. Data on the prevalence of Tourette syndrome in adults are scarce because most studies focus on children. Our data demonstrate a decreasing prevalence risk ratio for sex in adults compared to children. A diagnosis of Tourette syndrome is associated with lower education, income, and employment in adulthood. © 2016 International Parkinson and Movement Disorder Society. © 2016
Dinesh Kumar Dhanwal
Full Text Available Background: A previous hospital based study from Delhi revealed a high prevalence of hypothyroidism in pregnant women. Several other studies with small sample size also indicate a rising trend of prevalence of hypothyroidism during pregnancy in India. Objective: To assess prevalence of hypothyroidism in pregnant women from various states/cities across India. Materials and Methods: This was a cross-sectional multicenter study conducted at Allahabad (Uttar Pradesh, Bengaluru (Karnataka, Chennai (Tamil Nadu, Kolkata (West Bengal, Hyderabad (Telangana, Nasik (Maharashtra, Rohtak (Haryana, Pune (Maharashtra, New Delhi (Delhi, Srinagar (Kashmir, and Vizag (Andhra Pradesh enrolling 2599 pregnant women. Estimation of thyroid stimulating hormone (TSH, free T4, and antithyroid peroxidase (TPO antibodies was carried out using Roche modular kit using ECLIA technology in a central laboratory. Results: We found in our study population that 13.13% of pregnant women have hypothyroidism (n = 388, using a cutoff TSH level of 4.5 μIU/ml. This prevalence was much higher using the American Thyroid Association criteria. Anti-TPO antibodies were positive in 20.74% of all pregnant women (n = 613, whereas 40% (n = 155 of hypothyroid pregnant women were positive for anti-TPO antibodies. Conclusion: This study concludes that there is a high prevalence of hypothyroidism (13.13%, majority being subclinical in pregnant women during the first trimester from India and universal screening of hypothyroidism may be desirable in our country.
Chan-Yeung, M; Anthonisen, N R; Becklake, M R; Bowie, D; Sonia Buist, A; Dimich-Ward, H; Ernst, P; Sears, M R; Siersted, H C; Sweet, L; Van Til, L; Manfreda, J
Geographical variations in atopic sensitization in Canada have not been described previously. This study used the standardized protocol of the European Community Respiratory Health Survey-1 (ECRHS-1) to investigate the distribution and predictors of atopic sensitization in six sites across Canada and to compare the results with some ECRHS-1 centers. Adults aged 20-44 years in six study sites across Canada underwent allergy skin testing using 14 allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae) cat, cockroach, grasses (Timothy grass, Kentucky grass), molds (Cladosporium herbarium, Alternaria alternata, Aspergillus fumigatus, Penicillium), trees (tree mix, birch, Olea europea), and common ragweed. The overall prevalence of atopy (skin test over 0 mm to any allergen) was 62.7%. There was significant geographical variation in the prevalence of atopy in the six study sites (lowest 55.6% [95% C.I.51.3-59.9] in Prince Edward Island, highest 66.0 [61.7-70.3] in Montreal) and of sensitization to each of the allergens tested even after adjustment for confounders. When the first eight of the nine allergens in the ECRHS were used to estimate the prevalence of atopic sensitization, the prevalence of atopy in Canada was 57% compared with 35.2% overall for centers in the ECRHS. The prevalence of atopy in Vancouver (57% [52.3-61.8]) was close to that of Portland, Oregon (52.1% [46.2-58.0]). There was a significant variation in atopic sensitization among different study sites across Canada. The prevalence of atopic sensitization is relatively high in Canada compared with sites in the ECRHS and this may, in part, account for the high prevalence of asthma and asthma symptoms in Canada. © 2010 John Wiley & Sons A/S.
Hae Dong Woo
Full Text Available The prevalence of metabolic syndrome has been increasing in Korea and has been associated with dietary habits. The aim of our study was to identify the relationship between dietary patterns and the prevalence of metabolic syndrome. Using a validated food frequency questionnaire, we employed a cross-sectional design to assess the dietary intake of 1257 Korean adults aged 31 to 70 years. To determine the participants' dietary patterns, we considered 37 predefined food groups in principal components analysis. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III. The abdominal obesity criterion was modified using Asian guidelines. Prevalence ratios and 95% confidence intervals for the metabolic syndrome were calculated across the quartiles of dietary pattern scores using log binomial regression models. The covariates used in the model were age, sex, total energy intake, tobacco intake, alcohol consumption, and physical activity. The prevalence of metabolic syndrome was 19.8% in men and 14.1% in women. The PCA identified three distinct dietary patterns: the 'traditional' pattern, the 'meat' pattern, and the 'snack' pattern. There was an association of increasing waist circumference and body mass index with increasing score in the meat dietary pattern. The multivariate-adjusted prevalence ratio of metabolic syndrome for the highest quartile of the meat pattern in comparison with the lowest quartile was 1.47 (95% CI: 1.00-2.15, p for trend = 0.016. A positive association between the prevalence of metabolic syndrome and the dietary pattern score was found only for men with the meat dietary pattern (2.15, 95% CI: 1.10-4.21, p for trend = 0.005. The traditional pattern and the snack pattern were not associated with an increased prevalence of metabolic syndrome. The meat dietary pattern was associated with a higher prevalence of metabolic syndrome in Korean male adults.
Woo, Hae Dong; Shin, Aesun; Kim, Jeongseon
The prevalence of metabolic syndrome has been increasing in Korea and has been associated with dietary habits. The aim of our study was to identify the relationship between dietary patterns and the prevalence of metabolic syndrome. Using a validated food frequency questionnaire, we employed a cross-sectional design to assess the dietary intake of 1257 Korean adults aged 31 to 70 years. To determine the participants' dietary patterns, we considered 37 predefined food groups in principal components analysis. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III. The abdominal obesity criterion was modified using Asian guidelines. Prevalence ratios and 95% confidence intervals for the metabolic syndrome were calculated across the quartiles of dietary pattern scores using log binomial regression models. The covariates used in the model were age, sex, total energy intake, tobacco intake, alcohol consumption, and physical activity. The prevalence of metabolic syndrome was 19.8% in men and 14.1% in women. The PCA identified three distinct dietary patterns: the 'traditional' pattern, the 'meat' pattern, and the 'snack' pattern. There was an association of increasing waist circumference and body mass index with increasing score in the meat dietary pattern. The multivariate-adjusted prevalence ratio of metabolic syndrome for the highest quartile of the meat pattern in comparison with the lowest quartile was 1.47 (95% CI: 1.00-2.15, p for trend = 0.016). A positive association between the prevalence of metabolic syndrome and the dietary pattern score was found only for men with the meat dietary pattern (2.15, 95% CI: 1.10-4.21, p for trend = 0.005). The traditional pattern and the snack pattern were not associated with an increased prevalence of metabolic syndrome. The meat dietary pattern was associated with a higher prevalence of metabolic syndrome in Korean male adults.
Droogleever Fortuyn, H.A.; Swinkels, S.H.N.; Buitelaar, J.K.; Renier, W.O.; Furer, J.W.; Rijnders, C.A.T.; Hodiamont, P.P.G.; Overeem, S.
Study Objectives: To study the prevalence of and symptoms of eating disorders in patients with narcolepsy. Design: We performed a case-control study comparing symptoms of eating disorders in patients with narcolepsy versus healthy population controls, using the Schedules for Clinical Assessment
Droogleever Fortuyn, H.A.; Swinkels, S.H.N.; Buitelaar, J.K.; Renier, W.O.; Furer, J.W.; Rijnders, C.A.T.; Hodiamont, P.P.G.; Overeem, S.
STUDY OBJECTIVES: To study the prevalence of and symptoms of eating disorders in patients with narcolepsy. DESIGN: We performed a case-control study comparing symptoms of eating disorders in patients with narcolepsy versus healthy population controls, using the Schedules for Clinical Assessment in
Thapa, S.S.; Thapa, R.; Paudyal, I.; Khanal, S.; Aujla, J.; Paudyal, G.; van Rens, G.H.M.B.
Background: Vitreo-retinal diseases are among the leading causes of visual impairment and blindness worldwide. This study reports the prevalence and pattern of vitreo-retinal diseases in the Bhaktapur Glaucoma Study (BGS), a population based study conducted in Nepal. Methods. BGS was a population
Exarchou, Sofia; Lindström, Ulf; Askling, Johan; Eriksson, Jonas K; Forsblad-d'Elia, Helena; Neovius, Martin; Turesson, Carl; Kristensen, Lars Erik; Jacobsson, Lennart T H
Prevalence estimates of ankylosing spondylitis vary considerably, and there are few nationwide estimates. The present study aimed to describe the national prevalence of clinically diagnosed ankylosing spondylitis in Sweden, stratified according to age, sex, geographical, and socio-economic factors, and according to subgroups with ankylosing spondylitis-related clinical manifestations and pharmacological treatment. All individuals diagnosed with ankylosing spondylitis according to the World Health Organization International Classification of Disease codes, between 1967 and 2009, were identified from the National Patient Register. Data regarding disease manifestations, patient demographics, level of education, pharmacological treatment, and geographical region were retrieved from the National Patient Register and other national registers. A total of 11,030 cases with an ankylosing spondylitis diagnosis (alive, living in Sweden, and 16 to 64 years old in December 2009) were identified in the National Patient Register, giving a point prevalence of 0.18% in 2009. The prevalence was higher in northern Sweden, and lower in those with a higher level of education. Men had a higher prevalence of ankylosing spondylitis (0.23% versus 0.14%, P ankylosing spondylitis of 0.18%. It revealed phenotypical and treatment differences between the sexes, as well as geographical and socio-economic differences in disease prevalence.
Uña Orejón, R; Gisbert de la Cuadra, L; Garríguez Pérez, D; Díez Sebastián, J; Ureta Tolsada, M P
To assess the types of maintenance fluids used in our hospital, comparing their volume and composition to the standards recommended by the guidelines. Observational, cross-sectional study. Volume and type of fluid therapy administered during 24h to patients admitted to various hospital departments were recorded. Patients receiving fluid therapy because of water-electrolyte imbalance were excluded. Out of 198 patients registered, 74 (37.4%) were excluded because they did not meet the criteria for inclusion. Mean administered volume was 2,500cc/day. Mean daily glucose dose was 36g per 24h (SD: 31.4). The most frequent combination included normal saline solution (NSS) and glucose 5% (64.4%). Mean daily dose of sodium and chlorine was, respectively, 173mEq (SD: 74.8) and 168mEq (SD: 75), representing a surplus daily dose of +87.4mEq and +85mEq. Potassium, magnesium and calcium daily deficit was, respectively, -50mEq, -22mEq and -21mEq per day. Buffer administration was exceptional, bicarbonate (2.29%), acetate (1.29%), lactate (1.15%) and gluconate (1.10%) being the buffering agents most frequently used. NNS is the most frequently used solution. In contrast to excess doses of sodium and chlorine, there is a great deficit of other ions, buffering agents and caloric intake in the fluid therapy regimens that are usually prescribed. Copyright © 2017 Sociedad Española de Anestesiología, Reanimación y Terapéutica del Dolor. Publicado por Elsevier España, S.L.U. All rights reserved.
Mitchell, Kimberly J; Finkelhor, David; Jones, Lisa M; Wolak, Janis
To obtain national estimates of youth involved in sexting in the past year (the transmission via cell phone, the Internet, and other electronic media of sexual images), as well as provide details of the youth involved and the nature of the sexual images. The study was based on a cross-sectional national telephone survey of 1560 youth Internet users, ages 10 through 17. Estimates varied considerably depending on the nature of the images or videos and the role of the youth involved. Two and one-half percent of youth had appeared in or created nude or nearly nude pictures or videos. However, this percentage is reduced to 1.0% when the definition is restricted to only include images that were sexually explicit (ie, showed naked breasts, genitals, or bottoms). Of the youth who participated in the survey, 7.1% said they had received nude or nearly nude images of others; 5.9% of youth reported receiving sexually explicit images. Few youth distributed these images. Because policy debates on youth sexting behavior focus on concerns about the production and possession of illegal child pornography, it is important to have research that collects details about the nature of the sexual images rather than using ambiguous screening questions without follow-ups. The rate of youth exposure to sexting highlights a need to provide them with information about legal consequences of sexting and advice about what to do if they receive a sexting image. However, the data suggest that appearing in, creating, or receiving sexual images is far from being a normative behavior for youth.
Yon, Yongjie; Mikton, Christopher; Gassoumis, Zachary D; Wilber, Kathleen H
Elder abuse is an important public health and human rights issue, yet its true extent is not well understood. To address this, we will conduct a systematic review and meta-analysis of elder abuse prevalence studies from around the world. This protocol describes the methodological approach to be adopted for conducting this systematic review and meta-analysis. In particular, the protocol describes the search strategies and eligibility criteria to be used to identify and select studies and how data from the selected studies will be extracted for analysis. The protocol also describes the analytical approach that will be used to calculate pooled prevalence estimates and discusses the use of meta-regression to assess how studies' characteristics influence the prevalence estimates. This protocol conforms to the Preferred Reporting Items for Systematic reviews and Meta-Analysis - or PRISMA - guidelines and has been registered with the PROSPERO International Prospective Register of systematic reviews.
Full Text Available Objective: the main purpose of this research was to collect and survey the prevalence studies of drug misuse and psychotropic drugs that have been done sporadically among university students from 1374 to 1392. Up to now, no survey of data has been done on these researches. Method: library has been the method of this research and 37 researches were analyzed. Findings: methodologically, the results show that many prevalence studies are with major faults. Conclusion: the studies show that less attention has been given to prevalence studies of drug addiction and useful interventions in university students. However, the rate of addiction during 2 decades has been stable cigar, hookah take the first and second places and alcohol, opium, hashish and heroine come respectively.
Roy A. Wong-McClure
Full Text Available OBJECTIVE: To report the prevalence of metabolic syndrome (MetS as found by the Central American Diabetes Initiative (CAMDI study for five major Central American populations: Belize (national; Costa Rica (San José; Guatemala (Guatemala City; Honduras (Tegucigalpa; and Nicaragua (Managua. METHODS: Study data on 6 185 adults aged 20 years or older with anthropometric and laboratory determination of MetS from population-based surveys were analyzed. Overall, the survey response rate was 82.0%. MetS prevalence was determined according to criteria from the Adult Treatment Panel III of the National Cholesterol Education Program. The study's protocol was reviewed and approved by the bioethical committee of each country studied. RESULTS: The overall standardized prevalence of MetS in the Central American region was 30.3% (95% confidence interval (CI: 27.1-33.4. There was wide variability by gender and work conditions, with higher prevalence among females and unpaid workers. The standardized percentage of the population free of any component of MetS was lowest in Costa Rica (9.0%; CI: 6.5-11.4 and highest in Honduras (21.1%; CI: 16.4-25.9. CONCLUSIONS: Overall prevalence of MetS in Central America is high. Strengthening surveillance of chronic diseases and establishing effective programs for preventing cardiovascular diseases might reduce the risk of MetS in Central America.
Harper, C; Gold, J; Rodriguez, M; Perdices, M
In a prospective necropsy study, the prevalence of the Wernicke-Korsakoff syndrome (WKS) in Sydney, Australia was 2.1% of adults over the age of 15 years. The population studied encompassed a wide spectrum of socio-economic and cultural backgrounds. Abuse of alcohol appeared to be the major predisposing factor to the development of the WKS in cases which were adequately documented. This high prevalence rate is in line with other clinical and pathological Australian studies and provides additional support for the idea of prevention of the WKS by the use of thiamin supplements in the Australian diet in flour, bread and perhaps alcoholic beverages. PMID:2784828
Xambre, Pedro Augusto Oliveira Santos; Valerio, Claudia Scigliano; Alves Cardoso, Claudia Assuncoe; Custodio, Antonio Luis Neto; Manzi, Flavio Ricardo [Dept. of Oral Radiology, School of Dentistry, Pontifical Catholic University of Minas Gerais, Belo Horizonte (Brazil)
In the present study, we coined the term 'alveolar dome' and aimed to demonstrate the prevalence of alveolar domes through digital periapical radiographs. This study examined 800 digital periapical radiographs in regard to the presence of alveolar domes. The periapical radiographs were acquired by a digital system using a photostimulable phosphor (PSP) plate. The χ2 test, with a significance level of 5%, was used to compare the prevalence of alveolar domes in the maxillary posterior teeth and, considering the same teeth, to verify the difference in the prevalence of dome-shaped phenomena between the roots. The prevalence of alveolar domes present in the first pre-molars was statistically lower as compared to the other maxillary posterior teeth (p<0.05). No statistically significant difference was observed in the prevalence of alveolar domes between the maxillary first and second molars. Considering the maxillary first and second molars, it was observed that the palatal root presented a lower prevalence of alveolar domes when compared to the distobuccal and mesiobuccal roots (p<0.05). The present study coined the term 'alveolar dome', referring to the anatomical projection of the root into the floor of the maxillary sinus. The maxillary first and second molars presented a greater prevalence of alveolar domes, especially in the buccal roots, followed by the third molars and second pre-molars. Although the periapical radiograph is a two-dimensional method, it can provide dentists with the auxiliary information necessary to identify alveolar domes, thus improving diagnosis, planning, and treatment.
Raposo, Luís; Severo, Milton; Barros, Henrique; Santos, Ana Cristina
The PORMETS study was designed to estimate the prevalence of metabolic syndrome and its determinants in the overall and administrative regions of the Portuguese mainland. A cross-sectional study of a representative sample of non-institutionalized Portuguese adults selected from primary health care centres lists including 1695 men and 2309 women was conducted from February 2007 to July 2009. A structured questionnaire was administered, collecting information on personal medical history and socio-demographic and behavioural characteristics. Anthropometrics, blood pressure, and venous blood samples were obtained. Metabolic syndrome was defined according to three operational definitions. The prevalence ratios and their respective 95% confidence intervals were calculated using binomial generalized linear regression, with the log link function. The prevalence rates of metabolic syndrome in this sample of Portuguese adults were 36.5%, 49.6%, and 43.1%, using the Adult Treatment Panel III, International Diabetes Federation and Joint Interim Statement definitions, respectively. The most prevalent feature of metabolic syndrome in this sample was high blood pressure (64.3%) and the lowest was high fasting glucose (24.9%). After adjustment for age and gender, significant differences were observed for the 18 districts of the Portugal mainland. Additionally, metabolic syndrome was significantly more frequent in non-urban areas than in urban ones (p = 0.001). The prevalence of metabolic syndrome was significantly higher in women (p˂0.001) and older participants (p˂0.001), as well as in those who reported being housewives (p = 0.010), retired (p = 0.046) or unemployed (p = 0.024). This study showed that metabolic syndrome is highly prevalent in the Portuguese adult population. Regional differences in the prevalence of this syndrome were observed, and this condition was more common in non-urban areas and less favoured socio-economic categories.
Full Text Available INTRODUCTION: Obesity is a worldwide public health issue. Since the epidemiological behaviour of this disease is not well established in our country, the purpose of this study was to determinate its prevalence in the Maracaibo City, Zulia State- Venezuela. MATERIALS AND METHODS: A cross-sectional study was undertaken using the data set from the Maracaibo City Metabolic Syndrome Prevalence Study. The sample consists of 2108 individuals from both genders and randomly selected: 1119 (53.09% women and 989 (46.91% men. The participants were interrogated for a complete clinical history and anthropometric measurements. To classify obesity, the WHO criteria for Body Mass Index (BMI, and Waist Circumference (WC from the IDF/NHLBI/AHA/WHF/IAS/IASO-2009 (IDF-2009 and ATPIII statements were applied. RESULTS: For BMI, obesity had an overall prevalence of 33.3% (n = 701, and according to gender women had 32.4% (n = 363 and men had 34.2% (n = 338. Overweight had a prevalence of 34.8% (n = 733, Normal weight had 29.8% (n = 629, and Underweight had 2.1% (n = 45. Adding Obesity and Overweight results, the prevalence of elevated BMI (>25 Kg/m(2 was 68.1%. Using the IDF-2009 WC's cut-off, Obesity had 74.2% prevalence, compared to 51.7% using the ATPIII parameters. CONCLUSIONS: These results show a high prevalence of abdominal obesity in our locality defined by the WHO, IDF-2009 and ATPIII criteria, which were not designed for Latin-American populations. We suggest further investigation to estimate the proper values according to ethnicity, genetic background and sociocultural aspects.
