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Tissue Biopsies (tissue + biopsy)
Kinds of Tissue Biopsies Selected AbstractsChronic lymphocytic leukemia presenting as cutaneous and bone involvementINTERNATIONAL JOURNAL OF DERMATOLOGY, Issue 1 2001Maria P. Stefanidou MD An 84-year-old man had a 3-year history of a progressive, painless, papulonodular eruption, that was particularly prominent on the face and extremities. Physical examination revealed firm, bluish-red nodules and plaques, located on the tip of the nose, the cheeks, ears, and distal digits. Skin lesions produced a leonine facies (Fig. 1), deformities of the fingers and toes, finger clubbing, and onyxis. An identical lesion was seen on a postoperational scar on the left cheek. The mucous membranes were spared. The patient had anterior and posterior cervical and bilateral axillary lymphadenopathy and splenomegaly. Figure 1. Leonine facies On admission, the peripheral blood count revealed 260,000/mm3 leukocytes (lymphocytes 97%, neutrophils 2%, and monocytes 1%), a hemoglobin level of 9.5 g/dL, and platelet count of 100,000/mm3. Hypogammaglobulinemia with reduction of immunoglobulin G (IgG) and IgM was found. Radiography of the fingers showed multiple osteolytic lesions of the phalanges and phalangette destruction of the left median finger (Fig. 2a,b). Computed tomography of the chest and abdomen revealed bilateral axillary, mediastinal, and para-aortic lymphadenopathy and spleen enlargement. Figure 2. X-Ray of the hands: (a) ,multiple osteolytic lesions of the phalanges and (b) ,partial destruction of the left median phalangette Skin biopsy specimens from the ear and finger lesions showed a massive nonepidermal leukemic infiltration in the papillary and reticular dermis, with a grenz zone consisting of small lymphocytes (Fig. 3). Figure 3. Skin biopsy (hematoxylin and eosin, ×,250). Massive leukemic infiltration consisting of small lymphocytes. Subepidermally, a grenz zone of connective tissue is noted Biopsy of the enlarged cervical lymph node showed a diffuse infiltration with lymphocytes. Tissue biopsy from a finger lytic lesion revealed infiltration of bone trabecular and fibrous tissue with a dense population of small- and medium-sized lymphocytes. Immunohistochemical study of cutaneous and bone lesions showed that the infiltrate in both biopsies consisted mainly of B lymphocytes (CD20+, CD45R+, CD45Ro,, OPD4,). Peripheral blood smear had a B-cell phenotype (CD19 98%, CD20 97%, CD23 99%, CD25 40%, CD5 90%, HLA-DR 100%). Bone marrow smear and immunophenotyping surface marker analysis found a diffuse pattern of B-lymphocytic infiltration. A diagnosis of B-cell chronic lymphocytic leukemia stage C (Binet staging system), with specific cutaneous and bone lesions, was established. The patient received chemotherapy with chlorambucil and methylprednisolone, which resulted in improvement of the hematologic profile. Two years later, the cutaneous lesions showed partial remission. [source] Imiquimod in the treatment of eyelid basal cell carcinomaACTA OPHTHALMOLOGICA, Issue 5 2007Jari Leppälä ABSTRACT. Purpose:, To assess the efficacy and safety of topical imiquimod 5% cream in the treatment of eyelid basal cell carcinoma. Methods:, Four patients with eyelid basal cell carcinoma were treated with imiquimod once daily, 5 days per week, for 6 weeks. Tissue biopsy was taken and clinical examination with slit-lamp microscopy was performed at the beginning of the study and after a follow-up of 12 weeks. Photographic follow-up was performed from the baseline visit to 6, 12 and 26 weeks. Results:, In the 12-week follow-up after imiquimod treatment, histopathological tissue sample analysis showed no signs of basal cell carcinoma in any of the patients. Conclusions:, The results indicate that 5% imiquimod cream with topical administration may represent a new therapy option for eyelid basal cell carcinoma. [source] Current diagnostic approaches to invasive candidiasis in critical care settingsMYCOSES, Issue 5 2010Javier Pemán Summary For the specialist, the management of invasive candidiasis infections, from diagnosis to selection of the therapeutic protocol, is often a challenge. Although early diagnosis and treatment are associated with a better prognosis, apart from cases with positive blood cultures or fluid/tissue biopsy, diagnosis is neither sensitive nor specific, relying on many different factors, clinical and laboratory findings but there is certainly a need for the specific markers in this disease. Recently, new serodiagnostic assays as Candida albicans germ-tube antibodies or (1,3)-,- d -glucan detection and molecular techniques for the detection of fungal-specific DNA have been developed with controversial results in critical care setting. One of the main features in diagnosis is the evaluation of risk factor for infection, which will identify patients in need of preemptive or empirical treatment. Clinical scores were built from those risk factors. For these reasons, an approach to the new diagnosis tools in the clinical mycology laboratory and an analysis of the new prediction rules and its application situations has been made. Currently, the combination of prediction rules and non-culture microbiological tools could be the clue for improving the diagnosis and prognosis of invasive fungal infections in critically ill patients. [source] Clinical applications of laser scanning cytometryCYTOMETRY, Issue 3 2002Attila Tárnok Abstract This study reviews existing and potential clinical applications of laser scanning cytometry (LSC) and outlines possible future developments. LSC provides a technology for solid phase cytometry. Fluorochrome-labeled specimens are immobilized on microscopic slides that are placed on a conventional epifluorescence microscope and analyzed by one or two lasers. Data comparable to flow cytometry are generated. In addition, the position of each event is recorded, a feature that allows relocalization and visualization of each measured event. The major advantage of LSC compared with other cytometric methods is the combination of two features: (a) the minimal clinical sample volume needed and (b) the connection of fluorescence data and morphological information for the measured event. Since the introduction of LSC, numerous methods have been established for the analysis of cells, cellular compartments, and tissues. Although most cytometric methods use only two or three colors, the characterization of specimens with up to five fluorochromes is possible. Most clinical applications have been designed to determine ploidy and immunophenotype; other applications include analyses of tissue biopsies and sections, fluorescence in situ hybridization, and the combination of vital and nonvital information on a single-cell basis. With the currently available assays, LSC has proven its wide spectrum of clinical applicability in