Análise da associação entre a periodontite e polimorfismos de nucleotídeo único da interleucina-1beta / Analysis of the association between periodontitis and interleukin-1beta single nucleotide polymorphisms

A periodontite é uma doença inflamatória, que pode ser classificada em Periodontite Crônica (PC) ou Periodontite Agressiva (PA), desencadeada por um desequilíbrio na microbiota subgengival, que pode ser influenciado por diversos fatores, como polimorfismos genéticos. Dessa forma, o objetivo deste trabalho foi analisar, através de uma revisão da literatura, se há ou não associação entre a periodontite e polimorfismos genéticos de nucleotídeo único (SNPs). Os genes IL1B +3954(3), -511, -31 foram os alvos desta pesquisa, por serem os mais estudados e apresentarem boa plausibilidade biológica. Foi realizada uma busca no PubMed e ao final 24 artigos de estudos casos-controle foram selecionados. Na maioria dos estudos não foi encontrada associação positiva entre os SNPs +3954(3), -511, -31 da IL1B e a PA ou PC. Dessa forma, é possível concluir que não há associação positiva entre a periodontite e os SNPs IL1B +3954(3), -511, -31 e PA e PC. Todavia os resultados devem ser analisados com cautela, pois os estudos apresentam limitações. (AU)

Periodontitis is an inflammatory disease which is classified as chronic periodontitis (PC) or aggressive periodontitis (PA) and is initiated by an unbalance in subgengival microbiota which for their part can be influenced by a lot of factors, such as genetic polymorphism. Therefore, the aim of this study was to analyse if there is an association between periodontitis and single nucleotide polymorphisms (SNPs). The genes IL1B +3954(3), -511, -31 were chosen as the targets of this literature review, because they have good biologic plausibility and are the most studied in literature. A search was conducted on PubMed and the results analysed and 24 case-controls articles were chosen.Most of the studies did not find a positive association between the ILB1 SNPs +3954(3), -511, -31 and PA e PC. Therefore, the case-controls studies indicated that there is no positive association between SNPs IL1B +3954(3), -511, -31 and PA or PC. However, the results of this work must be analysed carefully as the studies used have limitations. (AU)

IL-1 Receptor Antagonist Reduced Chemical-Induced Keratinocyte Apoptosis through Antagonism to IL-1α/IL-1β

Extracellular interleukin 1 alpha (IL-1α) released from keratinocytes is one of the endpoints for in vitro assessments of skin irritancy. Although cells dying via primary skin irritation undergo apoptosis as well as necrosis, IL-1α is not released in apoptotic cells. On the other hand, active secretion has been identified in interleukin-1 receptor antagonist (IL-1ra), which was discovered to be a common, upregulated, differentially-expressed gene in a microarray analysis performed with keratinocytes treated using cytotoxic doses of chemicals. This study examined whether and how IL-1ra, particularly extracellularly released IL-1ra, was involved in chemically-induced keratinocyte cytotoxicity and skin irritation. Primary cultured normal adult skin keratinocytes were treated with cytotoxic doses of chemicals (hydroquinone, retinoic acid, sodium lauryl sulfate, or urshiol) with or without recombinant IL-1ra treatment. Mouse skin was administered irritant concentrations of hydroquinone or retinoic acid. IL-1ra (mRNA and/or intracellular/extracellularly released protein) levels increased in the chemically treated cultured keratinocytes with IL-1α and IL-1β mRNAs and in the chemically exposed epidermis of the mouse skin. Recombinant IL-1ra treatment significantly reduced the chemically-induced apoptotic death and intracellular/extracellularly released IL-1α and IL-1β in keratinocytes. Collectively, extracellular IL-1ra released from keratinocytes could be a compensatory mechanism to reduce the chemically-induced keratinocyte apoptosis by antagonism to IL-1α and IL-1β, suggesting potential applications to predict skin irritation.

