Elevated levels of anti-endotoxin antibodies in patients with bilateral idiopathic acute anterior uveitis.

PURPOSE: Endotoxins have been proved to be responsible for acute anterior uveitis (AAU) in animals in a well-established experimental model of endotoxin-induced uveitis (EIU). The purpose of our study was the detection of antibodies against endotoxins of selected enterobacteria in the serum of patients with idiopathic AAU and searching for correlations between the levels of these antibodies and the presence of HLA-B27 antigen as well as characteristic signs of EIU such as bilaterality and the absence of spontaneous recurrences of the disease. METHODS: Reactions of serum IgG antibodies with lipopolysaccharides (LPSs) of Escherichia coli O1, E. coli O10, E. coli O111, E. coli J5, and Klebsiella pneumoniae O3 were determined for 60 patients with idiopathic AAU and 40 healthy volunteers. The presence of HLA-B27 antigen in patients was determined. Documentation of the frequency of recurrences of AAU during a follow-up period of 8 years was collected. RESULTS: We have observed that the sera of patients with a first attack of AAU reacted stronger with the LPS of K. pneumoniae O3 than the sera of patients with relapse of the disease. Patients with bilateral AAU had markedly higher levels of antibodies against four of the five used LPSs than patients with one eye involved. A multiply comparison showed higher levels of IgG reacting with LPS of E. coli O111 in patients with bilateral eye inflammation admitted with the first attack of AAU comparing to controls. The incidence of recurrent form of AAU was significantly increased in HLA-B27-positive patients compared to HLA-B27-negative patients. However, we found in HLA-B27 carriers that those with the bilateral form of AAU had over three times smaller risk of recurrence and showed stronger immunization by endotoxins than patients with unilateral inflammation. CONCLUSION: Our results suggest a potential role of endotoxins in the aetiology of the nonrecurrent bilateral form of AAU. We suggest that not only HLA-B27 status but also determination of number of involved eyes may be useful to assess the risk of recurrence of the idiopathic AAU.

Proinflammatory and atherogenic activity of monocytes in type 2 diabetes.

UNLABELLED: Cytokines secreted by the monocyte-macrophage system play a key role in the progression of atherosclerotic lesions in Type 2 diabetes. The objectives of this study were to assess the influence of cytokine gene expression in monocytes from patients with Type 2 diabetes on direct markers of endothelial injury with regard to clinically manifest atherosclerosis. METHODS: Monocytes from 58 patients with Type 2 diabetes and from 22 age-matched healthy volunteers of a control group were isolated in order to assess expression of tumor necrosis factor alpha (TNFalpha), interleukin (IL)-6, IL-8 and IL-10 cytokines (RTPCR, Applied Biosystems). Thrombomodulin concentration was determined using a Diagnostica Stago Immunoenzymatic assay, and circulating endothelial cell numbers were assayed using immunofluorescence studies with CLB-HEC19 antibodies. RESULTS: In 28 patients, TNFalpha expression in monocytes was observed. In these patients, as compared to those with undetectable levels of this cytokine's expression, higher hemoglobin A(1c) (P=.012) and thrombomodulin (P=.005) concentrations were found. IL-8 expression was determined in 36 patients. Higher expression of TNFalpha (P=.048) and IL-8 (P=.049) was detected in patients with peripheral arterial disease in contrast to those free from this complication. CONCLUSION: TNFalpha and IL-8 play a significant role in the proatherogenic activity of monocytes in Type 2 diabetes. The TNFalpha-connected activity of monocytes may directly determine endothelial dysfunction and injury. The location of atherosclerosis should be taken into account in the assessment of the proinflammatory activity of peripheral blood monocytes.

Antibodies to 46-kDa retinal antigen in a patient with breast carcinoma and cancer-associated retinopathy.

Cancer-associated retinopathy (CAR) is a rare paraneoplastic syndrome usually associated with small-cell lung carcinoma and serum autoantibodies against recovering. We report the breast cancer woman with visual impairments and electrophysiological abnormalities characteristic of CAR. Her serum contained high-titer antibodies against alpha-enolase but not against other retinal proteins. This suggests that anti-enolase antibodies could be responsible for the development of CAR symptoms.

[The Pro12Ala polymorphism of the peroxisome proliferator-activated receptor gamma and immunological processes in patients with type 2 diabetes and insulin resistance]. / Polimorfizm Pro12Ala genu receptora aktywowanego przez proliferatory peroksysomów gamma a procesy immunologiczne w cukrzycy typu 2 wiklanej insulinoopornoscia. Doniesienie wstepne.

