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Generation of monocyte-derived dendritic cells from patients with renal cell cancer: modulation of their functional properties after therapy with biological response modifiers (IFN-alpha plus IL-2 and IL-12).

The combination of interferon-alpha (IFN-alpha) plus interleukin (IL-2) has been accepted in the treatment of metastatic renal cell carcinoma (MRCC), whereas vaccines based on IL-12 or dendritic cells (DCs) are still being investigated. Here the authors analyzed 1) the feasibility to generate functional monocyte-derived DCs (MDDCs) from patients treated with biological response modifiers (BRMs) who have MRCC, 2) the phenotypic modulations of these MDDCs during BRM treatment. Eight and 13 MRCC patients received IL-2 plus IFN-alpha or IL-12 immunotherapy, respectively. The adherent fraction of mononuclear cells from patients' blood drawn before, during, and after immunotherapy was incubated in clinically approved culture medium supplemented with 5% autologous serum, rhu granulocyte macrophage colony-stimulating factor, and rhuIL-4 for a week. At day 7 or 8 of culture, floating cells were examined in flow cytometric and functional assays (alloreactivity, proliferation assays in the presence of tetanus toxoid or tumor peptides, IL-12 secretion). In all patients except two, MDDCs could be generated but at a lower rate compared with healthy volunteers. Morphologic and phenotypical analyses revealed immature DCs with low levels of CD1a or CD83 expression throughout therapy with BRMs. Capacities in mixed leukocyte reactions were similar to those of healthy volunteers and stable during immunotherapy, whereas presentation of major histocompatibility complex class II tetanus toxoid peptide complexes was slightly enhanced during and after IL-12 therapy. IL-12 expression levels under IFN-gamma and CD40L stimulation were significantly lower in MDDC cultures from patients with MRCC compared with healthy volunteers. Overall, peripheral blood mononuclear cells from a cohort of 21 patients with metastatic disease who were treated with BRMs maintained their ability to differentiate into functional MDDCs with no selective quantitative or qualitative advantage.

Antibodies, Monoclonal↗

Impact of hospital volume on operative mortality for major cancer surgery.

CONTEXT: Hospitals that treat a relatively high volume of patients for selected surgical oncology procedures report lower surgical in-hospital mortality rates than hospitals with a low volume of the procedures, but the reports do not take into account length of stay or adjust for case mix. OBJECTIVE: To determine whether hospital volume was inversely associated with 30-day operative mortality, after adjusting for case mix. DESIGN AND SETTING: Retrospective cohort study using the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database in which the hypothesis was prospectively specified. Surgeons determined in advance the surgical oncology procedures for which the experience of treating a larger volume of patients was most likely to lead to the knowledge or technical expertise that might offset surgical fatalities. PATIENTS: All 5013 patients in the SEER registry aged 65 years or older at cancer diagnosis who underwent pancreatectomy, esophagectomy, pneumonectomy, liver resection, or pelvic exenteration, using incident cancers of the pancreas, esophagus, lung, colon, and rectum, and various genitourinary cancers diagnosed between 1984 and 1993. MAIN OUTCOME MEASURE: Thirty-day mortality in relation to procedure volume, adjusted for comorbidity, patient age, and cancer stage. RESULTS: Higher volume was linked with lower mortality for pancreatectomy (P=.004), esophagectomy (P<.001), liver resection (P=.04), and pelvic exenteration (P=.04), but not for pneumonectomy (P=.32). The most striking results were for esophagectomy, for which the operative mortality rose to 17.3% in low-volume hospitals, compared with 3.4% in high-volume hospitals, and for pancreatectomy, for which the corresponding rates were 12.9% vs 5.8%. Adjustments for case mix and other patient factors did not change the finding that low volume was strongly associated with excess mortality. CONCLUSIONS: These data support the hypothesis that when complex surgical oncologic procedures are provided by surgical teams in hospitals with specialty expertise, mortality rates are lower.

Aged↗

Heterotypic tetramer (A2D2) complexes of non-epidermal keratins isolated from cytoskeletons of rat hepatocytes and hepatoma cells.

