Home About us Contact
|
Home About us Contact | ||
|
|
||
Immunoperoxidase Staining (immunoperoxidase + staining)
Selected AbstractsDemonstration of aberrant T-cell and natural killer-cell antigen expression in all cases of granular lymphocytic leukaemiaBRITISH JOURNAL OF HAEMATOLOGY, Issue 6 2003William G. Morice Summary. The diagnosis of granular lymphocytic leukaemia (GLL) requires the presence of an immunophenotypically distinct T-cell (T-GLL) or natural killer-cell (NK-GLL) population. Flow cytometric immunophenotyping was performed on 21 T-GLL patients, 11 NK-GLL patients and 20 normal control subjects using antibodies to T and NK cell-associated antigens in order to accurately identify the distinguishing features of T-GLL and NK-GLL. The NK antigens evaluated included: CD16, CD57, CD94, CD161, and the killing inhibitory receptors (KIRs) CD158a, CD158b and CD158e (p70). Abnormal T-antigen expression was present in all T-GLL patients. CD57 was frequently expressed in T-GLL, however, one-third of patients showed partial CD57 expression similar to that seen in T cells from normal control subjects. Ten T-GLL were KIR positive; all expressed a single KIR isoform. All NK-GLL showed a distinctive, abnormal immunophenotype. Four NK-GLL expressed a single KIR isoform; the remaining seven patients lacked all tested KIRs, which is also a distinct, abnormal finding. Immunoperoxidase staining of bone marrow biopsy specimens from NK-GLL patients with antibodies to CD8, TIA-1 and granzyme B revealed the disease-specific distinctive staining patterns previously found in T-GLL. These studies delineate the unique immunophenotypic features diagnostic of T-GLL and provide strong evidence that NK-GLL, like T-GLL, represents a clonal lymphoproliferative disorder. [source] Cytomorphological study of soft tissue neoplasms: role of fluorescent immunocytochemistry in diagnosisCYTOPATHOLOGY, Issue 5 2005B. Rekhi Objectives:, Exact categorization of soft tissue tumours (STTs) on smears requires application of various ancillary techniques. This study was aimed at evaluating the role of fluorescent immunocytochemistry (FICC) in cyto-diagnosis of 30 STT cases. Methods:, Thirty cases of soft tissue tumours were included in the present study. All cases were subjected to routine Giemsa and Papanicolaou stain. Extra smears were made and kept for fluorescent immunostaining. A panel of cytoskeletal antibodies, tagged with FITC (Fluorescein isothyocynate), was employed in all these cases. Fluorescent immunostained smears were examined under Zeiss Confocal Laser scanning microscope, using double immunofluorescence (red-green). Finally, all cases were subjected to biopsy and again immunoperoxidase staining. Results:, Among the 30 cases in the present study, unaided cytological diagnoses ranged from ,spindle cell' tumour in four (13.3%) cases, benign and malignant spindle cell tumour in 17 (56.6%) cases, to malignant mesenchymal tumour in nine (30%) cases. FICC helped in further correct categorization of 25/30 (83.3%) cases viz. leiomyoma (three), benign neurogenic tumour (six), schwannoma (one), dermatofibrosarcoma protuberans (three), synovial sarcoma (two), rhabdomyosarcoma (two), malignant fibrous histiocytoma (five) and malignant peripheral nerve sheath tumour (three). Aggressive fibromatosis was found to be a missed diagnosis in two cases. Overall concordance between cyto-diagnosis with FICC, and histopathology results was 83.3% (P < 0.05). Conclusion:, Fluorescent immunocytochemistry is a significant ancillary technique for making a rapid and specific diagnosis of STT, as required for their timely management. Incorporation of a wide panel of antibody markers with clinico-cytological correlation is recommended in forming an exact diagnosis in these cases. [source] Effect of Laser Resurfacing on p53 Expression in Photoaged Facial SkinDERMATOLOGIC SURGERY, Issue 6 2007MOETAZ M. EL-DOMYATI MD BACKGROUND p53 overexpression has been reported in photoaged skin. Meanwhile, p53 gene mutations have been implicated as an important factor in the pathogenesis of ultraviolet (UV) light,induced skin cancer. OBJECTIVE The objective was to evaluate the effect of laser resurfacing on the epidermal thickness and expression of p53 in photoaged skin. METHODS Specimens were obtained from the facial skin of 10 patients before and after 3 months and 1 year of treatment using CO2 (five cases) and erbium (Er):YAG (five cases) lasers. Specimens were also obtained from six age-matched controls. These biopsies were used for routine histopathology, histometry, and p53 immunoperoxidase