INTRODUCTION: The peroxisome proliferator-activated receptor gamma (PPARgamma), a transcriptor factor, regulates immunological and metabolic processes, which are important for carbohydrate and lipid metabolism. Various polymorphic forms of PPARgamma may promote diabetes mellitus and diabetic complications. AIM OF THE WORK: The assessment of TNFalpha gene expression in peripheral blood monocytes, serum TNFalpha concentration and anti-GAD and ICA antibodies in relation to the polymorphism Pro12Ala in patients with 2 diabetes. PATIENTS AND METHODS: 58 patients with type 2 diabetes (average age 59.0 +/- 11 years) and 18 healthy people were examined. The Pro12Ala polymorphism of PPARy gene were assessed using mini-sequence technic SnaPshot [ABIPRISM-310]. The TNFalpha gene expression were estimated using real-time PCR [Applied Bio-systems]. The TNFalpha concentration [Quantikin Immunoassay, R&D Systems] and ICA and GAD antibodies [immunofluorescence method, DRG] were evaluated in venous blood. RESULTS: A heterozygotous genotype Pro12Ala was estimated in 32 patients and a homozygotous genotype Pro12Pro in 21. Only 6 patients were positive for GAD antibodies and only 6 patients for ICA antibodies. The TNFalpha concentration in serum and the TNFalpha gene expression in monocytes did not refer to the Pro12ala polymorphism of PPARy and neither to antibodies. CONCLUSION: 1) The TNFalpha concentration in serum and the TNFalpha gene expression in monocytes do not refer to the Pro12ala polymorphism of PPARgamma in patients with type 2 diabetes. 2) The Pro12Ala genotype do not influence autoimmunologic processes of diabetes.

Experimental uveitis induced by different uniform salts of enterobacterial lipopolysaccharides.

BACKGROUND: Injection of lipopolysaccharides/LPS/, the major component of the outer membrane of gram-negative bacteria, can induce inflammation in the eyes of susceptible animals. The LPS-induce ocular inflammation is termed endotoxin-induced uveitis/EIU/and is characterized by iris hyperemia, miosis, a rise in aqueous humor protein, and inflammatory cell infiltration into the anterior uvea and aqueous humor. Biological activities of endotoxin depend also on its molecular weight. OBJECTIVES: The aim of our study was to find the effect of different aggregation forms on the clinical and histopathological characteristics of the EIU. MATERIALS/METHODS: Lipopolysaccharides were electrodialyzed and neutralized by adding: triethylomine, sodium hydroxide, or calcium hydroxide. EIU was produced in Lewis rats by footpad injection of different enterobacterial LPS. Their eyes were examined for clinical signs of inflammation in slit lamp, protein and cells were measured in the aqueous hum RESULTS/CONCLUSIONS: The correlation between the physical parameters and biological activity is discussed. Our results have shown recently that monomeric form of endotoxin is more active than an aggregated form in induction of experimental uveitis.

Cancer-associated retinopathy in patients with breast carcinoma.

INTRODUCTION: Cancer-associated retinopathy (CAR) is a paraneoplastic neurological syndrome resulting in progressive loss of vision and clinical signs of retinal degeneration. It is associated with various types of cancer and is also considered to be an autoimmune disorder that involves cross-reaction between autoantibodies and retinal proteins. The aim of this study was to establish whether immunoreactivity to retinal antigens (RAs) observed in patients with breast cancer is accompanied by any visual impairments. MATERIALS AND METHODS: Sera of 295 patients with diagnosed breast cancer were screened for the presence of anti-RAs antibodies using immunoblotting. Cellular immunoreactivity to RAs present in retinal extracts and to purified recoverin and arrestin was determined by means of a lymphocyte proliferation assay. Six patients with high-titer antibodies to RAs then underwent ophthalmic and neurological examinations. RESULTS: Four serum samples contained high-titer antibodies to a 46-kDa protein, most probably retinal alpha-enolase, three had antibodies to a 48-kDa protein identified as retinal arrestin, while 56-, 43-, 41-, and 34-kDa antigens were recognized only by one serum sample each. Moreover, weak cellular response to all the RAs tested was observed in one patient and another patient responded only to retinal extract. Two of the examined patients displayed symptoms of CAR. CONCLUSIONS: Immunoreactivity to RAs in patients with breast cancer may also be present in cases without clinical signs of CAR.

Stimulation of the peroxisome proliferator-activated receptor gamma (PPAR gamma) and the expression of selected blood monocyte cytokine genes in diabetic macroangiopathy.