Cytoskeletal residues obtained after extraction of rat liver and cultured rat hepatoma cells (line MH1C1) were used to isolate cytokeratin subunit complexes by solubilization in low salt buffer containing 4 M-urea. Alternatively, the complexes were prepared by solubilization of total cytoskeletal proteins in 9.5 M-urea or 6 M-guanidinium hydrochloride (Gu . HCl), followed by separation using reversed phase high pressure liquid chromatography and dialysis first against either 9.5 M-urea or 6 M-Gu . HCl and then against buffers containing either 4 M-urea or 2 M-Gu . HCl, respectively. The complexes contained only two cytokeratin polypeptides in a 1 : 1 ratio as demonstrated by electrophoresis and isoelectric focusing, i.e. components A (Mr 55,000; isoelectric point in 9.5 M-urea, pH 6.4) and D (Mr 49,000; isoelectric point, pH 5.38) which were separated from each other at urea concentrations higher than 7 M. The complex had a sedimentation coefficient S25,w of 4.96 S in 2 M-Gu . HCl. Sedimentation equilibrium analysis gave an average Mr value of 207,000 which was interpreted as a tetramer containing two chains each of A and D. This complex was also directly demonstrated by gel electrophoresis under non-dissociating conditions. Using dimethyl suberimidate to cross-link the complex in solution of 4 M-urea or 2 M-Gu . HCl, we identified covalently linked heterodimers of A and D, and a tetrameric unit containing equal amounts of A and D which was the largest cross-link product obtained. This complex was similar to the tetrameric complex of rat and human vimentin formed under the same conditions. The constituents of the cross-linked products were identified by two-dimensional ("diagonal") gel electrophoresis, involving the cleavage of the bis(amidine) cross-links after the initial separation in the first dimension. Identical cross-link products were recognized when cytokeratin filaments were used. By electron microscopy the complexes appeared as threads of 2 to 3 nm diameter with a mean length of approximately 48 nm. On dialysis to low salt buffer, the complexes formed 2 to 3 nm protofilaments, intertwisted 3 to 4 nm protofilaments and typical 7 to 11 nm intermediate-sized filaments. Complexes formed from equivalent cytokeratins of other species such as man and cow, as well as heterologous recombinations such as human component A mixed with bovine component D and vice versa, showed the same characteristics.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

[Study on recombinant BCG].

The progress of study on recombinant BCG was stated briefly. And then our studies on recombinant BCG were mentioned. Recombinant BCG secreting alpha antigen-fused merozoite surface protein 1 (MSP 1) was prepared and tested for its ability to control infections of Plasmodium yoelii. Result turned out it controlled the infection better than recombinant MSP 1 mixed with Freund incomplete adjuvant did. Recombinant BCG secreting excess amounts of antigen 85 complex A controlled infection of Mycobacterium leprae. Addition of recombinant BCG secreting alpha antigen-fused IL-2 to peritoneal exudate cells induced IFN-gamma resulting in killing bladder cancer cells more efficiently than parental BCG did.

Adjuvants, Immunologic↗

Subfragments of the papain solubilized TL antigen.

TL antigen was solubilized from the tumor ASLI (TL. 1,2,3) by papain digestion. The subfragments of 125I-labeled TL were examined by two methods. The first involved immune precipitation followed by electrophoresis on SDS-acrylamide gels. This treatment yielded three bands of molecular weight 39,000 and 19,000, as well as material which migrated with the tracking dye. In the second procedure the papain digested material was partially purified on Sephadex G-200. The active fraction from G-200 was labeled with 125Iodine, mixed with alloantiserum and rechromatographed on G-200. The isolated immune complexes were boiled in SDS and 2-mercaptoethanol, then separated on a SDS-Sephadex G-150 column. Two radioactive peaks were eluted indicating an absence of the 19,000 m.w. component following the latter method of purification.

Animals↗

Diet, nutrition, and cancer.

As the second leading cause of death in the United States, cancer is a major public health problem today. Cancer incidence varies worldwide and tends to change with migration. These epidemiological observations have led to the concept that environmental factors may be important in carcinogenesis. Diet and nutrition are receiving increased attention and the National Cancer Institute, as mandated by the Nation Cancer Act Amendments of 1974, is playing a major leadership role in expanding research efforts in the areas of environmental carcinogenesis, and nutrition in relation to cancer. The subject of diet, nutrition, and cancer is complex. Different types of cancer are not necessarily affected by dietary components in the same manner. Although the development of certain neoplasms may be repressed by specific dietary deficiencies, other types, particularly those of the liver and upper gastrointestinal tract, are actually augmented or potentiated by such deficienceis. In extrapolating results from animal experiments to humans, caution must be exercised because of possible differences in species response to the same dietary stimulus and because spontaneous tumors may react differently from experimentally-induced tumors. Diet and nutrition are viewed more appropriately as modifiers, rather than initiators, of tumorigenesis. Caloric intake, type and amount of fat, protein, amino acids, vitamins, minerals, fiber, and other dietary constituents have been studied in regard to their influence on the development of neoplasms. Dietary components may have opposing effects on tumorigenesis, i.e., protective and predisposing, and the consequence to the host will depend on the balance between these opposing forces. Studies conducted to date indicate that the modifying effect of diet and nutrition may be exerted through specific effects on 1) intestinal bacteria and substrates for bacterial metabolism, 2) microsomal mixed-function oxidase system, 3) endocrine system, 4) immunological system, 5) availability of metabolites for cell proliferations, and 6) rate of carcinogen transfer and duration of exposure to the carcinogen. More research is needed to elucidate the interaction between diet and each of these factors and to test the validity of the mechanisms proposed to explain such interactions. These studies will lead not only to a better understanding of carcinogenesis itself but also to a new understanding of the influence of diet on human physiology and metabolism.

