Immunogenic Dendritic Cell Generation from Pluripotent Stem Cells by Ectopic Expression of Runx3.

Application of dendritic cells (DCs) to prime responses to tumor Ags provides a promising approach to immunotherapy. However, only a limited number of DCs can be manufactured from adult precursors. In contrast, pluripotent embryonic stem (ES) cells represent an inexhaustible source for DC production, although it remains a major challenge to steer directional differentiation because ES cell-derived cells are typically immature with impaired functional capacity. Consistent with this notion, we found that mouse ES cell-derived DCs (ES-DCs) represented less mature cells compared with bone marrow-derived DCs. This finding prompted us to compare the gene expression profile of the ES cell- and adult progenitor-derived, GM-CSF-instructed, nonconventional DC subsets. We quantified the mRNA level of 17 DC-specific transcription factors and observed that 3 transcriptional regulators (Irf4, Spi-B, and Runx3) showed lower expression in ES-DCs than in bone marrow-derived DCs. In light of this altered gene expression, we probed the effects of these transcription factors in developing mouse ES-DCs with an isogenic expression screen. Our analysis revealed that forced expression of Irf4 repressed ES-DC development, whereas, in contrast, Runx3 improved the ES-DC maturation capacity. Moreover, LPS-treated and Runx3-activated ES-DCs exhibited enhanced T cell activation and migratory potential. In summary, we found that ex vivo-generated ES-DCs had a compromised maturation ability and immunogenicity. However, ectopic expression of Runx3 enhances cytokine-driven ES-DC development and acts as an instructive tool for the generation of mature DCs with enhanced immunogenicity from pluripotent stem cells.

Value of carcinoembryonic antigen (CEA) and cholesterol assays of ascitic fluid in cases of inconclusive cytology.

AIM: To determine whether assays of carcinoembryonic antigen (CEA) and cholesterol in ascites add diagnostic value to cytology. METHODS: The additional diagnostic efficacy of the biochemical assays was studied in the ascitic fluid from 130 patients, of whom 57 had peritoneal carcinomatosis. All diagnoses were verified by subsequent necropsy and/or histology. RESULTS: CEA concentrations over 5 ng/ml indicated carcinomas, occasionally without peritoneal involvement of the tumour. However, increased values were significantly more common in cancer with peritoneal involvement (p < 0.01), giving a sensitivity of 51% and specificity of 97% for carcinomatosis. A cholesterol value exceeding 1.21 mmol/litre was found in 93% of cancers with peritoneal involvement, but it was not entirely specific (96%) for carcinomatosis. Simultaneous increases in CEA and cholesterol concentrations were specific for carcinomatosis and this combination increased the sensitivity for diagnosing carcinomatosis from 77% with cytology alone to 88%. The correct diagnosis could thus be made in five of 12 cases with inconclusive cytology. CONCLUSIONS: The measurements of both CEA and cholesterol concentrations in ascites give additional specific information about peritoneal carcinomatosis and can therefore be a useful adjunct to cytology-in particular, in inconclusive cases.

Histiocytic and t-cell rich b-cell lymphoma (TCRBCL) of the stomach.

Although stomach is a frequent site of extranodal lymphomas, histiocyte-rich TCRBCL has not yet been described there. Even histology of repeated gastrobiopsies of this uncommon, diffuse, large B-cell lymphoma may be inconclusive and partial gastrectomy cannot be avoided. It is only immunohistology (CD20, CD43, CD68) of the paraffin blocks from the resection specimen that can lead to the final diagnosis of intermediate grade malignant lymphoma.

New human Dirofilarioses in Hungary.

About ten cases of filariosis have recently been recorded in the Hungarian medical literature, six of them caused by Dirofilaria repens. Dirofilaria repensis a mosquito-transmitted filaroid worm in the subcutaneous tissue of dogs and cats in the temperate areas of the Old World. It accidentally infects man, too, and can remain unidentified due to physicians, poor knowledge of the parasite. In the last two years six new Dirofilaria repensinfections have been found in various parts of the country: five localised dermally and one in the deep tissues. Two of the cases might have been acquired in Italy during summer travels. Four patients, however, have never been abroad, these cases must be considered autochtonous infections. The thickness of the multilayered cuticle of the worm, diameter of the body and the size, form and number of the longitudinal ridges on its surface are used in the histological diagnosis of the the parasite.

