Natural human IgG inhibits the production of tumor necrosis factor-α and interleukin-1α through the Fc portion

The overproduction of cytokines such as the tumor necrosis factor- (TNF-) and interleukin-1 (IL-1) may cause further deterioration in the already critical condition of patients with shock, sepsis, and acute inflammation. The effectiveness of infusion therapy of natural human IgG to such patients is suggested to depend partly upon the inhibition of the productivity of these cytokines. In this study, we investigated the modulation effects of IgG and its fragments on the production of TNF- and IL-1, on human peripheral blood mononuclear cells (PBMC). The production of TNF- and IL-1 was found to be dose-dependently inhibited by IgG when stimulated by lipopolysaccharide (LPS), phytohemagglutinin (PHA), concanavalin A (Con A), and interleukin-2 (IL-2). However, no inhibition was seen when stimulated by phorbormyristate acetate (PMA). The F(ab)₂ fragment showed enhancing effects on cytokine production by LPS, while the Fc fragment showed not as much inhibitory effect as whole intact IgG. IgG showed no direct cytotoxic effect on PBMC. These data suggest that natural human IgG inhibits TNF- and IL-1 production by PBMC through the Fc portion. The results of this study led us to conclude that whole intact IgG may be the best form of therapeutic delivery.     

Growth fraction of human bladder tumors

Summary The growth fraction of bladder tumors was immunohistochemically assessed in situ using anti-DNA polymerase (Pol ) monoclonal antibody. This enzyme is known to be present in the nucleus of the cells in G1, S, and G2 phases. The percentage of labeled cells was expressed as the labeling index (LI). The average LI was 6.0% in normal epithelium and 17.8% in bladder tumors, this difference being significant. The labeled cells were distributed throughout both the basal and surface layers of bladder tumors. However, in some bladder tumors, the distribution of Pol -labeled cells varied from area to area. The higher fraction of labeled cells was found in high grade or invasive tumors. Papillary and nodular bladder tumors showed a greater rate of cell proliferation than papillary tumors. These findings suggest that Pol immunostaining could be a potent tool for easy and quick evaluation of proliferating cells in bladder tumors, there-by providing a supplement to conventional histological findings.     

Cytostatic effect of different strains of Bacillus Calmette-Guérin on human bladder cancer cells in vitro alone and in combination with mitomycin C and Interferon-α

Summary The cytostatic activity of five Bacillus Calmette-Guérin (BCG) strains (Pasteur, Evans, Tice, RIVM and Connaught) on human transitional cell cancer T24 cells was examined. A striking effect was noted even in 2-day cultures, and the effect was more pronounced when the cells were incubated for 5 days with different BCG strains alone. The concentrations needed were about the same as those used in clinical practice (10⁹ colonyforming units of Pasteur strain in 100 ml buffered saline solution). Combination with mitomycin C or interferon--2b potentiated the cytostatic effect. A slight difference in cytostatic activity between different BCG strains was found.     

Comparison of adrenoceptor subtype expression in porcine and human bladder and prostate

We have quantified and characterized ₁-, ₂-and -adrenoceptor subtypes in porcine bladder detrusor and bladder neck, human bladder detrusor, and porcine and human prostate. ₁-, ₂- and -adrenoceptor were identified in radioligand binding studies using [³H]prazosin, [³H]RX 821002 and [¹²⁵I]iodocyanopindolol, respectively, as the radioligands. In porcine male and female detrusor and bladder neck and male prostate, adrenoceptors were detected in the order of abundance > _{2 1} (not detectable), with no major differences between the sexes or between detrusor and bladder neck. In human detrusor and prostate the order of abundance was > _{2 1} (not detectable) and ₁ > ₂. respectively. The ₂-adrenoceptors in all tissues were homogeneously of the _2A-subtype as evidenced by competition binding studies with yohimbine, prazosin, ARC 239 and oxymetazoline. The -adrenoceptors represented a mixed population with a dominance of the ₂-subtype in all tissues as demonstrated by competition binding with ICI 118,551 and CGP 20,712A. We conclude that pigs may be a suitable model for studies of detrusor function with respect to adrenoceptor expression. They may be less suitable for studies of bladder neck or prostate function.     