Full Text Available Abstract Background The PORMETS study was designed to estimate the prevalence of metabolic syndrome and its determinants in the overall and administrative regions of the Portuguese mainland. Methods A cross-sectional study of a representative sample of non-institutionalized Portuguese adults selected from primary health care centres lists including 1695 men and 2309 women was conducted from February 2007 to July 2009. A structured questionnaire was administered, collecting information on personal medical history and socio-demographic and behavioural characteristics. Anthropometrics, blood pressure, and venous blood samples were obtained. Metabolic syndrome was defined according to three operational definitions. The prevalence ratios and their respective 95% confidence intervals were calculated using binomial generalized linear regression, with the log link function. Results The prevalence rates of metabolic syndrome in this sample of Portuguese adults were 36.5%, 49.6%, and 43.1%, using the Adult Treatment Panel III, International Diabetes Federation and Joint Interim Statement definitions, respectively. The most prevalent feature of metabolic syndrome in this sample was high blood pressure (64.3% and the lowest was high fasting glucose (24.9%. After adjustment for age and gender, significant differences were observed for the 18 districts of the Portugal mainland. Additionally, metabolic syndrome was significantly more frequent in non-urban areas than in urban ones (p = 0.001. The prevalence of metabolic syndrome was significantly higher in women (p˂0.001 and older participants (p˂0.001, as well as in those who reported being housewives (p = 0.010, retired (p = 0.046 or unemployed (p = 0.024. Conclusions This study showed that metabolic syndrome is highly prevalent in the Portuguese adult population. Regional differences in the prevalence of this syndrome were observed, and this condition was more common in non-urban areas and less
Full Text Available Abstract Background Several studies conducted during the 1990s indicated that childhood allergic diseases were increasing worldwide, but more recent investigations in some Western countries have suggested that the trend is stabilizing or may even be reversing. However, few data are available on the current status of allergic disease prevalence in Chinese children. The aim of the present study was to investigate the prevalence rates of asthma, allergic rhinitis, and eczema in children of three major cities of China, to determine the status of allergic diseases among Chinese children generally, and to evaluate the prevalence of allergic diseases in children of different ages. Methods We conducted a cross-sectional survey between October 2008 and May 2009 in three major cities of China (Beijing, Chongqing, and Guangzhou to evaluate the prevalence rates of childhood allergic diseases including asthma, allergic rhinitis, and eczema, using a questionnaire of the International Study of Asthma and Allergies in Childhood (ISAAC group. A total of 24,290 children aged 0-14 years were interviewed, using a multi-stage sampling method. To acquire data on children aged 3-14 years, we visited schools and kindergartens. To access children too young to attend school or kindergarten, we extended our survey to community health service centers. Each questionnaire was completed by a parent or guardian of a child after an informed consent form was signed. Results Of the 24,290 children in our study, 12,908 (53.14% were males and 11,382 (46.86% females; 10,372 (42.70% were from Beijing, 9,846 (40.53% from Chongqing, and 4,072 (16.77% from Guangzhou. Our survey indicated that in Beijing, Chongqing, and Guangzhou, the prevalence rates of asthma were 3.15%, 7.45%, and 2.09%, respectively; the rates of allergic rhinitis were 14.46%, 20.42%, and 7.83%; and the rates of eczema were 20.64%, 10.02%, and 7.22%. The prevalence of allergic diseases varied with age. Asthma was
Ibrahim, Ahmed K; Kelly, Shona J; Adams, Clive E; Glazebrook, Cris
Depression is a common health problem, ranking third after cardiac and respiratory diseases as a major cause of disability. There is evidence to suggest that university students are at higher risk of depression, despite being a socially advantaged population, but the reported rates have shown wide variability across settings. To explore the prevalence of depression in university students. PubMed, PsycINFO, BioMed Central and Medline were searched to identify studies published between 1990 and 2010 reporting on depression prevalence among university students. Searches used a combination of the terms depression, depressive symptoms, depressive disorders, prevalence, university students, college students, undergraduate students, adolescents and/or young adults. Studies were evaluated with a quality rating. Twenty-four articles were identified that met the inclusion and exclusion criteria. Reported prevalence rates ranged from 10% to 85% with a weighted mean prevalence of 30.6%. The results suggest that university students experience rates of depression that are substantially higher than those found in the general population. Study quality has not improved since 1990. Copyright © 2012 Elsevier Ltd. All rights reserved.
Ben Haider, Nour Y; Ziyab, Ali H
This cross-sectional study sought to estimate the prevalence of prediabetes and assess its association with obesity among young adults in Kuwait; a country with a high prevalence of obesity and diabetes. The estimated prevalence of prediabetes was 6.3% (95% CI: 4.8-8.1) and obesity was associated with elevated prediabetes prevalence. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Full Text Available Hypertension is one of the leading causes of disease burden across the world. In China, the latest nationwide survey of prevalence of hypertension was ten year ago, and data in rural areas is little known. More information about hypertension prevalence could help to improve overall antihypertensive health care. We aimed to estimate the pooled prevalence of hypertension in rural areas of China.Comprehensive electronic searches of PubMed, Web of Knowledge, Chinese Web of Knowledge, Wangfang, Weipu and SinoMed databases were conducted to identify any study in each database published from January 1, 2004 to December 31, 2013, reporting the prevalence of hypertension in Chinese rural areas. Prevalence estimates were stratified by age, area, sex, publication year, and sample size. All statistical calculations were made using the Stata Version 11.0 (College Station, Texas and Statsdirect Version 2.7.9.We identified 124 studies with a total population of 3,735,534 in the present meta-analysis. Among people aged 18 years old in Chinese rural areas, the summarized prevalence is 22.81% (19.41%-26.41%. Subgroup analysis shows the following results: for male 24.46% (21.19%-27.89%, for female 22.17% (18.25%-26.35%. For 2004-2006: 18.94% (14.41%-23.94%, for 2007-2009, 21.24% (15.98%-27.01% for 2010-2013: 26.68%, (20.79%-33.02%. For Northern region 25.76% (22.36%-29.32%, for Southern region 19.30%, (15.48%-24.08%.The last decade witnessed the growth in prevalence of hypertension in rural areas of China compared with the fourth national investigation, which has climbed the same level as the urban area. Guidelines for screening and treatment of hypertension in rural areas need to be given enough attention.
Ellervik, Christina; Kvetny, Jan; Christensen, Kaj Sparle
to describe the prevalence of antidepressants received by the respondents in the GESUS study and the correspondence to their subjective well-being on the WHO-5 questionnaire. Methods: To evaluate the validity (scalability) of the MDI and the WHO-5 in the GESUS study we performed the non-parametric Mokken...
Straaten, B. Van; Rodenburg, G.; Laan, J. van der; Boersma, S.N.; Wolf, J.R.; Mheen, D. Van de
BACKGROUND: Previous studies have shown that substance use among homeless people is a prevalent problem that is associated with longer durations of homelessness. Most studies of substance use among the homeless were carried out outside Europe and have limited generalizability to European countries.
Van Straaten, Barbara; Rodenburg, Gerda; Van der Laan, Jorien; Boersma, Sandra N; Wolf, Judith R L M; Van de Mheen, Dike
BACKGROUND: Previous studies have shown that substance use among homeless people is a prevalent problem that is associated with longer durations of homelessness. Most studies of substance use among the homeless were carried out outside Europe and have limited generalizability to European countries.
Siu, Angela F. Y.
Studies in Hong Kong indicated that there is a tendency for young children to use internalizing as a means to cope with their daily difficulties. Mother-child relationship has been seen as a factor affecting a child's adaptive coping skills. In this study, we explored the prevalence of internalizing problems among primary school children in Hong…
Cow’s milk allergy is a common disease in infancy. However, until recently accurate numbers were not available. To investigate how many infants suffered from cow’s milk allergy, the EuroPrevall study was designed. In this European, multicentre study, nine European hospitals followed over 12,000
A cross sectional study aimed at determining the prevalence and type of common Fasciola species in cattle was conducted in and around Bahir Dar from November 2008- March 2009. The study was based on post-mortem inspection of livers of slaughtered animals at Bahir Dar municipality abattoir and coprological ...
Quispel, R.; van Boxel, O. S.; Schipper, M. E.; Sigurdsson, V.; Canninga-van Dijk, M. R.; Kerckhoffs, A.; Smout, A. J.; Samsom, M.; Schwartz, M. P.
BACKGROUND AND STUDY AIMS: The first cases of squamous cell carcinoma in esophageal lichen planus were recently described. We performed a study to establish the prevalence of endoscopic and histopathologic abnormalities consistent with lichen planus and (pre-) malignancy in a cohort of patients with
N. Terzikhan (Natalie); K.M.C. Verhamme (Katia); A. Hofman (Albert); B.H.Ch. Stricker (Bruno); G.G. Brusselle (Guy); L. Lahousse (Lies)
textabstractCOPD is the third leading cause of death in the world and its global burden is predicted to increase further. Even though the prevalence of COPD is well studied, only few studies examined the incidence of COPD in a prospective and standardized manner. In a prospective population-based
In recent years, there has been a marked change in life-style of South Asian countries caused by economic growth, affluence, urbanization and dietary westernization. Few studies on the prevalence of obesity, hypertension and diabetes in the Indian population have been reported. However, there has been scarce literature on the study of prevalence of type 2 diabetes mellitus (DM) and Hypertension in overweight and obese people in India with criteria suggested by World Health Organization (WHO) for Asians. Information on such public health issues would provide evidence based data to develop guidelines and policies on this subject. The aim of this article is to determine the prevalence of hypertension and type 2 DM in overweight and obese people. A cross-sectional study consisted of people selected from the out-patient department and indoors of a large defense hospital in a semi urban area of Assam. Patients with overweight and obesity, reporting for consultation and medical examination were taken into the study. The data collected was analyzed using the criteria for overweight, obesity, diabetes and hypertension defined by WHO, Joint National Committee VII and International Diabetes Federation, American Diabetes Association. A descriptive statistical analysis has been carried out in the study. A total of 300 people were the subject population of this study. Among the subject population, there were 97 overweight and 203 obese. The 56 subjects were found to be diabetic. The prevalence of type 2 DM in overweight subjects was 15.5% and in obese was 20.2% and overall was 18.7%. Prevalence of hypertension in the overweight population was 8.2% and in obese was 22.2% and overall found to be 17.7%. The prevalence of type 2 DM, hypertension in the obese group of the study population were found to be 20.2%, 22.2% and in the overweight population were 15.5% and 8.2%, respectively. This indicates that the prevalence of type 2 DM and hypertension increases with increasing weight of
Full Text Available Context: In recent years, there has been a marked change in life-style of South Asian countries caused by economic growth, affluence, urbanization and dietary westernization. Few studies on the prevalence of obesity, hypertension and diabetes in the Indian population have been reported. However, there has been scarce literature on the study of prevalence of type 2 diabetes mellitus (DM and Hypertension in overweight and obese people in India with criteria suggested by World Health Organization (WHO for Asians. Information on such public health issues would provide evidence based data to develop guidelines and policies on this subject. Aim: The aim of this article is to determine the prevalence of hypertension and type 2 DM in overweight and obese people. Setting and Design: A cross-sectional study consisted of people selected from the out-patient department and indoors of a large defense hospital in a semi urban area of Assam. Materials and Methods: Patients with overweight and obesity, reporting for consultation and medical examination were taken into the study. The data collected was analyzed using the criteria for overweight, obesity, diabetes and hypertension defined by WHO, Joint National Committee VII and International Diabetes Federation, American Diabetes Association. A descriptive statistical analysis has been carried out in the study. Results: A total of 300 people were the subject population of this study. Among the subject population, there were 97 overweight and 203 obese. The 56 subjects were found to be diabetic. The prevalence of type 2 DM in overweight subjects was 15.5% and in obese was 20.2% and overall was 18.7%. Prevalence of hypertension in the overweight population was 8.2% and in obese was 22.2% and overall found to be 17.7%. Conclusions: The prevalence of type 2 DM, hypertension in the obese group of the study population were found to be 20.2%, 22.2% and in the overweight population were 15.5% and 8.2%, respectively
Saedi, Sedigheh; Oraii, Saied; Hajsheikholeslami, Farhad
Recurrent syncope of obscure etiology, particularly in those with structural heart diseases, is associated with higher mortality rates. There are insufficient and conflicting data on prevalence and etiology of syncope in the urban Middle Eastern population. Evaluating the etiologic basis of syncope can be an effective step in prevention of morbidities and sudden cardiac death in susceptible populations. The aim of our study was to determine the prevalence and etiology of syncope in an outpatient cardiology clinic in Tehran the capital of Iran. In this cross sectional study data was collected from patients' records whose first visits were from March 2006 to September 2007 and had undergone thorough examination for syncope (ECG, Holter monitoring, echocardiography, Tilt table testing, Electrophysiological study). All efforts were done to determine underlying heart diseases, physical exam and test abnormalities, final diagnosis and treatment in all selected patients. Overall prevalence of syncope was estimated to be 9%. The age-specific prevalence rates were 5-14 years: 4.14%, 15-44 years: 44.8%, 45-64 years: 31%, 65 years and Older: 20%. The most frequently identified cause (60%) was neurally-mediated (vasovagal) syncope. Mean age of patients was 44.9 years with a minimum of 5 years and maximum of 85 years. In our study, coronary heart disease had a high prevalence among participants (12.4%). Syncope is a common clinical problem. In this study prevalence rates peaked in 15-44 years age group. Considering that recurrent syncope is often disabling and may cause injury and the fact that heart diseases are more common in people affected by syncope, especial cautions should be taken while evaluating this group of patients.
Full Text Available More than 80% cardiovascular disease (CVD is preventable despite the fact that it is currently the ultimate cause of disability in the world. Assessment of the nationwide prevalence of dyslipidemia as a major CVD risk factor is essential to efficiently conduct prevention programs. We extracted data according to the cut-off points of dyslipidemia used in each study. All published papers on this topic in Iranian and international journals with affiliation of "Iran" were reviewed using standard keywords up to September 2011. We included all available population-based studies and national surveys conducted in individuals aged ≥ 15 years. We excluded studies with < 300 individuals, non-population-based studies, or duplicated citations. We analyzed by random effect method due to between-study heterogeneity. The estimated prevalence and 95% confidence intervals in 29 eligible articles and one un-published data for hypercholesterolemia (≥200 mg/dl, hypertriglyceridemia (≥150 mg/dl, high levels of low density lipoprotein cholesterol ([LDL-C] [ ≥ 130 mg/dl] and low levels of high density lipoprotein cholesterol ([HDL-C] <40 mg/dl in males, <50 mg/dl in females, in Iranian people were 41.6% (36.1-47.0, 46.0% (43.3-48.7, 35.5% (24.0-47.1 and 43.9% (33.4-54.4, respectively among both sexes and in both rural and urban areas. Hypercholesterolemia, high LDL-C and low HDL-C were more prevalent in women, whereas hypertriglyceridemia was more prevalent in men. All types of lipid component abnormalities were more prevalent in urban residents. Prevalence of dyslipidemia is considerable in Iran. It is necessary to enforce current measures of dyslipidemia control in the Iranian people to reduce CVD burden.
Sherwin, Justin C; Kelly, John; Hewitt, Alex W; Kearns, Lisa S; Griffiths, Lyn R; Mackey, David A
We aimed to determine the prevalence and associations of refractive error on Norfolk Island. Population-based study on Norfolk Island, South Pacific. All permanent residents on Norfolk Island aged ≥ 15 years were invited to participate. Patients underwent non-cycloplegic autorefraction, slit-lamp biomicroscope examination and biometry assessment. Only phakic eyes were analysed. Prevalence and multivariate associations of refractive error and myopia. There were 677 people (645 right phakic eyes, 648 left phakic eyes) aged ≥ 15 years were included in this study. Mean age of participants was 51.1 (standard deviation 15.7; range 15-81). Three hundred and seventy-six people (55.5%) were female. Adjusted to the 2006 Norfolk Island population, prevalence estimates of refractive error were as follows: myopia (mean spherical equivalent ≥ -1.0 D) 10.1%, hypermetropia (mean spherical equivalent ≥ 1.0 D) 36.6%, and astigmatism 17.7%. Significant independent predictors of myopia in the multivariate model were lower age (P refractive error were increasing age (P prevalence of myopia on Norfolk Island is lower than on mainland Australia, and the Norfolk Island population demonstrates ethnic differences in the prevalence estimates. Given the significant associations between refractive error and several ocular biometry characteristics, Norfolk Island may be a useful population in which to find the genetic basis of refractive error. © 2011 The Authors. Clinical and Experimental Ophthalmology © 2011 Royal Australian and New Zealand College of Ophthalmologists.
Full Text Available Abstract Background Hypospadias is a congenital displacement of the urethral meatus in male newborns, being either an isolated defect at birth or a sign of sexual development disorders. The aim of this study was to assess the prevalence rate of hypospadias in different Districts of Italy, in order to make a comparison with other countries all over the world. Methods We reviewed all the newborns file records (years 2001–2004 in 15 Italian Hospitals. Results We found an overall hypospadias prevalence rate of 3.066 ± 0.99 per 1000 live births (82.48% mild hypospadias, 17.52% moderate-severe. In newborns Small for Gestational Age (birthweight th percentile of any gestational age the prevalence rate of hypospadias was 6.25 per 1000 live births. Performing multivariate logistic regression analysis for different degrees of hypospadias according to severity, being born SGA remained the only risk factor for moderate-severe hypospadias (p = 0.00898 but not for mild forms (p > 0.1. Conclusion In our sample the prevalence of hypospadias results as high as reported in previous European and American studies (3–4 per 1000 live births. Pathogenesis of isolated hypospadias is multifactorial (genetic, endocrine and environmental factors: however, the prevalence rate of hypospadias is higher in infants born small for gestational age than in newborns with normal birth weight.
Varma, Rohit; Torres, Mina; McKean-Cowdin, Roberta; Rong, Fen; Hsu, Chunyi; Jiang, Xuejuan
To estimate the prevalence of refractive errors in adult Chinese Americans, and to evaluate factors associated with myopia and high myopia. A population-based, cross-sectional study. Chinese Americans 50 years and older residing in Monterey Park, California, were recruited. Noncycloplegic automated refraction with supplemental subjective refraction was performed. Myopia, high myopia, hyperopia, and high hyperopia were defined as a spherical equivalent of +0.5 D, and ≥+3.0 D, respectively. Astigmatism and high astigmatism were defined as a cylinder of >0.5 D and >2.25 D, respectively. Risk factor assessment was guided by a conceptual model. Data from 4144 participants were analyzed. The overall prevalence of myopia, high myopia, hyperopia, high hyperopia, astigmatism, and high astigmatism in the right eye was 35.1% (95% confidence interval, 33.6%-36.6%), 7.4% (6.6%-8.3%), 40.2% (38.7%-41.8%), 2.7% (2.2%-3.3%), 45.6% (44.1%-47.2%), and 3.7% (3.1%-4.3%), respectively. The prevalence of myopia and high myopia was lower among older individuals (P refractive errors (P prevalence of refractive errors, except for a higher prevalence of hyperopia among female subjects (P = .010). Age, acculturation, education, income, marital status, birth country, history of ocular disease, nonocular comorbidities, and recent eye examination were associated with prevalence of myopia. All of these factors, except for acculturation, were also associated with high myopia. Our data present the first population-based estimates of the prevalence of refractive errors among adult Chinese Americans. Compared with whites, Hispanics, and blacks, Chinese Americans have a higher burden of myopia, high myopia, and astigmatism. Copyright © 2016 Elsevier Inc. All rights reserved.
Varma, Rohit; Torres, Mina; McKean-Cowdin, Roberta; Rong, Fen; Hsu, Chunyi; Jiang, Xuejuan
Purpose To estimate the prevalence of refractive errors in adult Chinese Americans, and evaluate factors associated with myopia and high myopia. Design A population-based, cross-sectional study. Methods Chinese Americans 50 years and older residing in Monterey Park, California, were recruited. Noncycloplegic automated refraction with supplemental subjective refraction was performed. Myopia, high myopia, hyperopia, and high hyperopia were defined as a spherical equivalent of +0.5D, and ≥ +3.0D, respectively. Astigmatism and high astigmatism were defined as a cylinder of > 0.5D and > 2.25D, respectively. Risk factor assessment was guided by a conceptual model. Results Data from 4144 participants were analyzed. The overall prevalence of myopia, high myopia, hyperopia, high hyperopia, astigmatism, and high astigmatism was 35.1% (95% confidence interval, 33.6%–36.6%), 7.4% (6.6%–8.3%), 40.2% (38.7%–41.8%), 2.7% (2.2%–3.3%), 45.6% (44.1%–47.2%), and 3.7% (3.1%–4.3%), respectively. The prevalence of myopia and high myopia was lower among older individuals (Ps refractive errors (Ps prevalence of refractive errors, except for a higher prevalence of hyperopia among females (P = 0.010). Age, acculturation, education, income, marital status, birth country, history of ocular disease, non-ocular comorbidities, and recent eye exam were associated with prevalence of myopia. All of these factors, except for acculturation, were also associated with high myopia. Conclusions Our data present the first population-based estimates of the prevalence of refractive errors among adult Chinese Americans. Compared with whites, Hispanics, and blacks, Chinese Americans have a higher burden of myopia, high myopia, and astigmatism. PMID:27769895
Hou, Wen-Hsuan; Li, Chung-Yi; Chen, Lu-Hsuan; Wang, Liang-Yi; Kuo, Ken N; Shen, Hsiu-Nien; Chang, Ming-Fong
Population-based data for diabetic hand syndrome (DHS) are limited. The aim of the present epidemiological study was to estimate the overall and cause-specific prevalence and rate ratio (RR) of DHS in patients with diabetes. The present study was a cross-sectional study based on a random sample of 57 093 diabetics and matched controls, both identified from Taiwan National Health Insurance claims in 2010. The DHS analyzed in the present study included carpal tunnel syndrome (CTS), stenosing flexor tenosynovitis (SFT), limited joint mobility (LJM), and Dupuytren's disease (DD). The prevalence of overall DHS was estimated at 2472 per 10 5 for the diabetics, compared with 1641 per 10 5 for the controls, representing a prevalence RR of 1.51 (95 % confidence interval [CI] 1.39-1.64). Stratified analyses further revealed that the significantly increased prevalence of overall DHS was more evident in females than males (1.59 vs 1.36) and was only noted in diabetics aged ≥35 years. Cause-specific analysis suggested that patients with diabetes had the highest prevalence of CTS (1244 per 10 5 ), followed by SFT (1209 per 10 5 ), LJM (39 per 10 5 ), and DD (6 per 10 5 ). In addition, diabetes was only significantly associated with CTS (RR 1.34; 95 % CI 1.20-1.51) and SFT (1.74; 95 % CI 1.54-1.97). The prevalence of overall and certain cause-specific DHS was significantly elevated in patients with diabetes in Taiwan. © 2016 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and John Wiley & Sons Australia, Ltd.
Faghihi, Mohammad; Hashemi, Hassan; Nabovati, Payam; Saatchi, Mohammad; Yekta, Abbasali; Rafati, Shokoofeh; Ostadimoghaddam, Hadi; Khabazkhoob, Mehdi
To determine the prevalence of amblyopia and its determinants in a population-based study in Mashhad County, Iran. This cross-sectional, population-based study was conducted on the population of Mashhad County aged >1 year using randomized stratified cluster sampling. Examinations were performed after selection of the participants and their free transportation to the sampling site. The examinations included the measurement of uncorrected and corrected visual acuity, cycloplegic and non-cycloplegic refraction, cover testing, slit-lamp biomicroscopy, and ophthalmoscopy. In this study, amblyopia was defined as best corrected visual acuity (BCVA) of 20/30 or less or 2-line interocular optotype acuity differences with no pathology. After considering the exclusion criteria, the data of 2739 individuals, 65.6% of whom were women, were analyzed. The mean age of the participants was 29.5±17.5 years. The prevalence of amblyopia was 4.6% (95% CI: 3.77%-5.43%) in the total population. The lowest prevalence was 2.24% in the age group 5-15 years (95% CI: 0.99%-3.48%) and the highest prevalence was 7.14% in the age group 55-65 years (95% CI: 2.64%-11.56%). Anisometropic amblyopia was observed in 45.24% of the amblyopic participants. Isometropic, mixed (strabismic/anisometropic), and strabismic amblyopia were other common causes of amblyopia, with a prevalence of 24.6%, 16.67%, and 13.49% in amblyopic patients, respectively. The odds ratio (OR) of having amblyopia for each 1-year increase in age was 1.02 (95% CI: 1.01-1.03). Amblyopia was less common in people with better socioeconomic status. This study showed the prevalence of amblyopia in all age groups in a population-based study for the first time. The findings of this study regarding the relatively high prevalence of amblyopia in the older population and its lower prevalence in young people indicate attention to amblyopia in recent years.
