IgE rheumatoid factor

Summary A solid phase enzyme-linked immunosorbent assay for detecting class-specific rheumatoid factors (RF) has been devised. IgE RF, which has not been reported yet, was found in sera from 3 of approximately 100 rheumatoid patients.     

Action of D-penicillamine on immunocomplexes containing rheumatoid factor

Summary The affinity between purified rheumatoid factors (RF) and native or heat aggregated human IgG has been studied in vitro by polarization florescence in the presence and in the absence of D-penicillamine. The value of the dissociation constant was the same using native and heat aggregated IgG suggesting that binding to the aggregated protein is not dependent on the exposure of a new determinant lacking in the native molecule. The results obtained in the presence of D-penicillamine suggest that the concentration of the drug necessary to get a pronounced effect on the apparent dissociation constant of the immunocomplex between IgG and RF is not reached in vivo, in clinical situations.     

The humoral immune response to heat shock proteins.

Humoral immune reactions to heat shock proteins (hsp) from microorganisms are one aspect of microbial infections in humans. The production of antibodies which are specific to epitopes present on procaryotic hsp leads also to the appearance of cross-reactive serum antibodies in the host organism that react with human hsp. This article discusses the consequences of such autoreactive antibodies for the host in context with the development of immune tolerance and autoimmune diseases, especially rheumatoid arthritis (RA), and in experimental animal models for arthritis such as adjuvant arthritis in rats. On the basis of epitope cross-reactivity between hsp and other host proteins, a hypothesis is presented for the development of autoimmune disease following the production of hsp-specific antibodies.     

The humoral immune response to heat shock proteins

Humoral immune reactions to heat shock proteins (hsp) from microorganisms are one aspect of microbial infections in humans. The production of antibodies which are specific to epitopes present on procaryotic hsp leads also to the appearance of cross-reactive serum antibodies in the host organism that react with human hsp. This article discusses the consequences of such autoreactive antibodies for the host in context with the development of immune tolerance and autoimmune diseases, especially rheumatoid arthritis (RA), and in experimental animal models for arthritis such as adjuvant arthritis in rats. On the basis of epitope cross-reactivity between hsp and other host proteins, a hypothesis is presented for the development of autoimmune disease following the production of hsp-specific antibodies.     

Etiology of rheumatoid arthritis

Definite genetic associations with immunological cooperative HLA-D(R) antigens have been demonstrated for rheumatoid arthritis (RA). Microbial etiology has not been proven, but some hope for the supporters of this view is still given by small viruses, plasmids of enteric bacteria or perhaps oncogen-like DNA-sequences. Yet, electrophoretical analysis of membrane proteins or surface glycoproteins of RA synovial cells does not show any differences compared to reference cells. Autoimmunity to several tissue elements has been demonstrated, but most of it is of secondary nature. Antigenicities of type II and III collagens are probably only contributory factors for HLA-DR4 positive individuals. Proteoglycans or minor cartilage collagens have not been extensively studied, so far. Endocrine, dietary or psychological influences might be triggering events for otherwise 'preloaded' individuals.     

Etiology of rheumatoid arthritis

Summary Definite genetic associations with immunological cooperative HLA-D(R) antigens have been demonstrated for rheumatoid arthritis (RA). Microbial etiology has not been proven, but some hope for the supporters of this view is still given by small viruses, plasmids of enteric bacteria or perhaps oncogen-like DNA-sequences. Yet, electrophoretical analysis of membrane proteins or surface glycoproteins of RA synovial cells does not show any differences compared to reference cells. Autoimmunity to several tissue elements has been demonstrated, but most of it is of secondary nature. Antigenicities of type II and III collagens are probably only contributory factors for HLA-DR4 positive individuals. Proteoglycans or minor cartilage collagens have not been extensively studied, so far. Endocrine, dietary or psychological influences might be triggering events for otherwise preloaded individuals.     

Presence of hsp65 in bacterial extracts (OM-89): A possible mediator of orally-induced tolerance?