slide-based cytometry and can be introduced as a standard technology in multiple clinical settings. Cytometry (Clin. Cytometry) 50:133,143, 2002. © 2002 Wiley-Liss, Inc. [source] Comparison of p16INK4A and Hybrid Capture® 2 human papillomavirus testing as adjunctive tests in liquid-based gynecologic SurePathÔ preparationsDIAGNOSTIC CYTOPATHOLOGY, Issue 3 2008Aziza Nassar M.D., F.I.A.C. Abstract p16INK4a, cyclin-dependent kinase inhibitor, is functionally inactivated in many tumors, including cervical cancer. We compared p16INK4A immunocytochemical staining and Hybrid Capture® 2 (HCII) on SurePathÔ specimens using tissue biopsies (as the gold standard). Their utility in a spectrum of atypical and preneoplastic lesions, and their ability to accurately identify underlying lesions of CIN II or greater was assessed using biopsy follow-up data. One-hundred and seventeen residual SurePathÔ samples were collected: 43 atypical squamous cells of undetermined significance (ASCUS), 47 low-grade (LGSIL), and 27 high-grade (HGSIL) squamous intraepithelial lesions. Two slides were prepared from each sample; one stained with the SurePathÔ autocyte stain and one immunostained using the CINtecÔ p16INK4a Cytology Kit (Dakocytomation). High-risk HPV testing was performed using the HCII DNA test (Digene, Gaithersburg, MD). Available tissue biopsy follow-up data was retrieved. p16INK4a was positive in 32.6% (14/43) ASCUS, 46.8% (22/47) LGSIL, and 48.1% (13/27) HGSIL specimens. HCII DNA test was positive in 41.9% (18/43) ASCUS, 78.7% (37/47) LGSIL, and 96.3% (26/27) HGSIL samples. The sensitivity, specificity, positive (PPV) and negative (NPV) predictive values of p16INK4a and HCII were: 58.7% and 89.8%, 58.6% and 34.6%, 69.2% and 72.1%, 47.2% and 64.3%, respectively. In patients with cervical biopsies, the PPV of HCII (92.3%) results for a biopsy with CINII/III was significantly higher than the PPV of p16INK4a (52%) (P = 0.001). Using liquid-based cytology specimens, HCII is a more sensitive test than p16INK4a for detection of abnormal cytology. HCII has a higher PPV than p16INK4a for identifying CIN II/III. Diagn. Cytopathol. 2008;36:142,148. © 2008 Wiley-Liss, Inc. [source] Urinary bladder biopsy with denuded mucosa: Denuding cystitis,Cytopathologic correlatesDIAGNOSTIC CYTOPATHOLOGY, Issue 5 2004Anil V. Parwani M.D., Ph.D. Abstract Denuding cystitis is often encountered in tissue biopsies of bladder mucosa performed by either cold-cup forceps or wire loop electrocautery to evaluate hematuria or to rule out recurrent urothelial carcinoma. Lack of urothelium in these biopsies is often a frustrating experience, leading to a nonspecific interpretation. In this study, 151 cases of denuding cystitis were retrieved from the surgical pathology files of The Johns Hopkins Hospital over a 4-year period (1996,1999). Patients under the age of 40 years and outside consultation material were excluded. Of the 151 cases of denuding cystitis, 48 patients were identified who had concurrent urinary cytologic studies. Of these patients, 35 were male (73%) and 13 were female (27%). Patient ages ranged from 43 to 85 years (mean, 67). Twenty-six of these 48 patients (54%) had at least one concurrently positive urinary cytology, which was histologically confirmed. All except three cases were high-grade urothelial carcinoma with the following histologic subtypes: flat carcinoma in situ (n = 11), noninvasive papillary (n = 9), and invasive urothelial carcinoma (n = 3). We conclude that urinary cytology is a sensitive modality that detects exfoliated carcinoma cells in patients with a histologic diagnosis of denuding cystitis. An inconclusive diagnosis of denuding cystitis on tissue might be related to biopsy method and technique, small sample size, or biopsy of cystoscopically abnormal urothelium that is denuded. A cytologic diagnosis of high-grade urothelial carcinoma in these cases leads to a timely clinical intervention for optimal patient management. Diagn. Cytopathol. 2004;30:297,300. © 2004 Wiley-Liss, Inc. [source] Cover Picture: Electrophoresis 13'09ELECTROPHORESIS, Issue 13 2009Article first published online: 20 JUL 200 Issue 13 is a special issue on "CE and CEC of Amino Acids, Peptides and Proteins" assembling 19 papers on various topics including fast, high efficient and high sensitive "CE and CEC techniques for quality control and purity determination of native and (bio)synthetic amino acids, peptides and proteins, for monitoring of their synthesis, isolation, chemical derivatization and enzymatic digestion and also for investigation of their interactions with other molecules. New methodologies, such as electrodialysis for sample preparation, chiral ligand-exchange CE, immunoaffinity CE, affinity capillary isoelectric focusing, combination of transient isotachophoretic preconcentration with capillary zone electophoresis (CZE) analysis, two-dimensional CE-mass spectrometry (MS) separations and advances in high-sensitive CE-laser induced fluorescence (LIF) and CE-electrochemiluminescence detection schemes, are widely presented here. The applications of CE and CEC methods include chiral analysis of amino acids, determination of low abundant amino acids, peptides and proteins in complex matrices, such as human and animal body fluids and tissue biopsies, and profiling of cell lysates and recombinant proteins, e.g. birch pollen allergen and human interleukin 7. As can be seen from several contributions, preparation of new capillary coatings suppressing the adsorption of peptides and proteins to the fused silica capillary wall in their CZE analyses and/or increasing the selectivity of their open-tubular CEC separations remains a hot topic in the area of CE and CEC developments. In addition, it is shown that through the theoretical modelling of the CZE determined effective electrophoretic mobilities of proteins, the important parameters, such as charge, hydration and shape of their molecules, can be estimated." [source] Use of the 1-mm micro-probe for metabolic analysis on small volume biological samplesJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 8b 2009Natalie J. Serkova Abstract Endogenous metabolites are promising diagnostic end-points in cancer research. Clinical application of high-resolution NMR spectroscopy is often limited by extremely low volumes of human specimens. In the present study, the use of the Bruker 1-mm high-resolution TXI micro-probe was evaluated in the elucidation of metabolic profiles for three different clinical applications with limited sample sizes (body fluids, isolated cells and tissue biopsies). Sample preparation and 1H-NMR metabolite quantification protocols were optimized for following oncology-oriented applications: (i) to validate the absolute concentrations of citrate and spermine in human expressed prostatic specimens (EPS volumes 5 to 10 ,l: prostate cancer application); (ii) to establish