Full Text Available Background: Intellectual disability (ID is a permanent and highly disabling condition. The birth of a disabled child induces complex feelings in mother and other family members. This study was planned to investigate phenomenology of ID and the prevalence of depression in their mothers. Objective: To find prevalence, influence of various sociodemographic variables, and its clinical correlation with depression in mothers of ID children. Study Design: A cross-sectional study. Materials and Methods: A total of 100 patients diagnosed as ID were included in the study. Objective data were collected in a special Pro forma, and mothers of these patients were evaluated with the Beck's Anxiety Inventory and Beck's Depression Inventory. Results: The mean age of patients with ID was 11.52 years, had received an average of 3.01 years of schooling, mean age at diagnosis was 6.01 years, mean intelligence quotient was 45.17, and 79% had significant comorbidities. The prevalence of depression in mothers was 79%; it was more in mothers of female ID child, ID child with significant comorbidities, severer forms of retardation, and with higher levels of anxiety in the mother. Conclusions: The prevalence of depression in mothers of ID children in the present study seems to be much greater than those reported from the previous studies. The determination of predictors of depression among mothers of ID children may help health professionals in identifying mothers at risk. Regular screening of mothers of ID children should be included in the protocol for management.
Full Text Available Background: Wushu is one of the martial arts that combines explosive strength and speed movements with combat techniques. Most studies to date of wushurelated injuries have been published in Chinese languages. No published studies have reported the prevalence of these injuries in Iran. Therefore, the aim of the present study was to determine the prevalence of musculoskeletal injuries in male wushu players in Shiraz. Methods: Male wushu athletes aged 18 to 30 years were included in this cross sectional study if they had been practicing for 2 hours per session, twice a week during the past 3 years. The athletes were recruited by convenience sampling from 30 wushu clubs in Shiraz, Iran. The sample size was 165. The study was conducted from June to September 2016. Each participant was asked to complete an information questionnaire about his musculoskeletal injuries related to wushu during the previous year. Results: About two third (65.45% of wushu players had one or more injuries during the previous year. Severe injuries were most commonly located in the head/neck and knee/tibia areas, and inflammation was more prevalent than the other types of injuries. Conclusion: Wushu players in this study reported a high prevalence of injuries affecting different parts of the body.
Rodgers William H
Full Text Available Abstract Context. - Myeloid sarcoma (MS is a neoplasm of immature granulocytes, monocytes, or both involving any extramedullary site. The correct diagnosis of MS is important for adequate therapy, which is often delayed because of a high misdiagnosis rate. Objective. - To evaluate the lineage differentiation of neoplastic cells in MS by immunohistochemistry, and to correlate the results with clinicopathologic findings and cytogenetic studies. Design. - Histologic and immunohistochemical examinations were performed on formalin-fixed paraffin-embedded tissue samples from 13 cases of MS. They were classified according to the World Health Organization criteria. Chromosomal analysis data were available in 11 cases. Clinical, pathological, and cytogenetic findings were analyzed. Results. - The study included six male and seven female patients with an age range of 25 to 72 years (mean, 49.3 years and a male to female ratio of 1:1.2. MS de novo occurred in 4/13 (31% of cases examined. The most sensitive immunohistochemical markers were CD43 and lysozyme present in all cases with MS (13/13, 100%. All de novo MS showed a normal karyotype, monoblastic differentiation, and lack of CD34. The most common chromosomal abnormalities in MS associated with a hematopoietic disorder were trisomy 8 and inv(16 (2/11, 18%. Conclusion. - An immunohistochemical panel including CD43, lysozyme, myeloperoxidase (MPO, CD68 (or CD163, CD117, CD3 and CD20 can successfully identify the vast majority of MS variants in formalin-fixed paraffin-embedded tissue sections. The present report expands the spectrum of our knowledge showing that de novo MS has frequent monoblastic differentiation and frequently carries a normal karyotype.
Nesamony Maneksh Kumar
Full Text Available BACKGROUND Obesity is defined as having excess body fat due to ‘calorie imbalance’ -too few calories expended for the amount of calories consumed. Childhood obesity has both immediate and long-term effects on health and well-being. This study is aimed to highlight the burden of obesity among school children. The objectives of this study are to find the prevalence of obesity among school children aged 11-15 years and to compare prevalence of obesity among school children of government and private schools. MATERIALS AND METHODS School-based cross-sectional study conducted in 2 private and 2 government schools of a rural area in Trivandrum district. Total 800 students (200 from each school aged 11-15 years in 6 th to 10 th standards were selected by systematic random sampling and consent were obtained. Height (cm and weight (kg were measured using stadiometer and standardised weighing machine, respectively. Body Mass Index (BMI was calculated and categorised as underweight, normal, overweight and obese. The prevalence is expressed in percentages. RESULTS In this study, the total prevalence of overweight and obesity among the total 800 school children is found to be 4% and 1%, respectively. An alarming incidental finding is that the underweight prevalence is 61%, which is significantly high and should be addressed. The overall prevalence of overweight and obesity is 4.25% and 1.75%, respectively in private schools, whereas it is 3.5% and 1%, respectively, in government schools. There is not a significant difference between private and government school children. This may be due to the awareness about problems of obesity and its effects among school children and also timely health checkups in the schools. CONCLUSION This study done in a rural area found that there is a significant prevalence of obesity and overweight as well as a very high prevalence of underweight. It indicates that childhood obesity is an emerging health problem in rural areas
Ambika Gopalakrishnan Unnikrishnan
Full Text Available Background: Hypothyroidism is believed to be a common health issue in India, as it is worldwide. However, there is a paucity of data on the prevalence of hypothyroidism in adult population of India. Materials and Methods: A cross-sectional, multi-centre, epidemiological study was conducted in eight major cities (Bangalore, Chennai, Delhi, Goa, Mumbai, Hyderabad, Ahmedabad and Kolkata of India to study the prevalence of hypothyroidism among adult population. Thyroid abnormalities were diagnosed on the basis of laboratory results (serum FT3, FT4 and Thyroid Stimulating Hormone [TSH]. Patients with history of hypothyroidism and receiving levothyroxine therapy or those with serum free T4 5.50 μU/ml, were categorized as hypothyroid. The prevalence of self reported and undetected hypothyroidism, and anti-thyroid peroxidase (anti-TPO antibody positivity was assessed. Results: A total of 5376 adult male or non-pregnant female participants ³18 years of age were enrolled, of which 5360 (mean age: 46 ± 14.68 years; 53.70% females were evaluated. The overall prevalence of hypothyroidism was 10.95% (n = 587, 95% CI, 10.11-11.78 of which 7.48% (n = 401 patients self reported the condition, whereas 3.47% (n = 186 were previously undetected. Inland cities showed a higher prevalence of hypothyroidism as compared to coastal cities. A significantly higher ( P 5.50 μIU/ml. Anti - TPO antibodies suggesting autoimmunity were detected in 21.85% (n = 1171 patients. Conclusion: The prevalence of hypothyroidism was high, affecting approximately one in 10 adults in the study population. Female gender and older age were found to have significant association with hypothyroidism. Subclinical hypothyroidism and anti-TPO antibody positivity were the other common observations.
Yoo, Jun-Il; Ha, Yong-Chan; Lee, Young-Kyun; Hana-Choi; Yoo, Moon-Jib; Koo, Kyung-Hoi
Alcohol consumption is considered a risk factor for sarcopenia, but the association between alcohol consumption and the prevalence of sarcopenia has not been evaluated in detail. This study was to identify the relationship between alcohol drinking patterns and the prevalence of sarcopenia in the elderly Korean population. The cross-sectional study was performed using data from the Korea National Health and Nutrition Examination Survey. Participants were excluded if they were under the age of 65, or if data was not available regarding skeletal muscle mass or dietary intake. After these exclusions, a total of 4020 participants (men: 1698; women: 2322) were analyzed in the present study. Sarcopenia is defined according to the criteria for the Asia Working Group for Sarcopenia (AWGS). Binge drinking was defined as consuming ≥5 standard alcoholic drinks (≥4 drinks for women) consecutively on one occasion. This data was subcategorized into two groups based on presence of binge drinking: Social drinking (≤1 time/month) and binge drinking (>1 time/month). Women binge drinkers with weekly or daily consumption had 2.8 times higher prevalence of sarcopenia than social drinkers (Odds Ratio [OR] = 2.84; 95% Confidence Interval [CI] = 1.12-7.29). However, there were no associations between binge drinkers and sarcopenia in men. After adjusting for age, body mass index (BMI), energy intake, moderate physical activity, and energy intake, women binge drinkers with weekly or daily alcohol consumption had 3.9 times higher prevalence of sarcopenia than social drinkers (OR = 3.88; 95% CI = 1.33-11.36). The prevalence of sarcopenia in elderly women was related to binge drinking frequency and amounts of drinking after adjusting for covariates. Elderly Korean women who binge drink once or more per week may be associated with sarcopenia, as seen with the observed 3.9 times higher prevalence compared to social drinkers.
Bowles Heather R
Full Text Available Abstract Background Physical activity (PA is one of the most important factors for improving population health, but no standardised systems exist for international surveillance. The International Physical Activity Questionnaire (IPAQ was developed for international surveillance. The purpose of this study was a comparative international study of population physical activity prevalence across 20 countries. Methods Between 2002–2004, a standardised protocol using IPAQ was used to assess PA participation in 20 countries [total N = 52,746, aged 18–65 years]. The median survey response rate was 61%. Physical activity levels were categorised as "low", "moderate" and "high". Age-adjusted prevalence estimates are presented by sex. Results The prevalence of "high PA" varied from 21–63%; in eight countries high PA was reported for over half of the adult population. The prevalence of "low PA" varied from 9% to 43%. Males more frequently reported high PA than females in 17 of 20 countries. The prevalence of low PA ranged from 7–41% among males, and 6–49% among females. Gender differences were noted, especially for younger adults, with males more active than females in most countries. Markedly lower physical activity prevalence (10% difference with increasing age was noted in 11 of 19 countries for males, but only in three countries for women. The ways populations accumulated PA differed, with some reporting mostly vigorous intensity activities and others mostly walking. Conclusion This study demonstrated the feasibility of international PA surveillance, and showed that IPAQ is an acceptable surveillance instrument, at least within countries. If assessment methods are used consistently over time, trend data will inform countries about the success of their efforts to promote physical activity.
Ashina, Sait; Bendtsen, Lars; Lyngberg, Ann C; Lipton, Richard B; Hajiyeva, Nazrin; Jensen, Rigmor
We assessed the prevalence of neck pain in the population in relation to headache. In a cross-sectional study, a total of 797 individuals completed a headache interview and provided self-reported data on neck pain. We identified migraine, TTH or both migraine and TTH (M+TTH) groups. Pericranial tenderness was recorded in 496 individuals. A total tenderness score (TTS) was calculated as the sum of local scores with a maximum score of 48. The one-year prevalence of neck pain was 68.4% and higher in those with vs. without primary headache (85.7% vs. 56.7%; adjusted OR 3.0, 95% CI 2.0-4.4, pneck pain (56.7%) was significantly higher in those with M+TTH (89.3%), pure TTH (88.4%) and pure migraine (76.2%) (pneck pain had higher TTS than individuals without neck pain (15.1±10.5 vs. 8.4±8.0, pNeck pain is highly prevalent in the general population and even more prevalent in individuals with primary headaches. Prevalence is highest in coexistent M+TTH, followed by pure TTH and migraine. Myofascial tenderness is significantly increased in individuals with neck pain. © International Headache Society 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.
Mishra, Apurva; Pandey, Ramesh K
To determine the prevalence of molar incisor hypomineralization (MIH) in Indian children and to analyze the possible etiological factors. First permanent molars and all permanent incisors were examined in 1,369 children aged 8 to 12 years. Examinations were performed by two calibrated observers. The subjects were evaluated using judgment criteria proposed by Weerheijm et al in 2003. The parents accompanying children were given a questionnaire regarding pre- and postnatal history of the children. A total of 191 children were diagnosed with MIH with a prevalence of 13.9%. Chi-square/Fisher exact test was used to compare the dichotomous variables. The relative risk with its 95% confidence interval was calculated to find the risk of clinical infections, such as chicken pox, jaundice, renal disorders, cardiac disorders, and affected molars with sex and type of delivery. Pre- and postnatal history of infection in a child was significantly correlated with the prevalence of MIH. The prevalence of MIH was 13.9% in the age group of 8 to 12 years. Prenatal and postnatal infections play an important role in hypomineralization of molars and incisors. How to cite this article: Mishra A, Pandey RK. Molar Incisor Hypomineralization: An Epidemiological Study with Prevalence and Etiological Factors in Indian Pediatric Population. Int J Clin Pediatr Dent 2016;9(2):167-171.
May, O; Arildsen, H; Damsgaard, E M
OBJECTIVES: The aim of the study was to estimate the prevalence of silent ischaemia in diabetic subjects in the population, to compare the prevalence of silent ischaemia in diabetics and non-diabetics and to attempt to predict the presence of silent ischaemia in diabetic subjects. METHODS: A random...... was registered simultaneously with ECG evidence of ischaemia. Individuals with ischaemia, but without angina pectoris, were defined as persons with silent ischaemia. RESULTS: Seventy-four percent of the invited group were included. The observed prevalence of silent ischaemia in diabetics was 13.5% (95% CI = 8.......5-19.8%). No association was found between silent ischaemia and gender (P = 0.83) or diabetes type (P = 0.67). In the group of diabetics who had controls, the prevalence was 11.4%, and among the controls the prevalence was 6.4% (OR = 1.87, one-sided P = 0.079). Systolic blood pressure was highly predictive of silent...
Roux, Anne-Laure; Catherinot, Emilie; Ripoll, Fabienne; Soismier, Nathalie; Macheras, Edouard; Ravilly, Sophie; Bellis, Gil; Vibet, Marie-Anne; Le Roux, Evelyne; Lemonnier, Lydie; Gutierrez, Cristina; Vincent, Véronique; Fauroux, Brigitte; Rottman, Martin; Guillemot, Didier; Gaillard, Jean-Louis
We performed a multicenter prevalence study of nontuberculous mycobacteria (NTM) involving 1,582 patients (mean age, 18.9 years; male/female ratio, 1.06) with cystic fibrosis in France. The overall NTM prevalence (percentage of patients with at least one positive culture) was 6.6% (104/1,582 patients), with prevalences ranging from 3.7% (in the east of France) to 9.6% (in the greater Paris area). Mycobacterium abscessus complex (MABSC; 50 patients) and Mycobacterium avium complex (MAC; 23 patients) species were the most common NTM, and the only ones associated with fulfillment of the American Thoracic Society bacteriological criteria for NTM lung disease. The “new” species, Mycobacterium bolletii and Mycobacterium massiliense, accounted for 40% of MABSC isolates. MABSC species were isolated at all ages, with a prevalence peak between 11 and 15 years of age (5.8%), while MAC species reached their highest prevalence value among patients over 25 years of age (2.2%). PMID:19846643
Handler, Marc Z; Derrick, Kristina M; Lutz, Richard E; Morrell, Dean S; Davenport, Marsha L; Armstrong, April W
The absence of data on the prevalence of pilomatricoma among patients with Turner syndrome served as the catalyst for this multicenter investigation. To ascertain the prevalence of pilomatricoma among patients with Turner syndrome and to determine any association between the development of pilomatricomas and the use of exogenous hormones in patients with Turner syndrome. A retrospective medical record review from January 1, 2000, through January 1, 2010, was performed of all patients with Turner syndrome. Data on pilomatricomas and the use of hormone therapy were collected. University of California-Davis Medical Center, University of Nebraska Medical Center, and The University of North Carolina at Chapel Hill. Patients with a diagnosis of Turner syndrome. Prevalence of concomitant pilomatricoma and diagnosis of Turner syndrome. Secondary outcome measures included the use of the exogenous hormones estrogen or recombinant human growth hormone (rhGH). In total, 311 patients with Turner syndrome were identified from these 3 institutions. Among them, 8 patients (2.6%) were diagnosed as having pilomatricomas. Before the development of pilomatricomas, 5 patients had been treated with rhGH but not estrogen, 1 patient had received estrogen but not rhGH, and 2 patients did not receive either therapy. Although the prevalence of pilomatricoma among the general population is unknown, this study demonstrates a high prevalence (2.6%) of pilomatricomas among patients with Turner syndrome. No apparent relationship was noted among our patients or in the literature between the use of rhGH and the development of pilomatricomas.
Grammatikopoulou, Maria G; Pritsa, Agathi A; Badeka, Sophia; Aggelaki, Irini; Giantsiou, Ioanna; Houta, Assimina; Zeibekoglou, Vasiliki; Kyriazi, Maria; Papanastasiou, Polykseni; Perdiki, Eleni; Gkipatidou, Despoina; Tsigga, Maria
To report a pilot prevalence of maternal overweight, obesity and underweight in selected Greek counties. A total of 441 adult childbearing women were recruited from maternity clinics in 6 Greek counties for this cross-sectional study. Pre-gravid weight status was defined according to the WHO cut-offs and gravid weight status was diagnosed with the Mardones and Rosso weight gain chart. During gestation the majority of the participants were of normal body weight (BW) (34.0%), obesity was apparent in 25.6% of the sample, 23.8% of the participants were underweight, and the remaining 16.6% were overweight. Overall, pregnancy tripled the prevalence of underweight, increased the prevalence of obesity (by 388.0%) and decreased the number of participants in the normal BW category (p≤0.001 for all). The majority of participants classified in each pre-gravid weight-category remained in the same weight category during their gestation. All the pre-gravidly obese women were also obese during pregnancy. Underweight was more prevalent in Kavala (37.5%) and obesity was more frequent in Thessaloniki (30.8%). Women who were overweight prior to conception were highly likely to be overweight/obese during pregnancy (OR: 23.8, CI: 11.1-51.0). The results indicate a high prevalence of overweight, obesity and underweight among pregnant women in Greece. Copyright © 2012 SEEN. Published by Elsevier Espana. All rights reserved.
Full Text Available Multimorbidity is a major concern in primary care. Nevertheless, evidence of prevalence and patterns of multimorbidity, and their determinants, are scarce. The aim of this study is to systematically review studies of the prevalence, patterns and determinants of multimorbidity in primary care.Systematic review of literature published between 1961 and 2013 and indexed in Ovid (CINAHL, PsychINFO, Medline and Embase and Web of Knowledge. Studies were selected according to eligibility criteria of addressing prevalence, determinants, and patterns of multimorbidity and using a pretested proforma in primary care. The quality and risk of bias were assessed using STROBE criteria. Two researchers assessed the eligibility of studies for inclusion (Kappa= 0.86.We identified 39 eligible publications describing studies that included a total of 70,057,611 patients in 12 countries. The number of health conditions analysed per study ranged from 5 to 335, with multimorbidity prevalence ranging from 12.9% to 95.1%. All studies observed a significant positive association between multimorbidity and age (odds ratio [OR], 1.26 to 227.46, and lower socioeconomic status (OR, 1.20 to 1.91. Positive associations with female gender and mental disorders were also observed. The most frequent patterns of multimorbidity included osteoarthritis together with cardiovascular and/or metabolic conditions.Well-established determinants of multimorbidity include age, lower socioeconomic status and gender. The most prevalent conditions shape the patterns of multimorbidity. However, the limitations of the current evidence base means that further and better designed studies are needed to inform policy, research and clinical practice, with the goal of improving health-related quality of life for patients with multimorbidity. Standardization of the definition and assessment of multimorbidity is essential in order to better understand this phenomenon, and is a necessary immediate step.
Zagdańska, Marta; Kiejna, Andrzej
The aim of this study was to identify risk factors and estimate prevalence of depressive episodes among Wroclaw's universities students. Polish adaptation of the Composite International Diagnostic Interview (CIDI) was implemented to gather epidemiological data from 370 students of public universities in Wroclaw. Proportional stratified sampling was performed to obtain distinct, independent strata representing sex, year of study and educational profiles. Randomisation was ensured by recruitment procedures. Prevalence of depressive episodes among students in Wroclaw is high - 14.7% throughout life, 9.8% within 12 months prior to the interview. High prevalence of severe and moderate depressive episodes seems to be worrisome (respectively 5.1% and 6.6% throughout life, 3.8% and 3.9% within 12 months prior to the study). Year of study, profile and lack of partner relationship remain risk factor for depression. High prevalence of depressive episodes indicates the need for prevention and therapy based on epidemiological data and tailored to the students' needs. Depression among students requires further epidemiological studies.
Firth, Jill; Nelson, Elizabeth Andrea; Hale, Claire; Hill, Jacqueline; Helliwell, Philip
The aim of this review was to examine design and reporting issues that affect prevalence estimates of leg ulceration obtained using self-report and outline strategies to strengthen the validity and reliability of research in this area. We identified leg ulcer prevalence studies and evaluated them against the STROBE (Strengthening the Reporting of Observational Studies in Epidemiology) statement. The authors draw upon the wider literature and their own empirical work in discussing strategies to overcome design and reporting issues. Common deficiencies in the design and reporting of studies include wide variations in diagnostic criteria and age parameters for participant selection, a lack of description of efforts to address bias/study size rationale, and low participation rates in clinical examination stages. These factors and differences in statistical methods of analysis affect the validity and reliability of findings and hinder interpretation, making comparisons across populations difficult. Opportunities for subgroup analyses are frequently missed. Self-report is a valuable means of capturing leg ulcer prevalence, but the future design and reporting of studies need to be strengthened, including addressing weaknesses in validation strategies. Capture-recapture analysis or a multiple-methods approach has the potential to yield the most valid and reliable prevalence estimates. Copyright (c) 2010 Elsevier Inc. All rights reserved.
Maya Ramesh; R Shankar; Ramesh Krishnan; Narasimhan Malathi; Rita Mary Aruna
Background: Tamil Nadu is one of the 18 states affected by fluorosis in India. The maximum tolerance limit of fluoride in drinking water specified by the World Health Organization (WHO, 1984) is 1.5 mg/l while it is proved to be above in many areas of Salem. Aims and Objectives: The aim of the following study is to identify the prevalence of dental fluorosis among children in the district of Salem. To compare the prevalence of dental fluorosis between different sexes and age groups. Materials...
