Search

We conducted comprehensive clinical and molecular characterization of claudin 18.2 expression (CLDN18.2) in advanced gastric or gastroesophageal junction cancer (GC/GEJC).Patients with advanced GC/GEJC who received systemic chemotherapy from October 2015 to December 2019 with available tumor specimens were analyzed. We evaluated clinicopathological features of CLDN18.2 expression with four molecular subtypes: mismatch repair deficient, Epstein-Barr virus-positive, human epidermal growth factor receptor 2-positive, and others. In addition, programmed death-ligand 1 (PD-L1) combined positive score (CPS), genomic alterations, and the expression of immune cell markers were assessed. Clinical outcomes of standard first- or second-line chemotherapy and subsequent anti-programmed cell death protein 1 (anti-PD-1) therapy were also investigated according to CLDN18.2 expression.Among 408 patients, CLDN18.2-positive (moderate-to-strong expression in ≥75%) was identified in 98 patients (24.0%) with almost equal distribution in the four molecular subtypes or CPS subgroups. CLDN18.2-positive was associated with Borrmann type 4, KRAS amplification, low CD16, and high CD68 expression. Overall survival with first-line chemotherapy was not significantly different between CLDN18.2-positive and -negative groups [median 18.4 versus 20.1 months; hazard ratio 1.26 (95% confidence interval 0.89-1.78); P = 0.191] regardless of stratification by PD-L1 CPS ≥5. Progression-free survival and objective response rates of first- and second-line chemotherapy, and anti-PD-1 therapy also showed no significant differences according to CLDN18.2 status.CLDN18.2 expression in advanced GC/GEJC was associated with some clinical and molecular features but had no impact on treatment outcomes with chemotherapy or checkpoint inhibition. CLDN18.2-positive also had no impact on overall survival. This information could be useful to interpret the results from currently ongoing clinical trials of CLDN18.2-targeted therapies for advanced GC/GEJC and to consider a treatment strategy for CLDN18.2-positive GC/GEJC.

Background and Objective Tissue regeneration is affected by the porosity, chemical properties and geometric structure of graft materials. Regeneration of severe periodontal defects, such as one-wall intrabony defects, is difficult because of reduced tissue support, and bone grafts are commonly used in such cases. In the present study, a tunnel-structured β-tricalcium phosphate (tunnel β-TCP) graft material designed to stimulate bone formation was fabricated. The objective of this pilot study was to evaluate the effect of this graft material on periodontal regeneration in one-wall intrabony defects in dogs. Material and Methods Six male beagle dogs were used in this study. First, the mandibular second and third incisors were extracted. Experimental surgery was performed 12 wk after tooth extraction. Bilateral 4 × 8 mm (width × depth) one-wall intrabony defects were created in the mesial side of the mandibular canines. At the experimental sites, the defects were filled with tunnel β-TCP, whereas the control defects were left empty. Twelve weeks after surgery, qualitative and quantitative histological analyses were performed. Results There were no signs of clinical inflammation 12 wk after surgery. Coronal extension indicative of new bone formation was higher at the experimental sites than at the control sites, although the differences between both the sites in the newly formed cementum and connective tissue attachment were not significant. Newly formed periodontal ligament and cementum-like tissue were evident along the root surface at the experimental sites. The inner surface of the tunnels was partially resorbed and replaced with new bone. New blood vessels were observed inside the lumens of tunnel β-TCP. Conclusion Tunnel β-TCP serves as a scaffold for new bone formation in one-wall intrabony defects.

We have constructed a high resolution rice genetic map containing 1,383 DNA markers at an average interval of 300 kilobases (kb). The markers, distributed along 1,575 cM on 12 linkage groups, comprise 883 cDNAs, 265 genomic DNAs, 147 randomly amplified polymorphic DNAs (RAPD) and 88 other DNAs. cDNAs were derived from rice root and callus, analysed by single-run sequencing and searched for similarities with known proteins. Nearly 260 rice genes are newly identified and mapped, and genomic DNA and cloned RAPD fragments were also sequenced to generate STSs. Our map is the first significant gene expression map in plants. It is also the densest genetic map available in plants and the first to be backed up comprehensively by clone sequence data.