Heat shock proteins (HSP) have been implicated in rodent models of autoimmunity, particularly arthritis, and there is suggestive though inconclusive evidence that they may also play a role in human autoimmune disease. The simplest hypothesis is based on molecular mimicry due to the amino-acid sequence homology between mammalian and microbial HSP. Recently OM-89, an extract of several strains ofEscherichia coli, has shown some efficacy in the treatment of rheumatoid arthritis (RA) when taken orally. Using species-specific antibodies, we show here that OM-89 contains the 65 kDa HSP (hsp65), while hsp65 was not detected in another bacterial extract containing other microorganisms, includingStaphylococcus aureus (OM-85). We suggest that if the human homologue of hsp65 is a relevant target antigen in the human disease, the efficacy of the preparation could be due to induction of oral tolerance or to switching the Th1 response towards Th2. Alternatively, even if the human hsp65 is not a target molecule in RA joints, OM-89 may evoke bystander suppression of joint inflammation via induction of TGF-secreting effector cells. These hypotheses should be tested in further studies.     

Retinoic acid signaling and neurogenic niche regulation in the developing peripheral nervous system of the cephalochordate amphioxus

The retinoic acid (RA) signaling pathway regulates axial patterning and neurogenesis in the developing central nervous system (CNS) of chordates, but little is known about its roles during peripheral nervous system (PNS) formation and about how these roles might have evolved. This study assesses the requirement of RA signaling for establishing a functional PNS in the cephalochordate amphioxus, the best available stand-in for the ancestral chordate condition. Pharmacological manipulation of RA signaling levels during embryogenesis reduces the ability of amphioxus larvae to respond to sensory stimulation and alters the number and distribution of ectodermal sensory neurons (ESNs) in a stage- and context-dependent manner. Using gene expression assays combined with immunohistochemistry, we show that this is because RA signaling specifically acts on a small population of soxb1c-expressing ESN progenitors, which form a neurogenic niche in the trunk ectoderm, to modulate ESN production during elongation of the larval body. Our findings reveal an important role for RA signaling in regulating neurogenic niche activity in the larval amphioxus PNS. Although only few studies have addressed this issue so far, comparable RA signaling functions have been reported for neurogenic niches in the CNS and in certain neurogenic placode derivatives of vertebrates. Accordingly, the here-described mechanism is likely a conserved feature of chordate embryonic and adult neural development.     

Retinoic acid enhances the proliferation of smooth muscle cells

Retinoic acid (RA, 10(-5) - 10(-7) M) is shown to enhance the proliferation of cultured rat aortic smooth muscle cells (SMC). This effect is not connected with a synergistic action of RA together with serum mitogens. Moreover, the expression of L1, a surface antigen specific for modulated SMC entering the cell cycle, is amplified by RA treatment.     

Retinoic acid modulates gap junctional intercellular communication in hepatocytes and hepatoma cells

Gap junctional communication permits the direct exchange of small molecules and ions and has been implicated in tissue homeostasis/metabolite exchange. The lack of gap junctional intercellular communication (GJIC) plays important roles in the promotion and progression of carcinogenesis. In the present study, we demonstrate that treatment of human hepatoma Hep G2 cells with retinoic acid (RA) results in increased amounts and phosphorylation of connexins, their stabilisation in plasma membrane plaques and enhanced GJIC. In cultured fetal hepatocytes, which represent a non-transformed, proliferating and incompletely differentiated liver system, the effects of RA are limited to the establishment of connexin in areas of cell-cell contact and the improvement of GJIC. This suggests that modulation of cell-cell channel communication by RA occurs differently in these two experimental models: while RA is able to revert cell transformation in Hep G2 cells, in fetal hepatocytes it may induce the expression of a more differentiated phenotype. Received 19 June 2002; received after revision 29 July 2002; accepted 8 August 2002 RID="*" ID="*"Corresponding author.     

A synergistic factor of an insect granulosis virus agglutinates insect cells

Summary The synergistic factor (SF), a lipoprotein which enhances in vivo and in vitro baculovirus infections, occurs in the matrix of the occlusion body (capsule) of a granulosis virus of the armyworm,Pseudaletia unipuncta. The SF attaches to certain areas on the surfaces of cell plasma membranes of cultured insect cells, resulting in marked cellular agglutination. The minimal amount of SF detectable by agglutination is approximately 3 g/ml. Cultured cells of 6 out of 10 insect species in 3 orders are agglutinated by the SF, but not those of 2 mammalian species and the erythrocytes of 8 vertebrate species.We thank Dr M. Nagata and Dr T. Yamamoto for preparing the synergistic factor and R.T. Hess and E.M. Omi for technical assistance. We also thank the donors who provided the cell lines. This work was supported by the National Science Foundation under Grant No. PCM-8201247.     

