Renato Coda

Selective growth response to IL‐3 of a human leukaemic cell line with megakaryoblastic features

A new human leukaemic cell line (M‐O7) with the phenotypic characteristics of CFU‐mega is described. Its cells are positive for T200 leucocyte common antigen (LCA) and negative with MAbs recognizing T and B cells and mature myelomonocytic antigens. In contrast, they react with MAbs recognizing antigenic determinants common to multilineage (CD13, CD33, CD34) and to bipotent erythromegakaryoblastic (CD36, H25) haemopoietic precursors, and with MAbs specific for platelet glycoproteins (CD41w, CD42w). A small proportion (10%) of the cells were large and multinucleated, and on electron microscopy examination showed peripheral splitting of platelet‐like cytoplasm particles. When transferred to a serum‐free Iscove modified Dulbeccos medium supplemented with human insulin and transferrin, M‐O7 cells stop proliferating. Of the haemopoietic growth factors tested for their ability to restore the proliferative activity of this quiescent population, only rH IL‐3 proved effective. Moreover, it also increased the cloning efficiency in methylcellulose more than any other CSFs. The M‐O7 cell line may provide a valuable tool for the biological assay of IL‐3, and a model for biochemical studies of the megakaryocytic lineage.

Mediators of immune-complex-induced aggregation of polymorphonuclear neutrophils. II. Platelet-activating factor as the effector substance of immune-induced aggregation.

[目的]观察灭幽汤对幽门螺杆菌（helicobacter pylori,Hp）相关性胃炎脾胃湿热证模型小鼠TLR4、NF-κB65的蛋白及mRNA、TNF-α、IL-8表达的影响,探讨灭幽汤治疗Hp相关性胃炎脾胃湿热证的机制.[方法]将70只BALB/c小鼠随机分为对照组、模型组、高浓度灭幽汤组（高灭组）、低浓度灭幽汤组（低灭组）、胃三联组（替硝唑＋克拉霉素＋枸橼酸铋钾颗粒）,每组14只;采用复合因素（肥甘食物＋湿热环境＋幽门螺杆菌）建立BALB/c小鼠Hp相关性胃炎脾胃湿热证模型;造模成功后连续给药14 d,而后分别采用免疫组化检测TLR4、NF-κB65蛋白,qPCR检测TLR4 mRNA、NF-κB65 mRNA的表达情况,采用ELISA检测血清TNF-α、IL-8的表达.[结果]模型组TLR4、NF-κB65的蛋白及mRNA、血清TNF-α、IL-8含量表达均高于对照组,差异有统计学意义;通过药物治疗后,低灭组、高灭组、胃三联组TLR4、NF-κB65的蛋白及mRNA、血清TNF-α、IL-8含量表达均低于模型组,差异有统计学意义;高灭组TLR4、NF-κB65的蛋白及mRNA、血清TNF-α、IL-8含量表达虽低于胃三联组,但差异无统计学意义.[结论]灭幽汤可能通过干预TLR4/NF-κB65信号通路,抑制TLR4、NF-κB65及下游因子的表达以达到治疗Hp相关性胃炎脾胃湿热证的目的.

Mediators of Immune-Complex-Induced Aggregation of Polymorphonuclear Neutrophils

This study reports the results of in vitro investigations on the aggregation of polymorphonuclear neutrophils (PMN) induced by the C5a anaphylatoxin complement component as well as the cationic proteins (CP), which are released by challenging PMN with immune complexes (IC). The carboxy-peptidase-derived des-Arg fragments of CP and C5a; CPi and C5ai, inactive in terms of anaphylactic and chemotactic activity, nevertheless showed a more potent ability to aggregate PMN than CP and C5a. The process of PMN aggregation required metabolic energy and divalent cations, Ca++ and Mg++. The microtubular system and the subplasmalemmal microfilaments appeared to be of critical importance. Electron microscopic studies on aggregates of PMN obtained on stimulation with CP, C5a, CPi and C5ai showed parallel tracts of variable length of cell membranes at the points where cells were in contact with each other.

Localization of neutrophil cationic proteins and loss of anionic charges in glomeruli of patients with systemic lupus erythematosus glomerulonephritis

Abstract The localization of human neutrophil cationic protein (NCP) antigens in 14 patients with systemic lupus erythematosus (SLE) glomerulonephritis was investigated by immunofluorescence, using a rabbit anti-NCP serum. The specificity of the antiserum was demonstrated by its ability to bind exclusively to cytoplasmic structures of human polymorphonuclear neutrophils (PMN), but not of lymphocytes, monocytes, or platelets. Furthermore, the immunofluorescence reaction was abolished by absorption of the antiserum with purified NCP. In 7 patients with active SLE, severe proteinuria, and nephrotic syndrome, NCP were detected in glomerular capillary walls (GCW). The glomerular deposits of NCP were associated with a loss of glomerular polyanions (GPA), as revealed by colloidal iron and Alcian blue staining. The PMN, present in the peripheral blood of these patients, were markedly depleted of NCP. The depletion of NCP in circulating PMN, and the presence of NCP deposits in GCW with concomitant loss of GPA are consistent with the interpretation that NCP are released in vivo and that they directly contribute to the development of glomerular injury and proteimuria in patients with active SLE glomerulonephritis.

4-Aminobiphenyl-DNA adducts and p53 mutations in bladder cancer.