The purpose of this study was to develop a new biodegradable bone substitute materials consisting of synthesized nano-size hydroxyapatite (nano-HAp) and Type I biodegradable honeycomb collagen sponge (HCS) composites. Bone defects in rabbit mandibles were prepared by a drill, and the composites were implanted into the bone defects. The HCS only and the HCS/calcined hydroxyapatite (HAp) composite were used as comparative materials. The bone tissues reaction at the early stage within 3 weeks after implantaion was investigated histologically. Amounts of new bone formation were determined by NIH-image analysis software using the histological sections. The amounts of the new bone formation were largest in the nano HAp/HCS compared to the comparative materials. Within 2 weeks after implantation, the nano-HAp/HCS composite was more rapidly exchanged by new bone than the comparative materials. From these results it was considered that the nano-HAp/HCS composites can be used as an effective biodegradable bone substitutive material.

Previously we found that laser perforation of a collagen membrane (35 microm thickness, Koken Co., Tokyo) produced an effective bone morphogenetic protein (BMP) carrier, if the created pore sizes were larger than 0.5 mm. In this study we applied the same technique to create pores of 0.2 and 1.0 mm in a thicker (1.2 mm thickness) porous biodegradable membrane made of polylactic acid and an epsilon-caprolactone copolymer (PLA-CL) to obtain an effective membranous BMP carrier with higher mechanical strength. Pieces of PLA-CL (0.5 x 1.0 x 0.12 cm) combined with rhBMP-2 (5 microg) were implanted subcutaneously into rats and processed for analyses at 1-3 weeks. The laser-perforated PLA-CL membranes equipped with 1.0 mm pores induced mineralization beginning from the margins of the pores judging from the X-ray patterns, but bone formation seemed to proceed irregularly inside the pores. In the perforated PLA-CL membrane with 1.0-mm pores bone formation did not significantly increase compared with the nonperforated one. This was due to the fact that the PLA-CL membrane was already a porous structure (85% porosity). In contrast with laser-perforated PLA-CL 0.2 mm pores, bone was induced on the collagen fibers and fiber bundles inside the pores. The different patterns of bone formation between the PLA-CL membranes with 1.0 and 0.2 mm pores seemed to be related to the active formation of perpendicular collagen fibers through the 0.2 mm pores.

Platelet-derived growth factor (PDGF)-BB is a polypeptide growth factor which has been shown to stimulate periodontal regeneration. In this study, we investigated the time- and dose-dependent effect of PDGF-BB on the proliferation and collagen synthesis of human periodontal ligament (PDL) cells.For the proliferation assay, PDL cells were cultured in 0.01-10 ng ml(-1) of PDGF-BB for 12 or 24 h, and cell numbers were counted. For the collagen synthesis assay, PDL cells were cultured in 0.1-10 ng ml(-1) of PDGF-BB for 1 to 24 h. The ratio of collagen content in total protein was evaluated, and the gene expression of type I collagen was assessed quantitatively by Northern blotting analysis.PDGF-BB stimulated the proliferation of PDL cells in a time- and dose-dependent manner with the maximum effect at 10 ng ml(-1). PDGF-BB induced the collagen synthesis of PDL cells with the maximum effect for 24-h treatment, and 1 ng ml(-1) of PDGF-BB. PDGF-BB exhibits an inverse dose-dependent effect on proliferation and collagen synthesis by PDL cells. These findings suggest that PDGF-BB is one of the important regulators of the maintenance of the extracellular matrix in PDL, and may play an important role in the regeneration of PDL.