From carrot to clinic: an overview of the retinoic acid signaling pathway

Vitamin A is essential for the formation and maintenance of many body tissues. It is also important for embryonic growth and development and can act as a teratogen at critical periods of development. Retinoic acid (RA) is the biologically active form of vitamin A and its signaling is mediated by the RA and retinoid X receptors. In addition to its role as an important molecule during development, RA has also been implicated in clinical applications, both as a potential anti-tumor agent as well as for the treatment of skin diseases. This review presents an overview of how dietary retinoids are converted to RA, hence presenting the major players in RA metabolism and signaling, and highlights examples of treatment applications of retinoids. Moreover, we discuss the origin and diversification of the retinoid pathway, which are important factors for understanding the evolution of ligand-specificity among retinoid receptors.     

Retinoic acid enhances the proliferation of smooth muscle cells

Summary Retinoic acid (RA, 10^–5–10^–7 M) is shown to enhance the proliferation of cultured rat aortic smooth muscle cells (SMC). This effect is not connected with a synergistic action of RA together with serum mitogens. Moreover, the expression of L1, a surface antigen specific for modulated SMC entering the cell cycle, is amplified by RA treatment.     

Sekretorische Aktivität des Flexural- und Subcommissuralorgans bei frühen Entwicklungsstadien vonTriturus alpestris Laur

Summary The development of Reissner's fibre (RF) by secretory activity of the flexural and subcommissural organ was studied in early ontogenetical stages ofTriturus alpestris. In young stages RF arises from the flexural organ, which gradually reduces its secretory activity in the following stages. Now the secretion of the subcommissural organ becomes predominant and begins to form the RF. After the hatching of larvae, only the subcommissural organ produces the RF.     

Molecular basis for chemoprevention by sulforaphane: a comprehensive review

The consumption of cruciferous vegetables has long been associated with a reduced risk in the occurrence of cancer at various sites, including the prostate, lung, breast and colon. This protective effect is attributed to isothiocyanates present in these vegetables, and sulforaphane (SF), present in broccoli, is by far the most extensively studied to uncover the mechanisms behind this chemoprotection. The major mechanism by which SF protects cells was traditionally thought to be through Nrf2-mediated induction of phase 2 detoxification enzymes that elevate cell defense against oxidative damage and promote the removal of carcinogens. However, it is becoming clear that there are multiple mechanisms activated in response to SF, including suppression of cytochrome P450 enzymes, induction of apoptotic pathways, suppression of cell cycle progression, inhibition of angiogenesis and anti-inflammatory activity. Moreover, these mechanisms seem to have some degree of interaction to synergistically afford chemoprevention. Received: 10 November 2006; received after revision 15 January 2007; accepted 5 February 2007     

The role of renal hyperaemia and plasma oncotic pressure in proximal tubular reabsorption in the rat

Zusammenfassung Die Resorptionsfähigkeit (RF) des proximalen Konvolutes der Rattenniere wurde mit der Methode der getrennten Ölsäule und die Passagezeit (PZ) der Tubuliflüssigkeit mit Lissamingrün gemessen. Aus beiden Werten konnte die prozentuale Resorption (PR) des proximalen Konvolutes berechnet werden. Mittels künstlich gesetzter Nierenhyperämie und nach Infusionen isotoner NaCl-Bikarbonat-Lösung (mit und ohne Albumin) wurden die Veränderungen von RF, PZ und PR geprüft.     