Monocytes and macrophages play a key role in the progression of atheromatous changes. The peroxisome proliferator-activated receptor gamma (PPAR gamma) can limit macroangiopathy through the control of cytokine transcription. The objectives of this study were to examine the influence of PPAR gamma and its agonist (rosiglitazone) on the TNFalpha, IL-6, IL-8 and IL-10 gene expression in monocytes of patients with diabetic macroangiopathy and to analyse obtained results in context of selected atherogenic factors ant direct indicators of endothelial lesion. TNFalpha, IL-6, IL-8, IL-10 and PPAR gamma gene expression was assessed in peripheral blood monocytes in 45 patients with type 2 diabetes before and following 22 weeks of rosiglitazone therapy (real-time PCR [Applied Biosystems]). As indicators of endothelial lesion, concentration of thrombomodulin (immunoassay [Diagnostica Stago]) and amount of circulating blood endothelial cells (immunofluorescence method with MoAb CLB-HEC19) were determined. Following rosiglitazone therapy, a statistically significant downward tendency of TNFalpha (p=0.026) and IL-8 (p=0.008) gene expression was noted. Before and following rosiglitazone treatment, PPAR gamma, IL-6 and IL-10 gene expression was undetectable in studied monocytes in vivo. In conclusion, TNFalpha and IL-8 play an important role in monocyte atherogenic activity. Rosiglitazone reduces monocyte proinflammatory readiness by influencing the expression of selected atherogenic cytokines (PPAR gamma-independent pathway).

[RANTES expression in nasal polyps fibroblasts; spontaneously and after stimulation with lipopolisaccharides (LPS) and phytohemagglutinin (PHA)]. / Ekspresja chemokiny RANTES w fibroblastach pochodzacych z polipów nosowych; samoistna i po stymulacji lipopolisacharydami (LPS) i fitohematoksylina (PHA).

INTRODUCTION: According to the results of research carried out by a number of authors, one of the main mechanisms of the generation of polyps is local inflammatory processes accompanied by immune system disorders. It has recently been shown that a number of differentiation factors and inflammatory mediators may be involved in the growth of nasal polyps. RANTES is a eosinophil chemoattractant factor likely could play an important role in a chronic inflammatory response in the nasal tissue that subsequently leads to the development of nasal polyps. OBJECTIVE: The objective of this study was detection of the chemokine RANTES in nasal polyps fibroblasts and researching influence of stimulation with lipopolisaccharides and phytohemagglutinin for RANTES expression in cultured nasal fibroblasts in vitro. METHODS: Nasal polyps were obtained from 17 subjects (9 atopic and 8 nonatopic) during polypectomy. RANTES was measured by immunofluorescence method. RESULTS: Intensive granular luminescence was observed in all cytoplasm of cells with the exception of nucleus. Immunoreactive RANTES was found to be present in 70% of cells. We not find increase percentage of positive RANTES fibroblasts after stimulation with lipopolisaccharides and phytohemagglutinin. RANTES expression was similar in the both: atopic and nonatopic polyps. CONCLUSIONS: This study demonstrates that cultured fibroblasts derived from both atopic and nonatopic patients release RANTES spontaneously and after stimulation with lipopolisaccharides and phytohemagglutinin. This observation and the finding that RANTES is present in nasal polyps fibroblasts suggest that this chemokine may be an important mediator of eosinophil in both atopic and nonatopic nasal polyposis. More research needs to expand on chemotactic factors such as RANTES and their interplay with other local cytokines.

New isothiazole derivatives: synthesis, reactivity, physicochemical properties and pharmacological activity.

The synthesis and biological investigation of the series of amide and ester derivatives 10-20 of 5-(4-chlorobenzoyl)amino-3-methyl-4-isothiazolecarboxylic acid 5 are presented. Because the amide series of 5-benzoylamino-3-methyl-4-isothiazolecarboxylic acid 2 has been studied extensively and from this series denotivir (vratizolin) 4 became the antiviral drug. The influence of exchanging the N-benzoyl for a N-(4-chlorobenzoyl) group at position 5 of the isothiazole ring on the pharmacological activity of 5-benzoylamino-3-methyl-4-isothiazolecarboxylic acid 2 derivatives is dealt with here. The effect of structure modifications in the carboxylic group of the 5-(4-chlorobenzoyl)amino-3-methyl-4-isothiazolecarboxylic acid 5 series of derivatives on their biological activity is discussed. Some of the tested 5-(4-chlorobenzoyl)amino-3-methyl-4-isothiazolecarboxylamides revealed significant anti-inflammatory activity in carrageenan induced edema and air-pouch inflammation tests. Physicochemical properties of 6-(4-chlorophenyl)-3-methylisothiazolo[5,4-d]-4H-1,3-oxazin-4-one 6 are described. Its use in the synthesis of isothiazole derivatives and its reactivity are also presented.