Effect of tumor necrosis factor-α and interferon-γ on the growth of human prostate cancer cell lines

Human recombinant tumor necrosis factor- (rTNF-, 10^-12–10^-8 M) inhibited the proliferation of androgen-dependent LNCaP cells by 32–56%. In contrast, proliferation of androgen-independent PC-3 and JCA-1 cells was only slightly inhibited, or not inhibited at all, respectively. Human recombinant interferon- (rIFN-, 500 U/ml) decreased proliferation of PC-3 and JCA-1 cells by 35% and 53%, respectively, but had no effect on LNCaP cells. Interestingly, the combination of rIFN- and TNF- had greater antiproliferative effects on JCA-1 cells than treatment with either cytokine alone. However, the antiproliferative effects of this combination were similar to those observed for PC-3 or LNCaP cells treated with rIFN- or TNF- alone, respectively. These data suggest that some forms of androgen-independent prostate cancer may benefit from a combination therapy of IFN- and TNF-, while the use of IFN- alone may be more efficacious in others.     

121. Modifizierte Technik der „Ersatzklappen”-Operation an der V. poplitea bei postthrombotischem Syndrom

Zusammenfassung Die Technik der Ersatzklappen-Operation an der V. poplitea bei postthrombotischem Syndrom wurde vom Initiator durch die Verwendung einer 24cm langen Silicone-Sehne modifiziert. Die Silicone-Sehne wird zwischen A. und V. poplitea durchgezogen, ihr mediales Ende an die Gracilissehne und ihr laterales Ende um die Bicepssehne vernäht. Bei 31 durchgeführten Operationen konnte klinisch und phlebographisch bei allen eine regelrechte Ersatzklappen-Funktion und eine Besserung der venösen Hämodynamik des Beines festgestellt werden.     

Transgenic Mice Reveal Roles for TGFα and EGF Receptor in Mammary Gland Development and Neoplasia

Transforming growth factor-alpha (TGF)⁴ and/or the EGF receptor (EGFR) are frequently overexpressed by human and rodent breast tumors, as well as tumor-derived cell lines. Additionally, various observations suggest a role for TGF and the EGFR signaling system in normal mouse mammary gland development. Recently, several laboratories have established TGF transgenic mice with which to study the role of this growth factor in normal and neoplastic mammary biology. Examination of these mice revealed that overexpression of TGF has profound consequences for this tissue. Most strikingly, transgenic mice expressing TGF under the control of tissue-specific and nonspecific promoters stochastically developed focal mammary tumors with an incidence and latency that was markedly affected by pregnancy. Most TGF-induced tumors were well-differentiated adenomas/adenocarcinomas, although some were undifferentiated and locally invasive. Distant metastases were only occasionally observed. Administration of the genotoxic carcinogen, 7,12-dimethylbenzanthracene (DMBA), dramatically accelerated mammary tumorigenesis induced by the TGF transgene, raising the possibility that TGF acts as a promoter in this tissue. Mice harboring dual transgenes encoding TGF and either wild-type ERBB2 or c-myc displayed markedly accelerated tumorigenesis compared to mice carrying any of the single transgenes alone, indicative of potent cooperativity. Moreover, tumorigenesis in the bitransgenic mice was less dependent on pregnancy, and tumors were generally more malignant in appearance. Finally, TGF also affected mammary gland dynamics. TGF transgenic mice consistently displayed precocious alveolar development, were variably impaired with respect to lactation, and showed markedly reduced postlactional involution. As a result, the glands of multiparous females accumulated hyperplastic lesions that generally resembled milk-producing alveoli. Limited data support the hypothesis that these lesions were precursors to TGF-induced tumors. In summary, these various findings underscore the potential importance of TGF for cellular differentiation and transformation in the mammary gland. They also establish TGF transgenic mice as a powerful model with which to study the role of EGFR signaling molecules in this dynamic tissue.     

Potential uses of interferon α2 as adjuvant therapy in cancer

Background: The purpose of this study was to provide an overview of the potential uses of adjuvant interferon (IFN) therapy for resected solid tumors at high risk for postsurgical relapse. Methods: A MEDLINE search (1970–1994) of the English-language literature for original articles, reviews, and abstracts addressing IFN use in the adjuvant setting together with the authors' collective experience formed the basis for this review. Results: The use of adjuvant IFN- has been studied most extensively in conjunction with the treatment of melanoma. Fewer data are available on IFN- use for the treatment of other solid tumors. In melanoma, there is evidence from Intergroup trials (Eastern Cooperative Oncology Group and World Health Organization) that IFN-2a given for 1–3 years prolongs the interval to relapse and may have a survival benefit. Trials of adjuvant IFN, with and without chemotherapy, are ongoing in the treatment of renal cell carcinoma and colorectal adenocarcinoma. Its value in the treatment of osteosarcoma and high-grade astrocytoma is unknown. Conclusions: The use of IFN in the adjuvant setting is an exciting area of medical and surgical oncology and has the potential to prolong the time to relapse and to increase survival of patients with melanoma. Its role in the adjuvant therapy of other solid tumors remains to be defined.     