Central neurocytoma with malignant course. Neuronal and glial differentiation and craniospinal dissemination.

Central neurocytoma is a benign neuronal tumor of young adults in the lateral cerebral ventricles with characteristic X ray and light microscopic findings. In many respects typical central neurocytoma is reported below, with recurrence in the third month requiring reoperation. Death ensued in the fifth postoperative month. Subsequent histology proved progressive vascular proliferation and increasing, unusual glial differentiation of the neuronal tumor. At autopsy tumorous seeding blocked the liquor circulation. A thin tumorous layer covered the surface of all ventricles, the cerebellum and medulla oblongata. The GFAP positive cells out-numbered the synaptophysin positive ones. Increase of GFAP positivity and vascular proliferation of the central neurocytoma may be alarming signs suggesting a malignant course in addition to the other atypical features.

Immunodiffusion analysis of the antigenic structure of the transplantable MC 29 tumor.

By the use of specific anti-chicken liver sera it could be proved with immunodiffusion techniques that the transplantable MC 29 tumor line contains liver-specific antigens. Production of chicken serum-proteins by the tumor could also be demonstrated. Thus, the transplantable MC 29 tumor line can be regarded as a hepatoma.

epsilon-N-trimethyl-lysine: a natural cell component with mitogenic activity.

The blastogenic capacity of a new mitogenic agent, epsilon-N-trimethyl-lysine (TML) was examined on human peripheral blood lymphocytes. The changes in relative DNA and histone content after TML treatment testified that activated lymphocytes passed through the whole cell cycle. The comparison of the 3H-TdR incorporation rate into unseparated T-B lymphocyte and purified T lymphocyte cultures suggested a preferential effect on the thymidine incorporation of T lymphocytes. TML induced preferentially high responses in cases of blood cells taken from women. The possibility of DNA repair mechanism could be excluded. The results of combined TML-PHA treatment experiments suggested a difference in the trigger mechanism of these two mitogenic compounds.

Investigation of group-specific antigen during the culture of MC-29 hepatoma.

The short-time culture established from MC-29 chicken hepatoma was used for the analysis of cell morphology and expression of avian gs antigen. The content of gs antigen was not changed during the period of cultivation. Two morphologically different types of cells were present in tested tumour cells. Fluorescent antibody test showed the presence of gs antigen in both types of cells.

Human Dirofilaria repens infection in Hungary: a case in the spermatic cord and a review of the literature.

Orchiectomy was performed in a 37-year-old Hungarian man exhibiting a swelling in his right testicle. Histology revealed a nodule attached to the spermatic cord, consisting of a granulomatous tissue around sections of a nematode. The worm was identified as Dirofilaria repens, an uncommon parasite in Hungary. As the patient had been abroad only in Italy where cases of dirofilariosis in dogs and humans are relatively frequent, it is assumed that the infection might have been acquired in that country 5 years earlier. This is the fifth case, published so far in the world, of such a localization in a human. The human cases of dirofilariosis reported in Hungary are reviewed.

[Oncologic diagnosis of serous effusions in body cavities (cytologic study and evaluation of cholesterol and carcinoembryonic antigen levels)]. / A testüregi folyadékgyülemek daganatdiagnosztikája (A cytologia, a koleszterin és a carcinoembryonalis antigén vizsgálat együttes értékelése).

