Rheumatoid disease (RA) is one of the complex diseases that shewed multiple progression stages. At first infiarnmatory response is occurred by self antibodies, and then the proliferation of synoviocytes is abnormally promotcd, These synoviecytes invade into articular structures and fi nally articular cartilage is destroyed. In order to analyze growth-promoting factors in synovial cells from articular tissues ofRA. we performedDNAmicroarrayanalysisandcomparedtheexpressionpatternsoftheirrnRNAtothatofsynoviocytes from esteoarthrttis (OA), The results showed that thc expression of many genes was up-regulated ;n RA synoviecytes comparing with OA synoviocytes. Among these genes, IULPCR anaLysis revealed that Wnt-1induced secreted protein 2 (wrSP-2, CCN5) and protein 3 (WISP-3, CCN6) which are belong to the novel gene famity, CCN, were hjghly up-regulated in RA synoviocytes, jn addition, one truncated isofbrm of WISP-3 which may be related to the ongoing of some type ofRA, was detected in RA synoviocytes. Furthermere, immunohistochemical analysis showed that the products ofboth CCN genes were highly distributed in active synoviecytes of articuLar tissues of ILN, These resu lts indicate that both CCN genes and their products p[ay some reles in the progressien of RA, Key werds: Rheumateid arthritis,Synovial ceils, Microarray, WrSP gene, CCN farniiy lntroduction ruieumatoid arthritis (RA) is a chronic inflarnmatory disease characterized by intlamed synovial hyperpEasia with excessive inflammatory cell infiltration, leading te erosion of articular carti[age and marginal bone, with subsequent joint destruction. Althougli an exp[osion of infbrmation over the past two decades contributed to the better understanding of the mechanism of infi ammation, the pathogenesis of synovitis has not been fu11y elucidated i}. Abnormal proliferation ofsyneviocytes is promoted in RA, and these synoviocytes invade into articu]ar structures and finally destroy articu]ar cartilage. Molecular basis ofthese phenornena are poorly understood. Recent[y, contrlbution ofsome cytokines, receptors and transeription factors to this disease was suggested2'fi}. In addition, some genetical background that Correspondencc to Tbhr" Nakanishi,Department ofClinical Diagriosis, ShLijitsu(JniversityScltoolofPhaiTnacy,Okayama703-g5]6,Japaii,Teli gl-g6-271-8363,Fax:81-86-271-83631E-mail:torhoshi@shujitsu,ac,ip89 contribute to the generation ofthis disease was revealedi). Reccnt progress in genome science made it possible to analyze thc expressien ofthousands ofgenes at ene time. This micro aray (DNAchip)technologyS}revealestheditibrenceofgeneexpression between ditlerent types of ce[Is er cells in different conditions, Although it stitl has some technical problem, the use ef this technique has been widely develeping. This technique also contributes the detection ofgenetic mutation occurred in tumor cells and detection of genetie polymorphisms (SNPs)"). These resultscontribatetodevelopnoveltherapyie,ordermademedicine, The CCN fami]y ofgenes'") which stands fbr Cyr61 (cysteine rich protein)i'), Ctgf(connectiye tissue growth factor)t2}, and Nov (nephroblastoma overexpressed gene)iS) consists ef six distinet membcrs. CTGF and Cyr61 are thougbt to be positive regulators ofcett gro-th, whi]e Nov and resent three new members, WISP1 (Wnt-1-induced secreted protein 1) tElm-1"', WISP-2 (Wnt-1induced secreted protein 2)lrCop1tCTGF-3iS) and WISP-3 (WntThe Society of Hard Tissue Biology NII-Electronic Library Service heSociety fHard is ue iology Hiroki Mori et al: Expression