Cathepsin B in hepatic Kupffer cells regulates activation of TLR4-independent inflammatory pathways in mice with lipopolysaccharide-induced sepsis / 南方医科大学学报

OBJECTIVE@#To investigate the role of cathepsin B in hepatic Kupffer cells (KCs) in activating Toll-like receptor 4(TLR- 4)-independent inflammatory pathways in mice with lipopolysaccharide (LPS)-induced sepsis.@*METHODS@#Eighteen wild-type (WT) mice and 18 TLR4-knockout (TLR4) mice were both divided into 3 groups for intraperitoneal injections of a lethal dose (54 mg/kg) of LPS, LPS and CA-074(a cathepsin B inhibitor), or normal saline, and the survival of the mice were observed. Another 36 WT mice and 36 TLR4mice were also divided into 3 groups and subjected to intraperitoneal injections of normal saline, 20 mg/kg LPS, or LPS with CA-074 pretreatment.After the treatments, KCs were collected from the mice for assessing the protein level and activity of cathepsin B.The histopathological changes of the liver were observed with HE staining, and the serum levels of IL-1α, IL-1β, TNF-α and IL-18 were detected.@*RESULTS@#Compared with the WT mice,TLR4mice receiving the lethal dose of LPS had significantly longer survival time (up to 84 h) after the injection,but were still unable to fully resist LPS challenge.CA-074 pretreatment prolonged the survival time of WT mice and TLR4mice to 60 h and 132 h,respectively.In the mouse models of sepsis,20 mg/kg LPS induced significantly enhanced activity of cathepsin B without affecting its expression level in the KCs (<0.05) and increased the serum levels of the inflammatory cytokines.CA-074 pretreatment of the mice obviously lessened the detrimental effects of LPS in TLR4mice by significantly lowering cathepsin B activity in the KCs,alleviating hepatocyte apoptosis and reducing the serum levels of inflammatory cytokines.@*CONCLUSIONS@#Cathepsin B plays an important role in activating TLR4-independent inflammatory pathways in mice with LPS-induced sepsis.

Increased Skin Irritation by Hydroquinone and Rsetinoic Acid Used in Combination

BACKGROUND: Hydroquinone (HQ) is frequently combined with retinoic acid (RA) to enhance lightening efficacy, which may also affect skin irritancy. Although skin irritation leads to postinflammatory hyperpigmentation, little research has been performed to compare skin irritancy between each component and the combination. OBJECTIVE: This study was done to examine whether HQ-RA combination increased skin irritation induced by HQ or RA alone. METHODS: Patch testing was performed using maximum therapeutic and higher concentrations of HQ and RA in 10 volunteers, and then, it was performed using their popular therapeutic concentrations and combination in the other 20 volunteers. In vitro irritation was also assessed in primary cultured normal human keratinocytes treated with 80% and 50% cell survival doses of HQ, 80% cell survival dose of RA, and their combination. RESULTS: The combination in patch testing induced stronger erythema than the corresponding concentrations of HQ and RA, which was remarkable with use of combination of higher concentrations. In cultured keratinocytes, the RA combination significantly decreased cell viability, but increased cytotoxicity and extracellular interleukin 1 alpha release with corresponding doses of HQ. CONCLUSION: The results of patch tests and in vitro irritation assessment tests suggested that HQ and RA increased skin irritation when used in combination.

Relationship between hypertension and periapical lesion: an in vitro and in vivo study

Abstract The aim of this study was to compare potential aspects of periapical lesion formation in hypertensive and normotensive conditions using hypertensive (BPH/2J) and wild-type control (BPN/3J) mice. The mandibular first molars of both strains had their dental pulp exposed. At day 21 the mice were euthanized and right mandibular molars were used to evaluate the size and phenotype of apical periodontitis by microCT. Proteins were extracted from periapical lesion on the left side and the expressions of IL1α, IL1β and TNFα were analyzed by ELISA. Bone marrow stem cells were isolated from adult mice femurs from 2 strains and osteoclast differentiation was evaluated by tartrate-resistant acid phosphatase (TRAP) in vitro. The amount of differentiated osteoclastic cells was nearly double in hypertensive mice when compared to the normotensive strain (p < 0.03). Periapical lesion size did not differ between hypertensive and normotensive strains (p > 0.7). IL1α, IL1β and TNFα cytokines expressions were similar for both systemic conditions (p > 0.05). Despite the fact that no differences could be observed in periapical lesion size and cytokines expressions on the systemic conditions tested, hypertension showed an elevated number of osteoclast differentiation.