Friberg, Jens; Scharling, Henrik; Gadsbøll, Niels
Atrial fibrillation (AF) is the most frequently encountered cardiac arrhythmia. It is a risk factor for stroke and premature death. We studied the temporal changes in the prevalence of AF from 1976 to 1994 in a random population aged 50 to 89 years. The prevalence of AF, diagnosed from electrocar.......4% in 1976 to 1978 (odds ratio [OR] 1.0, reference) to 1.9% in 1981 to 1983 (OR 1.6, 95% confidence interval [CI] 1.1 to 2.1), and to 3.3% in 1991 to 1994 (OR 2.3, 95% CI 1.6 to 3.4, p...
Thapa, Suman S; Thapa, Raba; Paudyal, Indira; Khanal, Shankar; Aujla, Jaskirat; Paudyal, Govinda; Rens, Ger van
Background Vitreo-retinal diseases are among the leading causes of visual impairment and blindness worldwide. This study reports the prevalence and pattern of vitreo-retinal diseases in the Bhaktapur Glaucoma Study (BGS), a population based study conducted in Nepal. Methods BGS was a population based cross-sectional study involving 4800 subjects aged 40 years and over from Bhaktapur district. Subjects were selected using a cluster sampling methodology and a door-to-door enumeration. All subje...
Wong, G W K; Mahesh, P A; Ogorodova, L; Leung, T F; Fedorova, O; Holla, A D; Fernandez-Rivas, M; Clare Mills, E N; Kummeling, I; van Ree, R; Yazdanbakhsh, M; Burney, P
Very little is known regarding the global variations in the prevalence of food allergies. The EuroPrevall-INCO project has been developed to evaluate the prevalence of food allergies in China, India and Russia using the standardized methodology of the EuroPrevall protocol used for studies in the European Union. The epidemiological surveys of the project were designed to estimate variations in the prevalence of food allergy and exposure to known or suspected risk factors for food allergy and to compare the data with different European countries. Random samples of primary schoolchildren were recruited from urban and rural regions of China, Russia and India for screening to ascertain possible adverse reactions to foods. Cases and controls were then selected to answer a detailed questionnaire designed to evaluate the possible risk factors of food allergies. Objective evidence of sensitisation including skin-prick test and serum specific IgE measurement was also collected. More than 37 000 children from the three participating countries have been screened. The response rates for the screening phase ranged from 83% to 95%. More than 3000 cases and controls were studied in the second phase of the study. Further confirmation of food allergies by double blind food challenge was conducted. This will be the first comparative study of the epidemiology of food allergies in China, India, and Russia using the same standardized methodology. The findings of these surveys will complement the data obtained from Europe and provide insights into the development of food allergy.
Full Text Available Obesity is one of the main public health problems which underlie many chronic illnesses and socioeconomic difficulties. According to the literature review, there are limited data on the prevalence of obesity in different parts of Iran as well as its trend and prevalence among different age and gender groups. The aim of this study was to estimate the obesity prevalence in Iran using meta-analysis. All the corresponding articles published in the external and internal journals, final reports of research projects, articles of related congresses and the reference index of the correlated papers published between 1995 and 2010 were collected via the electronic research engines (PubMed, Scopus, SID, Magiran, IranMedex. Data were analyzed using meta-analysis (random effects model and meta-regression. A total of 144 articles with the sample size of 377858 people (134588 males and 164858 females were enrolled in the study. The prevalence of obesity in populations above the age of 18 was estimated as 21.7% (CI 95%: 18.5% - 25% and in populations below 18 as 6.1% (CI 95%: 6.8%-5.4%. Meta-regression analysis showed an ascending trend in the prevalence of obesity in Iran. The prevalence rates of obesity according to the BMI index, NCHC and percentile above 95 were 17.4%, 7.6% and7.4%, respectively. The BMI mean was 19.3 in populations below the age of 18 (CI 95%: 17-21.6 and 25.2 in those above the age of 18 (CI 95%: 27.1-23.3. Considering the increasing rate of obesity in Iran and its effects on the public health, corresponding health authorities should revise the obesity preventive programs and, using public health interventions, reduce the rate of obesity in the country.
Maio, Sara; Baldacci, Sandra; Carrozzi, Laura; Pistelli, Francesco; Angino, Anna; Simoni, Marzia; Sarno, Giuseppe; Cerrai, Sonia; Martini, Franca; Fresta, Martina; Silvi, Patrizia; Di Pede, Francesco; Guerriero, Massimo; Viegi, Giovanni
Few epidemiological surveys on general population samples estimated changes in prevalence of respiratory symptoms/diseases over a long time interval; our study aims to quantify the temporal changes in the prevalence rates of asthma, allergic rhinitis and Chronic Obstructive Pulmonary Disease (COPD) after 25 years from baseline. A general population sample participated in 3 cross-sectional surveys carried out in Central Italy (Pisa) in 1985-88 (n = 3865), 1991-93 (n = 2841), 2009-11 (n = 1620). 2276 (47%) subjects participated in at least 1 survey, 1723 (35.5%) in at least 2 surveys and 849 (17.5%) in all the 3 surveys. All subjects filled in a standardized questionnaire about health status and risk factors; a sub-sample performed spirometry. Chi-square test was used to compare adjusted prevalence rates of respiratory symptoms/diseases and descriptive characteristics among the surveys. Generalised estimating equations (GEE) were used to analyze the association between respiratory symptoms/diseases and risk factors. There was an increasing trend in prevalence rates of all respiratory symptoms/diseases throughout the surveys: current asthma attacks (1st-3rd survey prevalence: 3.4-7.2%), allergic rhinitis (16.2-37.4%), usual phlegm (8.7-19.5%) and COPD (2.1-6.8%) more than doubled. The GEE model confirmed these increasing trends, indicating higher risk of having respiratory symptoms/diseases in the second and third surveys. While asthma and allergic rhinitis increasing trends were confirmed, with respect to other international studies, also a COPD increasing prevalence rates was shown. Copyright © 2015 Elsevier Ltd. All rights reserved.
Conclusion: The present study reflects the high prevalence rates of tobacco use among various sections of society. Apart from mass against tobacco products, authorities should focus on strict enforcement of law and promote research for creating new and harmless replacement products.
Joylene Diana; Anjum; Sujaya V
Dysmenorrhea or pain during menstruation is the most common gynaecological complaint among adolescents and college students. A good majority of patients with dysmenorrhea also experience pre - menstrual symptoms ( PMS ) . The objective of this cross s ectional study was to assess prevalence of dysmennorhea , PMS , college absenteeism and ...
Background: Obesity is socially and culturally acceptable in Nigeria and therefore not usually recognized as a medical problem. This study is aimed at determining the prevalence of obesity using body mass index(BMI) criterion and its common primary comorbidities among adult Nigerians attending a semiurban Mission ...
Dydensborg, Stine; Toftedal, Peter; Biaggi, Matteo
Aim: To determine the prevalence and incidence of diagnosed coeliac disease (CD) in Danish children and adolescents and to describe trends over time. Methods: All children with a CD diagnosis registered in the Danish National Patient Registry (DNPR) were included in the study. Data were validated...
Impacted teeth are commonly observed in Dental clinics all over the country. However. people may live with impacted teeth to old age without problems. This study aimed at determining the prevalence and the pattern of the impacted third molars among patients seen in Dar es Salaam. A convenient sample of 1198 patients ...
Ursum, J.; Korevaar, J.C.; Twisk, J.W.R.; Peters, M.J.L.; Schellevis, F.G.; Nurmohamed, M.T.; Nielen, M.M.J.
Objective. Little is known about the presence of chronic morbidity in inflammatory arthritis (IA) patients at disease onset. Previous studies have been mainly performed in established IA patients or they focus on isolated co-morbid diseases. Our aim was to determine the prevalence of chronic
Johansson, S G O; Florvaag, E; Oman, H
The aim of this study was to test, on a multinational level, the pholcodine (PHO) hypothesis, i.e. that the consumption of PHO-containing cough mixtures could cause higher prevalence of IgE antibodies to PHO, morphine (MOR) and suxamethonium (SUX). As a consequence the risk of anaphylaxis...
Ali, Solomon; Haileamlak, Abraham; Wieser, Andreas; Pritsch, Michael; Heinrich, Norbert; Loscher, Thomas; Hoelscher, Michael; Rachow, Andrea
Tuberculosis (TB) is one of the major health problems in prisons. This study was done to assess the prevalence and determinants of active tuberculosis in Ethiopian prisons. A cross-sectional study was conducted from January 2013 to December 2013 in 13 zonal prisons. All incarcerated inmates underwent TB symptom screening according to WHO criteria. From identified TB-suspects two sputum samples were analyzed using smear microscopy and solid culture. A standardized questionnaire assessing TB risk factors was completed for each TB suspect. 765 (4.9%) TB suspects were identified among 15,495 inmates. 51 suspects were already on anti-TB treatment (6.67%) and 20 (2.8%) new culture-confirmed TB cases were identified in the study, resulting in an overall TB prevalence of 458.1/100,000 (95%CI: 350-560/100,000). Risk factors for active TB were alcohol consumption, contact with a TB case before incarceration and no window in prison cell. HIV prevalence was not different between TB suspects and active TB cases. Further, the TB burden in prisons increased with advancing distance from the capital Addis Ababa. The overall TB prevalence in Ethiopian prisons was high and extremely variable among different prisons. TB risk factors related to conditions of prison facilities and the impact of implemented TB control measures need to be further studied in order to improve TB control among inmates.
Godefrooij, Daniel A|info:eu-repo/dai/nl/413648990; de Wit, G Ardine|info:eu-repo/dai/nl/167546589; Uiterwaal, Cuno S|info:eu-repo/dai/nl/136603947; Imhof, Saskia M|info:eu-repo/dai/nl/151386439; Wisse, Robert P L|info:eu-repo/dai/nl/344481336
PURPOSE: To determine the age-specific incidence and prevalence of keratoconus in the modern era of diagnostics. DESIGN: Epidemiologic study. PARTICIPANTS: 4.4 million patients from a mandatory health insurance database. METHODS: Data were extracted from the largest health insurance provider in the
Objective: The study was carried out to assess the oral hygiene status and to determine the prevalence of gingival and periodontal diseases in 10-12-year-old school children in Sholapur City, India. Method: A total of 1045 children (560 boys and 485 girls) aged 10-12 years were evaluated by questionnaires related to oral ...
S. Dahaghin; S.M. Bierma-Zeinstra (Sita); A.Z. Ginai (Abida); H.A.P. Pols (Huib); J.M.W. Hazes (Mieke); B.W. Koes (Bart)
textabstractOBJECTIVE: To investigate the prevalence and pattern of radiographic osteoarthritis (ROA) of the hand joints and its association with self reported hand pain and disability. METHODS: Baseline data on a population based study (age >/=55 years) were used (n = 3906). Hand
Jongenelis, K; Eisses, AMH; Beekman, ATF; Kluiter, H; Ribbe, MW
Background: Depression is a common and disabling psychiatric disorder in later life. Particular frail nursing home patients seem to be at increased risk. Nursing home-based studies on risk indicators of depression are scarce. Methods: Prevalence and risk indicators of depression were assessed in 333
Dehoust, Maria Christina; Schulz, Holger; Härter, Martin; Volkert, Jana; Sehner, Susanne; Drabik, Anna; Wegscheider, Karl; Canuto, Alessandra; Weber, Kerstin; Crawford, Mike; Quirk, Alan; Grassi, Luigi; DaRonch, Chiara; Munoz, Manuel; Ausin, Berta; Santos-Olmo, Anna; Shalev, Arieh; Rotenstein, Ora; Hershkowitz, Yael; Strehle, Jens; Wittchen, Hans-Ulrich; Andreas, Sylke
Somatoform disorders have rarely been addressed in epidemiological and health care services studies of the elderly. The few existing studies vary considerably in their methodologies limiting comparability of findings. Data come from the MentDis_ICF65+ study, in which a total of 3142 community-dwelling respondents aged 65-84 years from six different countries were assessed by the Composite International Diagnostic Interview adapted to the needs of the elderly (CIDI65+). The 12-month prevalence rate for any somatoform disorders was found to be 3.8, whereby the prevalence for somatization disorder according to DSM-IV was 0%, the prevalence for abridged somatization was 1.7% and the rate for 12-months somatoform pain disorder was 2.6%. We found a significant variation by study centre (p < 0.005). There was a significant gender difference for pain disorder, but not for abridged somatization. Significant age-related effects revealed for both disorder groups. Somatoform disorders were found to be associated with other mental disorders [odds ratio (OR) anxiety =4.8, OR affective disorders 3.6], as well as with several impairments and disabilities. Somatoform disorders are prevalent, highly impairing conditions in older adults, which are often associated with other mental disorders and should receive more research and clinical attention. Copyright © 2017 John Wiley & Sons, Ltd.
Yu, J.; Li, J.; Cuijpers, P.; Wu, S.; Wu, Z.
Objective In China, the rapid socioeconomic transition and the consequential traditional culture change had significant influences on Chinese older-adult depression. In the present study, the prevalence, the potential risk, and the protective factors of depression in the Chinese older population
Fernández-Rivas, Montserrat; Barreales, L.; Mackie, A. R.; Fritsche, P.; Vázquez-Cortés, S.; Jedrzejczak-Czechowicz, M.; Kowalski, M. L.; Clausen, M.; Gislason, D.; Sinaniotis, A.; Kompoti, E.; Le, T. M.; Knulst, A. C.; Purohit, A.; De Blay, F.; Kralimarkova, T.; Popov, T.; Asero, R.; Belohlavkova, S.; Seneviratne, S. L.; Dubakiene, R.; Lidholm, J.; Hoffmann-Sommergruber, K.; Burney, P.; Crevel, R.; Brill, M.; Fernández-Pérez, C.; Vieths, S.; Clare Mills, E. N.; Van Ree, R.; Ballmer-Weber, B. K.
Background The EuroPrevall project aimed to develop effective management strategies in food allergy through a suite of interconnected studies and a multidisciplinary integrated approach. To address some of the gaps in food allergy diagnosis, allergen risk management and socio-economic impact and to
Fernández-Rivas, M.; Barreales, L.; Mackie, A. R.; Fritsche, P.; Vázquez-Cortés, S.; Jedrzejczak-Czechowicz, M.; Kowalski, M. L.; Clausen, M.; Gislason, D.; Sinaniotis, A.; Kompoti, E.; Le, T.-M.; Knulst, A. C.; Purohit, A.; de Blay, F.; Kralimarkova, T.; Popov, T.; Asero, R.; Belohlavkova, S.; Seneviratne, S. L.; Dubakiene, R.; Lidholm, J.; Hoffmann-Sommergruber, K.; Burney, P.; Crevel, R.; Brill, M.; Fernández-Pérez, C.; Vieths, S.; Clare Mills, E. N.; van Ree, R.; Ballmer-Weber, B. K.
The EuroPrevall project aimed to develop effective management strategies in food allergy through a suite of interconnected studies and a multidisciplinary integrated approach. To address some of the gaps in food allergy diagnosis, allergen risk management and socio-economic impact and to complement
Kerimova, Melek; Gunuc, Selim
The purpose of the present paper was to adapt Gunuc and Kayri's (2010) "Internet Addiction Scale," with show validity and reliability for many various sampling groups, into the Azerbaijani language. Another objective of the study is to determine the prevalence of Internet addiction among Azerbaijani adolescents and youth, which…
Maulik, Pallab K.; Mascarenhas, Maya N.; Mathers, Colin D.; Dua, Tarun; Saxena, Shekhar
Intellectual disability is an extremely stigmatizing condition and involves utilization of large public health resources, but most data about its burden is based on studies conducted in developed countries. The aim of this meta-analysis was to collate data from published literature and estimate the prevalence of intellectual disability across all…
Schiøtz, Michaela L; Stockmarr, Anders; Høst, Dorte; Glümer, Charlotte; Frølich, Anne
Prevalences of multimorbidity vary between European studies and several methods and definitions are used. In this study we examine the prevalence of multimorbidity in relation to age, gender and educational attainment and the association between physical and mental health conditions and educational attainment in a Danish population. A cross-sectional design was used to study the prevalence of multimorbidity, defined as two or more chronic conditions, and of comorbid physical and mental health conditions across age groups and educational attainment levels among 1,397,173 individuals aged 16 years and older who lived in the Capital Region of Denmark on January 1st, 2012. After calculating prevalence, odds ratios for multimorbidity and mental health conditions were derived from logistic regression on gender, age, age squared, education and number of physical conditions (only for odds ratios for mental health conditions). Odds ratios for having multimorbidity and mental health conditions for each variable were adjusted for all other variables. Multimorbidity prevalence was 21.6%. Half of the population aged 65 and above had multimorbidity, and prevalence was inversely related to educational attainment: 26.9% (95% CI, 26.8-26.9) among those with lower secondary education versus 13.5% (95% CI, 13.5-13.6) among people with postgraduate education. Adjusted odds ratios for multimorbidity were 0.50 (95% CI, 0.49-0.51) for people with postgraduate education, compared to people with lower secondary education. Among all population members, 4.9% (95% CI, 4.9-4.9) had both a physical and a mental health condition, a proportion that increased to 22.6% of people with multimorbidity. Physical and mental health comorbidity was more prevalent in women (6.33%; 95% CI, 6.3-6.4) than men (3.34%; 95% CI, 3.3-3.4) and approximately 50 times more prevalent among older persons than younger ones. Physical and mental health comorbidity was also twice as prevalent among people with lower
Ellervik, Christina; Kvetny, Jan; Christensen, Kaj Sparle; Vestergaard, Mogens; Bech, Per
The Danish General Suburban Population Study (GESUS), the objective of which is to facilitate epidemiological and genetic research, has included the Major Depression Inventory (MDI) and the WHO-Five Well-Being Index (WHO-5) among the medical health questionnaires. We were thus in a position to compare the 2-week prevalence of ICD-10 depression in the period from 2010 to 2012 with our previous Danish general population study from 2003, in which the MDI was also included. The aim of our analysis was not only to evaluate the point prevalence of ICD-10 depression but also to describe the prevalence of antidepressants received by the respondents in the GESUS study and the correspondence to their subjective well-being on the WHO-5 questionnaire. To evaluate the validity (scalability) of the MDI and the WHO-5 in the GESUS study we performed the non-parametric Mokken analysis. The scalability of the MDI and the WHO-5 was quite acceptable. In total, 14,787 respondents were available from a response rate of 50%. The 2-week prevalence of ICD-10 depression was 2.3%, which is rather similar to the 2.8% in our 2003 study. The rate of people receiving antidepressants increased consistently with increasing severity of ICD-10 depression. This study has confirmed that the use of the MDI to obtain an ICD-10 depression diagnosis gives rather conservative estimates of the 2-week prevalence of depression in the Danish general population. The prescription of antidepressants depends on the severity of the ICD-10 depression diagnosis.
Bell, Katy JL; Del Mar, Chris; Wright, Gordon; Dickinson, James; Glasziou, Paul
Prostate cancer screening may detect nonprogressive cancers, leading to overdiagnosis and overtreatment. The potential for overdiagnosis can be assessed from the reservoir of prostate cancer in autopsy studies that report incidental prostate cancer rates in men who died of other causes. We aimed to estimate the age-specific incidental cancer prevalence from all published autopsy studies. We identified eligible studies by searches of Medline and Embase, forward and backward citation searches and contacting authors. We screened the titles and abstracts of all articles; checked the full-text articles for eligibility and extracted clinical and pathology data using standardized forms. We extracted mean cancer prevalence, age-specific cancer prevalence and validity measures and then pooled data from all studies using logistic regression models with random effects. The 29 studies included in the review dated from 1948 to 2013. Incidental cancer was detected in all populations, with no obvious time trends in prevalence. Prostate cancer prevalence increased with each decade of age, OR = 1.7 (1.6–1.8), and was higher in studies that used the Gleason score, OR = 2.0 (1.1–3.7). No other factors were significantly predictive. The estimated mean cancer prevalence increased in a nonlinear fashion from 5% (95% CI: 3–8%) at age 79 years. There was substantial variation between populations in estimated cancer prevalence. There is a substantial reservoir of incidental prostate cancer which increases with age. The high risk of overdiagnosis limits the usefulness of prostate cancer screening. What’s new? Before symptoms of prostate cancer manifest clinically, many men die of other causes. Yet, prostate screening, particularly in older men, frequently turns out positive, resulting in overdiagnosis and overtreatment. This meta-analysis of published autopsy studies shows that incidental prostate cancer increases with age and with the use of sensitive screening strategies
Full Text Available Abstract Background Following reports of high prevalence of low back pain (LBP in young physiotherapists 171819202122, we investigated whether LBP was a problem for undergraduate physiotherapy students. Method Physiotherapy students enrolled in one Australian tertiary institution completed a validated self-administered questionnaire in April 2001, seeking information on LBP prevalence (lifetime, 12 month, one-month, one-week, and its risk factors. The survey incorporated the Nordic back questionnaire, questions on common risks for LBP, and purpose-built questions regarding educational exposures. Univariate logistic regression models were applied to test associations. Results and Discussion 72% students responded. LBP prevalence was 69% (lifetime, 63% (12-month, 44% (one-month, 28% (one-week. The risk of LBP increased significantly for students once they completed first year. Being aged 20 or 21 years (final year students was significantly associated with all measures of LBP, compared with the youngest students. Exposure to tertiary study of greater than two years was associated with lifetime, 12 month and one-month LBP prevalence. Spending more than 20 hours in the past month 'sitting looking down' was significantly associated with one-month LBP prevalence. Similar exposure to 'treating patients' was significantly associated with one-month and one-week LBP prevalence. Conclusions Physiotherapy students should be alerted to the likelihood of LBP and is potential causes during their training, so that they enter the workforce with reduced risk of LBP. The potential for other undergraduate students to suffer LBP should also be considered.
Ferreira, J J; Gonçalves, N; Valadas, A; Januário, C; Silva, M R; Nogueira, L; Vieira, J L M; Lima, A B
Portugal has been identified as one of the countries with a high prevalence of LRRK2-G2019S, considered to be the most frequent known cause of familial and sporadic Parkinson's disease (PD). The aim of this study was to evaluate the prevalence of PD in Portugal using a door-to-door methodology. A cross-sectional study was conducted in the Portuguese community-dwelling population; that is, elderly people living in the community on their own, aged ≥50 years and resident in mainland Portugal, in two phases: (i) a questionnaire was applied to screen potential cases of PD; and (ii) screened cases were evaluated by an expert in PD to confirm diagnosis. The adjusted prevalence of PD for the Portuguese community-dwelling population aged ≥50 years was 0.24%. The estimated total number of cases of PD for the Portuguese population is 180/100 000 inhabitants. The results of this study show that a geographical region with a high frequency of a causal mutation for PD does not automatically imply a high prevalence of patients with PD. © 2017 EAN.