f WISP-2 and WISP-3 in 1-inducedsecretedprotein3Lfi)arethoughttobenegativeregulators. Tn addition, these proteins plays fundamentally important processes in cell differentiation, attacliment, migration and are also involved in pathological process, such as fibrosis, tumorigenesis and tumor invasionM. Recently defined nomenclature designated Cyr 61 as CCN 1, CTGF as CCN2, NOV as CCN3, and WISP-1-3 as CCN4-6. WISP-2 is also designated as CTGF-LiS) that 1aeks characteristic C-terminal domain ofCCN family protein. These Wisp factors identified as Wnt-inducible secreted protein indicating that these faetors are important for morphogenesisanddeveloprnent. In this paper, we cempared the expression levels ofabout a theusand of genes bctween synovial cells frem RA and osteoartiiritis (OA) by using cDNA micrearrays, and identified the characteristic genes which were up-regulated or downregulated in en synoviocytes. Furthermore, we analyzed the distribution of[we ofsuch genes, WISP-2 and WISP-3 which are included in the CCN gene imily and thought to be candidates for future diagnosis and therapeutic targets in RA syioviocytes. Materials and methods isvlation and cutr"re ofsynoviat cells Fresh synoviaL tissues were minced and digested with collagenase and DNase I. Tissue debris was removed threugh a cell strainer, and ce[ls were washed twice with medium. The resultant single cells were dispended into the wells efa 24-well microtiter plate (Costar, Cambridge, MA) at a density of2 x 1OG cells tml in 2 ml ofDubecco 's modified Eagle's mediurn (DMEM; Life Technelogies, Gaitherburg, MD) supp[ementcd with 10% HEPES (Life Tlrchnologies), 1OO IUIml penicil[in and leO pgfml streptomycin, The plates were incubated at 37"C in ahumidified atmosphere containing 5% CO. SynoviaE tissue cells were split weekly ence prirnary cultures had reachcd confiucnce, and used after the third passage. These cells appeared to be hemogeneous fi brobaast-like cclls morphologically. They were cultured at a density of 1 x 1OS celLs /ml in 2 ml DMEM ",ith 1O% FCS in a 24wel[ microtiter p[ate and allowed to adhere ovcrnight. After the third passage ofcelL cuLture, celLs are harvested and RNA was isolated from cells using ISOGEN (Nippon (}ene, Tbkyo, Japan) according to the manufacturer's recommendations. cDINL4 micraarray ana(ltsis Human [.O MIcroarrays (Clontech, Mountain View, (IA) containing over 1,OOO known human genes, were used for hybridization studies. The fi]ters were prehybridized with 5,O pg ofdenatured cot-1 DNA, 5.0 pg poly-dA, and S.e ml MicroHyb hybrldization so[ution for 2h at 42"C, Total ccllular RNA from primary cu[tures was prepared as described above, Prebes were generated fi'om reverse transcribed total RNA (1.0 pg) incubated with 2.0 gg oligo-dT, 6.0 pl of5x first strand buder, 1.0 ml dithiothretol, 1.5 ml reverse transcriptase, 1,5 pl dNTP mixture90 Synovial Cells containing dlYl/g dGTP, dTTP at 20 rnmol/1, and 1 O ml 32P-dCTP (3000Ci!mmol) for 90 min at 370C, Labeled probes were added to the pre-hybridization solution and left to hybridize with the membranes for for 1 8hr at 420C. Wtashes were dene twice at 50oC in 2xSSC with 1% sodium dodecylsulfate (SDS) fbr 20 min and once at reom temperature in O.5xSSC with 1% SDS for 15 min. The membranes were aligned on a Imaging screen and exposed fbr 16h at room temperature. The screens were scanned using a Bioimaging analyzer (FLiji Film, Tokyo, Japan) and saved as digital images, Arraygage (Fuji Film) was used fbr data analysis. Differential gene analysis was perfbrmed for all pairwise