the metabolic profile of isolated human lymphocytes (total cell count 4 = 106: chronic myelogenous leukaemia application); (iii) to assess the metabolic composition of human head-and-neck cancers from mouse xenografts (biopsy weights 20 to 70 mg: anti-cancer treatment application). In this study, the use of the Bruker 1-mm micro-probe provides a convenient way to measure and quantify endogenous metabolic profiles of samples with a very low volume/weight/cell count. [source] CXCL12 chemokine up-regulates bone resorption and MMP-9 release by human osteoclasts: CXCL12 levels are increased in synovial and bone tissue of rheumatoid arthritis patientsJOURNAL OF CELLULAR PHYSIOLOGY, Issue 2 2004Francesco Grassi Chemokines are involved in a number of inflammatory pathologies and some of them show a pivotal role in the modulation of osteoclast development. Therefore, we evaluated the role of CXCL12 chemokine on osteoclast differentiation and function and we analyzed its expression on synovial and bone tissue biopsies from rheumatoid arthritis (RA) patients. Osteoclasts were obtained by 7 days in vitro differentiation with RANKL and M-CSF of CD11b positive cells in the presence or absence of CXCL12. The total number of osteoclast was analyzed by Tartrate-resistant acid phosphatase (TRAP)-staining and bone-resorbing activity was assessed by pit assay. MMP-9 and TIMP-1 release was evaluated by ELISA assay. CXCL12 expression on biopsies from RA patients was analyzed by immunohistochemistry. Osteoclasts obtained in the presence of CXCL12 at 10 nM concentration displayed a highly significant increase in bone-resorbing activity as measured by pit resorption assay, while the total number of mature osteoclasts was not affected. The increased resorption is associated with overexpression of MMP-9. Immunostaining for CXCL12 on synovial and bone tissue biopsies from both rheumatoid arthritis (RA) and osteoarthritis (OA) samples revealed a strong increase in the expression levels under inflammatory conditions. CXCL12 chemokine showed a clear activating role on mature osteoclast by inducing bone-resorbing activity and specific MMP-9 enzymatic release. Moreover, since bone and synovial biopsies from RA patients showed an elevated CXCL12 expression, these findings may provide useful tools for achieving a full elucidation of the complex network that regulates osteoclast function in course of inflammatory diseases. J. Cell. Physiol. 199: 244,251, 2004© 2003 Wiley-Liss, Inc. [source] Histopathological observations of human periimplantitis lesionsJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 5 2004Tord Berglundh Abstract Objective: The aim of the present study was to analyze some characteristics of advanced and progressive periimplantitis lesions in man. Material and methods: Soft tissue biopsies were obtained from 12 implants in six patients. The implants had been in function between 4 and 21 years and were, with one exception, located in the maxilla. The radiographic examination performed prior to biopsy revealed that all sites exhibited advanced bone loss. Further, clinical signs of severe inflammation, such as suppuration, swelling and/or fistula formation were detected in the majority of sites and seven of the 12 implants were found to be mobile at biopsy. Each biopsy was following fixation embedded in epoxy resin and sections were prepared for histometric and morphometric analysis. Results and conclusion: It was demonstrated (i) that all soft tissue units harbored large inflammatory cell infiltrates (ICT) that extended to a position apical of a pocket epithelium and (ii) that about 60% of the lesions were occupied by inflammatory cells, among which plasma cells dominated. Numerous amounts of PMN cells occurred not only in the pocket epithelium and adjacent connective tissue areas, but were also present in peri-vascular compartments in more central areas of the ICT. [source] Auto-transplantation of the uterus in the domestic pig (Sus scrofa): Surgical technique and early reperfusion eventsJOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 4 2006Caiza Almén Wranning Abstract Aim:, To develop a method for auto-transplantation of the uterus in the pig and to evaluate the early reperfusion events after short-term cold ischemia. Methods:, The bicornate uterus, with the cervix but without ovaries, was dissected and isolated with its bilateral feeding and draining vessels. The uterine arteries were cannulated in situ and the uterus was flushed with heparinized Ringer Acetate. It was stored at 4°C for 1,2 h during continuous flushing. The uterus was then placed in its original pelvic position and the uterine arteries and veins were anastomosed end-to-end to their origin. During approximately 100 min of reperfusion, blood samples and tissue biopsies were taken for monitoring of reperfusion events and detection of ischemia-reperfusion injuries. Results:, Out of 19 auto-transplanted pigs, seven were considered well flushed and were kept for cold ischemia. Of these seven, four showed satisfactory reperfusion judged by change in gross appearance and presence of appropriate venous blood flow. Analysis of blood-gas and metabolite parameters and histology indicated that at least two of these transplants were well reperfused, with no severe ischemia-reperfusion injuries. Conclusion:, In this first report ever on auto-transplantation of the pig uterus it is demonstrated that an acceptable reperfusion can be achieved. Furthermore, it is suggested that because of the large total size of the pig uterus with long uterine horns and the small size of the vessels available for re-anastomosis, the pig is a fairly difficult model for further studies on transplantation of the uterus. [source] Optimal methods for fluorescence and diffuse reflectance measurements of tissue biopsy samplesLASERS IN SURGERY AND MEDICINE, Issue 3 2002Gregory M. Palmer BS Abstract Background and Objective In developing fluorescence spectroscopy systems for the in vivo detection of pre-cancer and cancer, it is often necessary to perform preliminary testing on tissue biopsies. Current standard protocols call for the tissue to be immediately frozen after biopsy and later thawed for spectroscopic analysis, but this process can have profound effects on the spectroscopic properties of tissue. This study investigates the optimal tissue handling methods for in vitro fluorescence spectroscopy studies. Study Design/Materials and Methods The epithelial tissue of the Golden Syrian hamster cheek pouch was used in this study. Three specific experiments were carried out. First, the fluorescence properties of tissues in vivo and of frozen and thawed tissue biopsies were characterized at multiple excitation wavelengths spanning the ultraviolet-visible (UV-VIS) spectrum. Next, comparison of tissue fluorescence emission spectra in vivo, ex vivo (immediately after biopsy), and