Animals↗

Influence of BCG on the development of 256 Walker carcinosarcoma inoculated intratesticularly in the rat, in relation to thyroid status alerations.

The effect of nonspecific antitumoral immunity caused by BCG on intratesticular development of 256 Walker carcinosarcoma has been studied on male hyper- or hypothyroidectomized rats. Thyroid hormones level was altered by administration of T4. Alkiron or thyroidectomy. The time and route of BCG administration differed from one lot to another. Authors' results show that administration of BCG before or concomitantly with the graft stimulates tumor development, whereas administration of BCG mixed with a suspension of tumoral cells causes marked inhibition in the development of primary tumor and metastasis, an inhibition which is not influenced by the thyroid hormones level. In vitro proliferation of the tumoral cells as well as histopathologic examination of the tumors suggest the involvement of a complex process in achieving the nonspecific antitumoral immunity.

Animals↗

Immunohistochemical localisation of cytokeratin and vimentin intermediate filament proteins in canine mammary tumours.

Monoclonal antibodies to human stratifying keratin (StK), simple keratin (SK), broad spectrum keratin (BSK) and vimentin were applied to 47 canine mammary tumours (three benign hyperplasia, 26 benign mammary tumours, one malignant mixed tumour, two malignant complex tumours, seven lobular carcinomas, three papillary carcinomas and five squamous cell carcinomas). In benign hyperplasia the SK antibody reacted with acinar and duct epithelial cells; the StK and BSK antibodies reacted strongly with duct epithelial cells in the canine mammary tumours expressed mainly keratins of stratified epithelia rather than those of simple epithelia. Myoepithelial cells in complex and mixed tumours can express StK and vimentin. The finding might be helpful in the assessment of complex tumours where stromal reactions may be confused with myoepithelial neoplastic proliferation. Stromal mesenchymal tissues, including precartilage in complex tumours, and clearly differentiated myoepithelial cells always stained positively for vimentin. Occasional carcinoma cells stained positively for vimentin.

Animals↗

The effect of Ge, Si and Sn phthalocyanine photosensitizers on cell proliferation and viability of human oesophageal carcinoma cells.

The photodynamic activity of water soluble mixed sulfonated metallophthalocyanines complexes: GePcSmix, SnPcSmix and SiPcSmix on human oesophageal carcinoma (SNO) cells are reported, and compared with the activity of the unmetallated H2PcSmix and of the newly synthesized water soluble adjacently substituted binaphthalo phthalocyanine (complex 3). The alkaline phosphate (ALP) showed damage to the cell membrane in the presence of complex 3 without irradiation. The GePcSmix complex caused a relatively large increase in inflammation and a high intracellular ATP.

Cell Division↗

The effects of cyclophosphamide on in vitro correlates of tumor immunity.

The effects of cyclophosphamide (CY) on tumor immunity against isografts of rat sarcomas induced by polyoma virus were studied using in vitro techniques. Groups of sarcoma-bearing animals received CY (250 mg/kg intraperitoneally 11 days after isografting) CY (150 mg/kg IP 8 days after isografting), or IP injections of 0.9% NaCl. In control rats tumor growth was progressive. All CY-treated animals showed transient tumor regression of at least 50% of their pretreatment tumor volume. Despite a drastic depression in numbers of blood leukocytes as well as lymph-node cells following CY treatment, animals treated with 150 mg/kg CY were shown to have blood lymphocyte and lymph-node cell cytotoxicity in vitro against plated sarcoma target cells comparable to untreated sarcoma-bearing animals. Sera obtained from sracoma-bearing rats prior to CY treatment specifically blocked lymphocyte cytotoxicity against sarcoma target cells. After CY treatment serum blocking activity could not be demonstrated during the period of tumor regression, but reappeared in parallel with tumor regrowth. Antibodies cytotoxic to sarcoma target cells when homologous complement was added could not be demonstrated in sera obtained from animals before CY treatment, but were present after treatment during tumor remission. Following CY treatment, sera obtained from treated animals, when mixed with blocking sera from control animals, could counteract or unblock the blocking activity of tumor-bearer sera. Unblocking capacity was present only in sera obtained during CY-induced tumor remission. Serum IgG concentration was significantly and temporarily decreased after CY treatment.