A unique biomaterial, a mixture of DNA and collagen (DNA/collagen), was developed and its efficacy as a carrier matrix for bone morphogenetic protein (BMP) was evaluated histologically. The material was prepared as a composite of DNA from salmon milt and pepsin-digested type I collagen (atelocollagen) from bovine dermis. Phase-contrast and fluorescence microscopy showed atelocollagen fibres with DNA coating. The dose-response and time-course of bone induction by BMP in DNA/collagen (5 x 10 x 1 mm) in the subcutaneous tissue was investigated in 20 male Wistar rats. The BMP/DNA/collagen induced new bone in a dose-dependent manner (0, 25, 50 or 100 micrograms of BMP). Histological examination in the time-course study showed that the BMP (100 micrograms)/DNA/collagen induced bone formation, while the DNA/collagen alone resulted in the accumulation of fibroblasts. These results indicate that the DNA/collagen is effective as a carrier matrix for BMP. It provides a cell anchorage for differentiation of osteoblasts and is absorbed as bone matures.

Three different types of porous hydroxyapatite with pore sizes of 100-200 micrometer in diameter-porous particles of hydroxyapatite (PPHAP), porous blocks of hydroxyapatite (PBHAP), and honeycomb-shaped hydroxyapatite (HCHAP)-were compared in terms of their abilities to induce osteogenesis when implanted subcutaneously with recombinant human BMP-2 into rats and extracted at 1, 2, 3, and 4 weeks. Histologically, direct bone formation occurred in PPHAP and PBHAP while only endochondral ossification took place in HCHAP. Interestingly, cartilage in the central zones and bone in the orifice zones of the tunnels of the HCHAP were observed at 2 weeks. After 3 weeks, the cartilage disappeared and bone formation occurred throughout the inner surface of the tunnels of the HCHAP, always leaving space for capillaries within the tunnels. Alkaline phosphatase activity and osteocalcin content were the highest in HCHAP among the three hydroxyapatite implants. These results clearly indicate that BMP-induced bone formation is highly dependent on the geometry of the carrier, which provides feasible structural factors for vascularization.

We demonstrated that the transcriptional expression of MT1-MMP, MMP-9, cathepsin K, and TRAP in highly enriched osteoclasts were regulated by different matrix proteins that bind to integrin on osteoclast, such as collagen type I (CoI), fibronectin (FN), vitronectin (VN), osteopontin (OPN), and ivory. Results suggested that the OPN-integrin alphavbeta3 binding plays a more important role than CoI-alpha2beta1 binding in the regulation of osteoclast activity.

Bone marrow cells are multipotent cells. When bone marrow cells were cultured with type I collagen matrix gels, they showed high alkaline phosphatase activity, collagen synthesis, and formed mineralized tissues. Furthermore, cells expressed osteocalcin and bone sialoprotein genes, which are osteoblast-specific genes. These findings indicate that type I collagen matrix gels induce osteoblastic differentiation of bone marrow cells. Type I collagen interacts with the alpha 2 beta 1 integrin receptor on the cell membrane and mediates extracellular signals into cells. DGEA peptide is a cell-binding domain of type I collagen molecule. When collagen-integrin interaction was interrupted by the addition of Asp-Gly-Glu-Ala (DGEA) peptide to the culture, the expression of osteoblastic phenotypes of bone marrow cells was inhibited. Furthermore, anti-alpha 2 integrin antibody, which interacts with alpha subunit of integrin and blocks the binding of integrin with collagen, suppressed the expression of osteoblastic phenotypes. These findings imply that collagen-alpha 2 beta 1 integrin interaction is an important signal for the osteoblastic differentiation of bone marrow cells.

Bone matrix is composed of collagen and non-collagenous proteins. The collagen is mainly type I collagen. Characteristics of bone collagen are in posttranslational modifications and utilization of transcriptional elements in the promoter. The non-collagenous proteins are acidic Ca-binding proteins: bone Gla protein(BGP), bone sialoprotein (BSP), osteopontin, osteonectin etc. BGP and BSP are specific to bone, and other proteins are present also in non-mineralized tissues. BGP functions in suppression of excessive mineralization. BSP and osteopontin are sialoproteins containing a RGD cell-attachment sequence and poly(acidic amino acid) sequences. BSP is present in sites of bone formation. Osteopontin is involved in attachment of osteoclasts to bone surface.