after the freeze and thaw process were systematically carried out at the excitation wavelengths corresponding to the previously identified fluorescence peaks. Lastly, intensities at the excitation and emission wavelength pairs corresponding to the fluorescence peaks were measured as a function of time after biopsy. Diffuse reflectance measurements over the UV-VIS spectrum were also made to evaluate the effects of oxygenation, blood volume, and scattering on the tissue fluorescence at these different excitation,emission wavelengths. Results This study indicates that the freezing and thawing process produces a significant deviation in intensity and lineshape relative to the in vivo fluorescence emission spectral data over the entire UV-VIS range between 300 and 700 nm. By contrast, examination of ex vivo emission spectra reveals that it closely preserves both the intensity and lineshape of the in vivo emission spectra except between 500 and 700 nm. The observed deviations can be explained by the diffuse reflectance measurements, which suggest increased hemoglobin deoxygenation and wavelength dependent changes in scattering in ex vivo tissues, and increased total hemoglobin absorption in the frozen and thawed samples. Furthermore, it was found that over a time window of 1.5 hours, spectroscopic changes brought about by degradation of the tissue due to biopsy or other factors are significantly smaller (10,30% variations in intensity) than those associated with the freezing and thawing process (50,70% decrease in intensity). Conclusions It was found that the effects of freezing and thawing on the fluorescence properties of tissue are greater than any changes brought about by degradation of tissue over a time frame of 90 minutes after biopsy. Performing ex vivo fluorescence measurements within a reasonable time window has the advantage of more accurately reproducing the clinically relevant in vivo conditions in the case of the hamster cheek pouch tissue. Therefore, in tissue biopsy studies, the tissue sample should ideally be maintained in an unfrozen state prior to measurement. Lasers Surg. Med. 30:191-200, 2002. © 2002 Wiley-Liss, Inc. [source] Cerebral mycosis: 7-year retrospective series in a tertiary centerNEUROPATHOLOGY, Issue 3 2010Kirtee Raparia This study focuses on the epidemiology, clinical manifestations, risk factors, diagnosis and outcome of all cases of central nervous system (CNS) fungal infections in a tertiary center. Medical records of 18 patients of culture-proven CNS fungal infections were retrospectively reviewed from 2000 to 2007, including 12 isolated from the cerebrospinal fluid (CSF) and seven from tissue biopsy. Patient demographic data included 10 males and eight females. The mean age was 55 years (range: 24,89 years). All but one patient were immunocompromised. Fungal organisms isolated from CSF included: Cryptococcous neoformans (8 patients), Coccidioides immitis (3 patients), and Aspergillus versicolor (1 patient). Histopathology of seven biopsy cases revealed groups of pigmented golden-brown fungal forms in three cases; three cases showed septate fungi, two of which had melanin in their walls; and one case showed multiple round spherules. These cases on microbiological cultures grew Coccidioides immitis (1 patient), Aspergillus fumigatus (1 patient), Cladophialophora bantiana (2 patients), Fonsecaea monophora (1 patient) and Scedosporium apiospermum (2 patients). Five of the seven fungal organisms isolated from tissue biopsies were dematiaceous fungi. Twelve patients died after a period of a few weeks to months, two were lost to follow-up, and four are alive with severe neurological sequelae. CNS fungal infections in our cohort were more common in patients post-transplant and with hematologic malignancies. In our series, rare dematiaceous fungi are emerging agents for cerebral mycosis. The outcome of CNS fungal infections is poor despite vigorous antifungal therapy. [source] Immunohistochemical estimation of cell cycle entry and phase distribution in astrocytomas: applications in diagnostic neuropathologyNEUROPATHOLOGY & APPLIED NEUROBIOLOGY, Issue 5 2005Ian S. Scott An immunohistochemical method for assessing cell cycle phase distribution in neurosurgical biopsies would enable such data to be incorporated into diagnostic algorithms for the estimation of prognosis and response to adjuvant chemotherapy in glial neoplasms, without the requirement for flow cytometric analysis. Paraffin-embedded sections of intracerebral gliomas (n = 48), consisting of diffuse astrocytoma (n = 9), anaplastic astrocytoma (n = 8) and glioblastoma (n = 31), were analysed by immunohistochemistry using markers of cell cycle entry, Mcm-2 and Ki67, and putative markers of cell cycle phase, cyclins D1 (G1-phase), cyclin A (S-phase), cyclin B1 (G2-phase) and phosphohistone H3 (Mitosis). Double labelling confocal microscopy confirmed that the phase markers were infrequently coexpressed. Cell cycle estimations by immunohistochemistry were corroborated by flow cytometric analysis. There was a significant increase in Mcm-2 (P < 0.0001), Ki67 (P < 0.0001), cyclin A (P < 0.0001) and cyclin B1 (P = 0.002) expression with increasing grade from diffuse astrocytoma through anaplastic astrocytoma to glioblastoma, suggesting that any of these four markers has potential as a marker of tumour grade. In a subset of glioblastomas (n = 16) for which accurate clinical follow-up data were available, there was a suggestion that the cyclin A:Mcm-2 labelling fraction might predict a relatively favourable response to radical radiotherapy. These provisional findings, however, require confirmation by a larger study. We conclude that it is feasible to obtain detailed cell cycle data by immunohistochemical analysis of tissue biopsies. Such information may facilitate tumour grading and may enable information of prognostic value to be obtained in the routine diagnostic laboratory. [source] Tissue HHV6 and 7 determination in pediatric solid organ recipients , a pilot studyPEDIATRIC TRANSPLANTATION, Issue 6 2003M. Gupta Abstract:, Herpes virus infections remain a major challenge in solid organ transplantation. HHV6 and 7 blood viral load was associated with pathology after renal transplantation. Little is known about the significance of tissue HHV6 and 7 infections. A total of 18 tissue biopsies (13 kidney, three GI and two BAL) from nine pediatric transplant patients (five kidney, two liver, one combined liver and kidney and one bone marrow transplant) were subjected to blood HHV6 IgG and IgM testing. In addition, tissue HHV6 and 7 semi-quantitative PCR analysis with subsequent detection by ELISA and quantitative methods were applied to the same samples. We also studied four native kidney biopsies of children with other kidney disease. The results of the biopsies were correlated with clinical data. Of the transplant patients, 