Animals↗

The effects of adjusting for case mix on mortality and length of stay following radical cystectomy.

PURPOSE: Prior studies evaluating quality of care following radical cystectomy have been constrained by the use of retrospective reviews of single institutional series and limited ability to examine risk factors in a comprehensive manner. Characterization of these factors could enhance preoperative patient counseling and facilitate perioperative management, thereby improving the quality of patient care. MATERIALS AND METHODS: The National Surgical Quality Improvement Project is a prospective quality management initiative at 123 Veterans Affairs Medical Centers nationwide. The project collects preoperative clinical and intraoperative data, and outcomes on a wide variety of surgical procedures from multiple surgical disciplines. Since 1991, 2,538 radical cystectomies have been captured by the National Surgical Quality Improvement Project. Modeling using logistic regression was performed to identify preoperative risk factors associated with mortality and prolonged length of stay (greater than 90th percentile) after radical cystectomy. RESULTS: The 30 and 90-day mortality rates following cystectomy were 2.9% and 6.8%, respectively, and median hospital stay was 11 days (90th percentile 30). Robust preoperative factors associated with mortality and prolonged length of stay that uniformly increased risk were older patient age (OR 1.2 to 1.4), American Society of Anesthesiologists class 3 or greater (OR 1.5 to 3.3), dependent functional status (OR 1.7 to 2.0) and low serum albumin (OR 2.1 to 12.0). CONCLUSIONS: A defined set of preoperative risk factors is independently associated with greater mortality and hospital stay following radical cystectomy. The breadth of these factors suggests that complex case mix adjustment is mandatory when comparing outcomes. Implementation of novel processes directed toward minimizing patient risk has the potential to improve outcomes following cystectomy.

Age Factors↗

Clear cell melanoma of the bladder.

Primary malignant melanoma of the urinary bladder is a rare tumor. A case in a 34-year-old man who presented with gross hematuria is reported. Cystoscopic biopsy showed a mixed spindle cell and clear cell neoplasm with focal pigmentation. The tumor cells were immunoreactive for S-100 protein and HMB-45. The patient did not have a history of melanoma, and clinical and radiologic work-up did not reveal other lesions, so a partial cystectomy and pelvic lymphadenectomy were performed. The tumor formed an ulcerated mass that extended into the perivesicular fat. Microscopically, the tumor was composed predominantly of clear cells in sheets or small nests. A solitary metastatic focus was present in 1 of 25 pelvic lymph nodes. Ultrastructurally, the tumor cells were characterized by irregular nuclear contours, complex nucleoli, abundant cytoplasmic glycogen, and premelanosomes.

Adult↗

[A case of intracranial osteochondroma originating with psychomotor epilepsy].

A 28-year-old woman was admitted to our clinic due to psychomotor epilepsy. Craniogram, CT scan demonstrated a calcified lesion in the frontal, parasagittal region. MR proton images showed a mixed hyper-and hypodense lesion. The tumor originated from the frontal bone and was totally removed using airdrill safely. Histological findings revealed osteochondroma originating in the frontal bone is rare, and diagnostic procedures and surgical management of this tumor are discussed.

Adult↗

Assembling a mixed phthalocyanine-porphyrin array in aqueous media through host-guest interactions.

A stable 1:1 host-guest complex is formed between a silicon(IV) phthalocyanine conjugated axially with two permethylated beta-cyclodextrin units and a tetrasulfonated porphyrin. The complex exhibits a light-harvesting property and works as an efficient photosensitizing system, killing HT29 human colon adenocarcinoma cells with an IC50 value of 0.09 microM. [structure: see text].

Adenocarcinoma↗

Immunohistochemical reactivity of basal and luminal epithelium-specific cytokeratin antibodies within normal and neoplastic canine mammary glands.

