Hydrocarbon Deposition Attenuates Osteoblast Activity on Titanium.

Although the reported percentage of bone-implant contact is far lower than 100%, the cause of such low levels of bone formation has rarely been investigated. This study tested the negative biological effect of hydrocarbon deposition onto titanium surfaces, which has been reported to be inevitable. Osteogenic MC3T3-E1 cells were cultured on titanium disks on which the carbon concentration was experimentally regulated to achieve carbon/titanium (C/Ti) ratios of 0.3, 0.7, and 1.0. Initial cellular activities such as cell attachment and cell spreading were concentration-dependently suppressed by the amount of carbon on the titanium surface. The osteoblastic functions of alkaline phosphatase activity and calcium mineralization were also reduced by more than 40% on the C/Ti (1.0) surface. These results indicate that osteoblast activity is influenced by the degree of hydrocarbon contamination on titanium implants and suggest that hydrocarbon decomposition before implant placement may increase the biocompatibility of titanium.

Study of an improved Allyl Di-Glycol carbonate sheet for high energy proton detection.

An allyl di-glycol carbonate (ADC) sheet which has been utilised as a neutron detector for personal dosimetry has recently been studied for its application as a device for radiation exposure control for astronauts in space, where protons are the dominant radiation. It is known that the fabrication process, modified by adding some kind of antioxidant to improve the sensitivity of ADC to high energy protons, causes a substantial increase in false tracks, which disturb the automatic counting of proton tracks using the auto-image analyser. This made clear the difficulty of fabricating ADC sheets which have sufficient sensitivity to high energy protons, while maintaining a good surface. In this study, we have tried to modify the fabrication process to improve the sensitivity to high energy protons without causing a deterioration of the surface condition of ADC sheets. We have successfully created fairly good products.

Rebamipide prevents occurrence of gastric lesions following transcatheter arterial embolization in the hepatic artery.

BACKGROUND: Transcatheter arterial embolization (TAE) of the hepatic artery is a common treatment method for hepatocellular carcinoma (HCC), but it often induces gastric mucosal injury. We examined whether or not rebamipide administration, beginning 1 week before and ending 2 weeks afterTAE, can prevent worsening of gastric mucosal disorders. METHODS: The subjects were 73 chronic hepatitis C or type C liver cirrhosis patients who concomitantly had HCC and received TAE in our hospital. The patients were randomly allocated to the rebamipide group (oral, 300 mg/day for 3 weeks starting 1 week before TAE) or the non-rebamipide group. Gastric endoscopy was performed 1 week before and 2 weeks afterTAE and the presence of erythema, erosion and/or submucosal haemorrhagic spots was monitored. Based on the findings, gastric mucosal disorder before and after TAE was quantitatively evaluated using the modified Lanza score (MLS). RESULTS: Overall, MLS after TAE increased significantly (P< 0.05). However, in the rebamipide group, MLS did not change. The MLS after TAE increased significantly in patients who had either liver cirrhosis, oesophageal varices or gastropathy (P< 0.01 or < 0.05). In the non-rebamipide group, a significant increase in MLS after TAE was observed in patients who had one of the above-mentioned three diseases (P< 0.01 or < 0.05). CONCLUSIONS: Gastric lesions which were present before TAE were significantly worsened after TAE. Rebamipide administration prevents TAE-induced aggravation of gastric lesions.

[Serum protein fractions in patients with laryngeal cancer undergoing radiation therapy. Possibility as a prognostic factor].