Ethanol during adolescence decreased the BDNF levels in the hippocampus in adult male Wistar rats, but did not alter aggressive and anxiety-like behaviors / O etanol durante a adolescência reduziu os níveis de BDNF no hipocampo de ratos Wistar machos adultos, mas não alterou os comportamentos agressivos e a ansiedade

Objective:To investigate the effects of ethanol exposure in adolescent rats during adulthood by assesssing aggression and anxiety-like behaviors and measuring the levels of inflammatory markers.Methods:Groups of male Wistar rats (mean weight 81.4 g, n = 36) were housed in groups of four until postnatal day (PND) 60. From PNDs 30 to 46, rats received one of three treatments: 3 g/kg of ethanol (15% w/v, orally, n = 16), 1.5 g/kg of ethanol (12.5% w/v, PO, n = 12), or water (n = 12) every 48 hours. Animals were assessed for aggressive behavior (resident x intruder test) and anxiety-like behaviors (elevated plus maze) during adulthood.Results:Animals that received low doses of alcohol showed reduced levels of brain-derived neurotrophic factor (BDNF) in the hippocampus as compared to the control group. No significant difference was found in prefrontal cortex.Conclusions:Intermittent exposure to alcohol during adolescence is associated with lower levels of BDNF in the hippocampus, probably due the episodic administration of alcohol, but alcohol use did not alter the level agression toward a male intruder or anxiety-like behaviors during the adult phase.

Objetivo: Investigar os efeitos da exposição ao etanol em ratos adolescentes durante a idade adulta sobre os comportamentos agressivos e semelhantes à ansiedade, bem como sobre as medidas de níveis de marcadores inflamatórios.Métodos:Os grupos de ratos Wistar machos (peso médio de 81,4 g; n = 36) foram alojados em grupos de quatro até o dia pós-natal (DPN) 60. Entre os DPNs 30 e 46, os ratos receberam um dos três tratamentos: 3 g/kg de etanol (15% w/v, oralmente, n = 16), 1.5 g/kg de etanol (12,5% w/v, oralmente, n = 12), ou água (n = 12) a cada 48 horas. Os comportamentos agressivos (teste residente-intruso) e semelhantes à ansiedade (labirinto em cruz elevado) foram avaliados durante a idade adulta dos animais.Resultados:Os animais que receberam doses menores de álcool mostraram níveis reduzidos de fator neurotrófico derivado do cérebro (BDNF) no hipocampo quando comparados ao grupo controle. Nenhuma diferença significativa foi verificada no córtex pré-frontal.Conclusões:A exposição intermitente ao álcool durante a adolescência é associada com menores níveis de BDNF no hipocampo, provavelmente divido a administração episódica de álcool, mas o uso não alterou o nível de agressão contra o macho intruso ou os comportamentos semelhantes à ansiedade durante a fase adulta.

Therapeutic Effects of Umbilical Cord Blood Derived Mesenchymal Stem Cell-Conditioned Medium on Pulmonary Arterial Hypertension in Rats

BACKGROUND: Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) may have multiple therapeutic applications for cell based therapy including the treatment of pulmonary artery hypertension (PAH). As low survival rates and potential tumorigenicity of implanted cells could undermine the mesenchymal stem cell (MSC) cell-based therapy, we chose to investigate the use of conditioned medium (CM) from a culture of MSC cells as a feasible alternative. METHODS: CM was prepared by culturing hUCB-MSCs in three-dimensional spheroids. In a rat model of PAH induced by monocrotaline, we infused CM or the control unconditioned culture media via the tail-vein of 6-week-old Sprague-Dawley rats. RESULTS: Compared with the control unconditioned media, CM infusion reduced the ventricular pressure, the right ventricle/(left ventricle+interventricular septum) ratio, and maintained respiratory function in the treated animals. Also, the number of interleukin 1alpha (IL-1alpha), chemokine (C-C motif) ligand 5 (CCL5), and tissue inhibitor of metalloproteinase 1 (TIMP-1)-positive cells increased in lung samples and the number of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling technique (TUNEL)-positive cells decreased significantly in the CM treated animals. CONCLUSIONS: From our in vivo data in the rat model, the observed decreases in the TUNEL staining suggest a potential therapeutic benefit of the CM in ameliorating PAH-mediated lung tissue damage. Increased IL-1alpha, CCL5, and TIMP-1 levels may play important roles in this regard.