Human basal epithelium (myoepithelium)-specific (312C8-1) and luminal epithelium-specific (13H5) cytokeratin antibodies were applied to frozen sections of normal canine mammary tissues (seven), benign adenomas and hyperplasias (five), mixed tumors (12), and adenocarcinomas (18) to determine if epithelial subsets could be discriminated by the use of an avidin biotin peroxidase complex immunohistochemical procedure. The 312C8-1 and 13H5 antibodies were consistently reactive with basal and luminal epithelium, respectively, in the normal mammary gland (7/7) and in benign adenomas and hyperplasias (5/5). Mixed mammary tumors had similar basal and luminal epithelial reactivity and also had proliferating spindle-shaped stromal cells that were reactive with 312C8-1 (10/12) and 13H5 (4/12). The adenocarcinomas were subclassified into basal, luminal, and basal/luminal on the basis of 312C8-1 reactivity (4/18), 13H5 reactivity (2/18), and dual reactivity with mutually exclusive anatomic distribution (11/18), respectively. Those tumors with dual immunoreactivity were indicative of noninvasive carcinomas. Dogs with neoplasms that were reactive with 312C8-1 and nonreactive with 13H5 had local recurrence or distant metastasis within 2 weeks to 6 months after diagnosis. Other antibodies used for comparison were pan cytokeratin AE1/AE3, actin HHF35, and vimentin. 312C8-1 and 13H5 antibodies are specific for canine mammary basal and luminal epithelium, respectively, and by employing these antibodies, the origin and differentiation of canine mammary neoplasms can be determined more accurately than on the basis of hematoxylin and eosin-stained tissue alone.

Actins↗

Induction of immune cytolysis: tumor-cell killing by complement is initiated by covalent complex of monoclonal antibody and stable C3/C5 convertase.

The unique specificity of monoclonal antibodies (Ab) was combined with the cell-killing function of the cytolytic alternative pathway of complement. The functionally C3b-like glycoprotein of cobra venom was linked to a murine monoclonal Ab directed to a human melanoma-associated antigen by a disulfide bond, by using a heterobifunctional crosslinking reagent. The covalent monoclonal Ab-cobra venom factor (CVF) complex exhibited specific cytolytic activity in the presence of normal or C4-deficient serum. It induced killing of melanoma cells but not of LG-2 lymphoblastoid cells or P815 mastocytoma cells. The cytolytic action of the monoclonal Ab-CVF complex was selective in that it was limited to the melanoma cells when these were mixed with one of the two other cell lines. In absence of serum, the complex was noncytotoxic. Monoclonal Ab or CVF alone had no cytolytic activity with or without serum. It is concluded that CVF covalently linked to melanoma cell-bound AB forms the stable C3/C5 convertase with factors B and D of the alternative pathway, which in turn causes formation of the membrane attack complex and thereby cell death.

Animals↗

Cells of the anterior pituitary.

The anterior pituitary is made up of a number of cell types that are essential for such physiological processes as growth, development, homeostasis, metabolism, and reproduction. These include the hormonal cells corticotropes, thyrotropes, gonadotropes, somatotropes, lactotropes and a small population of mammosomatotropes, together with a non-hormonal cell type called the folliculo-stellate cells. The anterior pituitary hormonal cells are highly differentiated and are committed very early on during embryonic development. Their development is tightly regulated by both extrinsic signals as well as by endogenous gene expression. Many transcription factors that shape the development and functions of the anterior pituitary cells have been identified. Even after differentiation, pituitary cells continue to undergo mitosis and this process could be augmented under certain conditions in adulthood. Some anterior pituitary cells are multifunctional and exhibit mixed phenotypes. Pituitary tumors, which are mostly monoclonal in nature, are rather common. The molecular pathogenesis of pituitary tumorigenesis involves complex and diverse mechanisms. Aberrant intra- and extra-pituitary factors are involved. Mutations of some genes specific to pituitary tumors also play a role.

Animals↗

Oligomeric assembly and ligand binding of the members of protein family YER057c/YIL051c/YJGF.

Proteins UK114 and p14.5 are both members of the putative family of small proteins YER057c/YIL051c/YjgF. The biological role of these proteins is not understood very well, and in addition, their oligomeric structure in solution remains controversial. We therefore investigated the oligomeric structure of UK114 and p14.5 using a number of methods. Both proteins have exhibited a homotrimeric structure in solution. Indeed the trimeric structure of the two proteins appeared to be so similar that when protein subunits derived from different species were mixed, stable heterotrimeric complexes (monomer ratio of 1:2 and 2:1 of UK114 and p14.5, respectively) could be formed in vitro. Furthermore, the trimeric structure of both UK114 and p14.5 proved essential for the stoichiometric hydrophobic ligand, such as fatty acid binding activity of the two proteins.

Anilino Naphthalenesulfonates↗