The levels of 21 protein components of the sera of 45 patients with cancer of the larynx undergoing radiation therapy were determined by a single radial immunodiffusion method before and after radiation therapy. We examined the correlation between changes in serum protein fractions and the prognosis of the patients. The patients with cancer of the larynx were treated with external irradiation of 60Co gammer-rays. Total target doses were 60 Gy. The levels of the same protein components were also measured in 43 normal adult individuals as a control. All patients were observed for 5 years and 12 years following radiation therapy. In the pretreatment sera obtained from patients with cancer of the larynx, the concentrations of prealbumin (Prealb), antithrombin III (ATIII) and plasminogen (Pmg) were significantly lower than controls. However the concentrations of alpha 1-acid glycoprotein (alpha 1AG), alpha 1-antitrypsin (alpha 1AT), alpha 1-antichymotrypsin (alpha 1X), haptoglobin (Hp), fibrinogen (Fib) and hemopexin (Hx) were elevated. At the completion of radiation therapy, the alpha 1AG, alpha 1AT, alpha 1X, Hp, Fib and inter alpha trypsin inhibitor (I alpha I) were significantly elevated than those normal controls. In patients without recurrent cancer after radiation therapy, the alpha 1AT, ceruloplasmin (Cp), Fib, IgA and Hx levels measured before radiation therapy were significantly lower than those patients with recurrent cancer. In patients without recurrent cancer after radiation therapy, the alpha 1AT, Hp, Cp, IgG, and IgA levels measured after radiation therapy were reduced compared with those patients with recurrent cancer. In patients who had lived more than 5 years after radiation therapy, the alpha 1AT, Cp and Fib levels measured before radiation therapy reduced significantly compared with those who had died within 5 years. In those who had lived more than 5 years, the alpha 1AG, alpha 1AT, Hp, Fib, IgG, and IgA levels measured after radiation therapy were reduced significantly compared with those who died within 5 years. In cases of laryngeal cancer following a period of 5 to 12 years after radiation therapy, multiple regression analysis was used to determine whether increased concentrations of serum protein fractions were associated with good prognosis for the original disease. AT III, Prealb, alpha 1AG, albumin (Alb) and I alpha I before radiation therapy were positively correlated with survival, whereas Hx, Pmg, Cp, IgM, Cl inhibitor (ClInh), alpha 1AT and Fib showed negative correlations. After radiation therapy, transferrin (Tf), Cp, Prealb, AT III and I alpha I were found to be positively associated with survival, whereas IgA, IgM, Pmg, alpha 1X and alpha 1AG were negative factors. From the elevation levels of acute phase proteins (alpha 1AT, Cp) and Fib and immunoglobulins (IgA, IgM) in the serum and lower levels of Prealb and AT III before and after radiation therapy, we may predict a relatively poor prognosis in these patients of laryngeal cancer.

[Studies on serum protein fractions of patients with maxillary sinus cancer undergoing a combination of radiotherapy and chemotherapy: II. Relationship between changes in serum protein fractions and prognosis].

We examined the correlations between changes in serum protein fractions and the prognosis of the patients. The levels of 21 protein components of the sera of 36 patients with maxillary sinus cancer were determined by a single radial immunodiffusion method before and after radiation therapy. The patients with maxillary sinus cancer were treated with combined intra-arterial infusion of bleomycin and external irradiation of 60 Co gamma-rays, and were concurrently treated with 5-fluorouracil at 200 mg/day p.o. The levels of the same protein components were also measured in 34 normal adult as a control. All patients were observed 5 years and 12 years after radiation therapy. In patients who had survived at least 5 years after radiation therapy, the Alb, Tf, Hx, IgG and IgM levels measured before radiation therapy were elevated significantly compared with those who had died within 5 years. In those who had survived at least 5 years, the Alb, Tf, Hx, IgG, IgM, IgA and I alpha I levels measured after radiation therapy were elevated significantly compared with those who had died within 5 years, and AT III was reduced. In cases of maxillary sinus cancer following a period of 5 to 12 years after radiation therapy, multiple regression analysis was used to determine whether increased concentrations of serum protein fractions were associated with good prognosis for the original disease, alpha 2HS. IgM, HX, alpha 1AT and alpha 1X before radiation therapy were positively correlated with survival, whereas AT III, Pmg, Cp, IgA, and alpha 1AG showed negative correlations. After radiation therapy, Pmg, Hx, Cp, Cl inh and Fib were found to be positive factors of survival rate, whereas alpha 2M, alpha 2Pl, I alpha 1, IgA, alpha 1AG and C3 were negative factors.

[Studies on serum protein fractions of patients with uterine cervical cancer undergoing radiotherapy: relationship between changes in serum protein fractions and prognosis].