78% were HHV6 IgG positive. Six of nine had a positive IgM on at least one occasion, however, only two of nine transplant patients were symptomatic with a mixed CMV/EBV septic picture of multi-organ failure. Only these two patients had a significant tissue viral load for HHV6. Additionally, a very significant tissue viral load for HHV6 was detected in an immunocompromised patient 3 wk after a roseola-like febrile illness. The HHV6 copies were 31, 88 and 206 per 10 ,L of DNA, respectively. In the patient who also had the fourth positive ELISA for HHV6 PCR product, the Multiplex PCR and restriction enzyme assay on its PCR product revealed a significant contribution by HHV7, while the HHV6-B signal was rather weak. Significant tissue HHV6 loads can be found in tissue biopsies from organ recipients with significant illness and also in native kidneys after primary infection. This may explain the high prevalence of HHV6 in transplanted kidneys. Further studies on HHV6 and 7 using molecular techniques should be supported. [source] Differential detection of S100A8 in transitional cell carcinoma of the bladder by pair wise tissue proteomic and immunohistochemical analysisPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 2 2006Jonathan Abstract The search for novel molecular markers of tumor invasion is vital if strategies are to become more effective in the diagnostic and prognostic management of transitional cell carcinoma of the bladder. Up to 50% of tumors detected at stage,1 (pT1) progress to a higher grade even after endoscopic surgical resection, and there are currently no protein markers of this aggressive, invasive phenotype. We have combined SELDI-TOF-MS, ClinProt magnetic bead enrichment, Nano-LC-ESI-ion trap tandem mass spectrometry and immunohistochemical analysis to the study of 12,invasive bladder cancer tissue biopsies paired with normal bladder tissue samples obtained from the same patients for the definition and identification of proteins up-regulated in the tumors. We report the inflammation-associated calcium binding protein,S100A8 (MRP-8, calgranulin,A) to be highly expressed in tumor cells in contrast to normal urothelium in 50% of the samples, as well as two unidentified protein markers at 5.75 and 6.89,kDa that were differentially detected in 9/12 and 10/12,tumor samples, respectively. These new markers, when fully characterized, may contribute to new target proteins for the prediction of aggressive, invasive bladder tumors. [source] Protein profiling in pathology: Analysis and evaluation of 239 frozen tissue biopsies for diagnosis of B-cell lymphomasPROTEOMICS - CLINICAL APPLICATIONS, Issue 5 2010Corine Jansen Abstract Purpose: We determined the potential value of protein profiling of tissue samples by assessing how precise this approach enables discrimination of B-cell lymphoma from reactive lymph nodes, and how well the profiles can be used for lymphoma classification. Experimental design: Protein lysates from lymph nodes (n=239) from patients with the diagnosis of reactive hyperplasia (n=44), follicular lymphoma (n=63), diffuse large B-cell lymphoma (n=43), mantle cell lymphoma (n=47), and chronic lymphocytic leukemia/small lymphocytic B-cell lymphoma (n=42) were analysed by SELDI-TOF MS. Data analysis was performed by (i) classification and regression tree-based analysis and (ii) binary and polytomous logistic regression analysis. Results: After internal validation by the leave-one-out principle, both the classification and regression tree and logistic regression classification correctly identified the majority of the malignant (87 and 96%, respectively) and benign cases (73 and 75%, respectively). Classification was less successful since approximately one-third of the cases of each group were misclassified according to the histological classification. However, an additional mantle cell lymphoma case that was misclassified as chronic lymphocytic leukemia/small lymphocytic B-cell lymphoma initially was identified based on the protein profile. Conclusions and clinical relevance: SELDI-TOF MS protein profiling allows for reliable identification of the majority of malignant lymphoma cases; however, further validation and testing robustness in a diagnostic setting is needed. [source] Virtobot,a multi-functional robotic system for 3D surface scanning and automatic post mortem biopsyTHE INTERNATIONAL JOURNAL OF MEDICAL ROBOTICS AND COMPUTER ASSISTED SURGERY, Issue 1 2010Lars Christian Ebert Abstract Background The Virtopsy project, a multi-disciplinary project that involves forensic science, diagnostic imaging, computer science, automation technology, telematics and biomechanics, aims to develop new techniques to improve the outcome of forensic investigations. This paper presents a new approach in the field of minimally invasive virtual autopsy for a versatile robotic system that is able to perform three-dimensional (3D) surface scans as well as post mortem image-guided soft tissue biopsies. Methods The system consists of an industrial six-axis robot with additional extensions (i.e. a linear axis to increase working space, a tool-changing system and a dedicated safety system), a multi-slice CT scanner with equipment for angiography, a digital photogrammetry and 3D optical surface-scanning system, a 3D tracking system, and a biopsy end effector for automatic needle placement. A wax phantom was developed for biopsy accuracy tests. Results Surface scanning times were significantly reduced (scanning times cut in half, calibration three times faster). The biopsy module worked with an accuracy of 3.2 mm. Discussion Using the Virtobot, the surface-scanning procedure could be standardized and accelerated. The biopsy module is accurate enough for use in biopsies in a forensic setting. Conclusion The Virtobot can be utilized for several independent tasks in the field of forensic medicine, and is sufficiently versatile to be adapted to different tasks in the future. Copyright © 2009 John Wiley & Sons, Ltd. [source] Androgen receptor as a potential sign of prostate cancer metastasisTHE PROSTATE, Issue 15 2009Marie-Hélène Lévesque Abstract BACKGROUND Androgen receptor (AR) expression and its modulation through the carcinogenesis process have been investigated in several studies with conflicting results. MATERIALS AND METHODS In situ hybridization and immunocytochemistry were used to examine AR expression in prostatic needle core biopsies of benign, high grade prostatic intraepithelial neoplasia (HGPIN) and prostatic adenocarcinoma. RESULTS A significant increase in AR mRNA levels was found in the cancerous prostatic cells when compared with the benign tissue biopsies. AR abundance in HGPIN was found to be almost half-way between that observed in benign and in cancerous tissue. In the benign prostatic epithelium, the immunocytochemistry data show that AR is exclusively expressed in the nuclei of