Ventura, Aline; Ronsoni, Marcelo Fernando; Shiozawa, Maria Beatriz Cacese; Dantas-Corrêa, Esther Buzaglo; Canalli, Maria Heloisa Busi da Silva; Schiavon, Leonardo de Lucca; Narciso-Schiavon, Janaína Luz
Celiac disease is an autoimmune disorder with an average prevalence of 1% in Europe and the United States. Because of strong European ancestry in southern Brazil, this study aimed to evaluate the seroprevalence of celiac disease among autoimmune thyroiditis patients. Cross-sectional study in a public university hospital. This cross-sectional prevalence study included autoimmune thyroiditis patients who were tested for anti-endomysial and anti-transglutaminase antibodies between August 2010 and July 2011. Fifty-three patients with autoimmune thyroiditis were included; 92.5% were women, with mean age of 49.0 ± 13.5 years. Five patients (9.3%) were serologically positive for celiac disease: three of them (5.6%) were reactive for anti-endomysial antibodies and two (3.7%) for anti-transglutaminase. None of them exhibited anemia and one presented diarrhea. Endoscopy was performed on two patients: one with normal histology and the other with lymphocytic infiltrate and villous atrophy. The prevalence of celiac disease among patients with autoimmune thyroid disease was 9.3%; one patient complained of diarrhea and none presented anemia. Among at-risk populations, like autoimmune thyroiditis patients, the presence of diarrhea or anemia should not be used as a criterion for indicating celiac disease investigation. This must be done for all autoimmune thyroiditis patients because of its high prevalence.
Tehrani-Banihashemi, Arash; Davatchi, Fereydoun; Jamshidi, Ahmad-Reza; Faezi, Tahereh; Paragomi, Pedram; Barghamdi, Mozhgan
To estimate the prevalence of osteoarthritis (OA) of different joints in rural areas of Iran. From five villages of Tuyserkan County, 1565 individuals were randomly selected and were interviewed to complete the Community Oriented Programme for Control of the Rheumatic Diseases (COPCORD) Core Questionnaire. Among these cases 1192 cases with rheumatic complaints were examined by a rheumatologist and laboratory and radiology tests were performed if necessary for the diagnosis. Definition of OA in various joints, were based on American College of Rheumatology (ACR) criteria. About 20% of the study population had OA in at least one of their joints. Prevalence of OA in the knee joint was 19.34%, in hand joins was 2.66% and in the neck was 2.21%. The most common findings on physical examination of patients with knee OA, hand OA and neck OA were bony crepitus (88.9%), Heberden's nodes (73.2%) and pain on movement (59.9%), respectively. This study revealed that OA in rural areas of Iran was more frequent in comparison with urban areas of Iran. Moreover, the prevalence of OA in rural areas of Iran was higher in comparison with prevalence of OA in rural areas of other Asian countries. Similar to previous studies OA was more frequently detected in the knee joint. © 2014 Asia Pacific League of Associations for Rheumatology and Wiley Publishing Asia Pty Ltd.
Osuna-Pozo, Carmen María; Serra-Rexach, José Antonio; Viña, José; Gómez-Cabrera, María del Carmen; Salvá, Antoni; Ruiz, Domingo; Masanes, Ferrán; Lopez-Soto, Alfonso; Formiga, Francesc; Cuesta, Federico; Cruz-Jentoft, Alfonso
There are few systematic studies on the prevalence of sarcopenia using the new diagnostic criteria in different geriatric care settings. To estimate the prevalence of sarcopenia, using the European Working Group on Sarcopenia in Older People (EWGSOP) criteria in older subjects living in nursing homes and in those who attend geriatric outpatient clinics. A single country multicentre study in two samples of older subjects: patients cared for in outpatient geriatric clinics, and individuals living in nursing homes. Data collected will include demographic variables, medical history, medication, geriatric syndromes, functional status (assessment of basic and instrumental activities of daily living), mobility, cognitive status, comorbidity, quality of life, nutritional status, and laboratory parameters. For the diagnosis of sarcopenia, 4m walking speed, handgrip strength, and body composition measured by bioelectrical impedance analysis will be assessed. Using the EWGSOP algorithm, the prevalence of sarcopenia in an elderly Spanish population will be estimated. In addition, concordance and correlation between the three parameters included in the definition (muscle mass, muscle strength, and physical performance) will be analysed, using the different existing cut-off points, and examining the diagnostic accuracy of each. Finally, demographic, anthropometric and functional data that define subjects with sarcopenia will be investigated. The ELLI study should improve knowledge on the prevalence and characteristics of sarcopenia in older people in our population. Copyright © 2012 SEGG. Published by Elsevier Espana. All rights reserved.
Høeg, Tracy B; Moldow, Birgitte; Ellervik, Christina; Klemp, Kristian; Erngaard, Ditte; la Cour, Morten; Buch, Helena
To determine the prevalence of amblyopia in Denmark before and after the initiation of the Danish national preschool vision screening programme. In a population-based cross-sectional study, 3826 participants of the Danish General Suburban Population Study (GESUS) aged 20 years and older from a Danish rural municipality received a complete general health examination and an ophthalmological interview and examination. This study included a comprehensive ophthalmologic interview, measurement of best corrected visual acuity (BCVA) in each eye, Hirschberg's test for strabismus and two 45-degree retinal fundus photographs of each eye. A complete ophthalmologic examination was performed when indicated. The prevalence of monocular visual impairment (MVI) was 4.26% (95% CI, 3.66-4.95, n = 163). Amblyopia was the most common cause, accounting for 33%. The prevalence of amblyopia was 1.44% (95% CI, 1.01-1.81, n = 55), being higher among non-preschool vision screened persons compared to those who were offered (estimated 95% attendance) preschool vision screening (1.78%, n = 41, 95% CI 1.24-2.33 versus 0.44%, n = 2, 95% CI, 0.12-1.60, p = 0.024). The leading cause of amblyopia was anisometropia (45.5%, 25/55). Amblyopia was the most common cause of MVI. Following the initiation of the Danish national preschool vision screening programme, which has an approximate attendance rate of 95%, the prevalence of amblyopia decreased by fourfold. © 2014 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.
Full Text Available CONTEXT AND OBJECTIVE: Celiac disease is an autoimmune disorder with an average prevalence of 1% in Europe and the United States. Because of strong European ancestry in southern Brazil, this study aimed to evaluate the seroprevalence of celiac disease among autoimmune thyroiditis patients.DESIGN AND SETTING: Cross-sectional study in a public university hospital.METHODS: This cross-sectional prevalence study included autoimmune thyroiditis patients who were tested for anti-endomysial and anti-transglutaminase antibodies between August 2010 and July 2011.RESULTS: Fifty-three patients with autoimmune thyroiditis were included; 92.5% were women, with mean age of 49.0 ± 13.5 years. Five patients (9.3% were serologically positive for celiac disease: three of them (5.6% were reactive for anti-endomysial antibodies and two (3.7% for anti-transglutaminase. None of them exhibited anemia and one presented diarrhea. Endoscopy was performed on two patients: one with normal histology and the other with lymphocytic infiltrate and villous atrophy.CONCLUSION: The prevalence of celiac disease among patients with autoimmune thyroid disease was 9.3%; one patient complained of diarrhea and none presented anemia. Among at-risk populations, like autoimmune thyroiditis patients, the presence of diarrhea or anemia should not be used as a criterion for indicating celiac disease investigation. This must be done for all autoimmune thyroiditis patients because of its high prevalence.
Akune, T; Muraki, S; Oka, H; Tanaka, S; Kawaguchi, H; Nakamura, K; Yoshimura, N
The present cross-sectional study investigated the prevalence of sarcopenia and clarified its associated factors in 1,000 elderly participants of Japanese population-based cohorts. Exercise habit in middle age was associated with low prevalence of sarcopenia in older age, suggesting that it is a protective factor against sarcopenia in older age. The present study investigated the prevalence of sarcopenia using the European Working Group on Sarcopenia in Older People (EWGSOP) definition, and clarified the association of sarcopenia with physical performance in the elderly participants of Japanese population-based cohorts of the Research on Osteoarthritis/osteoporosis Against Disability (ROAD) study. We enrolled 1,000 participants (aged ≥65 years) from the second visit of the ROAD study who had completed assessment of handgrip strength, gait speed, and skeletal muscle mass measured by bioimpedance analysis. Presence of sarcopenia was determined according to the EWGSOP algorithm. Information collected included exercise habits in middle age. Prevalence of sarcopenia was 13.8 % in men and 12.4 % in women, and tended to be significantly higher according to increasing age in both sexes. Factors associated with sarcopenia, as determined by logistic regression analysis, were chair stand time (odds ratio [OR], 1.09; 95 % confidence interval [CI], 1.04-1.14), one-leg standing time (OR, 0.97; 95 % CI, 0.96-0.99), and exercise habit in middle age (OR, 0.53; 95 % CI, 0.31-0.90). Exercise habit in middle age was associated with low prevalence of sarcopenia in older age. Furthermore, linear regression analysis revealed that exercise habits in middle age were significantly associated with grip strength (P gait speed (P < .001), and one-leg standing time (P = .005) in older age. This cross-sectional study suggests that exercise habit in middle age is a protective factor against sarcopenia in older age and effective in maintaining muscle strength and physical performance
Full Text Available Abstract Background Uncontrolled hypertension (HT is an established risk factor for the development of vascular diseases. Prevalence varies in different communities and no such study has been conducted in the Parsi community living in Bombay, India. The objectives of this study were to determine the prevalence, awareness, compliance to medication and control of HT in this community. Method We used a 1 in 4 random selection of subjects who were ≥ 20 years of age. A questionnaire was administered and the blood pressure (BP was measured by a doctor. HT was defined as diastolic blood pressure (DBP ≥ 90 mm Hg ± systolic pressure (SBP ≥ 140 mm Hg. Isolated systolic hypertension (ISH was defined as SBP ≥ 160 mm Hg with DBP Results 2879 subjects ≥ 20 years of age were randomly selected of which 2415 (84% participated in the study. The overall prevalence of HT in the community was 36.4%, of whom 48.5% were unaware of their hypertensive status. Of those aware of having HT, 36.4% were non-compliant with their anti-hypertensive drugs and only 13.6% had optimally controlled HT. Prevalence of ISH using the present criteria was 19.5% and 73% of hypertensives ≥ 60 years had ISH. Conclusion This study shows that prevalence of HT in the Parsi community is high and nearly half are unaware of their hypertensive status. ISH is the dominant form of HT in the elderly. Compliance to treatment is poor and optimal BP control is achieved in only a small minority. The study highlights the need for regular screening coupled with educational programs to detect and optimally treat HT in the community.
Chang, Thashi; Gajasinghe, Seneth; Arambepola, Carukshi
Stroke is a leading cause of disability and death worldwide. In the absence of published population-based prevalence data, we investigated the prevalence and risk factors of stroke in a population of varying urbanization in Sri Lanka. A population-based, cross-sectional study was conducted among 2313 adults aged ≥18 years residing in Colombo, selected using a multistage, probability proportionate-to-size, cluster sampling technique. Data were collected using an interviewer-administered questionnaire. Ever diagnosis of stroke was confirmed by medical doctors based on World Health Organization criteria and corroborated by documental evidence. Of the total population (52.4% women; mean age, 44.2 years; SD, 16.6), the prevalence of stroke was 10.4 per 1000 (95% confidence interval, 6.3-14.5) with a 2:1 male:female ratio. Beyond the age of 65 years, the prevalence was higher by 6-fold among men and by 2-fold among women. Ninety two percent had developed hemiparesis, 58.3% had dysphasia, and 16.7% had loss of balance. Hypertension was the commonest risk factor (62.5%) followed by smoking (45.8%), excess alcohol (41.7%), diabetes mellitus (33.3%), and transient ischemic attack (29.2%); 79.2%, predominantly men, had ≥2 risk factors. A percentage of 58.3 had brain computed tomographic scans, of whom 85.7% had ischemic strokes. A percentage of 64.3 had to change or give up working because of stroke-related disability. Age-adjusted stroke prevalence in urban Sri Lanka lies between high-income and low-/middle-income countries. The prevalence of stroke and its risk factors were higher among men. © 2015 American Heart Association, Inc.
Tolmane, Ieva; Rozentale, Baiba; Keiss, Jazeps; Arsa, Frida; Brigis, Girts; Zvaigzne, Aivars
Chronic viral hepatitis C (VHC) is one of the most discussed infectious diseases worldwide. The number of infected persons worldwide is approximately 170 million, and in Europe, it exceeds 9 million. The aim of this study was to determine the prevalence of antibodies to hepatitis C virus (anti-HCV prevalence) and prevalence of HCV viremia (HCV-RNA prevalence) in Latvia. A multistage randomized selection was used. A total of 42 primary care physicians (PCPs) were randomly selected from the register of PCPs from different regions of Latvia. From each PCP register, 60 subjects were selected (1651 individuals in total) and invited for the anti-HCV test with a screening method (ELISA). In case of positive results, antibodies were confirmed by the Western blot test, and all these subjects were tested for HCV-RNA by polymerase chain reaction. Of the 1459 subjects tested, 57 were positive for anti-HCV (3.9%; 95% CI 3% to 5%); 35 of them were positive for anti-HCV with a confirmatory test (2.4%; 95% CI, 1.7% to 3.3%): 19 men and 16 women (3.8% and 1.7%, respectively; P=0.011). The results of HCV RNA test were positive in 25 subjects (1.7%; 95% CI, 1.2% to 2.5%): 15 men and 10 women (3% and 1% respectively, P=0.019). The prevalence of anti-HCV and HCV-RNA in Latvia was found to be 2.4% and 1.7%, respectively. The prevalence of anti-HCV and HCV-RNA was higher in men than women.
Pan, Chen-Wei; Zheng, Ying-Feng; Anuar, Ainur Rahman; Chew, Merwyn; Gazzard, Gus; Aung, Tin; Cheng, Ching-Yu; Wong, Tien Y; Saw, Seang-Mei
To determine the prevalence of refractive errors in a multiethnic Asian population aged over 40 years and to examine secular trends and racial differences. A total of 10,033 adults (3353 Chinese, 3400 Indians, and 3280 Malays) participated in this study. Refractive error was determined by subjective refraction. Ocular biometric parameters were determined by partial coherence interferometry. Myopia and high myopia were defined as spherical equivalent (SE) of less than -0.5 diopters (D) and -5.0 D, respectively. Hyperopia was defined as SE of more than 0.5 D. Astigmatism was defined as cylinders less than -0.5 D. The prevalence of myopia, high myopia, hyperopia and astigmatism in Singapore adults aged over 40 years was 38.9% (95% confidence interval [CI] 37.1, 40.6); 8.4% (95% CI 8.0, 8.9); 31.5% (95% 30.5, 32.5); and 58.8% (95% CI 57.8, 59.9), respectively. Compared with the Tanjong Pagar Survey 12 years ago, there was a significant increase in the prevalence of astigmatism and mean axial length (AL) in Chinese adults aged over 40 years in Singapore. Chinese were most likely to be affected by myopia, high myopia, astigmatism, and had the longest AL among the three racial groups. The prevalence of myopia in Singapore adults is lower compared with the younger "myopia" generation in Singapore. The prevalence of astigmatism and mean AL have been increasing significantly within the past 12 years in the Chinese population. Chinese adults had higher prevalence of myopia, high myopia, astigmatism, as well as the longer AL compared with non-Chinese adults in Singapore.
Al-Ebraheem, A; Dao, E; Desouza, E; McNeill, F E; Farquharson, M J; Li, C; Wainman, B C
Routine tissue sample preparation using chemical fixatives is known to preserve the morphology of the tissue being studied. A competitive method, cryofixation followed by freeze drying, involves no chemical agents and maintains the biological function of the tissue. The possible effects of both sample preparation techniques in terms of the distribution of bio-metals (calcium (Ca), copper (Cu) zinc (Zn), and iron (Fe) specifically) in human skin tissue samples was investigated. Micro synchrotron radiation x-ray fluorescence (μSRXRF) was used to map bio-metal distribution in epidermal and dermal layers of human skin samples from various locations of the body that have been prepared using both techniques. For Ca, Cu and Zn, there were statistically significant differences between the epidermis and dermis using the freeze drying technique (p = 0.02, p < 0.01, and p < 0.01, respectively). Also using the formalin fixed, paraffin embedded technique the levels of Ca, Cu and Zn, were significantly different between the epidermis and dermis layers (p = 0.03, p < 0.01, and p < 0.01, respectively). However, the difference in levels of Fe between the epidermis and dermis was unclear and further analysis was required. The epidermis was further divided into two sub-layers, one mainly composed of the stratum corneum and the other deeper layer, the stratum basale. It was found that the difference between the distribution of Fe in the two epidermal layers using the freeze drying technique resulted in a statistically significant difference (p = 0.012). This same region also showed a difference in Fe using the formalin fixed, paraffin embedded technique (p < 0.01). The formalin fixed, paraffin embedded technique also showed a difference between the deeper epidermal layer and the dermis (p < 0.01). It can be concluded that studies involving Ca, Cu and Zn might show similar results using both sample preparation techniques, however studies involving Fe would need more
Decisions in healthcare should be based on information obtained according to the principles of Evidence-Based Medicine (EBM). An increasing number of systematic reviews are published which summarize the results of prevalence studies. Interpretation of the results of these reviews should be accompanied by an appraisal of the methodological quality of the included data and studies. The critical appraisal tool for prevalence studies developed and tested by Munn et al. comprises 10 items and aims at targeting all kinds of prevalence studies. This comment discusses the pros and cons of different designs of quality appraisal tools and highlights their importance for systematic reviews of prevalence studies. Beyond piloting, which has been performed in the study by Munn et al., it is suggested here that the validity of the tool should be tested, including reproducibility and inter-rater reliability. It is concluded that studies as the one by Munn et al. will help to establish a critical understanding of methodological quality and will support the use of systematic reviews of non-intervention studies for health policy making.
Arellano-Morales, Leticia; Roesch, Scott C; Gallo, Linda C; Emory, Kristen T; Molina, Kristine M; Gonzalez, Patricia; Penedo, Frank J; Navas-Nacher, Elena L; Teng, Yanping; Deng, Yu; Isasi, Carmen R; Schneiderman, Neil; Brondolo, Elizabeth
Empirical studies examining perceived ethnic discrimination in Latinos of diverse background groups are limited. This study examined prevalence and correlates of discrimination in a diverse sample of U.S. Latinos (N=5,291) from the multi-site Hispanic Community Health Study/Study of Latinos (HCHS/SOL) and HCHS/SOL Sociocultural Ancillary Study. The sample permitted an examination of differences across seven groups (Central American, Cuban, Dominican, Mexican, Puerto Rican, South American, and Other/Multiple Background). Most participants (79.5%) reported lifetime discrimination exposure and prevalence rates ranged from 64.9% to 98% across groups. Structural Equation Models (SEM) indicated that after adjusting for sociodemographic covariates most group differences in reports of discrimination were eliminated. However, Cubans reported the lowest levels of discrimination, overall among all groups. Furthermore, regional effects were more important than group effects. Participants from Chicago reported the highest levels of discrimination in comparison to other regions. Group differences among Latinos appear to be primarily a function of sociodemographic differences in education, income, and acculturation. In addition, differences in exposure to discrimination may be tied to variables associated with both immigration patterns and integration to U.S. culture. Results highlight the importance of considering historical context and the intersection of discrimination and immigration when evaluating the mental health of Latinos.
Disease (BOLD foi desenvolvida para que a prevalÃªncia da DPOC possa ser avaliada com metodologia normalizada. O objetivo deste estudo foi estimar a prevalÃªncia da DPOC em adultos com 40 ou mais anos numa populaÃ§Ã£o alvo de 2Â 700Â 000 habitantes na regiÃ£o de Lisboa, de acordo com o protocolo BOLD. MÃ©todos: A amostra foi estratificada de forma aleatÃ³ria multifaseada selecionando-se 12 freguesias. O inquÃ©rito compreendia um questionÃ¡rio com informaÃ§Ã£o sobre fatores de risco para a DPOC e doenÃ§a respiratÃ³ria autoreportada; adicionalmente, foi efetuada espirometria com prova de broncodilataÃ§Ã£o. Resultados: Foram incluÃdos 710 participantes com questionÃ¡rio e espirometria aceitÃ¡veis. A prevalÃªncia estimada da DPOC na populaÃ§Ã£o no estÃ¡dio GOLD I+ foi de 14,2% (IC 95%: 11,1; 18,1 e de 7,3% no estÃ¡dio ii+ (IC 95%: 4,7; 11,3. A prevalÃªncia nÃ£o ajustada foi de 20,2% (IC 95%: 17,4; 23,3 no estÃ¡dio i+ e de 9,5% (IC 95%: 7,6; 11,9 no estÃ¡dio ii+. A prevalÃªncia da DPOC no estÃ¡dio GOLD II+ aumentou com a idade, sendo mais elevada no sexo masculino. A prevalÃªncia estimada da DPOC no estÃ¡dio GOLD I+ foi de 9,2% (IC 95%: 5,9; 14,0 nos nÃ£o fumadores versus 27,4% (IC 95%: 18,5; 38,5 nos fumadores com carga tabÃ¡gica de â¥Â 20 Unidades MaÃ§o Ano. Detetou-se uma fraca concordÃ¢ncia entre a referÃªncia a diagnÃ³stico mÃ©dico prÃ©vio e o diagnÃ³stico espiromÃ©trico, com 86,8% de subdiagnÃ³sticos. ConclusÃµes: O achado de uma prevalÃªncia estimada da DPOC de 14,2% sugere que esta Ã© uma doenÃ§a comum na regiÃ£o de Lisboa, contudo com uma elevada proporÃ§Ã£o de subdiagnÃ³sticos. Estes dados apontam para a necessidade de aumentar o grau de conhecimento dos profissionais de saÃºde sobre a DPOC, bem como a necessidade de maior utilizaÃ§Ã£o da espirometria nos cuidados de saÃºde primÃ¡rios. Keywords: COPD prevalence, BOLD study, Burden of Obstructive
Suzuki, Okihide; Eguchi, Hidetaka; Chika, Noriyasu; Sakimoto, Takehiko; Ishibashi, Keiichiro; Kumamoto, Kensuke; Tamaru, Jun-Ichi; Tachikawa, Tetsuhiko; Akagi, Kiwamu; Arai, Tomio; Okazaki, Yasushi; Ishida, Hideyuki
To clarify the prevalence and clinicopathologic/molecular characteristics of mismatch repair (MMR)-deficient colorectal cancer in the young Japanese population. Immunohistochemical analyses for MMR proteins (MLH1, MSH2, MSH6, and PMS2) were performed in formalin-fixed paraffin-embedded sections prepared from the resected CRC specimens of 119 consecutive patients aged <50 years old, who underwent resection of the primary tumor at our institution between 1996 and 2015. Analyses for somatic BRAF V600E mutation, somatic hypermethylation of the MLH1 promoter, and germline MMR gene mutations were undertaken where indicated. MMR protein loss was found in 10 patients (8.4%), 7 (5.9%) of whom were subsequently identified to have Lynch syndrome (LS). The remaining 3 patients were categorized as having sporadic MMR-deficient CRC (n = 2) or "possible LS (n = 1)". In multivariate logistic regression analysis, the presence of tumor-infiltrating lymphocytes (P < 0.01), right-sided location of the tumor (P = 0.01), and a history of LS-associated tumors in the first-degree relatives (P < 0.01) were identified as independent factors predictive of MMR-deficient CRC. These results are of value in the clinical management of patients with the early onset CRC under circumstances where universal tumor screening approaches for LS are still not available, like in Japan.