Conventional cytological evaluation of serous effusions often yields border line result: apart from positive (malignant) or negative (benign) diagnoses, a relatively large part of the findings are "suspicious for malignancy" (P3). In the present paper the authors have analysed to what extent contributes the determination of cholesterol and CEA levels of ascites and pleural effusion to the diagnostic accuracy of cytologically "suspicious" (P3) cases. In 155 histologically controlled cases, specificity, sensitivity and diagnostic efficiency were assessed on the basis of cytology as well as the determination of CEA and cholesterol levels. Statistical parameters were determined for each method separately and for their combined application. According to the findings, cholesterol and CEA levels over 1.16 mmol/l and 2.5 ng/ml, respectively, indicate malignancy. In 19 out of 29 cytologically "suspicious" cases (66%), which histologically proved to be positive, cholesterol and/or CEA levels were elevated. In all of the 12 non-neoplastic "suspicious" cases the two parameters were under the cutoff values. The application of an easy and inexpensive cholesterol test proves to be a sensitive technique for indicating carcinomatosis and it completes adequately the specific cytological evaluation. If clinical symptoms speak for tumor, but the cytology is negative, the evaluation of CEA level may prove to be useful as far as it can indicate cancer not yet accompanied by carcinomatosis.

[Experiences with a new vacuum-accelerated microwave histoprocessor]. / Tapasztalataink egy új, vákuummal felgyorsított mikrohullámú hisztoprocesszorral.

Methods of microwave and vacuum accelerated fixation, dehydration and paraffin embedding are described. They can be carried out in a new type of vacuum and temperature stabilizable microwave histoprocessor: MFX-800. The whole histoprocessing lasts 3.5-5.5 hours depending on the thickness of the tissue blocks. Preserved structural detail, intensive staining and good antigen preservation were achieved with various tissues. This new histoprocessing facility can be highly recommended for pathology laboratories interested in speeding up the processing procedure, as well as in immunohistochemistry.

Study on the "toxic oxygen effect" of Janus green B in mouse ascites tumour cells.

10(7) mouse ascites tumor cells/ml incubated at 37 degrees C in 0.5 to 1.0 X 10(-4) M Janus green B or in 1.0 X 10(-4) M phenazine methosulphate are destroyed in 100 per cent oxygen atmosphere but remain transplantable in nitrogen atmosphere. The "sensitizing" effect of oxygen can be substituted by SH inhibitors (iodoacetic acid, iodoacetamide and their spinlabelled variants) as well as by some nitroxide free radicals. The "oxygen effect" is blocked by mercaptoethanole or cooling. Compared with the spectrum of native cells a more symmetrical singlet of larger amplitude, approximately g = 2 value, arose in the ESR spectrum of Janus green B treated cells. The "oxygen effect" observed in the presence of Janus green B differs in several ways from the oxygen effect of ionizing radiation and from the "photodynamic" effect.

Large cell dysplasia of hepatocytes, is it a premalignant condition?

Incidence and character of large cell dysplasia (LCD) of liver were examined by histological, statistical and morphometric methods in an aged Middle European population. The necropsy material of 487 patients with micronodular cirrhosis and cirrhosis with primary liver tumour was studied. In addition, 50 primary tumours developed in non-cirrhotic livers, 34 metastatic tumours and 90 normal livers were examined. According to the findings, LCD often developed in aged patients. The high occurrence is presumably due to the high age. The frequency of LCD was significantly higher in active than in inactive cirrhosis. LCD could mostly be found around metastatic tumours, especially those with large, destructive foci. Sex differences occurred in a part of cases but an age dependence could not be demonstrated. Normal and dysplastic cells did not differ significantly in most morphometric parameters but there was a significant difference between tumorous and normal or dysplastic parameters. The data suggested that LCD is a secondary, reactive phenomenon occurring in various pathological processes.

The free radical signal of pigment gallstones.

Bile stones have been studied by Electron Spin Resonance (ESR) spectroscopy. Pigment containing stones show a 8.76 +/- 0.02 gauss wide free radical singlet at g approximately 2.003 region of the spectrum. Amplitude of this free radical signal linearly correlates with pigment content of the stone, which can be applied in the estimation of pigment content. The possible role of free radicals is suggested in the formation of pigment bile stones.

Electron spin resonance spectra of chicken liver and hepatoma tissue embedded in paraffin.

ESR spectra of chicken liver and hepatoma tissue embedded in paraffin display signals characteristic of free radicals and paramagnetic complexes. Owing to embedding the spectrum is altered as compared to that of the native tissue but remains characteristic of the respective tissues. The spectra of tissues embedded in paraffin allow no more than qualitative conclusions to be drawn concerning the spectra of the native tissues, even if appropriate controls are applied.