Trehalose 6,6'-dicorynomycolate (TDCM) was first characterized in 1963 as a cell surface glycolipid of Corynebacterium spp. by Ioneda and co-workers. TDCM shows potent anti-tumor activity due to its immunoadjuvant properties. Furthermore, the toxicity of TDCM in mice is much weaker than the related trehalose diester of mycolic acid; trehalose 6,6'-dimycolate (TDM, formerly known as cord factor). We have investigated the chemical modification of this class of compound to generate novel agents that display increased immunoadjuvant activity with minimal associated toxicity. During the course of this work we recently developed 6,6'-bis-O-(3-nonyldodecanoyl)-α,α'-trehalose (designated as vizantin). Our results show that vizantin exhibited a potent prophylactic effect on experimental lung metastasis of B16-F0 melanoma cells without a loss of body weight and death in mice. Furthermore, vizantin effectively stimulated human macrophages in an in vitro model, making it a promising candidate for a safe adjuvant in clinical applications. In order to elucidate the pharmacokinetics of vizantin, a probe molecule with similar activity was developed on the basis of a structure-activity relationship (SAR) study with vizantin. The distribution of the probe molecule after intravenous administration into a mouse was assessed by macro confocal microscopy, where it was found to accumulate in the lungs and liver.

Vizantin, 6,6'-bis-O-(3-nonyldodecanoyl)-α,α'-trehalose, was developed as a safe immunostimulator on the basis of a structure-activity relationship (SAR) study with trehalose 6,6'-dicorynomycolate (TDCM). It was possible to synthesize vizantin on a large scale more easily than in the case of TDCM, and the compound exhibited more potent prophylactic effect on experimental lung metastasis of B16-F0 melanoma cells. Because vizantin stimulated human macrophages, it is a promising candidate for clinical application.

Aneurysmal bone cysts account for less than 1% of primary bone tumors and have a predilection for the metaphyses of the long bones of the leg. Only 1% of all aneurysmal bone cysts occur in the patella. We report on a 24-year-old man with a primary aneurysmal bone cyst in the left patella treated with curettage. The defect was filled with autogenic and allogeneic cancellous bone graft. The patient experienced no pain or tenderness and had a full range of knee movement.

Lymph node metastasis is considered a factor in determining the prognosis of squamous cell carcinoma (SCC). Both oral and cervical SCC tumor cells prefer lymph vessels as the route of metastasis. D2-40 is a specific marker of lymphatic endothelial cells. This study clarifies the distribution and characteristics of lymphatic vessels in oral and cervical SCCs. Immunohistochemistry was performed in 20 oral and 20 cervical SCCs (10 non-metastatic and 10 metastatic to lymph nodes) using D2-40, CD31, CD34, CD105 and double staining with D2-40 and keratin. Lymphatic vessel density (LVD) was also determined morphologically. Results showed that lymphatic vessels in both types of SCCs were distributed mainly at the superficial region beneath the epithelium. The LVD in each tumor was significantly higher compared to the corresponding normal mucosa. Moreover, the LVD in lymph node metastasis in each tumor was significantly higher compared to their non-metastatic counterparts. Cancer cell invasion was observed in the lymphatic vessels suggesting the existence of lymph node involvement during metastasis. The new lymphatic vessels that proliferated around the cancer nests in both SCCs have endothelial cell characteristics inferred to be associated with early lymphatic development and initial dissemination of cancer cells.

Quorum sensing (QS) refers to the phenomenon characterized by the accumulation of signaling molecules enabling a cell to sense a bacterial population triggering a coordinated response. Enterococcus faecalis is a resistant microorganism commonly found in oral infections. Salvadora persica has been traditionally used in Middle East countries to clean the teeth due to its antimicrobial activity. The study investigated the anti-quorum sensing (AQS) ability of S. persica extracts on E. faecalis. Extracts from the bark, leaves, root and shoots were tested against Chromobacterium violaceum. AQS ability was determined using differential scanning fluorimetry (DSF) and elastolytic assay to detect the level of protease and quantitative polymerase chain reaction (QPCR) to detect cytolysin (cylR1) and gelatinase (gelE) virulence factors. Preliminary disk diffusion assays revealed AQS activity by inhibiting violacein pigment production in C. violaceum without affecting its growth. Furthermore, only negligible amount of proteases was detected in DSF and elastolytic assays. CylR1 and gelE gene expression revealed no detectable signals even from the lowest possible threshold. The results indicate that S. persica has AQS ability against E. faecalis. S. persica extract can be used as alternative or in combination with other anti-microbial agents against infections caused by E. faecalis.

Oral mucosal melanoma (OMM) is a fatal sarcoma of unknown etiology. Histological morphology and genetic events are distinct from those of its cutaneous counterpart. Mutation and up-regulation of c-kit has been identified in OMM which may activate downstream molecules such as RAS and RAF. These molecules are involved in the mitogen-activated protein kinase (MAPK) pathway leading to tremendous cell proliferation and survival. NRAS and BRAF mutation and protein expression have been studied in other melanoma subtypes. The purpose of this study was to determine RAS protein expression and NRAS and BRAF mutation in 18 primary OMM cases using immunohistochemistry and mutation analysis. Results showed that RAS is intensely expressed in both in situ and invasive OMMs. However, NRAS mutation was only observed in 2/15 polymerase chain reaction (PCR) amplified cases both of which were silent mutations. On the other hand, BRAF missense mutations were observed only in 1/15 cases with PCR amplification. NRAS and BRAF mutations were independent from previously reported c-kit mutations. The classical V600E BRAF mutation was not found; instead a novel V600L was observed suggesting that the oncogenic event in OMM is different from that in skin melanoma. The low frequency of NRAS and BRAF mutations indicate that these genes are not common, but probable events in OMM pathogenesis, most likely independent of c-kit mutation.