Effect of colon cancer cell-derived IL-1α on the migration and proliferation of vascular endothelial cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effect of colon cancer cell-derived interleukin-1α on the migration and proliferation of human umbilical vein endothelial cells as well as the role of IL-1α and IL-1ra in the angiogenesis process.</p><p><b>METHODS</b>Western blot was used to detect the expression of IL-1α and IL-1R1 protein in the colon cancer cell lines with different liver metastatic potential. We also examined how IL-1α and IL-1ra influence the proliferation and migration of umbilical vascular endothelial cells assessed by PreMix WST-1 assay and migration assay, respectively. Double layer culture technique was used to detect the effect of IL-1α on the proliferation and migration of vascular endothelial cells and the effect of IL-1ra on the vascular endothelial cells.</p><p><b>RESULTS</b>Western blot analysis showed that IL-1α protein was only detected in highly metastatic colon cancer HT-29 and WiDr cells, but not in the lowly metastatic CaCo-2 and CoLo320 cells.Migration assay showed that there were significant differences in the number of penetrated cells between the control (17.9±3.6) and 1 ng/ml rIL-1α group (23.2±4.2), 10 ng/ml rIL-1α group (31.7±4.5), and 100 ng/ml rIL-1α group (38.6±4.9), showing that it was positively correlated with the increasing concentration of rIL-1α (P<0.01 for all). The proliferation assay showed that the absorbance values were 1.37±0.18 in the control group, and 1.79±0.14 in the 1 ng/ml rIL-1α group, 2.14±0.17 in the 10 ng/ml rIL-1α group, and 2.21±0.23 in the 100 ng/ml rIL-1α group, showing a positive correlation with the increasing concentration of rIL-1α(P<0.01 for all). IL-1ra significantly inhibited the proliferation and migration of vascular endothelial cells (P<0.01). The levels of VEGF protein were (1.697±0.072) ng/ml, (3.507±0.064)ng/ml and (4.139±0.039)ng/ml in the control, HUVECs+ IL-1α and HUVECs+ HT-29 co-culture system groups, respectively, showing a significant difference between the control and HUVECs+ 10 pg/ml rIL-1α groups and between the control and HUVECs+ HT-29 groups (P<0.01 for both).</p><p><b>CONCLUSIONS</b>Our findings indicate that colon cancer cell-derived IL-1α plays an important role in the liver metastasis of colon cancer through increased VEGF level of the colon cancer cells and enhanced vascular endothelial cells proliferation, migration and angiogenesis, while IL-1ra can suppress the effect of IL-1α and inhibit the angiogenesis in colon cancer.</p>

Microarray Analysis for Genes Associated with Angiogenesis in Diabetic OLETF Keratocytes

The purpose of this study was to identify the differences in angiogenesis gene expression between normal and diabetic keratocytes stimulated with interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha). Primarily cultured normal and diabetic keratocytes were treated with 20 ng/mL of IL-1a and TNF-alpha for 6 hr. cDNA was hybridized to an oligonucleotide microarray. Microarray analysis was used to identify differentially expressed genes that were further evaluated by real-time polymerase chain reaction (RT-PCR). Diabetes keratocytes overexpressed vital components of angiogenesis including Agtr1, and under-expressed components related to the blood vessel maturation, including Dcn. Cytokine-treated diabetic keratocytes differentially expressed components of angiogenesis. OLETF keratocytes after treatment with IL-1alpha and TNF-alpha showed the newly expressed 15 and 14 genes, respectively. Newly and commonly under-expressed five genes followed by treatment with both IL-1alpha and TNF-alpha were also evident. RT-PCR showed results similar to the microarray results. Agtr1 and Itga1 showed an increased expression in diabetic keratocytes compared with normal corneal keratocytes, especially after TNF-alpha treatment. Il6 appeared strong expression after interleukin-1alpha treatment, but showed down expression after TNF-alpha treatment. Further studies to analyze and confirm the significance of the identified angiogenetic genes of diabetes are needed.