In this study, we demonstrated that bone marrow stromal cells maintained on type I collagen matrix induced bone in vivo. The formed bone contained bone marrow, and the process of bone formation occurred without cartilage formation. Bone marrow stromal cells differentiated into osteoblasts on type I collagen matrix in vitro, but types II, III, and V collagens did not possess this activity. These findings imply that type I collagen matrix offers a suitable environment for the induction of osteoblastic differentiation in vitro and osteogenesis in vivo.

FePt–(C4F8)n granular films have been prepared by facing targets sputtering method with plasma polymerizing reaction. The FePt alloy was deposited on fused quartz substrate as same as the (C4F8)n matrix was polymerized. Then, they were annealed at 600°C in reduced air. Their magnetic properties and fine structures were investigated. The films prepared at C4F8 partial pressures of 0.10 and 0.20 mTorr, which consisted of fine FePt particles with a size of 5–10 nm, had high coercivities of 4–8 kOe caused by the stress between FePt particles and fluorocarbon matrix.

The biochemical properties of collagens from the periarticular connective tissues of the human knee (ligaments, semitendinosus and gracilis tendons, and the iliotibial band) were investigated in subjects ranging from 20 to 70 years of age. Although the total collagen content of most tissues was more than 90% of the dry weight, the anterior cruciate ligament and the iliotibial band had relatively low collagen contents. There were no significant changes in the tissue collagen content with aging. However, the anterior cruciate ligament and the patellar tendon of the 20-year-old subject had a higher content of soluble collagen than the other ligaments and tendons. The iliotibial band of the 20-year-old subject contained more collagen that was soluble by a combination of salt, sodium citrate extractions and pepsin digestion. Dihydroxylysinonorleucine was the major reducible cross-link of collagen from all the ligaments. The amount of dihydroxylysinonorleucine in the anterior cruciate ligament of the 20-year-old subject was much higher than that in the other ligaments. In contrast, the tendons and the iliotibial band contained a large amount of histidino-hydroxymerodesmosine and hydroxylysinonorleucine, while the patellar tendon was the only tendon with a significant content of dihydroxylysinonorleucine. Hydroxypyridinium non-reducible cross-links were more abundant in collagens from ligaments than in collagens from the other tissues. The cross-link study and the analysis of collagen solubility showed that patellar tendon collagen more closely resembled the collagen from the anterior cruciate ligament than that from periarticular tendons. It was also shown that the anterior cruciate ligament contains relatively immature collagen compared with the other ligaments.(ABSTRACT TRUNCATED AT 250 WORDS)

A fibrous collagen membrane (FCM) made of crosslinked reconstituted collagen fibers was applied as a carrier of BMP. The effectiveness of FCM as a BMP carrier was compared with conventionally used insoluble bone matrix (IBM). Partially purified BMP was obtained from a guanidine HCl extract of bovine bone after a three-step chromatographic procedure. The BMP fraction was combined with FCM or IBM and implanted subcutaneously in the back of rats. Bone and cartilage formation were determined by radiographic, histologic, and biochemical analyses after removal at 1, 2, 3 and 4 weeks. After 2 weeks, bone and cartilage formation were evident histologically in IBM-BMP and FCM-BMP. In IBM-BMP, cartilage was formed in the area of presumptive bone. On the other hand, in FCM-BMP, cartilage formation occurred in the space between the fibers of fiber-bundles, while on the surface of the fibers bone formation started independently. Biochemically, the most noteworthy difference between FCM-BMP and IBM-BMP was the marked increase in type II collagen content in FCM-BMP, which contrasted with its decrease in IBM-BMP at 3 weeks. This study demonstrated that FCM definitely functioned as a BMP carrier that led to bone and cartilage formation at distinct sites. Therefore, we conclude that BMP could induce differentiation of immature cells preferentially into either osteogenic cells or chondrocytes depending upon the nature of the carrier which provides the environment for cell differentiation.