The levels of 23 protein components of the sera of 84 patients with uterine cervical cancer undergoing radiotherapy were determined by a single radial immunodiffusion method, before radiation therapy, after whole pelvic irradiation, and after intracavitary irradiation using a remote after roading system (RALS). We examined the correlations between changes in serum protein fractions and the prognosis of the patients. The patients with uterine cervical cancer were treated with combined external irradiation of 60Co gamma-rays or 10 MV X-rays, and RALS therapy. The levels of the same protein components were also measured in 21 normal adult women as a control. All patients were observed between 4 years and 8 years after radiation therapy. In the pretreatment sera obtained from patients with uterine cervical cancer, the concentrations of prealbumin (Prealb), alpha 2HS glycoprotein (alpha 2 HS), alpha 2-plasmin inhibitor (alpha 2PI), transferrin (Tf), plasminogen (Pmg), albumin (Alb), IgM and hemopexin (Hx) were significantly lower than those normal controls. However the serum concentrations of: alpha 1-antichymotrypsin (alpha 1 X), haptoglobin (Hp), C9, fibrinogen (Fib), ceruloplasmin (Cp), alpha 1-acid glycoprotein (alpha 1AG), alpha 1-antitrypsin (alpha 1AT) and C4 were elevated. At the completion of whole pelvic irradiation and RALS therapy, the Hp, C4, and Fib levels were significantly lower than those before radiation therapy, whereas Prealb, alpha 2HS and alpha 2PI were elevated. In patients who had lived between 4 years and 8 years after radiation therapy, the Cp, alpha 1 AG, Hp and C9 levels measured before radiation therapy were reduced significantly, while Tf was elevated compared with patients who died within 4 years. In those who had lived more than 4 years, the alpha 1 AT, Hp, alpha 1X, Cp and C9 levels measured after whole pelvic irradiation were reduced significantly; while the C4 level was elevated. In cases of uterine cervical cancer followed for a period of 4 to 8 years after radiation therapy, multiple regression analysis was used to determine whether increased concentrations of serum protein fractions were associated with good prognosis for the original disease. Tf, Pmg, and alpha 1AT before radiation therapy were positively correlated with survival, whereas AT III, Cp, C1Inh, IgA, alpha 1 AG and C9 showed negative correlations. After whole pelvic irradiation,Pmg, C4 Prealb, Alb,alpha 2M and Hp were found to be positively associated with survival, whereas Tf, alpha 2PI, AT III, alpha 1 AT,C1Inh, C9 and IgA were negative factors.

Soybean protein isolate and soybean lectin inhibit iron absorption in rats.

Inhibitory effects of soybean protein isolate (SPI) and soybean lectin on the intestinal absorption of nonheme iron were investigated by in vivo studies in rats. Rats fed the SPI-based diet absorbed significantly less iron than did control rats fed the casein-based diet. Supplementing the SPI diets with 8% D-galactose significantly increased the incorporation of iron into liver ferritin, although D-galactose did not significantly increase iron absorption. Heat treatment of SPI significantly increased iron absorption. Ascorbate did not enhance iron absorption in rats fed the SPI-based diet. The presence of lectin in an aqueous extract of SPI was suggested by hemagglutination activity as well as by immunoreactivity with soybean lectin antibody. Soybean lectin introduced into ligated segments of the upper small intestine of rats inhibited ferrous iron absorption. This inhibitory effect, especially in the mucosal uptake, was significantly improved by addition of N-acetyl-D-galactosamine to soybean lectin. Soybean lectin had no effect on ferric iron absorption. Our results suggest that a portion of the reduction in iron absorption in rats fed SPI may be due to lectins.

c-myc gene in a murine plasmocytoma without visible chromosomal translocations moves to chromosome 12F1 with Pvt-1 and rearranges with IgH enhancer-S mu sequences.

The DCPC 21 plasmocytoma lacks any of the MPC-associated chromosomal translocations. However, the c-myc gene has been transposed to the IgH locus on chromosome 12 by an Ig switch-region-mediated recombination mechanism. DNA sequencing analysis, further, revealed that this recombination is consistent with an insertion of the IgH enhancer (E mu)-S mu sequences, 2341 bp in length, into the c-myc 5'-flanking region, resulting in 5': c-myc 5'-flanking-E mu-S mu-c-myc 5'-flanking-c-myc exon-1: 3' segment. In situ molecular hybridization of DCPC 21 metaphase chromosome spreads using a Pvt-1 probe demonstrated that Pvt-1 has also moved to the F1 sub-band region of chromosome 12 where the IgH genes are located. These results indicate that the c-myc gene has been inserted into the IgH locus together with the Pvt-1, regardless of whether plasmocytoma has cytogenetically identifiable translocations. The possible interaction between c-myc activation and Pvt-1 in the development of MPCs is discussed.