staining. RESULTS Both CO2 and Er:YAG lasers were found to induce a significant decrease in p53 expression in biopsies obtained after 3 months (p=.0004 and .002, respectively) followed by gradual increase (p=.01 in both groups). A significant increase (p<.01) in epidermal thickness was also observed after 1 year of resurfacing. This increase, however, is inversely correlated with the level of p53 expression in such patients. CONCLUSION The decrease in epidermal p53 expression after CO2 and Er:YAG lasers may account for some of the benefits of resurfacing on the epidermis, as well as prevention of actinic neoplasia by adjusting any disturbance in the proliferation/apoptosis balance observed in photoaged facial skin. [source] Sentinel node biopsy in oral cavity cancer: Correlation with PET scan and immunohistochemistryHEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 1 2003Francisco J. Civantos MD Abstract Background. Lymphoscintigraphy and sentinel node biopsy (LS/SNB) is a minimally invasive technique that samples first-echelon lymph nodes to predict the need for more extensive neck dissection. Methods. We evaluated this technique in 18 oral cavity cancers, stages T1,T3, N0. Patients underwent CT and positron emission tomography (PET) of the neck, followed by LS/SNB, frozen section, immediate selective neck dissection, definitive histology, and immunoperoxidase staining for cytokeratin. Histopathology of the sentinel node was correlated with that of the neck specimen. Results. There were 10 true positives: 6 identified on frozen section; 2 on permanent histology; and 2 only on immunoperoxidase staining. In six, the sentinel node was the only positive node. There were seven true negatives and one false negative. Conclusions. Gross tumor replacement of lymph node architecture may obstruct and redirect lymphatic flow. Overall LS/SNB holds promise for oral cancer. © 2002 Wiley Periodicals, Inc. Head Neck 25: 000,000, 2003 [source] Pemphigus vegetans , immunopathological findings in a rare variant of pemphigus vulgarisJOURNAL DER DEUTSCHEN DERMATOLOGISCHEN GESELLSCHAFT, Issue 3 2010Babak Monshi Summary A patient with painful erosions of the oral cavity and the labia minora developed multifocal blisters in inter-triginous areas. These blisters eroded and evolved into papillomatous erosive vegetations. Histopathology and immunopathological investigations confirmed the diagnosis of pemphigus vegetans, mediated by IgG autoantibodies. The circulating IgG1 and IgG4 autoantibodies were exclusively directed against desmoglein 3, as shown by ELISA and indirect immunofluorescence studies. These IgG1 and IgG4 isotypes were also in vivo bound, as demonstrated with immunoperoxidase staining of perilesional skin. Our clinical, biochemical and immunopathological observations confirm the hypothesis that pemphigus vegetans is a variant of pemphigus vulgaris. [source] HHV-6 and HHV-7 antigenemia related to CMV infection after liver transplantationJOURNAL OF MEDICAL VIROLOGY, Issue 6 2006Maiju Härmä Abstract Background Betaherpesviruses human herpesvirus-6 and -7 (HHV-6, HHV-7), which are closely related to cytomegalovirus (CMV), have been reported in transplant patients. In this retrospective study, we investigated the occurrence of HHV-6 and HHV-7 antigenemia in relation to symptomatic CMV infection after liver transplantation. Methods: Sample material from 64 adult liver transplant recipients was included in the study. The patients were monitored weekly for CMV, HHV-6, and HHV-7. CMV infections were diagnosed by pp65-antigenemia and viral cultures. Concomitantly HHV-6 and HHV-7 antigens were demonstrated in peripheral blood mononuclear cells by monoclonal antibodies against both variants A and B and immunoperoxidase staining. Altogether 540 post-transplant blood specimens were analyzed. Results: Nineteen patients (30%) developed symptomatic CMV pp65 antigenemia during the first 3 months (mean 33 days, range 5,62 days) post-transplantation and were treated with intravenous ganciclovir. Concurrent HHV-6 antigenemia was detected in 16/19 (median 9 days, range 6,24 days) and HHV-7 antigenemia 15/19 patients (median 17 days, range 5,58 days) after transplantation. HHV-6 appeared before CMV in most cases (12/16), HHV-7 usually together with CMV. In those cases that HHV-6 preceded CMV antigenemia, it also was a possible cause of graft dysfunction. HHV-7 and CMV were so closely overlapping, that no symptoms could solely be linked with HHV-7. Conclusion: HHV-6 and HHV-7 antigenemia usually occurred together with symptomatic CMV infection after liver transplantation. HHV-6 preceded CMV, but HHV-7 appeared together with CMV. Further investigation of the clinical significance of HHV-6 and HHV-7 antigenemia in organ transplant patients is necessary. J. Med. Virol. 