Mucosal bromodomain-containing protein 4 mediates aeroallergen-induced inflammation and remodeling

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Evaluation of proliferation parameters in in vivo bromodeoxyuridine labelled lung cancers

In a series of 44 bronchial biopsies from patients suspected of having endobronchial lung carcinoma, the validity of proliferating cell nuclear antigen (PCNA) and Ki67 antigen as proliferative indicators was evaluated in ethanol fixed, paraffin embedded tissue. The percentages of cells positive for these markers were compared to the in vivo bromodeoxyuridine (BrdU) labelling index. A good correlation was found between PCNA immunoreactivity and BrdU labelling index, while Ki67-antigen expression showed a significant relation with BrdU labelling index and with PCNA expression. All three parameters showed a trend towards similar values for the individual cases. Based on the fact that Ki67 antigen is expressed in all cycling cells, whereas replicon-associated PCNA and BrdU only reflect the S-phase fraction, the differences between Ki67-antigen scores on the one hand and BrdU and PCNA scores on the other were smaller than expected. In order to determine the degree of concordance between immunohistochemically and flow cytometrically detected proliferation variables, BrdU incorporation was measured using both methods in duplicate bronchial specimens. Discrepancies in labelling indices were observed predominantly in DNA diploid samples, with consistently lower values in the flow cytometrically analysed specimens. In tumour specimens with an aneuploid DNA content, flow cytometric determination of proliferative activity yielded results similar to those obtained by tissue section examination. We conclude that the scores for PCNA and Ki67 antigen, immunohistochemically detected in ethanol fixed, paraffin embedded tissue reflect functional proliferative activity.     

人核受体hLRH-1不同变异体在多种肿瘤组织和细胞系中的表达

目的:初步分析人核受体hLRH-1不同变异体在多种肿瘤组织和细胞系中的表达。方法:常规RT-PCR法分析hLRH-1v1和hLRH-1在肝癌等多种肿瘤组织和HepG2等肿瘤细胞系中的表达情况，实时定量PCR法分析hLRH-1v1，hLRH-1和hOct4在HepG2，SMMC-7721和BEL-7402三株肝癌细胞系的表达水平。结果:hLRH-1v1的表达见于所有检测的12种类型肿瘤和8种恶性肿瘤细胞系，hLRH-1的表达则仅在12种类型肿瘤中的6种肿瘤组织中可检测到，但8种恶性肿瘤细胞系均为hLRH-1阳性表达；在HepG2，SMMC-7721和BEL-7402三株肝癌细胞系中hLRH-1v1和hOct4的表达呈正相关。结论:hLRH-1v1在肿瘤中广泛表达，可能在维持肿瘤干细胞的自我更新中发挥重要作用。     

Cell proliferation rate and nuclear morphometry in Roberts syndrome

Roberts syndrome (RS) is a rare autosomal recessive disorder characterized primarily by symmetric reduction anomalies of all limbs, growth retardation and craniofacial abnormalities. Most RS patients are reported to present a typical abnormality of their constitutive heterochromatin, accompanied by abnormal cytological growth characteristics. We present an extremely severe case of an RS fetus, karyotypically documented, with a clinical presentation including growth deficiency, tetraphocomelia, frontal meningocele, craniofacial abnormalities and penile enlargement with hypospadias. Nuclear morphometrical analysis in tissues of various organs revealed a reduced nuclear size in RS as compared to normal controls, and statistically significant differences in morphometric parameters related to the nuclear shape. Immunohistochemical study of the same organs showed a reduced expression of proliferating cell nuclear antigen in the presented case, thus indicating a decreased cell proliferation rate in RS. Our results reconfirm previously reported findings in cultured fibroblasts of RS cases, thereby reinforcing on a histologic level, the hypothesis that reduced cell proliferation may be involved in the growth retardation and the reduction abnormalities observed in RS.     