[Influence of polymorphism pro12Ala of PPARgamma gene on endothelium destruction in patients with diabetes mellitus t. 2]. / Polimorfizm pro12Ala genu receptora PPARgamma a procey destrukcyjne sródblonka u chorych na cukrzyce typu 2.

UNLABELLED: The peroxisome proliferator-activated receptor gamma (PPARgamma) influences wide on metabolism and atheromatosus processes in vessels. The common polymorphic form of PPARy, Pro12Ala, could promote diabetes mellitus and diabetic vascular complications. AIM OF WORK: The assessment of indicators of endothelium destruction in patients with diabetes mellitus t.2 in relation to the polymorphism Pro12Ala of PPARgamma. PATIENTS AND METHODS: Circulating blood endothelium cells (immunofluorescens method with MoAb CLB-HEC19), thrombomodulin (Asserchrom Immunoassay) and polymorphism Pro12Ala [minisequence technic SnaPshot (Applera); ABI+PRISM310] were investigated in 58 patients with diabetes mellitus typ 2 and 22 healthy persons. Fibrinogen, uric acid, lipids, HbA1c, glucose, insulin concentration, blood pressure, BMI and WHR were evaluated too. RESULTS: The significant higher systolic (137.92 +/- 15.88 vs 122.0 +/- 15,67 [mmHg]; p < 0.025) and diastolic (85.00 +/- 7.38 vs 75.50 +/- 7.61 [mmHg]; p < 0.011) was determined in the group of healthy people, who have got a homozygous genotyp Pro12Pro in comparison with heterozygous genotyp Pro12Ala. The significant higher value of HbA1c was determined in the patients with diabetes mellitus t.2, who have got genotyp Pro12Ala in comparison with genotyp Pro12Pro (7.01 +/- 1.54 vs 8.39 +/- 1.81 [%]; p < 0.006). There was any significant difference for others parameters. Among people, which have got genotyp Pro12Pro there was significant difference between healthy and patients for circulating blood endothelium cells (2.19 +/- 1.53 vs 0.78 +/- 0.09 [EC/ml]; p < 0.009). On the contrary among people with genotype Pro12Ala there was not significant difference between healthy and patients for circulating blood endothelium cells (2.95 +/- 1.64 vs 1.61 +/- 1.08; p = 0.077 [EC/ml]). CONCLUSIONS: 1) The polymorphism Pro12Ala is not connected with the endothelium destruction. 2) Other researches are necessary to estimate influence of mutated allele on the control of diabetes. 3) The genotyp Pro12Pro promotes higher blood pressure by healthy people.

Retinal antigens are recognized by antibodies present in sera of patients with multiple sclerosis.

Multiple sclerosis (MS) is frequently accompanied by visual symptoms including those related to retinal disorders. Since they may be a consequence of an autoimmune reaction, we examined whether sera of patients with diagnosed MS and changes in visual-evoked potentials contain antibodies against retinal antigens (retAgs). Immunoblot analysis revealed that MS sera recognized mainly a 46-kD antigen, a 41-kD antigen, retinal arrestin, to a smaller extent also 70-, 56-, 43-, and 36-kD proteins. Patients whose sera showed the highest reactivity with 41- and 46-kD antigens had deficiencies in visual acuity, visual fields, ophthalmoscopy, and electroretinograms. Our observation suggests that antibodies to these retAgs may play a role in the origin of ophthalmologic impairment in MS.

[The role of peroxisome proliferator-activated receptor gamma in the pathogenesis of diabetes and atheromatosis]. / Rola receptora aktywowanego przez proliferatory peroksysomów gamma w patogenezie cukrzycy i miazdzycy.

The high risik of cardiovascular diseases in diabetes is connected with wide and premature atheromatosis. It is caused by systemic metabolic disorders like hyperglycaemia, insulin resistance, dyslipidaemia, endothelium dysfunction. This review will discuss the role of peroxisome proliferator-activated receptor gamma (PPARgamma) in the pathogenesis diabetes and atheromatosous injury of vessels. PPARgamma is a nuclear transcript factor with a very wide spectrum of biological activities. It influences important risik factors of atheromathosis, especially by patients with metabolic syndrome in diabetes type 2. Thiazolidinediones, which is activators PPARgamma, could be a turning-point in the treatment of diabetic angiopathy.

[Histopathologic studies in experimental uveitis]. / Badania histopatologiczne w eksperymentalnym zapaleniu blony naczyniowej.