Cigarette Smoke-Induced Interleukin-1 Alpha May Be Involved in the Pathogenesis of Adult Acne

BACKGROUND: Lipid peroxide (LPO) in comedones, which are produced as a result of sebum oxidation, might potentially induce interleukin-1alpha (IL-1alpha) and exacerbate comedogenesis and inflammatory changes in comedones. OBJECTIVE: To investigate the relationship of proinflammatory cytokines and LPO levels in the extracts of comedones with the acne of clinical difference between smokers and non-smokers, and with the severity and distribution of the acne lesions. METHODS: Twenty-two non-smoking and 21 smoking adult acne patients were evaluated by comedone extraction and measurement of proinflammatory cytokines and LPO levels. Acne severity and distribution of the lesions were also analyzed. RESULTS: Relative to the non-smoking group, smokers had significantly higher levels of IL-1alpha and LPO in comedones. Their levels showed a positive correlation. However, there were no statistically significant difference between the severity or distribution of the disease and the levels of LPO and IL-1alpha in comedones. CONCLUSION: Smoking may be involved in the pathogenesis of adult acne by increasing the oxidative stress that results in subsequent accumulation of LPO in comedones.

Dynamic changes of a group of cytokines in phosgene-induced lung injury and the function of ulinastatin / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic changes of a group of cytokines in phosgene-induced lung injury and the function of different dose of ulinastatin through animal experiment.</p><p><b>METHODS</b>104 male SD rats were randomly assigned into the control group, ulinastatin control group, phosgene treatment groups and different dose of ulinastatin intervention groups, 8 rats each group. Treatment groups were dynamic constant exposure in phosgene, and immediately injected ulinastatin intraperitoneal, and then the experimental animal, the lung tissue biopsy, lung wet/dry ratio, RT-PCR detection, the plasma for detection of Bio-Plex 18 cytokines.</p><p><b>RESULTS</b>Compared with the control group, plasma concentrations of IL-1α, IL-6, GM-CSF, TNF-α, INF-γ, MIP-3α, VEGF were increased significantly first (2 h), and gradually decreased with the passage of time , the difference was statistically significant (P < 0.05). Plasma concentrations of IL-4, IL-10 were decreased earlier (2h, 6 h) and increased later (24 h) (P < 0.05). The change of plasma concentration of IL-13 was not obvious earlier (2 h) and still rising later (24h), the difference was statistically significant (P < 0.05). After drug intervention, the levels of pro-inflammatory cytokines declined and the levels of anti-inflammatory cytokines raise by different degrees at different times in ulinastatin intervention groups, the difference was statistically significant. The degree of lung injury was improved than the phosgene treatment groups and better in high dose of ulinastatin intervention group. The expression of IL-10, IL-4, IL-13-mRNA of tissue increased in accordance with plasma results.</p><p><b>CONCLUSION</b>A group of cytokines are dynamicly changed in phosgene-induced lung injury by time. High dose of ulinastatin can improved phosgene-induced lung injury, regulate the synthesis and release of inflammatory cytokines and inhibit inflammatory react in a dose-dependent manner.</p>

Cytotoxic effects of sodium dodecyl benzene sulfonate on human keratinocytes are not associated with proinflammatory cytokines expression / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Keratinocytes play a crucial role in the biological function of skin barrier. The relationship between sodium lauryl sulfate (SLS) and keratinocytes has been studied. However, the cytotoxicity and effects of sodium dodecyl benzene sulfonate (SDBS), a common detergent similar to SLS, on keratinocytes are still not known. This study aimed to investigate the effects of SDBS on cytotoxicity and expression of proinflammatory cytokines in cultured human keratinocytes.</p><p><b>METHODS</b>This study was carried out using the keratinocytes cell line, HaCaT cells. The cytotoxicity of SDBS on HaCaT cells was evaluated with cell counting kit-8 (CCK-8) and phase-contrast microscopy. After exposure to different concentrations of SDBS, the total RNA of the HaCaT cells was extracted for evaluating the relative mRNA expression of IL-1α, IL-6, IL-8, and TNF-α by qPCR. The supernatants of cells were collected for measuring the levels of IL-6 and IL-8 by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>SDBS at concentrations of 20 µg/ml and over showed direct cytotoxicity and induced morphological changes of the HaCaT cells. The mRNA expressions of IL-1α, IL-6, IL-8, and TNF-a in different concentrations of SDBS at different time were comparable with that of controls. SDBS at concentrations of 5, 10, and 15 µg/ml had no significant effects on IL-6 and IL-8 excretion from HaCaT cells after 24-hour exposure. Moreover, no significant effects on the IL-6 and IL-8 excretion were found after 10 and 15 µg/ml SDBS stimulations for 6, 12, and 24 hours, respectively.</p><p><b>CONCLUSION</b>SDBS at higher concentrations had cytotoxicity on HaCaT cells but had no effects on the mRNA expression of IL-1α, IL-6, IL-8, and TNF-a, that was different from SLS.</p>