78:800,805, 2006. © 2006 Wiley-Liss, Inc. [source] Dilinoleoylphosphatidylcholine Reproduces the Antiapoptotic Actions of Polyenylphosphatidylcholine Against Ethanol-Induced Hepatocyte ApoptosisALCOHOLISM, Issue 6 2003Ki M. Mak Background: Polyenylphosphatidylcholine (PPC), a mixture of polyunsaturated phosphatidylcholines extracted from soybeans, attenuates hepatocyte apoptosis induced by ethanol feeding of rats. Our aims were to evaluate whether dilinoleoylphosphatidylcholine (DLPC), the main component of PPC, reproduces the antiapoptotic actions of PPC against alcohol-induced apoptosis and to identify the apoptotic proteins that are affected by PPC and DLPC. Methods: Rats were fed Lieber-DeCarli liquid diets containing ethanol (35% of energy) or an isocaloric amount of carbohydrate for 4 weeks. Another group of rats were given the ethanol diet supplemented with PPC (3 g/liter) or DLPC (1.5 and 3 g/liter). Hepatocyte apoptosis was assessed by terminal transferase-mediated dUTP nick end labeling staining and by caspase 3 enzyme activity. Activity of caspases 3 and 9 was assayed by using fluorogenic peptide substrates. Cytochrome c was quantified by enzyme-linked immunosorbent assay. The protein contents of cytochrome c, procaspase 3, caspase 3, Bcl-xL, and Bax were analyzed by Western blot and quantified by densitometry. Lobular localization of active caspase 3 was examined by immunoperoxidase staining. Results: PPC and DLPC decreased ethanol-induced increases in hepatocyte apoptosis, cytosolic cytochrome c, and caspase 3 content and its activity. Caspase 3 activity correlated with the number of apoptotic hepatocytes. Active caspase 3 was present predominantly in perivenular hepatocytes, and ethanol feeding extended it to lobular hepatocytes; this ethanol effect was reduced by PPC and DLPC. Ethanol significantly decreased Bcl-xL in homogenate, mitochondria, and cytosol, and there was a trend for increased Bcl-xL in these fractions after PPC and DLPC supplementation. Microsomal Bcl-xL did not differ between treatment groups. Bax was detected in homogenate and cytosol, and its level was not affected by ethanol. Conclusions: DLPC, at a dose contained in PPC, reproduces the antiapoptotic actions of PPC through a reduction in cytosolic cytochrome c concentration and caspase 3 activity, possibly in association with up-regulation of Bcl-xL expression. Because DLPC is a pure and well defined compound, it may be more suitable than PPC for intervention against alcohol-induced apoptosis. [source] Aldose Reductase Inhibition Counteracts Diabetes-Induced Retinal Oxidative Injury, Glial Activation, and ApoptosisACTA OPHTHALMOLOGICA, Issue 2007IG OBROSOVA Purpose: To access the role for increased aldose reductase (AR) activity in oxidative injury, glial activation, and apoptosis in retinae of diabetic rats and high glucose-exposed cultured retinal pericytes and endothelial cells. Methods: Control (C) and STZ-diabetic (D) rats were treated with/without the AR inhibitor fidarestat (F, 16 mg/kg/g, for 10 wks after 2 wks without treatment). The rate of apoptosis was assessed in flat-mounted retinas by TUNEL assay with immunoperoxidase staining, and nitrotyrosine (NT), poly(ADP-ribose) [PAR, a marker of poly(ADP-ribose) polymerase activation] and glial fibrillary acidic protein (GFAP) expression in retinal sections by immunohistochemistry. Primary bovine retinal pericytes and endothelial cells were cultured in 5 mM or 30 mM glucose with/without F (10 microM) for 3-14 d. Apoptosis was assessed by TUNEL assay, NT and PAR by immunocytochemistry, and Bax and Bcl-2 expression by Western blot analyses. Results: The number of TUNEL-positive nuclei (Mean ± SEM) was increased ~4-fold in D (207 ± 33 vs 49 ± 4 in C, p < 0.01), and this increase was partially corrected in D+F (106 ± 34, p < 0.05 vs D). The apoptotic cell number increased with prolongation of exposure of both pericytes and endothelial cells to high glucose. F counteracted high glucose-induced apoptosis, and NT and PAR accumulation in both cell types. Antiapoptotic effect of F in high glucose-exposed retinal pericytes was not associated with inhibition of Bax or increase in Bcl-2 expression (endothelial cell studies are in progress). Conclusions: AR inhibition with fidarestat conteracts diabetes-associated retinal oxidative injury, glial activation, and apoptosis. [source] | ||||||||||