Elevated concentrations of the small stress protein HSP27 in rat renal tumors

The expression of two small stress proteins, B crystallin and the 27-kDa heat shock protein (HSP27), was studied quantitatively and immunohistochemically in normal kidney and renal tumors in rats. Levels of B crystallin in renal cell tumors tended to be higher than in normal kidney (P = 0.07), but with a wide range of values, whereas they were significantly lower in mesenchymal tumors (P < 0.0001). In contrast, HSP27 concentrations in both renal cell (mean ± SD: 1790 ± 940 ng/mg protein,n = 15) and mesenchymal (1260 ± 1080 ng/mg protein,n = 10) tumors were significantly higher than the normal kidney value (142 ± 30 ng/mg protein,n = 10,P < 0.0001). A positive correlation was found between B crystallin and HSP27 levels limited to the renal cell tumor case (Pearson's correlation coefficient,r = 0.68,P < 0.01). Immunohistochemistry revealed the loops of Henle to be positive for B crystallin, whereas HSP27 staining was positive in glomerular and interstitial vascular walls and epithelial cells of proximal and distal tubules. Positive immunostaining for B crystallin was demonstrated in six of nine renal cell tumors (67%) studied and for HSP27 in all of the nine cases (100%).     

Role of the E-cadherin/α-catenin complex in modulating cell-cell and cell-matrix adhesive properties of invasive colon carcinoma cells

Background: The clinical behavior of colorectal cancer depends on its ability to invade and metastasize. Metastatic cells must dissociate from other cells and invade through basement membrane and stroma. Cell-cell adhesion in epithelial cells is mediated by the cell surface protein E-cadherin in association with - and -catenin, which link E-cadherin to the cytoskeleton. Decreased cell-cell adhesion and increased motility on laminin have been correlated with more poorly differentiated and aggressive carcinomas. Methods: In this study, the RKO cell line, previously shown by us to lack E-cadherin expression, was transfected with the complementary DNA for E-cadherin. The transfectants were selected for high levels of surface expression by sequential FACS and examined in functional assays. Results: In comparison to control transfectants, the E-cadherin transfectants exhibited a more epithelial-like morphology, a 30% increase in Ca²⁺-dependent cell-cell aggregation, and a markedly reduced motility on the matrix proteins, collagen I and laminin. Conclusions: These data demonstrate that correction of a defect in the cadherin/catenin cell-cell adhesion complex, often found in poorly differentiated and highly invasive tumors, facilitates increased cell-cell adhesion and retards tumor cell migration on basement membrane and stromal proteins.The results of this study were presented at the 47th Annual Cancer Symposium of The Society of Surgical Oncology, Houston, Texas, March 17–20, 1994.     

The role of the coagulation cascade in brain edema formation after intracerebral hemorrhage

Summary The coagulation cascade has a potential role in brain edema formation due to intracerebral hemorrhage. In this study blood and other solutions were injected stereotactically into the right basal ganglia in rats. Twenty-four hours following injection, brain water and ion contents were measured to determine the amount of brain edema. Intracerebral blood resulted in an increase in brain water content. The amount of brain edema surrounding the intracerebral hematoma was reduced by a thrombin inhibitor Na-(2-Naphthalenesulfonylglycyl)-4-amidino-DL-phenylalaninepiperidide, (-NAPAP) infused into the hematoma after the clot had been allowed to solidify. The inhibitor did not alter the actual size of the clot mass. An artificial clot composed of fibrinogen, thrombin, and styrene microspheres also produced brain edema. A fibrin clot led to edema formation even in the absence of mass effect provided by the microspheres. The single component responsible for production of brain edema in all these models was thrombin. The edema was formed in response to a fibrinogen-independent pathway. These results indicate that the coagulation cascade is involved in brain edema that develops adjacent to an intracerebral hematoma.     

Surgical remodeling of the silhouette by aspiration lipolysis or selective lipectomy

The author, a pioneer in originating the technique of suction lipectomy, discusses in detail the pathophysiology of the fat cell and fat metabolism. In addition, he describes the surgical means by which, since 1977, he has popularized this innovative new approach to the treatment of unsightly fat deposits in many sites in the body, including the thighs, buttocks, abdomen, hips, arms, ankles, breasts, and submental regions.     