Full Text Available Background. Atopic allergy is effected by a number of environmental exposures, such as dry air and time spent outdoors, but there are few estimates of the prevalence in populations from sub-arctic areas. Objective. To determine the prevalence and severity of symptoms of food, inhalation and skin-related allergens and coeliac disease (CD in the sub-arctic region of Sweden. To study the correlation between self-reported allergy and allergy test results. To estimate the heritability of these estimates. Study design. The study was conducted in Karesuando and Soppero in Northern Sweden as part of the Northern Sweden Population Health Study (n=1,068. We used a questionnaire for self-reported allergy and CD status and measured inhalation-related allergens using Phadiatop, food-related allergens using the F×5 assay and IgA and IgG antibodies against tissue transglutaminase (anti-tTG to indicate prevalence of CD. Results. The prevalence of self-reported allergy was very high, with 42.3% reporting mild to severe allergy. Inhalation-related allergy was reported in 26.7%, food-related allergy in 24.9% and skin-related allergy in 2.4% of the participants. Of inhalation-related allergy, 11.0% reported reactions against fur and 14.6% against pollen/grass. Among food-related reactions, 14.9% reported milk (protein and lactose as the cause. The IgE measurements showed that 18.4% had elevated values for inhalation allergens and 11.7% for food allergens. Self-reported allergies and symptoms were positively correlated (p<0.01 with age- and sex-corrected inhalation allergens. Allergy prevalence was inversely correlated with age and number of hours spent outdoors. High levels of IgA and IgG anti-tTG antibodies, CD-related allergens, were found in 1.4 and 0.6% of participants, respectively. All allergens were found to be significantly (p<3e–10 heritable, with estimated heritabilities ranging from 0.34 (F×5 to 0.65 (IgA. Conclusions. Self-reported allergy
Full Text Available Aim: To identify the prevalence of Hydatidosis in buffaloe-calves, heifers and adults slaughtered in slaughter house of Mirha Exports Pvt. Ltd. between March 2010 to Feb. 2011.Materials and Methods: 2,09,615 buffaloes slaughtered were included in this study and were categorized into three groups (A, B and C based on their age at the time of ante-mortem examination. Group Aconsisted of calves (>1 year, accounting for 5100 out of the total number of animals. Group B consisted of heifers (2-3 years accounting for 7840. Group C consisted of adult buffaloes (3-7 years accounting for 1, 96675. Results:Our study revealed an overall prevalence of hydatidosis as 50.96%. Furthermore, the group wise analysis showed the prevalence as 3.52%, 5.58% and 54% in calves, heifers and adult buffaloes, respectively.Conclusion: Based on the factors considered, we conclude that the higher prevalence of infections in buffaloes slaughtered in the Mirha Exports is likely due to an overpopulation of stray dogs in the areas from which the plant gets its supply of animals.
Jeppesen, Charlotte; Maindal, Helle T; Kristensen, Jette K
AIMS: The prevalence of gestational diabetes mellitus (GDM) is increasing worldwide. We studied the prevalence of GDM from 2004 to 2012 in Danish women aged 15-49 years using registries with records of the diagnosis of GDM at delivery. METHODS: We conducted a national register-based study of 12......,538 women with a diagnosis of GDM during a pregnancy leading to a live birth in the period 2004-2012. The diagnosis of GDM was taken from the National Patient Registry and combined with the total number of births at the national level from Statistics Denmark. Prevalence estimates were reported as crude...... and age-standardized using Danish and international data for women aged 15-49 years. RESULTS: A total of 566,083 live births was registered in Denmark from 2004 to 2012. The age-standardized prevalence of GDM increased from 1.7% (1095/63,465) of the total births in 2004 to 2.9% (1721/56,894) of the total...
Sallout, Bahauddin I.; Al-Hoshan, Manal S.; Attyyaa, Rehman A.; Al-Suleimat, Abdelmane A.
The exact antenatal prevalence of congenital anomalies in Saudi society is unknown. Early antenatal diagnosis of congenital anomalies is crucial for early counseling, intervention and possible fetal therapy. The objective of this study was to evaluate the antenatal frequency of major congenital anomalies and malformations patterns in our hospital population and to evaluate the outcome and perinatal mortality rates for major congenital anomalies. This was a prospective study of the antenatal diagnosis of major fetal congenital anomalies conducted in the ultrasound Department of the Women's Specialized Hospital at King Fahd Medical City from for 7762 patients and 5379 babies delivered in our institution. We diagnosed 217 cases of fetal anomalies. The antenatal prevalence of congenital anomalies was 27.96 per 1000. The median maternal age at diagnosis was 27.5 years. Te median gestational age at diagnosis was 31 weeks. Genitourinary and cranial anomalies were the commonest; for 186 patients delivered in our institution, the birth prevalence was 34.57 per 1000 births. The median gestational age at delivery was 38 weeks. The perinatal mortality arte was 34.9% (65/186), including all cases of intrauterine fetal and neonatal deaths. The prevalence of major congenital anomalies in our population appears to be similar to international figures. Major congenital anomalies are a major cause of perinatal mortality. (author)
Wahab Kolawole W
Full Text Available Abstract Background Obesity is assuming an epidemic dimension globally. It is important to appreciate factors associated with the disease so that a holistic approach can be taken in tackling the rising burden. The objective of this study was to determine the prevalence of overweight and obesity and the factors independently associated with obesity in an urban Nigerian population. Methods A cross-sectional study of 300 healthy adult subjects was conducted in the urban city of Katsina, northern Nigeria. Relevant sociodemographic and clinical information were obtained. Screening for obesity was done using the Body Mass Index while relevant laboratory investigations were conducted. Univariate and multivariate logistic regression analyses were performed to determine the predictors of obesity. Results Overweight and obesity was found in 53.3% and 21.0% respectively with a significantly higher prevalence in females compared to males (overweight: 62.0% vs 41.9%, p Conclusion There is a high prevalence of obesity in northern Nigeria and women are significantly more affected. The high prevalence is independently associated with female sex, hypercholesterolaemia and hyperuricaemia. Public health education is urgently needed in order to reduce this burden and prevent other non-communicable cardiovascular disorders.
Sajid, Saraj-Uddin; Sajid, Mahum; Hashmi, Ramiz Iqbal
Salmonella is an important zoonotic pathogen and its prevalence in the chicken meat and eggs acts as a continuous threat to human population. The current studies covering a time period of three years, was carried out to report the isolation of salmonellae from the chicken tissues, eggs and feed ingredient. A total of 1747 random samples from twelve different sources and 56 locations in Islamabad and Northern Punjab area of Pakistan, were screened for isolation studies according to the already published established protocols. The analysis of 1747 random samples comprising of 1069 (61.19%) chicken organs and 678 (38.81%) allied sources including eggs and feed ingredients, showed that a total of 162 (9.27%) were positive for salmonellae. Isolation prevalence in various chicken organs and allied sources was 86 (8.04%) and 76 (11.20%) respectively. The maximum isolation prevalence was recorded in meat meal (19.35%), followed by fish meal (17.54%), hatchery fluff (14.63%), livers (13.17%), poultry litter (10.89%), and eggs (9.64%). The range of Salmonella isolated varied from 19.35% to 4.72% in various organs and allied sources. Our findings highlighted a potential public health hazard and emphasized the significance of continuous surveillance system in the country to understand the ever changing epidemiological pattern of Salmonella serovers. The endemic prevalence of various serovars can cause outbreaks of human salmonellosis due to the consumption of contaminated meat and eggs as has already been reported worldwide.
Do, Loan Minh; Tran, Toan Khanh; Eriksson, Bo; Petzold, Max; Ascher, Henry
A plateau in childhood overweight and obesity has been reported in some developed countries while in almost all developing countries this problem is on the rise. The aim of this paper is to describe the changes in prevalence of overweight and obesity within a cohort of preschool children followed for 3 years, and to estimate and compare the incidences in urban and rural children of Hanoi, Vietnam. A longitudinal study of a cohort of 2677 children aged 3 to 6 years old at the beginning of the study was conducted in urban DodaLab and rural FilaBavi, Hanoi, Vietnam. Overall, 2602 children, 1311 urban and 1291 rural, were followed for 3 years with identical measurements of weight and height in 2013, 2014 and 2016. Standard methods were used to estimate prevalence and incidence as well as confidence intervals. During the three-year follow-up, the overall estimated prevalence of overweight increased from 9.1% to 16.7%. For the urban children, the increase was considerably higher. The overall prevalence of obesity decreased from 6.4% to 4.5% with less decrease in the urban children. In the group of children who were overweight and obese at the start of the study, 41.4% and 30.7%, respectively, remained in the same state three years later. The incidence of overweight and obesity during the three years were 12.4% and 2.7%, respectively. Boys were more likely to develop obesity than girls. Already in preschool age, the prevalence of overweight is high and it continues to increase with age, especially in the urban area. Prevention and intervention programs need to start at early preschool age and actions in urban areas deserve priority.
Ambrish Sanjay Dharmadhikari
Full Text Available Background: Epilepsy is one of the most common chronic neurological disorders. In children, it has long debilitating course and is associated with comorbidities including psychiatric comorbidity. To tackle this burden of comorbidities, we must know the extent of problem. Hence, there is a need for estimation of prevalence of psychiatry disorder in children with epilepsy. Aim: The present study was aimed at measuring the prevalence of various psychiatry disorders among children suffering from epilepsy. Settings and Design: Cross-sectional chart review. Methodology: We reviewed case record files of all patients with a diagnosis of epilepsy in the age group of 9–17 years. Chart review was done for 5 years, May 1, 2007, to April 30, 2012. A total of 718 patients record were included in the study after satisfying inclusion criteria and excluding nonepilepsy diagnosis. Statistical Analysis: Statistics was done using Statistical Package for Social Sciences (SPSS 18.0. Descriptive statistics were used to calculate the result, Chi-square and Mann–Whitney U-test used wherever applicable. Results: The prevalence of childhood psychiatric disorder among children with epilepsy found to be 31.2%. We also found that having a partial component (73.21%, n = 164 in seizure has more chances of psychopathology in comparison to generalized seizure (8.1%, n = 18. Among them, those having a partial component with generalization (66.96%, n = 150 had a greater prevalence of psychopathology. Mental retardation was most common psychiatric disorder among psychopathology followed by manic/depressive illness (unipolar followed by unspecified nonorganic psychosis. Conclusion: From our study, we demonstrate the significant mental health needs of children with epilepsy. The evident high prevalence of psychiatry disorder emphasizes the need for psychopathology assessment and treatment as a part of any comprehensive epilepsy clinic.
Full Text Available Although adult asthma is attributable to occupational factors, few reports are available on asthma prevalence among health care workers in Japan. The objective of this study was to estimate the prevalence of asthma and wheeze among Japanese nursing professionals. A cross-sectional study was conducted by postal survey using a translated version of the European Community Respiratory Health Survey questionnaire from April to June 2013. The analysis included 4634 nursing professionals (257 men and 4377 women and the overall response rate was 84.8%. The prevalence of current asthma and wheeze were 10.7% (95% confidence interval (CI, 9.9%–11.7% and 15.6% (95% CI, 14.5%–16.6%, respectively. More than one year of work experience as a nursing professional and more than one year of experience with bed-making tasks were associated with odds ratios (ORs of 1.95 (95% CI, 1.12–3.39 and 1.64 (95% CI, 1.15–2.23 for wheeze, respectively. Current smoking was significantly associated with the presence of wheeze, with ORs of 2.27 for men (95% CI, 1.11–4.64 and 2.01 for women (95% CI, 1.54–2.64. Among female nurses, latex allergy was associated with wheeze (OR, 1.87; 95% CI, 1.56–2.23, as was body mass index ≥30 (OR, 2.76; 95% CI, 1.65–4.62. This study has provided the prevalence of asthma and wheeze among Japanese nursing professionals. Employment period, bed-making tasks, latex allergy, obesity, and smoking may be risk factors for prevalent wheeze among nursing professionals.
Beatriz, Elizabeth D; Salhi, Carmel; Griffith, John L; Molnar, Beth E
Despite indications that there are differences in rates of child maltreatment (CM) cases in the child protection system between urban and rural areas, there are no published studies examining the differences in self-reported CM prevalence and its correlates by urbanicity. The present study aimed to: (1) identify the distribution of self-reported childhood experiences of maltreatment by urbanicity, (2) assess whether differences by urbanicity persist after adjusting for known risk factors, and (3) explore whether the associations between these risk factors and CM are modified by urban-rural designation. Using nationally representative data from waves I and III of the National Longitudinal Study of Adolescent to Adult Health, the prevalence of six maltreatment outcomes was estimated for rural, minor urban, and major urban areas (N = 14,322). Multivariable logistic models were estimated identifying if risk associated with urbanicity persisted after adjusting for other risk factors. Interactions between urbanicity and main effects were explored. Prevalence estimates of any CM, poly-victimization, supervision neglect, and physical abuse were significantly higher in major urban areas. Those from major urban areas were more likely to report any maltreatment and supervision neglect even after adjusting for child and family risk factors. The association between race/ethnicity, welfare receipt, low parental educational attainment, and disability status and CM were modified by urbanicity. Significant differences in the prevalence and correlates of CM exist between urban and rural areas. Future research and policy should use self-reported prevalence, in conjunction with official reports, to inform child maltreatment prevention and intervention. Copyright © 2018 Elsevier Ltd. All rights reserved.
Full Text Available Objective The present study was undertaken to determine the prevalence of periodontal diseases among adults visiting the dental department of a hospital based in Jorpati. Methods Four hundred and seventeen patients were randomly taken from a dental hospital situated in Jorpati during the time period of March 2013 to August 2013. All patients visiting the dental department who fulfilled the inclusion criteria were included in the study. The data included patients overall information along with their chief complaint, gingival bleeding on probing (BOP, probing depth (PD, frequency of brushing, adverse habits and the diagnosis of the patients examined. Results We found that 52.5% suffered from gingivitis and 47.5% suffered from periodontitis. Also 28.3% suffered from localized and 18% suffered from generalized form of periodontitis. There was no statistically significant difference in the gender when the prevalence of periodontal disease was compared. 51.4% of male and 44.4% of female was seen to be affected with periodontitis. Also habits like smoking and intake of smokeless tobacco was seen to be associated with periodontitis. Regarding the age group more number of patients in the age group of >50 years were seen to be suffering from periodontitis (84.3% as compared to age group of <35 years (25.9%. Conclusion There is high prevalence of periodontitis and gingivitis in the studied population. Periodontitis was seen to be more prevalent as age advanced which can be attributed mainly to untreated gingivitis. However, no significant difference was seen in the prevalence of periodontitis with respect to gender. DOI: http://dx.doi.org/10.3126/jcmsn.v10i1.12762 Journal of College of Medical Sciences-Nepal, 2014, Vol.10(1; 11-16
Full Text Available Mirna Waked1, George Khayat2, Pascale Salameh31Saint George Hospital University Medical Center, Beirut, Lebanon; 2Faculty of Medicine, Hotel-Dieu de France Hospital, Saint-Joseph University, Beirut, Lebanon; 3Faculties of Pharmacy and Public Health, Lebanese University, Beirut, LebanonBackground: Chronic obstructive pulmonary disease (COPD continues to increase worldwide. The objective of this study was to determine the prevalence of COPD in Lebanese adults.Methods: A cross-sectional study was carried out using a multistage cluster sample from all over Lebanon. Residents aged 40 years and over were enrolled. Subjects underwent baseline spirometry and answered a questionnaire. After an albuterol + ipratropium bromide bronchodilator, a posttest was performed.Results: Of 2201 individuals, only 33.3% had never smoked. The prevalence of COPD by the Global Initiative for Chronic Obstructive Lung Disease definition, was 9.7% (95% confidence interval [CI]: 8.5%–10.9%. According to the 5% lower limit of normal definition of COPD, the prevalence was 12.5% (95% CI: 11.2%–13.9%. A total of 20.2% were already diagnosed by a physician. No differences in symptoms across stages of COPD were found, but there was a significant trend for a higher number of visits to the emergency room and to the doctor (P < 0.001, and a higher number of hospitalizations (P < 0.001. Older individuals had an increased risk of COPD (adjusted odds ratio [ORa] = 1.05; so did “ever” cigarette smokers (ORa = 4.88 and water-pipe smokers (ORa = 2.53.Conclusion: This is the first epidemiological study in Lebanon that determined COPD prevalence and the link with water-pipe smoking.Keywords: COPD, prevalence, water-pipe smoking
Thapa, Suman S; Thapa, Raba; Paudyal, Indira; Khanal, Shankar; Aujla, Jaskirat; Paudyal, Govinda; Rens, Ger van
Vitreo-retinal diseases are among the leading causes of visual impairment and blindness worldwide. This study reports the prevalence and pattern of vitreo-retinal diseases in the Bhaktapur Glaucoma Study (BGS), a population based study conducted in Nepal. BGS was a population based cross-sectional study involving 4800 subjects aged 40 years and over from Bhaktapur district. Subjects were selected using a cluster sampling methodology and a door-to-door enumeration. All subjects underwent a detailed ocular examination at the base hospital which included log MAR visual acuity, refraction, applanation tonometry and a dilated fundus examination. Fundus photography, optical coherence tomography and fundus fluorescein angiography were performed where indicated. Complete data was available for 3966 (82.62%) out of the total of 4800 enumerated subjects. The mean age was 55.08 years (SD 11.51). The overall prevalence of vitreo-retinal disorders was 5.35% (95% CI, 4.67 - 6.09). Increasing age was associated with a higher prevalence of vitreo-retinal disorders (P < 0.001). The prevalence of diabetes mellitus was 7.69% (95% CI, 6.88 - 8.56). Age-related macular degeneration (AMD) was the most common vitreo-retinal disorder with a prevalence of 1.50% (95% CI, 1.15 - 1.94), increasing significantly with age. The prevalence of diabetic retinopathy among the study population was 0.78% (95% CI, 0.53 - 1.11) and among the diabetic population 10.16% (95% CI, 7.01 - 14.12). The population prevalence of other retinal disorders were hypertensive retinopathy 0.88%, macular scar 0.37%, retinal vein occlusion 0.50%, macular hole 0.20%, retinitis pigmentosa 0.12%. and retinal detachment 0.10%.The prevalence of low vision and blindness due to vitreo-retinal disorders was 1.53% (95% CI, 1.18 - 1.97) and 0.65% (95% CI, 0.43 - 0.96), respectively. The prevalence of low vision and blindness was 28.77% (95% CI, 22.78-35.37) and 12.26% (95% CI, 8.17-17.45), respectively among cases with vitreo
Karpov On Behalf Of Participants Of The Prometheus Study, Yu A
The main purpose of the study was to estimate prevalence of subjects with hypertriglyceridemia (HTG) in Russia. Secondary objectives were to explore HTG prevalence by levels, age and sex, and to assess correlation between glycated hemoglobin (HbA1C) and triglyceride (TG) level. Additionally, we analyzed geographical differences in HTG prevalence in regions of Russia. This was a cross-sectional, retrospective, observational study using database of results of lipid profile determination in 357,072 subjects from 254 Russian cities during the 3-year period from 2011 to 2013. Altogether, 29.2% (95% confidence interval [CI] 29.1-29.4%) of Russian individuals had HTG (serum TG more or equal 1.7 mmol/L). The percentage of patients with very high (TG more or equal 5.6 mmol/L) and severe HTG (TG more or equal 10.0mmol/L) was low (0.01% and 0.011%, respectively). At the same time, the portion of subjects with mixed hyperlipidemia (total cholesterol [TC] more or equal 5.2mmol/L, low-density lipoprotein cholesterol [LDL-C] more or equal 3.4 mmol/L, TG more or equal 1.7 mmol/L) was 19% of the study population. Men had 1.25 (95% CI, 1.24-1.26) times higher risk of HTG than women. Prevalence of HTG increased with age: in women TG level was maximal in the age group 60-69 years (34%), whereas in men TG level was maximal in the age group 40-49 years (43%). Prevalence of HTG increased from 2011 to 2013 from 28 to 30% (p<0.0001). Risk of HTG was 1.69 times greater when high HbA1C more or equal 6.5% was present, and vice versa, risk of HbA1C more or equal 6.5% was 2.04 times higher in individuals with HTG. Distribution of HTG and dyslipidemia by regions of Russia had large variability being higher in the south and lower in the northern regions of European part of Russia. Almost a third of Russian population has HTG. Men have higher risk of HTG than women. Prevalence of HTG increases with age and reaches its peak in age groups 60-69 years in (women) and 40-49 years (men). There is a
Ayesha S Al Dhaheri
Full Text Available Metabolic syndrome (MetS is a growing problem in the United Arab Emirates (UAE. Moreover, the prevalence of overweight and obesity is rapidly increasing in the UAE especially among young females. However, few studies have evaluated the prevalence of MetS among young female adults in the UAE. This study determined the prevalence of MetS in Emirati females aged 17-25 years and its relation to overweight and obesity.In total, 555 Emirati female college students were enrolled in a cross-sectional study, conducted during 2013-2014 at United Arab Emirates University in Al Ain, UAE. Anthropometric measurements, blood pressure and biochemical measurements were collected. MetS was defined according to the harmonised International Diabetes Federation criteria.Of the 555 participants enrolled, 23.1% were overweight and 10.4% were classified as obese. The overall prevalence of MetS was 6.8%. MetS prevalence was highest among obese participants (34.5%, as compared with normal-weight (1.7% and overweight (10.1% participants. MetS was significantly associated with overweight (adjusted odds ratio [aOR] = 3.8, 95% confidence interval [CI]; 1.15-12.52 and obesity (aOR = 11.2, 95% CI; 3.1-40.9, as compared with normal-weight. Waist-hip ratio ≥ 0.8 (aOR = 3.04, 95% CI; 1.10-8.44 was significantly associated with MetS, as compared with waist-hip ratio <0.8. The odds of MetS were 22 fold higher in participants with glycated haemoglobin (HbA1c ≥ 6.5% (aOR = 22.5, 95% CI; 6.37-79.42 compared to HbA1c <6.5%. This difference was 9 fold higher when HbA1c between 5.6%-6.4% was compared to HbA1c <5.6% (aOR = 8.9, 95% CI; 3.4-23.5.The prevalence of MetS among obese Emirati female students was significantly higher than overweight and normal weight students. The high prevalence of MetS highlights the importance of regular screening and intervention programmes targeting weight reduction.