Adipose tissue dysfunction has been associated with diabetogenic effects. The effects of repeated Cd exposure on adipocytes remain largely unknown. We administered Cd at doses of 0, 5, 10, and 20μmol/kgbw sc for 2 weeks (3.5times/week) to mice and assessed the possible alteration of epididymal white adipose tissue (WAT), including histological difference, adipocyte differentiation and functional capacity. Whereas hepatic weight did not differ between the control and Cd-exposed groups, WAT weight, as well as adipose cell mass, significantly decreased in a dose-dependent manner in Cd-treated mice. The Cd concentration in WAT significantly increased in Cd-treated groups after 2 weeks of exposure. Next, we examined the effects of Cd on adipocyte differentiation and hypertrophy. Cd exposure significantly decreased the paternally expressed gene 1/Mesoderm-specific transcript mRNA expression levels. Both peroxisome proliferator-activated receptor γ2 and CCAAT/enhancer-binding protein α mRNA expression levels in WAT tended to decrease in the Cd-treated groups. Next, we determined the effects of Cd exposure on the mRNA expression levels of adipose-derived hormones, such as adiponectin and resistin. The adiponectin mRNA expression level in WAT decreased after both 6h and 2 weeks of exposure to a high dose of Cd, and the reduction in resistin mRNA expression levels was observed after 2 weeks of exposure. These results suggest that Cd exposure causes abnormal adipocyte differentiation, expansion, and function, which might lead to development of insulin resistance, hypertension, and cardiovascular disease.

Rhabdomyolysis is a disorder of striped (skeletal) muscle and is caused by several factors, and acute renal failure associated with myoglobinuria is the most serious complication of rhabdomyolysis. However, the mechanism of tubule damage caused by myoglobin is still unclear. The early histopathological changes were not examined. Myoglobin reabsorption by the proximal renal tubules has not been reported to contribute to cellular damage. We performed histological analysis in a rat rhabdomyolysis model with administration of cytochalasin B (CB), which is an inhibitor of motile cell motile processes. CB inhibits the endocytosis of myoglobin in the proximal renal tubules and the increase in the BUN level, and decreased the percentage of area that had undergone tubular necrosis. On the other hand, initial degenerative changes (shortening of microvilli and protrusion of the cytoplasm) were observed on the cell surface of the proximal tubules with glycerolinduced cellular injury, and were not inhibited by the administration of CB. In conclusion, initial glycerolinduced cellular injury of the renal proximal tubules may be caused by free iron, which is isolated from myoglobin in urine. Further, the combination of the free iron in urine and the myoglobin reabsorbed by tubular cells may lead to cell necrosis.

A patient with Richter's syndrome developed rapid generalized lymph node enlargement with a decrease of peripheral blood lymphocytes after recombinant human granulocyte colony-stimulating factor (rhG-CSF) therapy for neutropenia induced by chemotherapy. The lymphadenopathy subsided spontaneously following discontinuation of rhG-CSF medication. Reinstitution of rhG-CSF therapy was followed by the same response as during initial therapy. Histopathologically, the lesions were characteristic of diffuse large cell lymphoma (DLL) with no evidence of myeloid cell involvement. No spontaneous contraction of enlarged lymph nodes followed withdrawal of the second course, but the enlargement subsided with chemotherapy. The patient died of myocardial infarction. All residual tumors examined post mortem presented microscopic features of small lymphocytic lymphoma (SLL), and G-CSF receptor was demonstrated on these neoplastic cells by Northern blot hybridization analysis. This observation indicates that some B cell malignancies may retain G-CSF receptor and respond to G-CSF.