Mineralization of dentin is a highly organized process that is controlled by many kinds of modulators. These modulators affect the nucleation and crystal growth of the mineral phases. Phosphophoryn, a highly phosphorylated protein of dentin [1], is a typical example of such modulators. Phosphophoryn binds calcium ions with modest affinity [2–6] and changes its conformation and molecular dynamics [2, 7]. Binding a large number of calcium ions, this protein aggregates into granules [8,9] and finally precipitates [3].

We investigated the condylar position by transcranial radiography which can be put daily clinical use easily. The patients are 8 anterior crossbite at the mixed dentition (4 functional crossbite and 4 skeletal crossbite). We observed their condylar positions before treatment (average age is 9.3 years old), after correction of overbite (average age is 10.0 years old) and 3-4 years after correction of overbite (average age is 14.0 years old). The results were as follows: 1. Condylar position of 7 cases changed after correction of overbite. The differences between right and left condylar position of six in seven cases had been recognized before treatment decreased after correction of overbite. Their condylar position were stable during the observation period of 3-4 years after correction of overbite. The condylar position of the other one case closed to fossa and then relapsed 3-4 years later. 2. Condylar position of only one case hasn't changed during the observation. 3. In functional crossbite cases, there were some differences between right and left condylar position before treatment, but no differences after crossbite correction. In skeletal crossbite cases, there was no tendency of the changing on their condylar position through treatment. 4. The changing to same direction was observed in the decrease of differences on condylar position and in the result of frontal cephalogram analysis. 5. There wasn't special relationship between changing of condylar position and orthodontic appliances. 6. TMJ dysfunction was observed in the case which has significant difference in its condylar positions before treatment and in the other case in which difference was not corrected after treatment. We recognized the usefulness of transcranial radiography for orthodontic treatment.

14569 Background: Allergic reaction to oxaliplatin is a chronic adverse event in the treatment of cancer. Patients with colorectal cancer were given FOLFOX4, using preventive approaches composed of two steps. Methods: Step I included 40 mg famotidine, 8 mg dexamethasone from the first cycle and 50 mg of diphenhydramine after cycle 4. In step II, for the patients who had a reaction but not a severe one in step I, a 20 mg dose of dexamethasone and four-hour oxaliplatin infusion were administered. Results: Forty-eight patients (15.1%) presented with allergic reaction, divided into group A receiving step I except diphenhydramine and B undergoing all of step I. Significant delay on occurrence was observed in group B. Thirty patients underwent step II. Prevention success (well-prevented) was defined as no reaction in two or more treatments. Allergic reaction appeared again in eleven patients (not well-prevented, 36.7%), but most well-prevented patients got four months longer treatment time with no reaction than not well-prevented patients. Conclusions: In our preventive approach, administration of H-1RA was effective in delaying the occurrence of allergic reaction to oxaliplatin, and increased dose of dexamethasone 20 mg and prolongation of oxaliplatin infusion from 2 to 4 hours was succeeded in continuing the FOLFOX4. No significant financial relationships to disclose.

Thin films of CoPtCr and CoPtCr–SiO2, perpendicular magnetic media materials, were investigated by synchrotron radiation grazing-incidence x-ray diffraction. The analysis of diffraction peaks in a reciprocal space provided the variation of stacking faults as functions of Pt content in these materials. It was found that stacking faults start increasing at 15–20 and 25–30at.% Pt for CoPtCr–SiO2 and CoPtCr films, respectively. These results can well explain the phenomenon whereby the magnetic anisotropy of CoPtCr–SiO2 increases with an increasing Pt content and decreases above 20at.% Pt, whereas that of CoPtCr increases up to 30at.% Pt and then decreases.

A CoPtCr-SiO2 film is a potential material for high-density perpendicular recording media due to their high magnetic anisotropy constant Ku. However, the value of Ku decreases in high levels of Pt content. In this study, this phenomenon was analyzed by synchrotron radiation grazing incidence x-ray diffraction and transmission electron microscopy techniques. Although the CoPtCr-SiO2 films contained almost no fcc phase at 20at.% Pt or less, it increased remarkably at 30at.% Pt. This result coincided with the variations of Ku with respect to the Pt content for the CoPtCr-SiO2 films. We consider that the fcc phase formation at Pt content levels in the vicinity of 30at.% is an influential factor of the Ku reduction. Our techniques proved to be sensitive for the detection of fcc regions in the CoPtCr-SiO2 films.