[Immunochemical analysis of two peak M proteinemia derived from structural differences of IgG Fc region].

We found M-proteins with two peaks by agarose electrophoresis in the serum of a myeloma patient. The M-proteins were identified as both IgG 1-kappa type, and classified as IgG-F (fast mobility) and IgG-S (slow mobility). 1) The possibility that the two M-proteins were derived from the post translational differences of sugar moieties of the same IgG molecule was unlikely, because no migration changes were observed in IgG-F and IgG-S after the treatment with 4 different sugar enzymes. 2) Fab fractions of IgG-F and IgG-S were analyzed. After papain or pepsin digestion, western blotting with anti-Fab antiserum revealed that the Fab fraction of IgG-F and IgG-S had identical mobility by agarose electrophoresis. However the Fc fractions of IgG-F and IgG-S analyzed by the same procedures with anti-Fe antiserum, were different. 3) Anti-idiotype antiserum prepared in rabbits against IgG-S, or -F, and absorbed by normal IgG and normal human serum showed a fused precipitin line with IgG-F and IgG-S. These findings suggest that two M-proteins with both IgG 1 and kappa type, have the same VH and VL regions but have different constant regions of heavy chain. Since one copy of IgG 1 constant gene is found in each human haploid gene. It is speculated that the switching of the rearranged VDJ gene to constant region gene occurred not only between cis chromosome but also between trans chromosome.

Interference of immunoglobulins in the radioimmunoassay of human beta-endorphin.

The interference of immunoglobulins in the radioimmunoassay (RIA) of human beta-endorphin was investigated. Human IgM showed no cross-reactivity. Human IgA showed a weak cross-reaction, but the dilution curve of IgA did not show parallelism with the standard curve of beta-endorphin, thus indicating its antigenic difference. The dilution curves of human IgG showed 0.18% displacement with respect to the human beta-endorphin standard curve, with good parallelism. Moreover, five patients with multiple myeloma of the IgG type showed falsely elevated beta-endorphin levels. We investigated the possibility that certain IgGs may be responsible for the displacement of [125I]beta-endorphin in the beta-endorphin kit. The apparent beta-endorphin level of plasma from multiple myeloma patients was markedly decreased after affinity chromatography of the serum on protein A-Sepharose. In another 3 patients with multiple myeloma, we examined IgG interference by measuring the beta-endorphin levels in their lyophilized IgG diluted with saline. The results demonstrated high values of 20.2, 25.5 and 21.2 pmol/l respectively, also showing good parallelism. These immunological parallels to human beta-endorphin verify that a part of the amino acid sequence of human IgG is similar to that of human beta-endorphin. Consequently, in the measurement of beta-endorphin with polyclonal antibody, the results may sometimes be spuriously high due to cross-reaction with IgG, e.g., in patients with IgG myeloma. To avoid IgG interference, a specific monoclonal antibody to synthetic beta-endorphin should be used rather than polyclonal antibodies.

Functional character and augmentation of lymphocytes in regional lymph nodes of patients with lung cancer.

It appears that lymph node metastases are more frequent in lung cancer than in other cancers because of impaired defensive mechanisms in the regional lymph nodes. However, little is known about the immunologic function of regional lymph node lymphocytes (RLNL) in patients with lung cancer. We have studied the immunologic properties of RLNL in comparison with peripheral blood lymphocytes (PBL). We measured the natural killer (NK) cell activity of RLNL and PBL in patients with lung cancer and found that the NK activity was significantly more depressed in the RLNL than in the PBL. In contrast, interleukin-2 (IL-2) production was markedly higher in the RLNL than in the PBL. The cytotoxic effect of RLNL in nonmetastatic lymph nodes on target cells (such as K562 cells) or PC-3 and PC-10 cells (NK-resistant, human lung cancer of adenocarcinoma and epidermoid carcinoma, respectively) was significantly enhanced by in vitro incubation with recombinant IL-2 (rIL-2). Furthermore, we clarified that both rIL-2 and OK-432, which is a biologic response modifier and IL-2 inducer as well, augmented the cytotoxicity of RLNL and that these effector cells were lymphokine-activated killer (LAK) cells. The depletion of lymphocyte subsets by pretreatment with specific monoclonal antibody showed that the LAK activity in RLNL was mediated by CD3+ and CD8+ cells, whereas the lymphocyte subsets contributing the LAK activity in PBL were CD3+ and CD16+ cells. It was concluded that a majority of the effector cells in RLNL were LAK cells of the cytotoxic T cell population.