FSL-1, a Toll-like Receptor 2/6 Agonist, Induces Expression of Interleukin-1alpha in the Presence of 27-hydroxycholesterol

We investigated the question of whether cholesterol catabolite can influence expression of inflammatory cytokines via Toll-like receptors (TLR) in monocytic cells. Treatment of THP-1 monocytic cells with 27-hydroxycholesterol (27OHChol) resulted in induction of gene transcription of TLR6 and elevated level of cell surface TLR6. Addition of FSL-1, a TLR6 agonist, to 27OHChol-treated cells resulted in transcription of the IL-1alpha gene and enhanced secretion of the corresponding gene product. However, cholesterol did not affect TLR6 expression, and addition of FSL-1 to cholesterol-treated cells did not induce expression of IL-1alpha . Using pharmacological inhibitors, we investigated molecular mechanisms underlying the expression of TLR6 and IL-1alpha. Treatment with Akt inhibitor IV or U0126 resulted in significantly attenuated expression of TLR6 and IL-1alpha induced by 27OHChol and 27OHChol plus FSL-1, respectively. In addition, treatment with LY294002, SB202190, or SP600125 resulted in significantly attenuated secretion of IL-1alpha . These results indicate that 27OHChol can induce inflammation by augmentation of TLR6-mediated production of IL-1alpha in monocytic cells via multiple signaling pathways.

Effect of methotrexate on serum levels of IL-1alpha and IL-8 in rheumatoid arthritis

Rheumatoid arthritis is a chronic systemic inflammatory disease affects many tissues and organs, but principally attacks flexible [synovial] joints. Methotrexate is the most commonly used disease-modifying antirheumatic drug for the treatment of rheumatoid arthritis. The aim of this study was to evaluate the effect of methotrexate on serum levels of IL-1alpha and IL-8 in rheumatoid arthritis. Blood samples were collected from 50 patients with rheumatoid arthritis [25 patients without treatment and 25 patients are received methotraxate] and from 30 healthy age and sex matched individuals served as controls. Serum IL-1alpha and IL-8 were measured by means of enzyme-linked immune-sorbent assay. The present results showed that serum levels of IL-1alpha and IL-8 were significantly higher in RA patients than in healthy controls [P<0.01], furthermore, level of IL-1alpha was significantly decrease in patients treated with methotraxate as compared to those patients who have received no treatment [P<0. 01]. On the other hand serum level of IL-8 didn't showed any significant differences between patients treated with methotraxate and those patients without treatment [P>0. 05]. These finding demonstrate that methotrexate turns out to be a good inhibitor for IL-alpha production. In addition, IL-1alpha and IL-8 may have a significant role in the pathogenesis of rheumatoid arthritis, and could be use as.

Removal of oral Prevotella intermedia Endotoxin by octyl phenyl polyoxyethylene ether extraction method / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate an effective purification method for removing endotoxin from Prevotella intermedia.</p><p><b>METHODS</b>The main protein ingredients of bacteria prepared from ammonium sulfate precipitation were further treated with octyl phenyl polyoxyethylene ether (Triton X-114), and then processed at 4°C, 37°C and 25°C. The obtained aqueous phase after at least two more cycle repeated operations was assayed for endotoxin by Western blotting, LAL-clotting method, in vitro cell stimulation and in vivo animal experiments.</p><p><b>RESULTS</b>Western blotting and LAL-clotting method demonstrated that the reduction in endotoxin level was greater than 99.99% and recovery of the proteins after endotoxin removal was greater than 90% with Triton X-114 treatment for 3 cycles. The cytokines expression level was lower in both in vitro cell stimulation and in vivo animal experiments than in untreated group (P < 0.05).</p><p><b>CONCLUSIONS</b>The extraction method provides a new choice for endotoxin removal from large volumes of the oral Prevotella intermedia.</p>

Allele, genotype, and composite genotype effects of IL-1A +4845 and IL-1B +3954 polymorphisms for chronic periodontitis in an Indian population.