Switching field distributions (SFDs), are presented for longitudinal media of ∼40 Gbit/in.2 and the influence of thermal agitation of magnetization is discussed. Two remanence curves were measured at different sweep rates of applied field of ∼10 and ∼108 Oe/s, respectively, and SFD curves were obtained by differentiating the remanence curves. The SFD becomes significantly narrower at the higher field sweep rate. A SFD curve without the effect of thermal agitation was calculated using Sharrock’s equation. The full width at half maximum of the SFD curve at 10 Oe/s, ΔHr/Hr, is 0.45 for an isotropic medium, while that estimated without the thermal agitation effect ΔH0/H0 is 0.24, indicating that the SFD at the recording frequency is nearly half that measured at vibrating sample magnetometer (VSM) time scales. The ΔH0/H0 of the ∼40 Gbit/in.2 medium is much smaller than that of a 15 Gbit/in.2 medium used for comparison, although the ΔHr/Hr is ∼1.4 times larger than that of the 15 Gbit/in.2 medium due to thermal...

More than 60% of brain chondroitin sulfate proteoglycans were extracted from 10-day-old rat brains by homogenization in ice-cold phosphate-buffered saline containing protease inhibitors. Although the soluble proteoglycan preparation was a mixture of chondroitin sulfate proteoglycans with a different hydrodynamic size as well as a different molecular density, each subfraction of the proteoglycans contained chondroitin sulfate side chains with virtually identical molecular weight (approximately 15,000) and chondroitin sulfate disaccharide composition (high content of 4-sulfate unit). Digestion of the purified proteoglycan preparation with protease-free chondroitinase ABC produced five core proteins with Mr = 250,000 (designated as 250K protein), 220,000 (220K), 150,000 (150K), 130,000 (130K), and 93,000 (93K). All these core proteins were obtained from chondroitin sulfate proteoglycan preparations extracted from various regions of the brain, but their composition varied among different brain regions. Analysis for amino acid composition of these core proteins and two-dimensional mapping of their proteolytic peptides revealed that three major core proteins (250K, 220K, and 150K proteins) were structurally different. These observations indicate that at least three distinct types of chondroitin sulfate proteoglycan occur in the developing rat brain.

Cross-linking patterns of the collagen from bovine gingiva, periodontium, and dental pulp were analyzed chromatographically. The ratios of two main cross-links, dihydroxylysinonorleucine to hydroxylysinonorlecuine, were 0.18, 0.31, and 0.49 for the bovine gingiva, periodontium, and dental pulp collagen, respectively. These ratios are similar to that of skin collagen rather than that of bone and dentin collagen.

The age-dependent changes in reducible collagen cross-links, Schiff bases and their precursors were followed for human bone and articular cartilage throughout the whole life span from 3 to 89 years of age. With aging, the reduced Schiff bases dihydroxylysinonorleucine remarkable decreased, wheras hexitollysine peaks increased significantly in both collagens. Dihydroxynorleucine, the precursor of the cross-link, was found to increase slightly with age. But the changes in hydroxylysinonorleucine and lysinonorleucine were comparatively small. These changes were discussed in relation to the increased insolubility observed in both collagens at higher age.

Deciduous teeth affected by dentinogenesis imperfecta were obtained from two patients with osteogenesis imperfecta. Electronmicroscopy of the dentin revealed some structural alterations. The striation of the collagen fibrils was not clear, and the crystals were less dense than in normal dentin. The amino acid analysis of dentin collagen was slightly different from normal dentin. There were slight increases in the amounts of hydroxylysine. serine and acidic acids. On the other hand, lysine and arginine were reduced. The elevation of hexosyllysine content and an increase of carbohydrate were also observed. These results indicate that either dentin collagen is altered, or noncollagenous matrices associated with collagen are increased in dentinogenesis imperfecta dentin.