[Biosynthesis and significance of acute phase proteins].

Acute phase proteins (APP) increase by the chemical receptors of macrophage via stimulations of either Fc or complement or other chemical receptors. No differences of APP pattern were observed by the different stimulants, because the effects of stimulation are changed by many processing steps in the macrophage as well as in the liver cell thereafter. Whether the APP increase results in good prognosis for original sickness or not are examined by multiple regression analysis in 38 cases of maxillary and 51 cases of laryngeal cancer with the observation for over five years. Alpha 1X contributes positively with length of their life, whereas alpha 1AG and ceruloplasmin contribute negatively. We could say that not all components of APP increase are advantageous for the diseases.

Serum amyloid P component regulation by sex steroids in rats.

Serum amyloid P component (SAP) has been designated as a female protein in hamsters. To investigate the effects of sex steroids on SAP concentration in rats, SAP was purified from Wistar rats by affinity chromatography. Anti-SAP was raised. Sample sera were obtained from 150 young and old rats, after which, they were injected with either estradiol (E2), testosterone (T), or dehydroepiandrosterone (DHA). Sera were serially obtained from the tail vessels until the 8th day after injection. Serum levels of SAP were measured by micro single radial immuno-diffusion with a standard pooled serum as 100%. As the rats aged, the SAP levels in untreated female rats increase with age from 93% at 11 weeks to 346% at 58 weeks. In 28-week-old rats, the SAP levels in females (135 +/- 40%) were significantly (p less than 0.01) higher than those in males (80 +/- 32%). When old female rats (56 wk) were injected with DHA (3.5 mg), the SAP decreased significantly (p less than 0.01) to 70.8% of the preinjection level on the 8th day after injection. When young male rats (11 wk) were jnjected with E2 (0.5 mg), the SAP levels increased rapidly to 188% of the prelevel on the 5th day. Serum E2 levels reached a peak on the 2nd day. After the injection of T, the SAP levels did not change.

Isolation of two new proteins from bovine colostrum which stimulate epidermal growth factor-dependent colony formation of NRK-49F cells.

Defatted bovine colostrum and decaseinated whey contain biological activity similar to that of TGF-beta in that they stimulate the anchorage-independent growth of NRK-49F cells in the presence of EGF. Two new proteins with such activity, termed BC-1 and BC-2, were isolated from decaseinated colostrum by a sequence of DEAE-Sephacel chromatography, Sephadex G-100 gel filtration in 1 M acetic acid, Sephadex G-50 re-gelfiltration in 8 M urea-1 M acetic acid and reverse-phase FPLCs. The two proteins showed distinctly different molecular weight from that of TGF-beta 1. BC-1 was a small Mr peptide of 8.5 kD. BC-2 had molecular mass of 46 kD, which yielded peptides of 46,40, and 6 kD on reduction. In contrast to TGF-beta, both proteins did not lose their biological activity on reduction. Both BC-1 and BC-2 suppress DNA synthesis in concanavalin A-stimulated thymocytes. These results suggest that BC-1 and BC-2 belong to a new class of mitogen/inhibitors, though their biological activities resemble those of TGF-beta.

C-myc-S mu rearrangement in a murine plasmacytoma without visible chromosomal translocations.