epithelial cells. However, in 72% of examined cancer biopsies, AR was expressed in both the cytoplasm and nuclei. After examination of medical records of 100 patients diagnosed with prostate cancer, it was found that the AR was expressed in both cellular compartments of cancer cells in 81% of cases when cancer was found to have metastasized outside the prostate. In contrast, when the cancer was organ-confined, AR was localized in both the nuclei and cytoplasm in only 66% of cases. Moreover, when the AR was expressed in the cytoplasm of cancerous cells, consecutive serial sections immunostained with the mitochondrial marker suggest that AR is localized in the mitochondria. CONCLUSIONS AR mRNA expression is significantly higher in prostate cancer when compared to benign prostatic tissue. Prostate 69: 1704,1711, 2009. © 2009 Wiley-Liss, Inc. [source] Concomitant Endothelin-1 Overexpression in Lung Transplant Donors and Recipients Predicts Primary Graft DysfunctionAMERICAN JOURNAL OF TRANSPLANTATION, Issue 3 2010M. Salama Primary graft dysfunction (PGD) causes significant morbidity following lung transplantation (LTX). Mortality is high in PGD and therapeutic strategies are limited. To investigate whether endothelin-1 (ET-1) that mediates increased vascular permeability and edema formation in lung grafts can predict PGD, ET-1 mRNA expression was examined in lung tissue biopsies of 105 donors and recipients obtained shortly before LTX. Serum ET-1 concentration was assessed by ELISA. PGD grade was diagnosed and scored by oxygenation and radiological characteristics according to ISHLT guidelines. PGD grade 3 developed in 11% of patients. ET-1 mRNA expression was significantly increased in both donor (p < 0.0001) and recipient (p = 0.01) developing PGD as compared to no PGD group. Pretransplant ET-1 serum concentrations were elevated in recipients with PGD as compared to no PGD group (p < 0.0001), although serum ET-1 was not different between donors whose grafts developed PGD grades 0,3. In regression analysis, concomitant elevated donor tissue ET-1 and recipient serum ET-1 predicted PGD grade 3. This study indicates that pretransplant ET-1 mRNA overexpression in donors associated with elevated pretransplant serum ET-1 in recipients contribute to PGD development and that their assessment might be beneficial to predict PGD and to identify recipients who could benefit from a targeted ET-1 blockade. [source] Herpes simplex virus type-2 in Egyptian patients with bladder cancer or cystitisAPMIS, Issue 1 2010HALA BADAWI Badawi H, Ahmed H, Aboul Fadl L, Helmi A, Fam N, Diab M, Ismail A, Badawi A, Saber M. Herpes simplex virus type-2 in Egyptian patients with bladder cancer or cystitis. APMIS 2010; 118: 37,44. The present study was designed to investigate the prevalence of herpes simplex virus type-2 (HSV-2) in Egyptian patients with bladder cancer or cystitis and to evaluate the performance of different diagnostic HSV-2 assays. The study included 50 patients: 27 with bladder cancer (group I), 23 with cystitis (group II) and 20 subjects as controls (group III). HSV-2 DNA was detected using polymerase chain reaction (PCR) on bladder tissue and buffy coat cells (BCC). Electron microscopic studies (EMS) on BCC and ELISAs for IgM, IgG and specific glycoprotein G-2 (gG-2) IgG were performed. HSV-2 DNA was detected by PCR on bladder tissue biopsies in 29.6% and 21.7% of group I and II respectively and it was also detected by PCR on BCC in 22.2% and 21.7% of group I and II respectively. EMS revealed HSV like particles in 16.6% of cases. IgG, specific gG-2 IgG and IgM were detected in 30%, 16% and 6% of cases respectively. The different assays were evaluated in relation to PCR on bladder tissue biopsies. The gG-2-based ELISA and EMS on BCC were found to be highly specific (97.3% and 100% respectively), with similar low sensitivity of ,54%. PCR on BCC was the most sensitive assay. The association of HSV-2 with bladder cancer is suggested especially in schistosomal patients. [source] Cyclooxygenase-2 expression on urothelial and inflammatory cells of cystoscopic biopsies and urine cytology as a possible predictive marker for bladder carcinomaAPMIS, Issue 1 2009MONA MOUSSA Cyclooxygenase-2 (COX-2) is a key inducible enzyme involved in the production of prostaglandins. It contributes to human carcinogenesis by various mechanisms. The aim of the current study was to elucidate the possible involvement of COX-2 in human bladder carcinoma by examining its expression on both urothelial and inflammatory cells in tissue biopsies and urine cytology samples of different urinary bladder lesions. A total of 65 patients were included in the study and were selected from cases admitted to Urology Department, Theodor Bilharz Research Institute (TBRI), Giza, Egypt. They represented seven control cases with almost normal-looking bladder tissue; pure chronic cystitis (n=12); premalignant lesions (18) in the form of squamous metaplasia (n=8) or urothelial dysplasia (n=10) as well as transitional cell carcinoma (TCC) (n=18), and squamous cell carcinoma (SqCC) (n=10). Immunohistochemistry of formalin-fixed, paraffin-embedded tissue sections and urine cytology samples was performed for all cases using COX-2 (H-62): sc-7951, a rabbit polyclonal antibody. The study revealed positive COX-2 expression on the urothelial and inflammatory cells of cystoscopic biopsies from all cases of pure chronic cystitis, squamous metaplasia and SqCC compared with 42.8% and 71.4% of normal controls, respectively. The score of urothelial COX-2 expression was sequentially up-regulated from normal to chronic cystitis (either pure or associated with premalignant changes) (p<0.05) to malignant changes (p<0.05). However, the inflammatory cellular expression was down-regulated with malignant transformation compared with chronic cystitis (p<0.05). In TCC, COX-2 was over-expressed on both urothelial and inflammatory cells in advanced tumors. Urine cytology samples were positive for COX-2 in a comparable manner to that observed in cystoscopic biopsies. Accordingly, the results of the current study have provided new information in two aspects: First, is the possibility of using the differential COX-2 expression on both inflammatory and urothelial cells as markers for premalignant or malignant transformation; second, besides cystoscopy, urine cytology was found to have a high sensitivity for COX-2 expression and hence proved to be valuable in malignancy as a non-invasive substitute for cystoscopy. [source] C-type natriuretic peptide in prostate cancerAPMIS, Issue 1 2009SOEREN JUNGE NIELSEN C-type natriuretic peptide (CNP) is expressed in the male reproductive organs in pigs. To examine whether the human prostate also expresses the CNP gene, we measured CNP and N-terminal proCNP in prostate cancer tissue extracts and performed immunohistochemical biopsy staining. Additionally, proCNP-derived