Introduction: The pro-inflammatory cytokine interleukin-1 (IL-1) is a key modulator of host responses to microbial infection and a major modulator of extracellular matrix catabolism and bone resorption, and polymorphisms in the IL-1 gene cluster have been associated with an increased risk of developing severe adult periodontitis. A case control study was performed to determine the role of IL-1A+4845 and IL-1B+3954 polymorphisms in the predisposition to chronic periodontitis. Materials and Methods: The study was conducted with 103 unrelated participants recruited from Manipal College of Dental Sciences, Manipal, which included 51 chronic periodontitis patients and 52 normal periodontally healthy individuals. Extensive clinical data were collected, bone loss was the major outcome variable and smokers and diabetics were excluded from the study to eliminate the influence of these risk factors. Genomic DNA was isolated from the blood samples of participants for genotyping IL-1A+4845 and IL-1B+3954 polymorphisms by polymerase chain reaction-restriction fragment length polymorphism and the data statistically analyzed. Results: Allele 2 of the IL-1A+4845 polymorphism was carried by 38% of all participants; of these only 6 were homozygous for the allele. Allele 2 of the IL-1B+3954 was carried by 21% of the subjects; only 1 was homozygous for allele 2. The composite genotype was carried by 31% of the cases and by 38% of the controls. Overall, 35% participants carried the composite IL-1 genotype. No statistically significant association was found for the distributions. Conclusions: The distribution of the IL-1 positive composite genotype is in concordance with the frequencies reported in the Caucasians. Association was not found for the effect of allele, genotype, composite genotype, and haplotypes of IL-1A+4845 and IL-1B+3954 polymorphisms with periodontitis. Its utility as a risk marker in this population was not borne out by the study.

Interleukin- 8- 251T > a, interleukin -1α -889C > t and apolipoprotein e polymorphisms in Alzheimer's disease

An inflammatory process has been involved in numerous neurodegenerative disorders such as Parkinson's disease, stroke and Alzheimer's disease (AD). In AD, the inflammatory response is mainly located in the vicinity of amyloid plaques. Cytokines, such as interleukin-8 (IL-8) and interleukin-1α (IL-1α), have been clearly involved in this inflammatory process. Polymorphisms of several interleukin genes have been correlated to the risk of developing AD. The present study investigated the association of AD with polymorphisms IL-8 -251T > A (rs4073) and IL-1α-889C > T (rs1800587) and the interactive effect of both, adjusted by the Apolipoprotein E genotype. 199 blood samples from patients with AD, 146 healthy elderly controls and 95 healthy young controls were obtained. DNA samples were isolated from blood cells, and the PCR-RFLP method was used for genotyping. The genotype distributions of polymorphisms IL-8, IL-1α and APOE were as expected under Hardy-Weinberg equilibrium. The allele frequencies did not differ significantly among the three groups tested. As expected, the APOE4 allele was strongly associated with AD (p < 0.001). No association of AD with either the IL-1α or the IL-8 polymorphism was observed, nor was any interactive effect between both polymorphisms. These results confirm previous studies in other populations, in which polymorphisms IL-8 -251T > A and IL-1α-889C > T were not found to be risk factors for AD.