UNLABELLED: Experimental uveitis is one of the main models in the diseases of autoimmunological background. The purpose of this paper was to analyze the quantitative histological changes in the experimental uveitis, induced by different types of homogenous endotoxin salts of Havnia alvei. We studied 74 eyes of Lewis rats (males) divided into 4 groups. Each group received a homogenous salt of Havnia alvei in a single subcutaneous injection. In the 1 group-LPS Ca++, in 2 group-LPS Na++, in 3 group-LPS 981, in 4 group physiological salt (control group). The histologic and immunocitochemical examinations were performed after 24, 48 hours, and after 4 and 7 days following the injection. The histologic changes were analyzed (he intensity of inflammatory reaction) using a Highly Optimazed Microscope Enviroment system. The most intensive inflammation was observed in experimental group after 24 hours (n the LPS 981D group in 5 rats out of 6, in LPS Ca++ group in 3 rats out of 6). After 48 hours the intensity of inflammatory reaction visibly decreased. On the fourth day the inflammation revealed a minimal intensity and after 7 days was practically absent. In the control group minimal inflammation was observed only in a few rats. Cilliary body hypermia was present for 48 hours in most of the experimental rats. Only a few of them had hyperemia on the fourth day. In the posterior segment minimal inflammation was noted at the end of the first day (I and II-nd group after 24 hours); this process continued until the fourth day, on the 7th day disappeared. CONCLUSIONS: The most intensive inflammation of the anterior and posterior choroidal segment is caused by homogenous salts of Havnia alvei 981.

Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid.

The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor alpha and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells.

[The influence of nitric synthase inhibitor (L-NAME) on the intraocular pressure in endotoxin induced uveitis in rats]. / Wplyw inhibitora syntazy tlenku azotu (L-NAME) na cisnienie sródgalkowe u szczurów w przebiegu zapalenia blony naczyniowej indukowanego endotoksyna.

We studied the influence of the nitric synthase inhibitor (L-NAME) on the induction of uveitis in Wistar rats. The inflammation was induced by a footpad injection of endotoxin in rat. The clinical signs were evaluated in a slit-lamp, the intraocular pressure was measured using Tono Pen tonometer. We showed, that in endotoxin-induced uveitis the intraocular pressure elevation was observed and that L-NAME application reduced the inflammation and the level of the intraocular pressure.

Biological activity of alpha-galactoside preparations from Lupinus angustifolius L. and Pisum sativum L. seeds.

Biological activity tests were performed on alpha-galactoside preparations obtained from Lupinus angustifolius L. cv. Mirela (alkaloid-rich) and Pisum sativum L. cv. Opal seeds. The studies included the following tests: acute toxicity, cytotoxic test, delayed type hypersensitivity (DTH), plaque-forming cell number (IgM-PFC), and influence on the growth of bifidobacteria and coliform presence in rat colon. Results of these studies showed that alpha-galactosides from lupin and pea seeds were essentially nontoxic. Their acute toxicity (LD(50)) in mice was >4000 mg kg(-1) of body weight. alpha-galactoside preparations were not cytotoxic for mouse thymocytes in vitro. The in vitro test shows that oligosaccharides from lupin and pea are utilized by selected beneficial colon bacterium strains. The in vivo experiment demonstrated that alpha-galactosides from legume significantly influenced the growth of bifidobacteria in rats colon. Simultaneously, the decrease of the coliform presence was observed. The chemical composition of the tested preparations had no significant effect on their biological activity.

[Potential use of cysteine endopeptidase inhibitors in radiodiagnostics of laryngeal and tongue neoplasms]. / Badania nad wykorzystaniem inhibitorów endopeptydazy cysteinowej w radiodiagnostyce nowotworów jezyka i krtani.

The methods applied up to now do not allow to define the border between the tumor and healthy tissue especially in advanced cases. Microlaryngoscopy facilitates the evaluation of the infiltration superficially, but does not allow for estimation its deeper penetration. CT and MRI often do not answer the question concerning the tumor spread as well. That is why the monoclonal antibodies marked with isotopes directed against specific antibodies of different neoplasms are applied in diagnostic. In the neoplasm development the great role play endopeptidases. They degrade the intercellular matrix and basement membrane and by this way made possible the neoplasm invasion. The high level of the endopeptidase in patients with neoplasm is a bad prognosis. The disposition of endopeptidase in neoplasm tissue is characteristic and gives opportunity to localise the border of healthy tissue thanks their inhibitors application. The inhibitor of cysteine endopeptidase obtained from hen egg and marked by J 125 was used in our study. The reaction between the cysteine endopeptidase inhibitor and malignant cells received directly from patients with tongue and laryngeal carcinoma and cells cultivated in vitro from the carcinoma.