peptides were quantitated in plasma from patients with prostate cancer. Blood was collected from healthy controls and patients before surgery for localized prostate cancer. Tissue extracts were prepared from tissue biopsies obtained from radical prostatectomy surgery. N-terminal proCNP, proCNP (1,50) and CNP were measured in plasma and tissue extracts. Biopsies were stained for CNP-22 and N-terminal proCNP. Tissue extracts from human prostate cancer contained mostly N-terminal proCNP [median 5.3 pmol/g tissue (range 1.0,12.9)] and less CNP [0.14 pmol/g tissue (0.01,1.34)]. Immunohistochemistry demonstrated the presence of the peptides in prostatic epithelial cells. The N-terminal proCNP concentrations in plasma were marginally lower in patients with localized prostate cancer compared with control subjects [13.8 pmol/l (11.0,17.2) vs. 15.1 pmol/l (10.4,23.2), p=0.002] but not enough to justify the use of N-terminal proCNP as a cancer marker. Further research is needed to establish whether measurement of proCNP-derived peptides may offer clinical information. [source] Homeoprotein Cdx2 and nuclear PTEN expression profiles are related to gastric cancer prognosis,APMIS, Issue 12 2007ZHIGANG BAI The aim of the study was to analyze the expression of Cdx2 and nuclear PTEN in relation to clinicopathological features of gastric cancer tissue biopsies in order to determine the value of a combined analysis of Cdx2 and nuclear PTEN expression in distinguishing histological types and prognosis of gastric cancers. The expression of Cdx2 and nuclear PTEN was studied using immunohistochemistry of paraffin-embedded tumor specimens from 99 patients who underwent radical D2 gastrectomy between 1999 and 2001. Cdx2 and nuclear PTEN expression were detected in 39.6% (36 of 91) and 70.3% (64 of 91) of gastric cancer cases, respectively. There was a negative correlation between Cdx2 expression and Lauren classification (p=0.032), and between nuclear PTEN expression and lymph node metastasis (p=0.049). Patients with Cdx2-positive, or nuclear PTEN-positive expression had higher survival rates than those with Cdx2-negative or nuclear PTEN-negative expression (p<0.001 and p=0.003, respectively). Co-expression of Cdx2 and nuclear PTEN showed significantly lower levels in diffuse- or mixed-type cancers than in intestinal-type cancers (p=0.005). Multivariate analysis revealed that Cdx2 expression was an independent prognostic indicator of gastric cancer (p=0.014). These data suggest that combined analysis of Cdx2 and nuclear PTEN expression can have significant value in distinguishing histological types of gastric cancer and assessing prognosis in patients with gastric cancer. [source] Granulomas in common variable immunodeficiency: A diagnostic dilemmaAUSTRALASIAN JOURNAL OF DERMATOLOGY, Issue 1 2004Karyn R Lun SUMMARY A 60-year-old man with common variable immunodeficiency presented with a 7-year history of violaceous plaques and papules on the thighs, arms and trunk. In the preceding 2 years he had developed new lesions on both hands. He had been previously diagnosed with sarcoidosis on the basis of skin and visceral histology, but subsequent opinion was that these were sarcoid-like granulomas rather than being representative of true sarcoidosis. Biopsy of the hand lesions showed necrotizing granulomas, and a single acid-fast bacillus (AFB) was identified on Wade,Fite stain. Subsequent repeat tissue biopsies for histology, culture and polymerase chain reaction testing failed to confirm the presence of mycobacterial organisms and it was felt that the organism was a contaminant introduced during tissue processing. The hand lesions responded well to intralesional injections of triamcinolone acetonide 10 mg/mL and oral tetracycline 500 mg b.d. was later introduced with a good clinical response. The diagnostic dilemma of finding granulomatous inflammation in a patient with common variable immunodeficiency, and the significance of a single AFB on histology are discussed. The treatment of sarcoid-like granulomas with tetracycline therapy is also commented on. [source] Distinctive E-cadherin and epidermal growth factor receptor expression in metastatic and nonmetastatic head and neck squamous cell carcinomaCANCER, Issue 1 2008Predictive, prognostic correlation Abstract BACKGROUND. The authors investigated whether coexpression and localization of E-cadherin (E-cad) and epidermal growth factor receptor (EGFR) had predictive and/or prognostic correlations with lymph node metastasis and/or survival in patients with squamous cell carcinoma of the head and neck (SCCHN). METHODS. Immunohistochemistry (IHC) of archival tissue was performed to measure expression of EGFR and E-cad in surgical specimens of SCCHN (n = 143) that included primary tumors (PTs) with positive lymph nodes (Tu+Met) and their paired lymph node metastases (LnMet), PTs with negative lymph nodes (Tu,Met), and benign tissue biopsies as normal controls. IHC staining was quantified as a weighted index and as the ratio of membrane to cytoplasmic staining. Correlative expression between EGFR and E-cad also was examined in SCCHN cell lines by immunoblotting and immunofluorescence analyses. RESULTS. Three distinct expression patterns of EGFR and E-cad were observed. Membrane localization of E-cad was significantly lower in the Tu+Met group than in the Tu,Met group (P = .01) and was associated inversely with lymph node status (P = .009). Wilcoxon analysis of the combined markers demonstrated that expression and/or membrane localization of EGFR and E-cad were correlated with disease-free survival and overall survival in patients with SCCHN. The study of SCCHN cell lines demonstrated that cells with positive but low EGFR expression and with negative E-cad expression were relatively resistant to the EGFR tyrosine kinase inhibitor erlotinib. CONCLUSIONS. The current study suggested that examining not only the expression but also the localization of EGFR and E-cad simultaneously may have clinical relevance in predicting lymph node metastasis, patient survival, and response to EGFR-targeted therapy in patients with SCCHN. Cancer 2008. © 2008 American Cancer Society. [source] Response measurement after intraarterial chemoradiation in advanced head and neck carcinomaCANCER, Issue 8 2006Magnetic resonance imaging, evaluation under general anesthesia? Abstract BACKGROUND The objectives of this prospective trial were to evaluate the diagnostic accuracy and predictive value of magnetic resonance imaging (MRI) and to use MRI evaluation under general anesthesia (EGA) 6 to 8 weeks after chemoradiation to determine local control. METHODS Eighty-two consecutive patients with advanced-stage squamous cell carcinoma of the oral cavity, oropharynx, hypopharynx, or supraglottic larynx were treated with selective targeted chemoradiation. All patients who completed treatment and survived