The histologic hallmark distinguishing desmoplastic ameloblastoma (DA) from conventional ameloblastoma (CA) is its pronounced stromal desmoplasia, and this formed the basis of this investigation. To elucidate the stromal characteristics, localization patterns of oxytalan fibers, type III collagen and BMP family in DA (n=8) was compared with CA (n=24), and periodontal ligament (PL) (n=8). Oxytalan fibers formed apico-occlusal bundles in PL, thick radial bundles around tumor nests in DA, and as scanty fibers in CA. Type III collagen was identified in PL, strongly expressed in DA stroma, but weakly in CA. BMP-2, -3, -4 and -7 expression patterns in tumor epithelium and stroma were more pronounced in DA (including sites of bone formation), than CA. No immunoreactivity for BMP-5 and -6 were detected. Current findings suggest that the stroma in DA is neoplastic and derived from odontogenic ectomesenchyme, and recommends its reclassification as an odontogenic epithelial-ectomesenchymal neoplasm.

Wnt genes encode a large family of secreted cysteine-rich signaling molecules involved in cell growth, differentiation and tumorigenesis. Wnt5a, a non-transforming member of the Wnt family behaves as a putative oncogene in many cancers including melanomas. The aim of our study was to determine Wnt5a expression in primary oral mucosal melanomas (OMM) and correlate it with tumor thickness. Archival tissues from 18 OMM cases were subjected to immunohistochemical detection of Wnt5a by the streptavidin-biotin method. These were categorized into tumors of <4 mm (thin and intermediate thickness lesions) and >4 mm (thick lesions) thickness. Most OMM cases (17/18; 94.4%) stained positive for Wnt5a, though heterogeneously. Seven thick (7/11; 64%) and one intermediate thickness (1/7, 14%) OMM demonstrated strongly positive Wnt5a staining (P<0.05). The only Wnt5a-negative case was a thick OMM without local recurrence after treatment. Strong Wnt5a expression at tumor advancing sites suggests a role in local tumor spread. Identification of pleomorphic epithelioid and spindle cells as melanoma cell populations with the most pronounced Wnt5a staining suggests that Wnt5a overexpression influences cellular phenotype. These results taken together suggest that Wnt5a is up-regulated in OMM and may play a role in tumor progression.

Human bone marrow mesenchymal stem cells (hMSCs), one of the tissue stem cells, have the ability to differentiate into adipocytes, osteoblasts or chondrocytes, leading to the attempt to apply these stem cells to regeneration of bone and cartilage. To analyze characteristics of these cells and explain pluripotency of hMSCs, we analyzed gene expression profiles of hMSC clones by using DNA chips that contain over 10,000 genes per chip. We also analyzed their proliferation rates and cell surface markers. As a result, these hMSC clones showed various proliferation rates indicating that hMSCs contains multiple types of cells in their population. Furthermore, analysis of surface markers, especially analysis of CD34 and CD44 showed that these stem cells are not hematopoietic stem cells but mesenchymal origin. Profiling of gene expression by using DNA chip showed that several genes were specifically expressed in hMSCs, and the highest expression was observed in clone no.12 showing that this clone possesses unique characters as a MSC applicable to regeneration research and regenerative medicine.

We studied the toxicity dose and toxicity mechanism of rhubarb, a Chinese traditional medicine, by administering different doses to rats and examining the effects on the liver. Kunming rats were given intragastric administration of large, medium, small and minimum doses of rhubarb twice daily for five consecutive days. Then the general status of the rats, the hepatoenzymological data, histopathology of liver, and TNF mRNA expression were examined. The following results were obtained. 1. The general status of rats deteriorated with the increase in rhubarb dosage. The physical status of rats in the control group and minimum dose group were normal, but the status in other groups gradually deteriorated along with the increase in rhubarb dose. 2. Regarding the changes of hepatoenzymology profile, alanine aminotransferase and gamma-glutamyltransferase were elevated with increase in rhubarb dose; while the difference between normal control group and minimum dose group was unremarkable, the differences among other groups were obvious. 3. Examination of H&E-stained slides showed fatty degeneration of the liver starting from the minimum dosage group. This phenomenon is aggravated with increase in rhubarb dosage. 4. TNF expression was elevated significantly with increase in rhubarb dosage, beginning from the minimum dosage group. The present results showed that even a small dose of rhubarb given to rats by the intragastric route for five consecutive days was toxic to the liver, and the toxic effect was enhanced with increase in rhubarb dosage. The toxic effect of excessive intake of rhubarb to the liver is mainly manifested as fatty degeneration. TNF , an injury factor leading to fatty degeneration of cells, was probably involved in the hepatotoxicity by excessive intake of rhubarb.