Effect of Apocynin on Acute Lung Injury in Rats Given Interleukin-1alpha Intratracheally / 결핵및호흡기질환

BACKGROUND: Based on the assertion that apocynin diminishes acute lung injury (ALI) by inhibition of NADPH oxidase, the effect of apocynin was tested in interleukin-1alpha (IL-1)-induced ALI in rats. METHODS: IL-1 was insufflated into the trachea of Sprague-Dawley rats to induce ALI, and apocynin (8 mg/kg) was given intravenously for inhibition of NADPH oxidase. In addition, we determined whether apocynin inhibited generation of superoxide anions from isolated human neutrophils. Five hours after IL-1 instillation, lung injury parameters, expression of cytosolic phospholipase A2 (cPLA2) by cells from bronchoalveolar lavage (BAL), an index of oxidative stress in lung tissues (gamma-glutamyltranspeptidase, activity), and ultrastructure of alveolar type II (AT II) cells were evaluated. RESULTS: Apocynin decreased the generation of free radicals from phorbol myristate (PMA)-activated neutrophils in vitro, but did not ameliorate ALI. IL-1 induced enhancement of the expression of cPLA2 on neutrophils was not altered by apocynin. CONCLUSION: Apocynin induced suppression of the generation of superoxide anions from neutrophils by inhibition of NADPH oxidase does not attenuate IL-1-induced ALI in rats.

Role of IL-1alpha in Cisplatin-Induced Acute Renal Failure in Mice

BACKGROUND/AIMS: For unknown reasons, caspase-1 -/- mice, protected against cisplatin-induced acute renal failure (ARF), are deficient in interleukin (IL)-1alpha. We thus asked whether IL-1alpha deficiency underlies the mechanism of protection against cisplatin-induced ARF in these mice. METHODS: Cisplatin (30 mg/kg) was injected intraperitoneally into wild-type C57BL/6 mice to produce a cisplatin-induced model of ARF. IL-1alpha was measured in control vehicle- and cisplatin-treated wild-type animals. We also examined whether IL-1alpha -/- mice were similarly protected against cisplatin-induced ARF. Additionally, infiltration of CD11b- and CD49b-positive cells, as markers of macrophages, natural killer, and natural killer T cells (pan-NK cells), was investigated in wild-type and IL-1alpha -/- mice. RESULTS: Compared with vehicle-treated mice, renal IL-1alpha increased in cisplatin-treated wild-type mice beginning on day 1. IL-1alpha -/- mice were shown to be protected against cisplatin-induced ARF. No significant difference in the infiltration of neutrophils or CD11b- and CD49b-positive cells were observed between wild-type and IL-1alpha -/- mice. CONCLUSIONS: Mice deficient in IL-1alpha are protected against cisplatin-induced ARF. The lack of IL-1alpha may explain, at least in part, the protection against cisplatin-induced ARF observed in caspase-1 -/- mice. Investigation of the protective mechanism (s) in IL-1alpha -/- mice in cisplatin-induced ARF merits further study.

Development of an Intervertebral Disc Degeneration Model using Newzealand White Rabbits

STUDY DESIGN: An experimental animal study. OBJECTIVES: To create a more appropriate disc degeneration model which shows how Interleukin 1alpha may induce the activation of metalloproteinases within the nucleus pulposus. SUMMARY OF LITERATURE REVIEW: There are few disc degeneration models wherein there is activation of metalloproteinases within the nucleus pulposus without structural destruction of the intervertebral disc. MATERIALS AND METHODS: Three consecutive intervertebral discs in New Zealand White Rabbits were exposed. Each disc was injected with 0.1ml of saline (Saline group), 0.1ml of 1microg/ml (IL-1 group), 0.1ml of 10microg/ml (IL-10 group) of IL-1alpha through a 30-gauge needle. The lumbar spine was harvested 12 weeks after operation. We then analyzed radiographic findings and histological changes. RESULTS: There was no difference in the radiological disc height index among the three groups; 0.071 in saline group, 0.045 in IL-1 group and 0.058 in IL-10 group (p=0.194). The histological cellularity of the nucleus pulposus revealed a decrease in the number of cells (p=0.0001, 1.42 in saline group vs. 3.00 in IL-10 group; p=0.001, 2.00 in IL-1 group and 3.00 in IL-10). The histological matrix of the nucleus pulposus was 1.42 in saline group and 2.42 in IL-10(p=0.007), which meant that there had been condensation of the extracellular nucleus pulposus matrix. CONCLUSIONS: The results of this study demonstrate that interleukin-1alpha may contribute to degradation of the nucleus pulposus. This is useful for future study into the effects of the cytokine inhibitor on matrix regeneration and cellularity in the nucleus pulposus in intervertebral disc disease.