Wolfram, Christian; Höhn, René; Kottler, Ulrike; Wild, Philipp; Blettner, Maria; Bühren, Jens; Pfeiffer, Norbert; Mirshahi, Alireza
To study the distribution of refractive errors among adults of European descent. Population-based eye study in Germany with 15010 participants aged 35-74 years. The study participants underwent a detailed ophthalmic examination according to a standardised protocol. Refractive error was determined by an automatic refraction device (Humphrey HARK 599) without cycloplegia. Definitions for the analysis were myopia +0.5 D, astigmatism >0.5 cylinder D and anisometropia >1.0 D difference in the spherical equivalent between the eyes. Exclusion criterion was previous cataract or refractive surgery. 13959 subjects were eligible. Refractive errors ranged from -21.5 to +13.88 D. Myopia was present in 35.1% of this study sample, hyperopia in 31.8%, astigmatism in 32.3% and anisometropia in 13.5%. The prevalence of myopia decreased, while the prevalence of hyperopia, astigmatism and anisometropia increased with age. 3.5% of the study sample had no refractive correction for their ametropia. Refractive errors affect the majority of the population. The Gutenberg Health Study sample contains more myopes than other study cohorts in adult populations. Our findings do not support the hypothesis of a generally lower prevalence of myopia among adults in Europe as compared with East Asia.
Probert, C S; Jayanthi, V; Hughes, A O; Thompson, J R; Wicks, A C; Mayberry, J F
The family history of patients identified during incidence studies in Leicestershire were investigated and the prevalence and comparative risks calculated; 1254 patients aged 15 to 80 years were sent a questionnaire about their family history. All cases with a positive family history were reviewed and confirmed cases included in the study. In Europeans the standardised prevalence of Crohn's disease was 75.8/10(5) and that of ulcerative colitis 90.8/10(5). The prevalence of Crohn's disease amo...
Sarokhani, Diana; Delpisheh, Ali; Veisani, Yousef; Sarokhani, Mohamad Taher; Manesh, Rohollah Esmaeli; Sayehmiri, Kourosh
Introduction. Depression is one of the four major diseases in the world and is the most common cause of disability from diseases. The aim of this study is to estimate the prevalence of depression among Iranian university students using meta-analysis method. Materials and Methods. Keyword depression was searched in electronic databases such as PubMed, Scopus, MAGIran, Medlib, and SID. Data was analyzed using meta-analysis (random-effects model). Heterogeneity of studies was assessed using ...
Ahmadi, Rahil; Ramazani, Nahid; Nourinasab, Rahmatollah
Objective The aim of this study was to investigate the prevalence of molar incisor hypomineralization (MIH) and its relationship with systemic conditions in a group of Iranian children. Methods The study population comprised of 433 7-9 year olds, from four schools in Zahedan, Iran. Subjects were evaluated clinically by one examiner, and at a separate session, their mothers completed a coded medical history questionnaire. Hypo-mineralized molars and incisors were recorded based on DDE (develop...
Aoun, Georges; Berberi, Antoine
Objective: Chronic erythematous candidiasis also known as denture-related stomatitis refers to inflammatory changes of the denture-bearing mucosa. The aim of this study was to evaluate the prevalence of chronic erythematous candidiasis in a Lebanese population using clinical and microbiological examinations. Materials and Methods: Ninety-eight patients wearing full acrylic maxillary denture (50 women and 48 men) were included in this study. A clinical oral assessment and a microbiological exa...
Mohammadi, Khadijeh; Rahnama, Parvin; Montazeri, Ali
Background Multiple sclerosis is increasingly becoming a major health problem among women worldwide. The aim of the present study was to estimate prevalence of depression in women with multiple sclerosis and also to identify risk factors contributing to its development. Methods This was a cross-sectional study of depression in a sample of 226 women with multiple sclerosis. The sample was recruited from an outpatient clinic in Tehran, Iran. Depression was assessed using the Beck Depression Inv...
Fida, Hashim R; Abdelmoneim, Ismail
Background This study was conducted to examine the prevalence of smoking and the smoking habits among male secondary school students in Jeddah, Kingdom of Saudi Arabia and to assess their knowledge and attitudes towards smoking. Methods A cross-sectional study was conducted in Jeddah, using a two-stage cluster sample that randomly selected four schools from 85 public secondary schools for males. Data were obtained through a self-administered questionnaire containing questions on personal back...
Del R?o N?jera, Danyella; Santana, Natalia; Pel?ez-Ballestas, Ingris; Gonz?lez-Ch?vez, Susana A.; Qui?onez-Flores, Celia M.; Pacheco-Tena, C?sar
This study aimed to determine the prevalence of musculoskeletal (MSK) pain and rheumatic diseases in the Raramuri population (also known as Tarahumaras) who are an indigenous group in the northern state of Chihuahua in Mexico. We used the Community-Oriented Program for Control of Rheumatic Diseases (COPCORD) methodology. An analytical cross-sectional study was conducted including indigenous Raramuri aged ?18?years from communities settled in Chihuahua City. Subjects with positive MSK pain wer...
Luby Stephen P
Full Text Available Abstract Background Measuring recurrent infections such as diarrhoea or respiratory infections in epidemiological studies is a methodological challenge. Problems in measuring the incidence of recurrent infections include the episode definition, recall error, and the logistics of close follow up. Longitudinal prevalence (LP, the proportion-of-time-ill estimated by repeated prevalence measurements, is an alternative measure to incidence of recurrent infections. In contrast to incidence which usually requires continuous sampling, LP can be measured at intervals. This study explored how many more participants are needed for infrequent sampling to achieve the same study power as frequent sampling. Methods We developed a set of four empirical simulation models representing low and high risk settings with short or long episode durations. The model was used to evaluate different sampling strategies with different assumptions on recall period and recall error. Results The model identified three major factors that influence sampling strategies: (1 the clustering of episodes in individuals; (2 the duration of episodes; (3 the positive correlation between an individual's disease incidence and episode duration. Intermittent sampling (e.g. 12 times per year often requires only a slightly larger sample size compared to continuous sampling, especially in cluster-randomized trials. The collection of period prevalence data can lead to highly biased effect estimates if the exposure variable is associated with episode duration. To maximize study power, recall periods of 3 to 7 days may be preferable over shorter periods, even if this leads to inaccuracy in the prevalence estimates. Conclusion Choosing the optimal approach to measure recurrent infections in epidemiological studies depends on the setting, the study objectives, study design and budget constraints. Sampling at intervals can contribute to making epidemiological studies and trials more efficient, valid
Francesco Chirico; Giuseppe Ferrari; Giuseppe Taino; Enrico Oddone; Ines Giorgi; Marcello Imbriani
Introduction: Over the past two decades, numerous studies on indoor air and the Sick Building Syndrome (SBS) have been conducted, mostly in office environments. However, there is little knowledge about SBS in police officers. This study was aimed to fill this gap. Methods: A cross-sectional questionnaire survey was conducted in 2016 at the Triveneto Penitentiary Center, Northern Italy. Chi-square was used to test the difference of prevalence between office workers (OWs) and correctional of...
Full Text Available Abstract Background Overweight and obesity are considered a serious health problem. There are little data on the prevalence of overweight and obesity among the Yi ethnic group in China. This study aimed to investigate the epidemiologic features of overweight/obesity among Chinese Yi nationality. Methods A cross-sectional study, including 1255 subjects aged 20-75 years, was carried out in Liangshan Yi Autonomous Prefecture of Sichuan province from 2007 to 2008. Overweight/overall obesity was defined by World Health Organization (WHO or the Working Group on Obesity in China. Results Overall, the prevalence of overweight and obesity was 19.0% and 2.9%, respectively, based on the WHO definition, while it was 21.0% and 7.4%, respectively, according to the Working Group on Obesity in China, which is similar to data reported in the 2002 Chinese National Nutrition and Health Survey. Urban residents had a significantly higher prevalence of obesity (WHO criteria: 4.3% vs 1.7% p = 0.008; China criteria: 11.4% vs 3.7%, p p p Conclusions The prevalence of overweight/obesity in the Yi nationality is similar to that in Chinese adults 5 years ago. However, urban residents have a much higher prevalence of overweight/obesity than their rural counterparts. Lifestyle and diet patterns associated with socioeconomic status may explain the difference between urban and rural residents. The prevention of overweight/obesity among urban inhabitants deserves more attention in national health education programs.
Ostadimoghaddam, Hadi; Fotouhi, Akbar; Hashemi, Hassan; Yekta, Abbasali; Heravian, Javad; Rezvan, Farhad; Ghadimi, Hamidreza; Rezvan, Bijan; Khabazkhoob, Mehdi
Refractive errors are a common eye problem. Considering the low number of population-based studies in Iran in this regard, we decided to determine the prevalence rates of myopia and hyperopia in a population in Mashhad, Iran. Cross-sectional population-based study. Random cluster sampling. Of 4453 selected individuals from the urban population of Mashhad, 70.4% participated. Refractive error was determined using manifest (age > 15 years) and cycloplegic refraction (age ≤ 15 years). Myopia was defined as a spherical equivalent of -0.5 diopter or worse. An spherical equivalent of +0.5 diopter or worse for non-cycloplegic refraction and an spherical equivalent of +2 diopter or worse for cycloplegic refraction was used to define hyperopia. Prevalence of refractive errors. The prevalence of myopia and hyperopia in individuals ≤ 15 years old was 3.64% (95% CI: 2.19-5.09) and 27.4% (95% CI: 23.72-31.09), respectively. The same measurements for subjects > 15 years of age was 22.36% (95% CI: 20.06-24.66) and 34.21% (95% CI: 31.57-36.85), respectively. Myopia was found to increase with age in individuals ≤ 15 years and decrease with age in individuals > 15 years of age. The rate of hyperopia showed a significant increase with age in individuals > 15 years. The prevalence of astigmatism was 25.64% (95% CI: 23.76-27.51). In children and the elderly, hyperopia is the most prevalent refractive error. After hyperopia, astigmatism is also of importance in older ages. Age is the most important demographic factor associated with different types of refractive errors. © 2011 The Authors. Clinical and Experimental Ophthalmology © 2011 Royal Australian and New Zealand College of Ophthalmologists.
Valentina Forni Ogna
Full Text Available Background. Previous observations found a high prevalence of obstructive sleep apnea (OSA in the hemodialysis population, but the best diagnostic approach remains undefined. We assessed OSA prevalence and performance of available screening tools to propose a specific diagnostic algorithm. Methods. 104 patients from 6 Swiss hemodialysis centers underwent polygraphy and completed 3 OSA screening scores: STOP-BANG, Berlin’s Questionnaire, and Adjusted Neck Circumference. The OSA predictors were identified on a derivation population and used to develop the diagnostic algorithm, which was validated on an independent population. Results. We found 56% OSA prevalence (AHI ≥ 15/h, which was largely underdiagnosed. Screening scores showed poor performance for OSA screening (ROC areas 0.538 [SE 0.093] to 0.655 [SE 0.083]. Age, neck circumference, and time on renal replacement therapy were the best predictors of OSA and were used to develop a screening algorithm, with higher discriminatory performance than classical screening tools (ROC area 0.831 [0.066]. Conclusions. Our study confirms the high OSA prevalence and highlights the low diagnosis rate of this treatable cardiovascular risk factor in the hemodialysis population. Considering the poor performance of OSA screening tools, we propose and validate a specific algorithm to identify hemodialysis patients at risk for OSA for whom further sleep investigations should be considered.
Dedov, Ivan; Shestakova, Marina; Benedetti, Massimo Massi; Simon, Dominique; Pakhomov, Iakov; Galstyan, Gagik
To estimate type 2 diabetes mellitus (T2DM) prevalence in Russian adults. NATION is a national, epidemiological, cross-sectional study, conducted in Russia. In adults (aged 20-79 years), recruitment was stratified by age, sex, geographic region and settlement type to obtain a representative sample. Recruitment was in public areas with high numbers of people. T2DM was diagnosed by glycated haemoglobin A1c (HbA1c) levels (diabetes: HbA1c ≥6.5% [≥48mmol/mol]; pre-diabetes: HbA1c ≥5.7 to diabetic. T2DM prevalence increased with age (up to 70 years) and was higher among females than males (6.1% vs. 4.7%, pdiabetes and T2DM tended to increase with increasing body mass index. T2DM prevalence was higher in rural versus urban populations (6.7% vs. 5.0%, pdiabetes, T2DM prevalence was 5.4%, and 54% of subjects with diabetes were previously undiagnosed. These results may help to develop a new T2DM predictive, preventative and management programme in Russia. Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.
Vilppula, Anitta; Kaukinen, Katri; Luostarinen, Liisa; Krekelä, Ilkka; Patrikainen, Heikki; Valve, Raisa; Mäki, Markku; Collin, Pekka
Celiac disease may emerge at any age, but little is known of its appearance in elderly people. We evaluated the prevalence of the condition in individuals over 55 years of age, and determined the incidence of biopsy-proven celiac disease (CDb) and celiac disease including seropositive subjects for anti-tissue transglutaminase antibodies (CDb+s). The study based on prevalence figures in 2815 randomly selected subjects who had undergone a clinical examination and serologic screening for celiac disease in 2002. A second screening in the same population was carried out in 2005, comprising now 2216 individuals. Positive tissue transglutaminase antibodies were confirmed with small bowel biopsy. Within three years the prevalence of CDb increased from 2.13 to 2.34%, and that of CDb+s from 2.45 to 2.70%. Five new cases were found among patients previously seronegative; two had minor abdominal symptoms and three were asymptomatic. The incidence of celiac disease in 2002-2005 was 0.23%, giving an annual incidence of 0.08% in this population. The prevalence of celiac disease was high in elderly people, but the symptoms were subtle. Repeated screening detected five biopsy-proven cases in three years, indicating that the disorder may develop even in the elderly. Increased alertness to the disorder is therefore warranted.
Full Text Available Abstract Background Celiac disease may emerge at any age, but little is known of its appearance in elderly people. We evaluated the prevalence of the condition in individuals over 55 years of age, and determined the incidence of biopsy-proven celiac disease (CDb and celiac disease including seropositive subjects for anti-tissue transglutaminase antibodies (CDb+s. Methods The study based on prevalence figures in 2815 randomly selected subjects who had undergone a clinical examination and serologic screening for celiac disease in 2002. A second screening in the same population was carried out in 2005, comprising now 2216 individuals. Positive tissue transglutaminase antibodies were confirmed with small bowel biopsy. Results Within three years the prevalence of CDb increased from 2.13 to 2.34%, and that of CDb+s from 2.45 to 2.70%. Five new cases were found among patients previously seronegative; two had minor abdominal symptoms and three were asymptomatic. The incidence of celiac disease in 2002–2005 was 0.23%, giving an annual incidence of 0.08% in this population. Conclusion The prevalence of celiac disease was high in elderly people, but the symptoms were subtle. Repeated screening detected five biopsy-proven cases in three years, indicating that the disorder may develop even in the elderly. Increased alertness to the disorder is therefore warranted.
Bouzid, Maha; Halai, Kapil; Jeffreys, Danielle; Hunter, Paul R
Giardia has a wide range of host species and is a common cause of diarrhoeal disease in humans and animals. Companion animals are able to transmit a range of zoonotic diseases to their owners including giardiasis, but the size of this risk is not well known. The aim of this study was to analyse giardiasis prevalence rates in dogs and cats worldwide using a systematic search approach. Meta-analysis enabled to describe associations between Giardia prevalence and various confounding factors. Pooled prevalence rates were 15.2% (95% CI 13.8-16.7%) for dogs and 12% (95% CI 9.2-15.3%) for cats. However, there was very high heterogeneity between studies. Meta-regression showed that the diagnostic method used had a major impact on reported prevalence with studies using ELISA, IFA and PCR reporting prevalence rates between 2.6 and 3.7 times greater than studies using microscopy. Conditional negative binomial regression found that symptomatic animals had higher prevalence rates ratios (PRR) than asymptomatic animals 1.61 (95% CI 1.33-1.94) in dogs and 1.94 (95% CI 1.47-2.56) in cats. Giardia was much more prevalent in young animals. For cats >6 months, PRR=0.47 (0.42-0.53) and in dogs of the same age group PRR=0.36 (0.32-0.41). Additionally, dogs kept as pets were less likely to be positive (PRR=0.56 (0.41-0.77)) but any difference in cats was not significant. Faecal excretion of Giardia is common in dogs and slightly less so in cats. However, the exact rates depend on the diagnostic method used, the age and origin of the animal. What risk such endemic colonisation poses to human health is still unclear as it will depend not only on prevalence rates but also on what assemblages are excreted and how people interact with their pets. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.
Kari A O Tikkinen
Full Text Available BACKGROUND: In earlier studies, one in six adults had overactive bladder which may impair quality of life. However, earlier studies have either not been population-based or have suffered from methodological limitations. Our aim was to assess the prevalence of overactive bladder symptoms, based on a representative study population and using consistent definitions and exclusions. METHODOLOGY/PRINCIPAL FINDINGS: The aim of the study was to assess the age-standardized prevalence of overactive bladder defined as urinary urgency, with or without urgency incontinence, usually with urinary frequency and nocturia in the absence of urinary tract infection or other obvious pathology. In 2003-2004, a questionnaire was mailed to 6,000 randomly selected Finns aged 18-79 years who were identified from the Finnish Population Register Centre. Information on voiding symptoms was collected using the validated Danish Prostatic Symptom Score, with additional frequency and nocturia questions. Corrected prevalence was calculated with adjustment for selection bias due to non-response. The questionnaire also elicited co-morbidity and socio-demographic information. Of the 6,000 subjects, 62.4% participated. The prevalence of overactive bladder was 6.5% (95% CI, 5.5% to 7.6% for men and 9.3% (CI, 7.9% to 10.6% for women. Exclusion of men with benign prostatic hyperplasia reduced prevalence among men by approximately one percentage point (to 5.6% [CI, 4.5% to 6.6%]. Among subjects with overactive bladder, urgency incontinence, frequency, and nocturia were reported by 11%, 23%, and 56% of men and 27%, 38%, and 40% of women, respectively. However, only 31% of men and 35% of women with frequency, and 31% of subjects of both sexes with nocturia reported overactive bladder. CONCLUSIONS/SIGNIFICANCE: Our results indicate a prevalence of overactive bladder as low as 8% suggesting that, in previous studies, occurrence has been overestimated due to vague criteria and selected study
Stevens, Jennifer P.
Background Dyspnea (breathing discomfort) can be as powerfully aversive as pain, yet is not routinely assessed and documented in the clinical environment. Routine identification and documentation of dyspnea is the first step to improved symptom management and it may also identify patients at risk of negative clinical outcomes. Objective To estimate the prevalence of dyspnea and of dyspnea-associated risk among hospitalized patients. Design Two pilot prospective cohort studies. Setting Single academic medical center. Patients Consecutive patients admitted to four inpatient units: cardiology, hematology/oncology, medicine, and bariatric surgery. Measurements In Study 1, nurses documented current and recent patient-reported dyspnea at the time of the Initial Patient Assessment in 581 inpatients. In Study 2, nurses documented current dyspnea at least once every nursing shift in 367 patients. We describe the prevalence of burdensome dyspnea, and compare it to pain. We also compared dyspnea ratings with a composite of adverse outcomes: 1) receipt of care from the hospital’s rapid response system, 2) transfer to the intensive care unit, or 3) death in hospital. We defined burdensome dyspnea as a rating of 4 or more on a 10-point scale. Results Prevalence of burdensome current dyspnea upon admission (Study 1) was 13% (77 of 581, 95% CI 11%-16%). Prevalence of burdensome dyspnea at some time during the hospitalization (Study 2) was 16% (57 of 367, 95% CI 12%-20%). Dyspnea was associated with higher odds of a negative outcome. Conclusions In two pilot studies, we identified a significant symptom burden of dyspnea in hospitalized patients. Patients reporting dyspnea may benefit from a more careful focus on symptom management and may represent a population at greater risk for negative outcomes. PMID:27070144
Seoane-Mato, Daniel; Sánchez-Piedra, Carlos; Silva-Fernández, Lucía; Sivera, Francisca; Blanco, Francisco J; Pérez Ruiz, Fernando; Juan-Mas, Antonio; Pego-Reigosa, José M; Narváez, Javier; Quilis Martí, Neus; Cortés Verdú, Raúl; Antón-Pagés, Fred; Quevedo Vila, Víctor; Garrido Courel, Laura; Del Amo, Natividad Del Val; Paniagua Zudaire, Inmaculada; Añez Sturchio, Gustavo; Medina Varo, Fermín; Ruiz Tudela, María Del Mar; Romero Pérez, Antonio; Ballina, Javier; Brandy García, Anahy; Fábregas Canales, Dolores; Font Gayá, Teresa; Bordoy Ferrer, Carolina; González Álvarez, Beatriz; Casas Hernández, Laura; Álvarez Reyes, Fátima; Delgado Sánchez, Mónica; Martínez Dubois, Cristina; Sánchez-Fernández, Simón Ángel; Rojas Vargas, Luisa Marena; García Morales, Paula Virginia; Olivé, Alejandro; Rubio Muñoz, Paula; Larrosa, Marta; Navarro Ricos, Noemí; Graell Martín, Eduard; Chamizo, Eugenio; Chaves Chaparro, Lara; Rojas Herrera, Sara; Pons Dolset, Jordi; Polo Ostariz, Miguel Ángel; Ruiz-Alejos Garrido, Susana; Macía Villa, Cristina; Cruz Valenciano, Ana; González Gómez, María Luisa; Morcillo Valle, Mercedes; Palma Sánchez, Deseada; Moreno Martínez, María José; Mayor González, Marta; Atxotegi Sáenz de Buruaga, Joana; Urionagüena Onaindia, Irati; Blanco Cáceres, Boris Anthony; Díaz-González, Federico; Bustabad, Sagrario
To describe the methodology of the EPISER 2016 (study of the prevalence of rheumatic diseases in adult population in Spain), as well its strengths and limitations. The aim of this study is to estimate the prevalence of rheumatoid arthritis (RA), psoriatic arthritis (PsA), ankylosing spondylitis (AS), systemic lupus erythematosus (SLE), Sjögren's syndrome (SS), osteoarthritis (knee, hip, hands, and cervical and lumbar spine), fibromyalgia, gout and clinical osteoporotic fracture. Population-based, multicenter, cross-sectional study, with the participation of 45 municipalities in the 17 Spanish autonomous communities. The reference population will consist of adults aged 20 years and over residing in Spain. A computer-assisted telephone interview (CATI) system will be used for data collection. Diagnostic suspicions and diagnoses received by the participants will be studied by rheumatologists in the referral hospitals in the selected municipalities. the prevalence of the rheumatic diseases will be calculated using estimators and their 95% confidence intervals. Weights will be calculated in each of the sampling stages in accordance with the probability of selection. The distribution of the population in Spain will be obtained from the Spanish Statistics Institute. Sociodemographic and lifestyle changes over the last 16 years justify EPISER 2016. This study will provide current data about the prevalences of RA, AS, PsA, SLE, SS, osteoarthritis, fibromyalgia, gout and clinical osteoporotic fracture. The results will allow comparisons with studies from other countries and EPISER 2000. Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Reumatología y Colegio Mexicano de Reumatología. All rights reserved.