Enhanced polymer one-step staining (EPOS) for proliferating cell nuclear antigen (PCNA) and Ki-67 antigen: Application to intra-operative frozen diagnosis

Enhanced polymer one-step staining (EPOS) is a novel, highly sensitive one-step immunostaining method. This simple and rapid technlque was applied to intra-operattve frozen diagnosis. The markers of choice were proliferating cell nuclear anmen (PCNA) and Ki-67 antigen. These cell prollferation markers were both identifiable in fresh frozen see tions of the human tonsil In approximately 7 min. The suitable staining sequences are as follows. Frozen sections prepared using 3-aminopropyitimethoxysilane-cpated glass slides are immediately fixed, without air drying, for 15s in a mixture of 50% formalin and 50% methanol for PCNA, and in 10% formalln for Ki-67 antigen. After a brief rinse in phosphate-buffered saline (PSS), sections are incubated with the EPOS antibody for 3 min, followed by PBS rinse for 1 min. The peroxidase activity is visualized in diaminobenzidine-H₂O₂ solution containing 10mmol/L imidazole for 2 min. After a light rinse in tap water, the nuclei are briefly counterstained with 5% methyl green. When necessary, endogenous peroxi-dase blockage in 1% periodic acid solution for 1 min is added before the EPOS antibody incubation. This procedure is applicable to frozen sections of gastric cancers, malignant lymphomas, and brain, liver and peritoneal lesions in which differential diagnosis between benignancy and malignancy was required.     

T suppressor lymphocytes reverse ongoing acute allograft rejection

(LEW X BN)F1 cardiac allografts are rejected within 8 days in untreated LEW recipients. At the critical time point of 5 days after transplantation, the obviously rejecting grafts are enlarged and maximally infiltrated by host cells as shown by 111In-labeled lymphocyte tracer studies. However, when such hearts were retransplanted back to naive (LEW X BN)F1 secondary hosts, they survive indefinitely, showing that even late rejection is reversible in the absence of sustained host immunological drive. Attempts were then made to abrogate this advanced immune responsiveness using Cyclosporine (CsA). CsA therapy (15 mg/kg/day for 7 days) starting from day 5 produced indefinite graft survival, similar as if initiated at the time of operation. Addition of exogenous IL-2, which drives the proliferation of Tc, could not reverse this effect. Serial changes in phenotype of lymphocyte subpopulations infiltrating both acutely rejecting and indefinitely functioning cardiac allografts in unmodified and CsA treated hosts, respectively, were then studied. Ratio of Th:Tc/s cells in acutely rejecting grafts was 1.6 by day 3; it inverted abruptly to 0.7 by day 5-6, suggesting predominance of Tc/s during the later stages of allograft rejection. Similarly, treatment with CsA produced a transient depression of Th, with recovery of original Th:Tc/s ratio during the next 2-3 weeks. Adoptive transfer experiments were then performed to investigate the functional significance of these findings.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative study of the prognostic value of nuclear grade and silver-binding nucleolar organizer region in renal cell carcinomas.

Silver-binding nucleolar organizer region (AgNOR) and nuclear grade were histologically assessed in 112 renal cell carcinomas. Nuclear grade was better capable of stratification than AgNOR in terms of survival prediction. It also had prognostic significance when total metastatic rates were evaluated. Although higher mean AgNOR numbers were constantly associated with higher nuclear grades, two arbitrary classifications of AgNOR (less than 1.5, 1.5-3.5, greater than 3.5; less than 2, 2-3, greater than 3) failed to resolve the survival difference.     

Barium blocks cell membrane and tight junction conductances inNecturus gallbladder epithelium

The site and concentration dependence of the blocking effect of Ba²⁺ onNecturus gallbladder epithelium has been investigated. A new approach was used which combines time-dependent electrical cell coupling analysis with intermittently performed measurements of transepithelial and apparent intracellular impedance. From the coupling pulse data the sum of apical and basolateral membrane conductances is obtained, which is then held constant during fitting of the impedance data. This combination technique yields more reliable estimates of apical and basolateral membranes resistances (R _a,R _bl) and of tight junction resistance (R _j) than our previous impedance analysis technique. Using the new approach we have found that luminal Ba²⁺ concentrations between 0.5 and 1.0 mmol/l increaseR _a with saturation-type kinetics without affectingR _bl andR _j, while higher luminal Ba²⁺ concentrations progressively increaseR _j. Corresponding effects were observed under serosal Ba²⁺. The results validate the new impedance analysis approach and demonstrate that millimolar concentrations of Ba²⁺ block tight junction conductances. Accordingly, Ba²⁺ can no longer be considered a tool to exclusively alter cell membrane resistances in epithelia.     