We report here that one murine plasmacytoma (DCPC 21) arising in a Balb/c mouse implanted i.p. with a Millipore diffusion chamber lacks any known chromosomal translocations. In situ molecular hybridization analysis of the metaphase spreads of DCPC 21, however, revealed that the c-myc gene has moved to the IgH locus on the chromosome 12. Further, an Ig switch region-mediated recombination was detected. The DNA sequencing analysis demonstrated that the c-myc gene has juxtaposed directly to S mu 5'-flanking sequence in a 'head-to-tail' orientation. The results strongly support our notion that the c-myc rearrangement is an essential step in the genesis of murine plasmacytomas, whether or not they have visible chromosomal translocations.

The role of fibronectin in the development of experimental amyloidosis. Evidence of immunohistochemical codistribution and binding property with serum amyloid protein A.

Azocasein-induced amyloid A (AA) amyloidosis in CBA/K1Jms mice was investigated to elucidate a preference of serum amyloid A (SAA) deposition in the spleen. By indirect immunofluorescence using anti-SAA/AA antibodies the initial deposition of SAA/AA was recognized in the marginal zone of spleen at 20 days after azocasein injection. Indirect immunofluorescence using anti-fibronectin antibodies also showed meshwork positivity in the corresponding area more intensely than that in controls. Immunoelectron microscopy using anti-SAA/AA revealed the presence of positively stained flocculent materials on cell surfaces of macrophages in the marginal area in addition to amyloid fibril. The tissue fibronectin rapidly increased in the spleen and maintained 10 times more than that of controls until the 20th day. Binding assay of SAA on frozen sections revealed the presence of SAA-binding substances in the perifollicular area. Affinity chromatographic assay showed fibronectin have a binding capacity to SAA1 and SAA2. By binding assay on the microtiter plate, SAA had more affinity to fibronectin than those of heparan sulfate, collagen type I, or serum amyloid P component. These results indicate that fibronectin plays an important role in the development of amyloidosis by working as a linking protein between SAA and the cell surface of macrophages.

Chromosomal assignment of woodchuck hepatitis virus (WHV) DNA integration sites in a woodchuck hepatocellular carcinoma-derived cell line (WH257GE10).

The chromosomal sites of woodchuck hepatitis virus (WHV) DNA integration were identified in a woodchuck hepatocellular carcinoma-derived cell line (WH257GE10) by the in situ hybridization technique using 3H-labelled WHV whole genome (WHV 2) as a probe. The G-banded chromosome spreads from WH257GE10 were identified and diagrammed schematically according to their band patterns. WHV DNA was integrated into 2 sites: 33 region of the long arm of chromosome 6 (6q33) and 31 region of the long arm of chromosome 8 (8q31). Chromosomal sites of WHV DNA integration were stable during successive passage periods analyzed in vitro.

The H-2 class II genes and the susceptibility to the development of pulmonary adenoma in mice.

To determine the locus in the H-2 complex that affects susceptibility to the development of pulmonary adenomas in mice, H-2 congenic and recombinant strains of mice with A/Wy, BALB/c, C3H, and B10 backgrounds were subjected to treatment with urethane. The average number and the incidence of adenoma foci were recorded five months after the treatment. In H-2 congenic strains on the A/Wy background, the average number of adenoma foci per mouse was significantly higher in mice of the A/Wy, A/J, and A-Tlab (H-2a) strains than in A.BY (H-2b) mice. In BALB/c and C3H congenic strains, the strains carrying the H-2k haplotype were more susceptible than those carrying the H-2b haplotype. In H-2 congenic strains on the B10 background, the average number and incidence of foci was also higher in haplotypes a, h2, k, and j than in haplotypes b, s, f, d, r, h4, i3, i5, and t4. The average numbers of adenoma foci in (A/J X A.BY)F1 (H-2a/H-2b) and (B10 X B10.A)F1 (H-2b/H-2a) were intermediate between the numbers in the parental strains. In [B10.A(4R) X B10.A (3R)]F1 (H-2h4/H-2i3) and [B10.A(4R) X B10.A(5R)]F1 (H-2h4/H-2i5), the numbers of adenoma foci were higher than in resistant parental recombinants. These patterns of response to urethane matched the patterns of the immune response to lactate dehydrogenase-B (LDH-B) and immunoglobulin gamma 2a (IgG2a) proteins. These differences between mice in their susceptibility to the development of pulmonary adenomas is probably due to the polymorphism of the class II genes in the H-2 complex.