Aenumulapalli, Ashok; Kulkarni, Manoj Mohan; Gandotra, Achleshwar Ramnarain
Flat foot (pes planus) is a commonly observed disorder in clinical practice. The height of Medial Longitudinal Arch (MLA) determines the degree of pes planus. Majority of the previous studies on prevalence of flexible flat foot were done among children. The present study was undertaken to determine the prevalence of flexible flat foot among 18-21-year-old Indian adults using Navicular Drop Test (NDT) which is regarded as reliable and valid method to characterise MLA. The present study was a cross-sectional study in which Brody's Navicular Drop Test was performed in five hundred healthy subjects (250 males and 250 females) aged 18-21-year-old. Navicular Drop (ND) of ≥ 10 mm was regarded as flexible flat foot. Statistical analysis was done using SPSS version 23.0. The prevalence of flexible flat foot was 13.6% (for males-12.8%; for females-14.4%). The median with Inter Quartile Range (IQR) for ND among males was 6 mm (4-8) and 6 mm (4-9) for right and left foot respectively. The median with IQR for ND among females was 6mm (4-10) and 7mm (3-8) for right & left respectively. The correlation of Right Side Navicular Drop (RND) with height and weight was significant, but insignificant correlation was found between RND and BMI. The correlation of Left Side Navicular Drop (LND) with height, weight and BMI of individuals was statistically not significant. The difference between the ND of males and female group was statistically not significant. The present study quantified the prevalence of flexible flat foot and gender wise normative values of ND among 18-21-year-old Indian adults. The information obtained by this study will be useful in the field of orthopaedics.
Shu, Shang-an; Chang, Christopher; Leung, Patrick S C
Global and regional studies on the prevalence of food allergies are plagued by inconsistent methodologies, variations in interpretation of results, and non-standardized study design. Hence, it becomes difficult to compare the prevalence of food allergies in different communities. This information would be useful in providing critical data that will enhance research to elucidate the nature of food allergies, and the role of gene-environment interactions in the sensitization of children and adults to foods. Testing methodologies range from questionnaires to objective in vitro and in vivo testing, to the gold standard, double-blind placebo-controlled food challenge (DBPCFC). Although considered the most accurate and reliable method in detecting the prevalence of food allergy, DBPCFC is not always practical in epidemiological studies of food allergy. On the other hand, multiple logistic regression studies have been done to determine predictability of the outcome of food challenges, and it appears that skin prick testing and in vitro-specific serum IgE are the best predictors. Future studies directed towards confirming the validity of these methods as well as developing algorithms to predict the food challenge outcomes are required, as they may someday become accessory tools to complement DBPCFC.
Evensen, O.; Lorenzen, Ellen
An immunohistochemical method is described for the detection of Flexibacter psychrophilus in formalin-fixed, parafiin-wax-embedded fry of rainbow trout. Rabbit antiserum as well as rainbow trout hyperimmune serum were used in the study. The distribution and tissue localization of the bacterium...... was compared in naturally and experimentally (intraperitoneal injections) infected fry by use of immunohistochemistry. This study showed that F. psychrophilus could be detected in paraffin-wax-embedded tissue of rainbow trout fry by immunohistochemistry. The principal immunohistochemical findings in naturally...... and experimentally infected fry showed that there was a localization of bacteria in the monocyte-macrophage system, in skin lesions, and in the retina and the choroid gland of the eye. The dermal changes included superficial or deep ulcers extending to the subcutaneous tissue or the musculature accompanied...
Gustavson, Kristin; Knudsen, Ann Kristin; Nesvåg, Ragnar; Knudsen, Gun Peggy; Vollset, Stein Emil; Reichborn-Kjennerud, Ted
Mental disorders often have onset early in life, contribute substantially to the global disease burden, and may interfere with young people's ability to complete age-relevant tasks in important developmental periods. However, knowledge about prevalence and course of mental disorders in young adulthood is sparse. The aim of the current study was to estimate prevalence and stability of mental disorders from the twenties to the thirties/forties. DSM-IV mental disorders were assessed with the Composite International Diagnostic Interview in two waves (1999-2004 and 2010-2011) in 1623 young adult Norwegian twins (63.2% women, aged 19-29 years in wave 1). In wave 1, the 12-month prevalence of any mental disorder among people in the twenties was 19.8% (men) and 32.4% (women), anxiety disorders: 9.6% (men) and 26.7% (women), anxiety disorders excluding specific phobias: 2.5% (men) and 6.9% (women), major depressive disorder (MDD): 4.4% (men) and 7.2% (women), and alcohol use disorder (AUD): 8.7% (men) and 4.4% (women). The prevalence of any mental disorder decreased from the twenties to the thirties/forties. This was due to a decrease in AUD and specific phobias. Anxiety disorders in the twenties predicted anxiety disorders and MDD ten years later, even when controlling for the association between these disorders in the twenties. MDD in the twenties predicted MDD ten years later. At both ages, two-week and 12-month prevalence estimates differed markedly for MDD - indicating an episodic course. Common mental disorders are highly prevalent among young adults in the twenties, and somewhat less prevalent in the thirties/forties. Those who suffer from one mental disorder in the twenties are at considerably increased risk for suffering from a disorder ten years later as well. This may have significant implications for young people's ability to attain education, establish a family, and participate in occupational life.
Mo, Jingfu; Wang, Lu; Au, William; Su, Min
Nowadays, coal workers' pneumoconiosis (CWP) is still believed to be the main occupational disease in China. However, information on the exact prevalence of the disease is not available. Therefore, the aims of our investigation were to provide the missing information in China by conducting a systematic evaluation of published data from 2001 to 2011 and to compare the prevalence of CWP with those in other countries. Published reports about the prevalence of CWP were searched from PudMed(English language databases), Foreign Medical Journal Full-Text Service Database (FMJS, English language databases), Chinese Journal Full-Text Database (CJFD, Chinese language databases), Chongqing VIP Chinese Science and Technology Journals Database (VIP, Chinese language databases), Chinese Biomedical Literature Database (CBM, Chinese language databases) and Chinese Medical Association Journals Database (CMAJ, Chinese language databases). The quality of identified reports was strictly evaluated using predetermined inclusion and exclusion criteria. Based on these criteria, 11 reports were selected. Then, the content of these reports were reviewed and the needed information was extracted. Meta-analysis was performed on the extracted data. The R2.15.1 software was applied for statistical analysis. The total populations from these reports were 173,646 and 10,821 for dust-exposed coal workers and patients with CWP, respectively. The pooled prevalence of CWP was 6.02% (95% CI: 3.43-9.26%) and the pooled rate of CWP patients combined with tuberculosis was 10.82% (95% CI: 8.26-13.66%). The prevalence was analyzed according to the geographic areas of the study, years of the investigation, duration of dust exposure, coal rank, stages of CWP, types of work and coal-mining categories, etc. Among them, the prevalence of CWP in locally owned mines (9.86%; 95% CI: 1.25-25.17%) was significantly higher than that of state-owned mines (4.83%; 95% CI: 2.35-8.13%) (P0.05). It was concluded that the
Engels, Thomas; Baglione, Quentin; Audibert, Martine; Viallefont, Anne; Mourji, Fouzi; El Alaoui Faris, Mustapha
Background Stroke is a growing public health concern in low- and middle- income countries. Improved knowledge about the association between socioeconomic status and stroke in these countries would enable the development of effective stroke prevention and management strategies. This study presents the association between socioeconomic status and the prevalence of stroke in Morocco, a lower middle-income country. Methods Data on the prevalence of stroke and stroke-related risk factors were collected during a large population-based survey. The diagnosis of stroke in surviving patients was confirmed by neurologists while health, demographic, and socioeconomic characteristics of households were collected using structured questionnaires. We used Multiple Correspondence Analysis to develop a wealth index based on characteristics of the household dwelling as well as ownership of selected assets. We used logistic regressions controlling for multiple variables to assess the statistical association between socioeconomic status and stroke. Findings Our results showed a significant association between household socioeconomic status and the prevalence of stroke. This relationship was non-linear, with individuals from both the poorest (mainly rural) and richest (mainly urban) households having a lower prevalence of stroke as compared to individuals with medium wealth level. The latter belonged mainly to urban households with a lower socioeconomic status. When taking into account the urban population only, we observed that a third of poorest households experienced a significantly higher prevalence of stroke compared to the richest third (OR = 2.06; CI 95%: 1.09; 3.89). Conclusion We conclude that individuals from the most deprived urban households bear a higher risk of stroke than the rest of the population in Morocco. This result can be explained to a certain extent by the higher presence of behavioral risk factors in this specific category of the population, which leads in
Viviane Gabriela Nascimento
Full Text Available CONTEXT AND OBJECTIVE: Brazil is undergoing a period of epidemiological transition associated with demographic and nutritional changes. The prevalence of obesity is also increasing in children and is causing numerous health problems that are becoming public health issues. The aim here was to evaluate the prevalence of overweight among children of two and three years of age. DESIGN AND SETTING: Cross-sectional study in municipal day care centers in Taubaté, state of São Paulo, Brazil. METHODS: Weight and height measurements were made on 447 preschool children forming a probabilistic randomized sample. Their body mass index (BMI was calculated. Their nutritional status was classified using the World Health Organization reference cutoff points (2006. Their mean weight, height and BMI were compared according to their age and sex. RESULTS: The mean values for the final sample (n = 447 were as follows: mean age: 38.6 months (± 3.5 and Z scores for: weight/height (W/H: 0.50 (± 1.22; height/age: -0.03 (± 1.07; weight/age (W/A: 0.51 (± 1.23; and BMI: 0.51(± 1.23. The prevalence of overweight children (BMI > 1 z was 28.86%, while the prevalence of underweight children (BMI < -2 z was 0.89%. There were no differences in mean BMI among the two and three-year age groups (P = 0.66. CONCLUSION: A high prevalence of overweight was observed in the sample of two and three-year-old children, with practically no malnutrition, thus showing that a significant nutritional transition may already be occurring, even in medium-sized cities of developing countries.
Full Text Available BACKGROUND: Stroke is a growing public health concern in low- and middle- income countries. Improved knowledge about the association between socioeconomic status and stroke in these countries would enable the development of effective stroke prevention and management strategies. This study presents the association between socioeconomic status and the prevalence of stroke in Morocco, a lower middle-income country. METHODS: Data on the prevalence of stroke and stroke-related risk factors were collected during a large population-based survey. The diagnosis of stroke in surviving patients was confirmed by neurologists while health, demographic, and socioeconomic characteristics of households were collected using structured questionnaires. We used Multiple Correspondence Analysis to develop a wealth index based on characteristics of the household dwelling as well as ownership of selected assets. We used logistic regressions controlling for multiple variables to assess the statistical association between socioeconomic status and stroke. FINDINGS: Our results showed a significant association between household socioeconomic status and the prevalence of stroke. This relationship was non-linear, with individuals from both the poorest (mainly rural and richest (mainly urban households having a lower prevalence of stroke as compared to individuals with medium wealth level. The latter belonged mainly to urban households with a lower socioeconomic status. When taking into account the urban population only, we observed that a third of poorest households experienced a significantly higher prevalence of stroke compared to the richest third (OR = 2.06; CI 95%: 1.09; 3.89. CONCLUSION: We conclude that individuals from the most deprived urban households bear a higher risk of stroke than the rest of the population in Morocco. This result can be explained to a certain extent by the higher presence of behavioral risk factors in this specific category of the population
Mohanty, D; Colah, R B; Gorakshakar, A C; Patel, R Z; Master, D C; Mahanta, J; Sharma, S K; Chaudhari, U; Ghosh, M; Das, S; Britt, R P; Singh, S; Ross, C; Jagannathan, L; Kaul, R; Shukla, D K; Muthuswamy, V
The population of India is extremely diverse comprising of more than 3,000 ethnic groups who still follow endogamy. Haemoglobinopathies are the commonest hereditary disorders in India and pose a major health problem. The data on the prevalence of β-thalassemias and other haemoglobinopathies in different caste/ethnic groups of India is scarce. Therefore the present multicentre study was undertaken in six cities of six states of India (Maharashtra, Gujarat, West Bengal, Assam, Karnataka and Punjab) to determine the prevalence of haemoglobinopathies in different caste/ethnic groups using uniform methodology. Fifty-six thousand seven hundred eighty individuals (college students and pregnant women) from different caste/ethnic groups were screened. RBC indices were measured on an automated haematology counter while the percentage of HbA(2), HbF and other abnormal Hb variants were estimated by HPLC on the Variant Hemoglobin Testing System. The overall prevalence of β-thalassemia trait was 2.78 % and varied from 1.48 to 3.64 % in different states, while the prevalence of β-thalassemia trait in 59 ethnic groups varied from 0 to 9.3 %. HbE trait was mainly seen in Dibrugarh in Assam (23.9 %) and Kolkata in West Bengal (3.92 %). In six ethnic groups from Assam, the prevalence of HbE trait varied from 41.1 to 66.7 %. Few subjects with δβ-thalassemia, HPFH, HbS trait, HbD trait, HbE homozygous and HbE β-thalassemia as well as HbS homozygous and HbS-β-thalassemia (India.
Michael Lynge Pedersen
Full Text Available Background: Within the last 20 years, the prevalence of gestational diabetes mellitus (GDM has been reported to be increasing worldwide in correlation with ethnic and geographic variations. The actual prevalence of GDM throughout all of Greenland remains unknown. Objective: The aim of this study was to estimate the prevalence of GDM among Greenlanders and non-Greenlanders living in Greenland and to estimate the efficacy of testing for GDM. Design: This study was performed as an observational, cross-sectional study including all women with permanent address in Greenland who had given birth to a singleton during 2014. The prevalence of GDM was calculated as the proportion of all pregnant women tested with a 75-g 2-h glucose tolerance test who had a 2-h capillary whole-blood glucose value of 8.5 mmol/l or above. Testing efficacy was calculated as the proportion of women who fulfilled the testing criteria who were actually tested in Greenland in 2014. Results: A total of 794 women (727 Greenlanders and 67 non-Greenlanders were included in the study. The prevalence of GDM among tested women was 3.3% (confidence interval, CI: 0.9–5.6 among Greenlanders and 12.5% (CI: 0–25.7 among non-Greenlanders, corresponding, respectively, to 1.0% (CI: 0.3–1.3 and 4.5% (CI: 0–9.4 of all singleton pregnancies in Greenland in 2014. The overall testing efficacy was 69.0% among all eligible residents of Greenland and 85.1% among eligible residents in the capital city, Nuuk. Conclusion: In conclusion, the prevalence of GDM seems quite low in Greenland. Although diagnostic testing activity has improved within the last 6 years, still around one-third of all pregnant women in all Greenland fulfilling the testing criteria were not tested. Universal testing for GDM may be needed to improve testing of GDM in Greenland.
Hooff, Miranda Van; McFarlane, Alexander C.; Davies, Christopher E.; Searle, Amelia K.; Fairweather-Schmidt, A. Kate; Verhagen, Alan; Benassi, Helen; Hodson, Stephanie E.
Background The Australian Defence Force (ADF) Mental Health Prevalence and Wellbeing Study (MHPWS) is the first study of mental disorder prevalence in an entire military population. Objective The MHPWS aims to establish mental disorder prevalence, refine current ADF mental health screening methods, and identify specific occupational factors that influence mental health. This paper describes the design, sampling strategies, and methodology used in this study. Method At Phase 1, approximately half of all regular Navy, Army, and Air Force personnel (n=24,481) completed self-report questionnaires. At Phase 2, a stratified sub-sample (n=1,798) completed a structured diagnostic interview to detect mental disorder. Based on data from non-responders, data were weighted to represent the entire ADF population (n=50,049). Results One in five ADF members met criteria for a 12-month mental disorder (22%). The most common disorder category was anxiety disorders (14.8%), followed by affective (9.5%) and alcohol disorders (5.2%). At risk ADF sub-groups were Army personnel, and those in the lower ranks. Deployment status did not have an impact on mental disorder rates. Conclusion This study has important implications for mental health service delivery for Australian and international military personnel as well as contemporary veterans. PMID:25206944
P N Jain
Full Text Available Background and Aims: Acute neuropathic pain (ANP is an under-recognised and under-diagnosed condition and often difficult to treat. If left untreated, it may further transform into persistent post-operative chronic pain leading to a disability. Aims: This prospective study was undertaken on 300 patients to identify the prevalence of ANP in the post-operative period by using a neuropathic pain detection questionnaire tool. Methods: This is an open-label study in which patients with six different types of cancer surgeries (Thoracic, gastro-intestinal, gynae/urology, bone/soft-tissue, head and neck and breast subgroups-50 each were included for painDETECT questionnaire tool on the 2 nd and 7 th day surgery. Results: This study found a 10% point prevalence of ANP. Analysis showed that 25 patients had ′possible′ ANP, the maximum from urological cancer surgery (6 followed by thoracic surgery (5. Five patients were found to have ′positive′ ANP including 2 groin node dissection, 2 hemipelvectomy and 1 oesophagectomy. Conclusion: Significant relationship between severity of post-operative pain was found with the occurrence of ANP in the post-operative period requiring a special attention to neuropathic pain assessment. Larger studies are required with longer follow-up to identify accurately the true prevalence and causative factors of ANP after surgery.
Full Text Available Introduction: Children’s obesity is strong predictor of obesity in adulthood, which increases the incidence of related diseases such as metabolic syndrome. Fatty liver is an abnormality related to metabolic syndrome with higher prevalence in obese children according to some previous studies. This study investigates the presence of Sonographic Fatty Liver (SFL in Iranian obese children in comparison with normal and overweight children. Methods: This was a cross-sectional study on 962 randomly selected children between the ages of 6 to 18 years. The subjects were divided into three groups of normal, overweight and obese based on body mass index (BMI. A questionnaire including demographic and anthropometrical characteristics was filled for each case. To detect the presence of SFL, all the subjects underwent assessment with ultrasonography by a radiologist who was not aware of their BMI. The incidence of SFL was determined on the basis of ultrasonographic diagnosis criteria. Results: The prevalence of SFL in obese children was 54.4% that was significantly higher than overweight (10.5% and normal children (1%. There was no significant difference in prevalence of SFL between sexes. Conclusion: The high rate of detected SFL in obese children in this study suggests that Iranian obese children are at risk of metabolic syndrome. It is therefore necessary to develop some plan to control obesity and its late complications.
Gui, Xin-Yu; Yu, Xiao-Ling; Jin, Hong-Zhong; Zuo, Ya-Gang; Wu, Chao
Psoriasis, a chronic autoimmune skin disorder, is believed to contribute to cardiovascular diseases and metabolic syndrome. Psoriasis's association with the components of metabolic syndrome has been reported previously. However, large-scale cross-sectional studies about psoriasis and metabolic syndrome are rare in China. We assessed the prevalence of metabolic syndrome in Chinese psoriasis patients and controls. A total of 859 psoriasis patients and 1,718 controls were recruited in an age- and sex-matched cross-sectional study. Metabolic syndrome occurred in 14.3% of the psoriasis patients as opposed to 10.0% of the control participants (P = 0.001). Psoriasis patients had a higher prevalence of overweight/obesity, hyperglycemia and dyslipidemia when compared with controls. Meanwhile, psoriasis patients with metabolic syndrome were older, and had an older age of onset and a longer disease duration when compared with those without metabolic syndrome. The prevalence of metabolic syndrome is higher in the Chinese psoriatic population, which can favor cardiovascular events. The present study strengthens the value of treating psoriasis patients not only dealing with the skin lesions, and we suggest appropriate screening and relevant health education be carried out in the treatment of psoriasis patients. © 2017 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd.
Full Text Available Objective: The purpose of the present study was to investigate the prevalence of behavioral disorder among of primary school students in Abhar. Materials & Methods: 600 primary school students (300 boys and 300 girls of public primary schools in Abhar, Iran were used as the sample for this study. Their behavioral disorder scores were obtained by means of Rutter's teachers' questionnaire. Results: Using a cutpoint of 9, the prevalence rate of behavioral disorder was 43.3% among subjects, with boys showing higher rates of behavioral disorder (44.7% than girls (42% but no significant differences were found by gender on rates of behavioral disorders. 6.3 percent of the subjects had severe behavioral disorder. Conclusion: Academic achievement was negatively related to behavioral disorder. Significant differences were found by mothers' education on children’s rates of behavioral disorder. Significant differences were found by fathers' occupation on children’s rates of behavioral disorder.
Cecilie Schou Andreassen