Epidemiologic studies have suggested that smokers of air‐cured tobacco (rich in arylamines) are at higher risk of bladder cancer than smokers of flue‐cured tobacco. The risk has been shown to be modulated by the N‐acetyltransferase genotype. We analyzed the biopsies of 45 patients with bladder cancer. p53 mutations were sought by direct sequencing, and 4‐aminobiphenyl‐DNA adducts were measured by negative ion gas chromatography‐mass spectrometry. 4‐Aminobiphenyl‐DNA adducts were higher in smokers of air‐cured tobacco and in current smokers, but no relationship with the number of cigarettes smoked was found. Adducts were higher in more advanced histologic grades of tumors. No pattern was evident for p53 mutations. Seven of 9 mutations occurred in grade 3 tumors. No association was found between 4‐ABP adducts and GSTM1 or NAT2 genetic polymorphisms. Int. J. Cancer 75:512–516, 1998.© 1998 Wiley‐Liss, Inc.

Glomerular findings in mixed IgG-IgM cryoglobulinemia

SummaryA case of immune-complex glomerulonephritis with membrano-proliferative patterns in a patient with mixed IgG-IgM cryoglobulinemia has been investigated; particular attention has been payed to histological, immunofluorescence, histochemical and electron microscopical findings in order to elucidate the nature of the intraluminal thrombi. The so called thrombi, strongly positive for IgG and IgM with the immunofluorescence, appeared to be clusters of mononuclear cells which, for their ultrastructural and histochemical (nonspecific esterase activity) patterns have been considered as monocytes. They showed cytoplasmic vacuoles containing microtubular structures; the latter have been detected free in the capillary lumina and in the basement membranes as well. These structures have been considered as the ultrastructural counterpart of the IgG-IgM immune-complexes responsible of the nephropathy. Monocytes have been hypotesized to play a pathogenetical role, acting as cleaners of the cryoprecipitate from the capillary lumina.

Dual secretory and myoepithelial differentiation in the transplantable R3230AC rat mammary carcinoma

SummaryThe hormone-responsive R3230AC mammary carcinoma, serially transplantable in Fisher rats, shows striking functional and morphological similarities to the normal mammary gland. We have studied its cellular composition by both light and electron microscopy, employing markers of myoepithelial and epithelial cells. We identified two cell types: the major cellular component corresponded to epithelial milk-protein secreting cells, while a second component showed immunocytochemical and ultrastructural characteristics of the myoepithelial cells. These cells were positive with a monoclonal antibody detecting alpha smooth muscle actin. The dual differentiation which normally occurs in breast ducts is therefore reproduced in a malignant experimental tumor. The coexistence of neoplastic cell populations, divergent in morphology and function, that persist in a tumor despite many transplant generations, leads to reconsideration of the relationship between cellular differentiation and malignant transformation.

Human platelet cationic proteins bind to rat glomeruli, induce loss of anionic charges and increase glomerular permeability

The binding of human platelet cationic proteins (HuPlt CP) to rat renal cortexin vitro andin vivo, the loss of glomerular polyanions (GPA) and the increase in glomerular permeability were studied. HuPlt CP were purified by sequential cation-exchange chromatography and chromatofocusing, by which these proteins were shown to be highly cationic in nature (pI 10.5) and mainly composed of three molecular species of 55.60 kD, 40.45 kD, and 10 kD as studied by gel permeation in high pressure liquid chromatography and SDS-polyacrylamide gel electrophoresis. Binding of HuPlt CP to glomerular capillary walls (GCW), mesangium and to peritubular capillaries of the rat renal cortex was demonstrated by immunofluorescence, using a specific goat anti-HuPlt CP antiserum, after incubation of the sections with HuPlt CPin vitro and after injection of HuPlt CPin vivo. This interaction was ionic in nature, since treatment of sections with heparin abrogated the binding of HuPlt CP to glomerular structures. The glomerular deposits of HuPlt CP were associated with the loss of GPA as revealed by colloidal iron staining (light microscopy) in bothin vitro andin vivo experiments and by ruthenium red staining (electron microscopy) inin vivo studies. After the injection of native ferritin, the increase in glomerular permeability produced by an infusion of HuPlt CP was observed by the increased ratio of counted particles within the glomerular basement membrane with respect to controls. The binding of HuPlt CP to GCW and the loss of GPA was consistent with the interpretation that HuPlt CP may increase glomerular permeability due to the neutralization of GPA.

Histological chronic prostatitis and high‐grade prostate intra‐epithelial neoplasia do not influence urinary prostate cancer gene 3 score

Study Type – Diagnostic (case series) Level of Evidence 4 Whats known on the subject? and What does the study add? While the relationship between PCA3 score and clinical or histological prostatitis was quite proven even in small patient groups, conversely the relationship between PCA3 score and HG‐PIN is still under debate. We demonstrated in a large series (432 patients) that histologically documented chronic prostatitis and HG‐PIN have similar PCA3 scores to patients with BPH and/or normal parenchyma at biopsy.

A combined glucagonoma and VIPoma syndrome. First pathologic and clinical report.

We report a case of pancreatic tumour metastatic to the liver in a patient with insulin‐treated diabetes, anaemia, cheilitis, necrolytic migratory erythema, hypokalemia and chronic watery diarrhea, a picture suggesting combined glucagonoma and VIPoma syndromes. Immunocytochemistry of a biopsied hepatic metastatic nodule revealed both glucagon and vasoactive intestinal peptide (VIP) positive cells. Increased plasma glucagon and VIP levels were detected (values of 900 pmol/1 and 277 pmol/1 respectively). This is the first reported case showing not only immunocytochemical, but also clinical evidence of the combined secretion of these hormones.