had a minimum follow-up of 3 years. MRI and EGA were performed from 6 to 8 weeks after treatment. Posttreatment MRI findings were compared with pretreatment MRI findings and were graded for risk of local recurrence/residual disease on a 4-point scale. The diagnosis of treatment failure was based on tissue biopsies, which were obtained during EGA or later during follow-up. The predictive value of MRI was analyzed by using a Cox proportional hazards model. RESULTS Only 1 patient with MRI Grade 0 or 1 findings (discrete mass < 10 mm; n = 62 patients) had residual disease 6 to 8 weeks after treatment that was detected during EGA. In 5 patients with MRI findings of Grade 2a and 2b (mass > 10 mm; n = 20 patients), residual disease was detected. After 2 years, 23 patients had a local failure (28%). Twelve local failures were found among 62 patients who had MRI findings of Grade 0 and 1. Posttreatment MRI emerged as an independent predictive factor (hazard ratio, 3.0; P = .014) for local control. CONCLUSIONS Posttreatment MRI studies provided predictive information on local control in addition to pretreatment predictors. In patients with focal masses < 10 mm, the combination of response evaluation under general anesthesia and posttreatment MRI from 6 to 8 weeks after chemoradiation hardly provided more information on the local control than posttreatment MRI alone. Cancer 2006. © 2006 American Cancer Society. [source] Optical Coherence Tomography: A Noninvasive Method to Assess Wound ReepithelializationACADEMIC EMERGENCY MEDICINE, Issue 5 2007Adam J. Singer MD BackgroundAccurate assessment of wound healing may require invasive tissue biopsies, limiting its clinical usefulness in humans. Optical coherence tomography (OCT) is a novel, high-resolution method using light reflection to obtain noninvasive cross sectional imaging of biological tissues. ObjectivesTo evaluate the utility of OCT for assessing wound reepithelialization in a porcine model. MethodsThe authors conducted an animal study with two domestic pigs. Excisional cutaneous wounds were created over the ventral surface of the animals using an electric dermatome set at a depth of 600 ,m. The wounds were excised two or three days later and precisely marked to guide initial OCT and subsequent tissue slicing and microscopy. Comparing hematoxylin and eosin,stained histologic sections and the corresponding OCT images from each tissue sample permitted identification of the correlative micromorphology. Scatter and Bland,Altman plots were used to present the data. The primary measure of agreement was the standard deviation of the pairwise differences in percent reepithelialization between OCT and histology together with a 95% confidence interval. ResultsIn normal skin, the epidermis was characterized by a thin, bright layer indicating a high degree of light scattering on OCT. The dermis below was characterized by a thicker, darker area indicating less scattering of light. All fresh excisional wounds lacked an outer bright layer of epidermis immediately after injury. At days 2 and 3, the wounds were partially reepithelialized. A new bright layer with intense light scattering was present on OCT corresponding to the neoepidermis on hematoxylin and eosin,stained sections. The correlation between percent reepithelialization measured with OCT and histology was 0.66 (p < 0.001), and the standard deviation of the differences was 11.0% (95% confidence interval = 8.4% to 16.1%). ConclusionsOCT accurately detects the presence or absence of the epidermal layer of skin, allowing noninvasive tracking of wound reepithelialization. [source] Differential cytokine expressions affect the severity of peri-implant diseaseCLINICAL ORAL IMPLANTS RESEARCH, Issue 5 2009Poliana Mendes Duarte Abstract Objective: This study assessed gene expression by quantitative polymerase chain reaction of inflammatory- [interleukin (IL)-12, tumor necrosis factor-, (TNF-,), IL-4, and IL-10] and osteoclastogenesis-related factors [receptor activator of NF-,B ligand (RANKL) and osteoprotegerin (OPG)] in sites exhibiting different severities of peri-implant disease. Material and methods: Peri-implant soft tissue biopsies (n=48) were harvested from healthy implant (HI), mucositis (MC), initial peri-implantitis (IP) and severe peri-implantitis (SP) sites. Results: IL-12 and TNF-, mRNA levels were higher in SP, followed by IP and MC (P <0.05). IL-4 was higher in HI, followed by MC, SP and IP (P <0.05). IL-10 was the lowest in HI, while no differences were detected among the diseased groups (P>0.05). OPG mRNA levels were higher in HI, followed by IP, SP and MC, whereas RANKL was increased as the peri-implantitis severity increased (P<0.05). The highest OPG/RANKL ratio was observed in HI and the lowest in SP (P<0.01). Conclusion: These findings suggest that expressions of inflammatory- and osteoclastogenesis-related factors may play an important role in the onset and severity of the peri-implant diseases. [source] Systematic review of methods to diagnose infection in foot ulcers in diabetesDIABETIC MEDICINE, Issue 4 2006S. O'Meara Abstract Aim, To undertake a systematic review of the diagnostic performance of clinical examination, sample acquisition and sample analysis in infected foot ulcers in diabetes. Methods, Nineteen electronic databases plus other sources were searched. To be included, studies had to fulfil the following criteria: (i) compare a method of clinical assessment, sample collection or sample analysis with a reference standard; (ii) recruit diabetic individuals with foot ulcers; (ii) present 2 × 2 diagnostic data. Studies were critically appraised using a 12-item checklist. Results Three eligible studies were identified, one each on clinical examination, sample collection and sample analysis. For all three, study groups were heterogeneous with respect to wound type and a small proportion of participants had foot ulcers due to diabetes. No studies identified an optimum reference standard. Other methodological problems included non-blind interpretation of tests and the time lag between index and reference tests. Individual signs or symptoms of infection did not prove to be useful tests when assessed against punch biopsy as the reference standard. The wound swab did not perform well when assessed against tissue biopsy. Semiquantitative analysis of wound swab might be a useful alternative to quantitative analysis. The limitations of these findings and their impact on recommendations from relevant clinical guidelines are discussed. Conclusion, Given the importance of this topic, it is surprising that only three eligible studies were identified. It was not possible to describe the optimal methods of diagnosing infection in diabetic patients with foot ulceration from the evidence identified in this systematic review. Diabet. Med. 23, 341,347 (2006) [source] | ||||||||||||||||||||||||||||||||||