Scanning Electrochemical Microscopy: Detection of Human Breast Cancer Cells by Redox Environment

Scanning electrochemical microscopy (SECM) can be used to measure the redox activity of individual human breast cells. A chemical mediator (e.g. quinone) that rapidly crosses the membrane participates in intracellular redox reactions that are recorded on a microsecond timescale by an ultramicroelectrode positioned close to the membrane. Measurements of redox reactivity yield rate constants that are different for cancerous and non-transformed human breast cells. With non-transformed or metastatic cells, rate constants are modulated by altered expression or activity of protein kinase C, an enzyme involved in the mechanism of cell metastasis. When used in two-dimensional scanning, SECM produces a spatially resolved redox map of an individual cell or field of cells and can detect individual breast cancer cells in a field of non-transformed cells. These studies identify a new technology for cancer detection and establish a framework for future analysis of malignant cells in human breast tissues and biopsies.     

Identification of α1-adrenoceptor subtypes in the dog prostate

Summary The ₁-adrenoceptor subtypes of dog prostate were characterized in binding and functional experiments. In saturation experiments, [³H]prazosin bound to ₁-adrenoceptors with high affinity. In the displacement experiments, unlabelled prazosin and WB4101 biphasically inhibited the binding of 400 pM [³H]prazosin, suggesting the presence of at least two distinct affinity sites for prazosin or WB4101. The proportion of high-affinity sites was approximately 10%. HV723 also recognized two distinct affinity sites but the proportion of high-affinity sites was approximately 20%. From these results the presence of three distinct ₁-adrenoceptor subtypes was suggested: presumably subtypes _1A (high affinity for prazosin and WB4101), _1N (high affinity for only HV723) and _1L (low affinity for the three antagonists) according to the recently proposed ₁-adrenoceptor subclassification. The density of subtype _1L was much higher than that of subtypes _1A and _1N subtypes. In the functional experiments, prazosin, WB4101 and HV723 competitively antagonized the contractile response to noradrenaline with low affinities close to those estimated for the _1L subtypes. These results suggest that the contractile response to noradrenaline in the dog prostate is mediated predominantly through _1L subtype -adrenoceptors.     

Adhesion,growth, and matrix production by osteoblasts on collagen substrata

Summary A number of studies have demonstrated the pivotal role of collagen molecules in modulating cell growth and differentiation. In order to analyze the direct effects of collagen type I on the osteoblastic phenotype, we have devised an in vitro culture system for studying the interactions between bovine collagen type I and Saos-2 cells, a human osteoblastic cell line. Saos-2 cells were cultured both on top of collagen-coated culture dishes as well as inside a three-dimensional collagen network. Plating on dishes treated with collagen induced maximal adhesion of Saos-2 cells after 24-hour incubation. Cells cultured on collagen gel matrix expressed about 2.5-fold more alkaline phosphatase when compared with untreated plastic dishes. On collagen-coated dishes the responsiveness of Saos-2 cells to parathyroid hormone was decreased, whereas no modifications were observed in the effect of vasoactive intestinal peptide on these cells. Using a microfluorimetric measurement of DNA, an increase of proliferation was observed in Saos-2 cells cultured on collagen gel Saos-2 cells were also able to colonize collagen sponges and in this three-dimensional network they were able to synthesize osteocalcin, as assessed both by immunocytochemistry and radioimmunoassay. In this study we have demonstrated that bovine collagen type I exhibits favorable effects on attachment and functional and growth activities of a human osteoblastic cell line, encouraging its use as a bone graft material.     

α-Adrenoceptor function before and after chemical sympathectomy in human and feline detrusor muscles

Summary Isolated bladder segments from man and cat were treated with 6-hydroxydopamine (6-OHDA) in vitro. Chemical sympathectomy was evaluated with fluorescence microscopy and found to be very similar to the effect of 6-OHDA administered in vivo to cats. Isometric smooth muscle contractile responses were induced by field stimulation (FS). The amplitude of the responses increased after denervation. The effects of -adrenoceptor agonists and antagonists on the FS-induced contractile responses were compared before and after treatment with 6-OHDA. The reduction in the contractile responses after the addition of noradrenaline to the feline bladder strips was more pronounced after treatment. Phentolamine induced an increase in contractile responses before treatment, an effect not seen afterwards in human bladder strips but which persisted in feline bladder strips. Selective -adrenoceptor agonists did not alter the contractile responses in denervated strips. It is suggested that the function of the -adrenoceptors in the detrusor is to inhibit neuronally mediated contractile responses of smooth muscle.