Rheumatoid disease (RA) is one of the complex diseases that showed multiple progression stages. At first inflammatory response is occurred by self antibodies, and then the proliferation of synoviocytes is abnormally promoted. These synoviocytes invade into articular structures and finally articular cartilage is destroyed. In order to analyze growth-promoting factors in synovial cells from articular tissues of RA, we performed DNA microarray analysis and compared the expression patterns of their mRNA to that of synoviocytes from osteoarthritis (OA). The results showed that the expression of many genes was up-regulated in RA synoviocytes comparing with OA synoviocytes. Among these genes, RT-PCR analysis revealed that Wnt-1-induced secreted protein 2 (WISP-2, CCN5) and protein 3 (WISP-3, CCN6) which are belong to the novel gene family, CCN, were highly up-regulated in RA synoviocytes. In addition, one truncated isoform of WISP-3 which may be related to the ongoing of some type of RA, was detected in RA synoviocytes. Furthermore, immunohistochemical analysis showed that the products of both CCN genes were highly distributed in active synoviocytes of articular tissues of RA. These results indicate that both CCN genes and their products play some roles in the progression of RA.

We investigated the expression of HSP70 in acute lung injury (ALI) induced by endotoxin (lipopolysaccharide: LPS) and the protective mechanisms of rhubarb against ALI. LPS was injected into rats via the internal jugular vein to induce ALI and rhubarb extract was given by intraperitoneal injection. The rats were divided into 4 groups: saline control group; LPS-induced ALI group; ALI treated with prophylactic rhubarb group; and ALI treated with therapeutic rhubarb group. The rats were sacrificed 2 hours after injection of LPS or saline. Physiological and pathological examinations were conducted, including arterial gas analysis, lung coefficient, hematoxylin-eosin staining, and HSP70 expression (immunohistochemistry and Western blotting). In the LPS-induced ALI group, interstitial edema, neutrophil infiltration, plasma exudation in alveoli, and damaged vascular endothelium were observed. HSP70 levels were slightly increased in LPS-treated rats, while HSP70 expression was enhanced significantly and overall lung function was improved in rhubarb-treated groups compared to ALI model rats. These data confirm that rhubarb protects rats from LPS-induced ALI and upergulates HSP70 expression. Thus, the protective effect of rhubarb may be mediated by the HSP70 signaling pathway.

Fluoride as a supplement can affect the structural integrity of bone. Fluoride that is incorporated in the mineral, substitutes for the hydroxyl group producing hydroxyfluorapatite crystals and presumed to increase bone strength by preventing resorption. Because of this, fluoride therapy has been carried out in clinical trials for the treatment of osteoporosis. Although fluoride renders the mineralized tissues resistant to resorption, the effects of fluoride on bone strength and cortical bone mass specially that of the developing bone has not yet been characterized. The purpose of the study was to analyze the effect of fluoride on developing teeth and bone in young rats through histomorphometry. Sixteen wistar rats were used and divided into two groups; non-fluoride supplemented (control) and fluoride-supplemented groups. Four weeks after, the rats were sacrificed and the mandible and femur were subjected to histological and histomorphometric analyses. No clinical difference was observed between the two groups. The amount of mineralized cartilage was higher in fluoride-supplemented group compared to the control group. However, the amount of new bone was lesser in fluoride-supplemented group compared to the control group. This histomorphometric analysis showed that during endochondral ossification, 10 ppm fluoride administration caused some changes in calcified cartilage and bone mineralization in young rats.

A new calcium silicate/vermiculite composite for building material has been prepared from unfired vermiculite, calcium hydroxide, silica and pulp by autoclaving at 187°C at a pressure of 1.08 MPa. The product exhibited the vermiculite crystals dispersed uniformly in a definite direction among the matrix of calcium silicate. The composite showed marked plastic deformation caused by interlayer sliding of vermiculite. The building material was fixed by nails and screws and also possible to cut and saw like artificial wood. The board had superior humidity control originated from chemical adsorption of water molecules at the interlayer of vermiculite, and relative humidity in an experimental house was maintained to be 50%. Formaldehyde was chemically adsorbed to the composite, and there was no release of formaldehyde at elevated temperature.