Functional histology of the human thymus

Summary The thymus develops from a paired epithelial anlage in the neck. This review considers how ectoderm (vesicula cervicalis) and endoderm (third pharyngeal pouch) contribute to the epithelial stroma of the thymus. Stromal elements of mesodermal origin are capillaries, septae and perivascular spaces and single invading cells. These elements separate the thymus into pseudolobuli. The thymus epithelial space and the perivascular spaces are always separated from each other by a closed, flat epithelial cell layer, with a basal lamina which contributes to the blood-thymus barrier. From the 9th gestational week, prethymic precursor cells from hemopoietic centers, begin to invade the thymus anlage. There they finally mature to committed post-thymic T cells. The thymus microenvironment of postnatal thymus is composed of six different types of epithelial cells and several stromal cells of mesodermal origin. The location of these diverse stationary cells is described, and their functional significance is discussed. Obviously these stromal cell types have a special function in providing the proper environment for T-cell maturation. The function of the thymus includes the maturation and/or selection of antigen specific T-cells. The main issue of intra-thymic T-cell differentiation is the development and expression of T-cell-antigen receptors. The great diversity of these receptors is generated by a rearrangement of the T-cell-receptor-genes in order to furnish the host with a mature T-cell repertoire that is capable of recognizing the world of extrinsic antigens. In a synopsis the manyfold interrelationships between the thymus microenvironment and the developing thymocytes are summarised.Abbreviations BALT Bronchus Associated Lymphoid Tissue - CD Cluster of Differentiation - cCD3 cytoplasmic CD3 - mCD3 membranous CD3 - C.R. Crown Rump - GALT Gut Associated Lymphoid Tissue - HLA Human Leucocyte antigen - HLA-DR Gene product of the MHC-class II (this antigen is a surface molecule of numerous stationary and free cells of the immune system) - IDC Interdigitating Cell - IL1 Interleukin 1 (cytokine derived mainly from makrophages, but also from other cell types) - IL2 and IL4 Interleukin 2 and 4 (cytokines derived mainly from T-cell, but also from other cell types) - IL4R Interleukin 4-Receptor - Mab Monoclonal antibody - MHC Major Histocompatibility Complex - p.c. post conception - TCR T-Cell-Receptor - TNC Thymic Nurse Cell - TdT Terminal deoxynucleotidyl Transferase     

Numerical chromosomal aberrations in prostate cancer: correlation with morphology and cell kinetics

Eleven routinely processed radical prostatectomy specimens were studied for the presence of numerical chromosomal aberrations by means of in situ hybridization with nucleic acid probes specific for chromosomes 7, 10, 17, X, and Y. Cytogenetic information was correlated with morphology, tumour stage and volume as well as with cell kinetics, the latter being assessed by immunohistochemistry with antibodies raised against the proliferative cell nuclear antigen (PCNA) and against a formalin-resistant epitope of the Ki-67 antigen, MIB 1. In 5 of 11 cases, numerical aberrations of at least one chromosome were found. The cases with normal chromosome numbers were those with the smallest volumes of Gleason grade 4 and/or 5 tumour (mean 0.5 cm³) and represented tumours restricted to the prostate. Tumours with aberrations in the number of detected chromosomes showed advanced stages and large volumes of high-grade tumour (mean 12.5 cm³). All 4 tumours with positive surgical margins were recruited from a group with marked local heterogeneity in chromosome numbers. Immunostaining with MIB 1 and PCNA was most intense in areas of high-grade tumour and was positively correlated with the emergence of chromosomal aberrations. The data suggest that the appearance of numerical chromosomal aberrations in prostate cancer coincides with aggressive tumour behaviour and could be used as an additional prognostic marker.This work is part of E.K.'s doctoral thesis