Antitumoral effects of 9-cis retinoic acid in adrenocortical cancer

The currently available medical treatment options of adrenocortical cancer (ACC) are limited. In our previous meta-analysis of adrenocortical tumor genomics data, ACC was associated with reduced retinoic acid production and retinoid X receptor-mediated signaling. Our objective has been to study the potential antitumoral effects of 9-cis retinoic acid (9-cisRA) on the ACC cell line NCI-H295R and in a xenograft model. Cell proliferation, hormone secretion, and gene expression have been studied in the NCI-H295R cell line. A complex bioinformatics approach involving pathway and network analysis has been performed. Selected genes have been validated by real-time qRT-PCR. Athymic nude mice xenografted with NCI-H295R have been used in a pilot in vivo xenograft model. 9-cisRA significantly decreased cell viability and steroid hormone secretion in a concentration- and time-dependent manner in the NCI-H295R cell line. Four major molecular pathways have been identified by the analysis of gene expression data. Ten genes have been successfully validated involved in: (1) steroid hormone secretion (HSD3B1, HSD3B2), (2) retinoic acid signaling (ABCA1, ABCG1, HMGCR), (3) cell-cycle damage (GADD45A, CCNE2, UHRF1), and the (4) immune response (MAP2K6, IL1R2). 9-cisRA appears to directly regulate the cell cycle by network analysis. 9-cisRA also reduced tumor growth in the in vivo xenograft model. In conclusion, 9-cisRA might represent a promising new candidate in the treatment of hormone-secreting adrenal tumors and adrenocortical cancer.     

Effects of retinoic acid on embryonic development of mice in culture

The effects of all-trans-retinoic acid (RA) (tretinoin) on the craniofacial development of mouse embryos were examined using whole embryo culture. In day 8 embryos cultured for 48 h, embryonic growth was inhibited concentration-dependently by all-trans-RA treatment. Most of the treated embryos exhibited hypoplasia of the primary palatal processes and a reduction in the development of the first visceral arches. In day 10 embryos cultured for 48 h, although embryonic growth was not inhibited at any concentrations of all-trans-RA, median cleft lip (93%), hypoplasia of the primary palatal processes (37%) and limb reduction deformities (48%) occurred commonly. Furthermore, RA treatment greatly reduced the size of the secondary palatal processes. The incorporation of 3H-thymidine in the treated maxillary processes was decreased to 65% of the control value at 1.0 x 10(-7) M all-trans-RA. These findings indicate that all-trans-RA is teratogenic in mouse whole embryo culture.     

新一代无线电平台数字射频核心技术研究与工程实践

无线电平台数字射频是提高现代无线电系统复杂电磁环境适应能力、机动能力、传输能力和综合集成能力等基础性能的关键.本文分析了射频技术的发展需求和国内外数字射频领域的研究进展,以及模拟射频核心单元——模拟射频功放主要指标相互矛盾的机理,并将数字射频概念扩展到综合数字射频,重点论述了数字射频核心技术——数字射频功放和全数字发信机的基本原理、优势和关键技术,提出了基于系统与工艺相结合的数字射频设计方法、数字射频功放的物理模型和一种全数字发信机体系结构以及我国自主可控发展数字射频的技术路线建议.本文认为在当今时代背景下,数字射频对于新一代无线电平台具有革命性和可行性;如果不能自主可控发展该项核心技术,我国又将出现一个新的受制于人的脆弱点.     

Isolation of human epidermal stem cells by adherence and the reconstruction of skin equivalents

The isolation of human epidermal stem cells is critical for their clinical applications. In the present study, we isolated three populations of epidermal keratinocytes according to their ability to adhere to collagen type IV: i.e., rapidly adhering (RA), slowly adhering (SA), and non-adhering (NA) cells. The aim of this study was to characterize RA cells and to investigate the possibility of using these cells for epidermis reconstruction. To identify RA cells, flow cytometric analysis was performed using anti-₆ integrin and anti-CD71 antibodies. RA cells express high levels of ₆ integrin and low levels of CD71, which are considered as markers of an epidermal stem cell nature. Furthermore, electron microscopy showed that RA cells are small and have a high nuclear to cytoplasmic ratio, whereas SA and NA cells have well-developed cellular organelles and abundant tonofilaments. Western blot analysis showed that RA cells are slow cycling and express p63, a putative epidermal stem cell marker, whereas SA and NA cells express c-Myc, which is known to regulate stem cell fate. To compare epidermal regenerative abilities, skin equivalents (SEs) were made using RA, SA, and NA cells. The epidermis constructed from RA cells was well formed compared to those formed from SA or NA cells. In addition, only SEs with RA cells expressed ₆ integrin and ₁ integrin at the basal layer. These results indicate that RA cells represent epidermal stem cells and are predominately comprised of stem cells. Therefore, the isolation of RA cells using a simple technique offers a potential route to their clinical application, because they are easily isolated and provide a high yield of epidermal stem cells.Received 2 July 2004; received after revision 20 August 2004; accepted 10 September 2004