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It is well known that mechanical loading influences the endochondral bone formation essential for the growth and development of longitudinal bones. The question was, however, asked whether the effect of mechanical loading on the chondrocyte metabolism is dependent on the loading frequency. This study was aimed at evaluating the effect of tensile loadings with various frequencies on the proliferation of growth plate chondrocytes and extracellular matrix synthesis. The chondrocytes obtained from rib growth plate cartilage of 4-week-old male Wistar strain rats were cultured by day 4 and day 11 and used as proliferating and matrix-forming chondrocytes, respectively. Intermittent tensile stresses with different frequencies were applied to each stage chondrocyte. DNA syntheses were examined by measuring the incorporation of [3H]thymidine into the cells. Furthermore, the rates of collagen and proteoglycan syntheses were determined by measuring the incorporation of [2,3-3H]proline and [35S]sulfate into the cells, respectively. At the proliferating stage, intermittent tensions with the frequencies of 30 cycles/min and 150 cycles/min significantly (p

The ability of FeCl3 to extract Cd from three paddy soils was compared with that of various irons, manganese, and zinc salts to elucidate the extraction mechanism. Manganese, zinc and iron salts (including FeCl3) extracted 4-41%, 8-44% and 24-66% of total Cd, respectively. This difference reflected the pH of the extraction solution, indicating that the primary mechanism of Cd extraction by FeCl3 is proton release coupled with hydroxide generation, as iron hydroxides are insoluble. Washing with FeCl3 led to the formation of Cd-chloride complexes, enhancing Cd extraction from the soils. FeCl3 effectively extracted Cd from all of the three soils compared to HCl that is a conventional washing chemical, when the concentrations of the two washing chemicals were between 15 and 60mM(c) (at above extraction pH 2.4), while the corresponding extraction pH of FeCl3 was slightly higher than HCl. As HCl is the strong acid of complete dissociation, if excess amount of HCl was added to soil, it is possible to give the dissolution of clay minerals in soils. In contrast, proton release from FeCl3 is controlled by the chemical equilibrium of hydroxide formation. While soil fertility properties were affected by a bench-scale soil washing with 45mM(c) FeCl3, adverse effects were not crucial and could be corrected. The bench-scale test confirmed the effectiveness of FeCl3 for removal of soil Cd. The washing had no negative effect on rice yield and lowered the Cd concentration of rice grain and rice straw in a pot experiment.

We conducted a pot experiment to evaluate the effect of soil washing with CaCl(2) on Cd absorption by two soybean cultivars. The results were as follows: (1) Soybean growth was not significantly different in washed and unwashed soils, but the seed Cd concentration for both cultivars decreased significantly, up to 25%, in the washed soils compared with the unwashed soils. (2) In the washed soils, the Cd concentration in the soil solution indicated an obviously lower value from sowing to the flowering stage; however, the change in Cd speciation was not evident in the CaCl(2)-washed soil solution. Consequently, the effect of soil washing using CaCl(2) on Cd-contaminated paddy soils can be expected to continue after a CaCl(2)-washed paddy field is converted to an upland field.

We developed a new, three-step soil-wash method to remediate Cd-contaminated paddy fields. The method comprises (1) chemically washing the field soil with a CaCl 2 solution; (2) washing the treated soil with water to eliminate residual Cd and CaCl 2 ; and (3) on-site treatment of wastewater using a portable wastewater treatment system. Cd concentrations in the treated water were below Japan's environmental quality standard (0.01 mg Cd L −1 ), and the removal of Cd from the exchangeable fraction was 55% and from the acid-soluble fraction 15%. While soil fertility properties were affected by the soil washing, adverse effects were not crucial and could be corrected. The washing had no affect on rice growth, and reduced the average Cd concentration in rice grains by about two-thirds compared to a control plot. These results confirmed the effectiveness of the soil-wash method in remediating Cd-contaminated paddy fields.

Vascular endothelial growth factor A (VEGF-A) plays an essential role in tumor progression through stromal neovascularization in malignant solid tumors. Neuropilin (NRP) is considered to be the specific receptor for limited types of VEGF-A isoform, VEGF165. The clinicopathological implications of NRP are not well understood in colon cancer, while almost all colon cancers overexpressed VEGF-A. We examined the expression levels of NRP1 and NRP2 genes in 54 colon cancer cases and paired extraneoplastic tissue with quantitative real-time polymerase chain reaction. The gene expression levels of NRP1 in the tumor (0.431+/-0.583) were significantly decreased compared to those in the extraneoplastic tissue (0.754+/-0.799) (paired t-test, p=0.0208). On the other hand, the gene expression levels of NRP2 in the tumor (0.763+/-0.791) were not decreased compared to those in the extraneoplastic tissue (0.508+/-0.386) (paired t-test, p=0.0511). Twenty cases, with preserved expression of the NRP1 gene in the tumor, showed a better prognosis as compared to the 34 cases with decreased NRP1 expression (p=0.0258, log-rank test). No significant relationship was noted between NRP2 gene expression and prognosis. The results suggested that preserved NRP1 expression provides colon cancer patients with a better prognosis.

The efficiencies of neutral salts, strong acids, and chelates were tested for extracting cadmium (Cd) from three paddy soils. The higher the selectivity of the cations of the added neutral salts toward soil adsorption sites, the lower the pH in the extracts and the more soil Cd could be extracted. In addition, soil carbon and nitrogen contents and mineral composition were closely associated with the amount of Cd extracted. Calcium chloride and iron(III) chloride were selected as wash chemicals to restore Cd-contaminated paddy soils in situ. Washing with calcium chloride led to the formation of Cd chloride complexes, enhancing Cd extraction from the soils. The washing also substantially decreased soil levels of exchangeable and acid-soluble Cd, which are the major forms of bioavailable Cd for rice (Oryza sativa L.). The optimum conditions for in situ soil washing were also determined for calcium chloride.

A 10-month-old infant with chronic myelomonocytic leukemia (CMML) of 5 months' duration, who had been treated only with transfusion, displayed leukemic transformation characterized by lymphoid morphology, PAS positivity, and myeloperoxidase negativity. Surface marker analysis of blast cells revealed expression of lymphoid-associated antigens (CD10 and CD19) but not myeloid-associated antigens (CD13, CD14, and CD33). These findings suggest that some cases of infantile CMML are clonal disorders arising in a pluripotent stem cell that can also differentiate along the lymphoid cell lineage.

It is unclear how the cytokine network contributes to the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) in non-small cell lung cancer (NSCLC). We examined the gene expression of GM-CSF and interleukin (IL)-10 in 78 surgical specimens and 16 xenografts of NSCLC by reverse transcription-polymerase chain reaction. Forty-four of the 78 (56.4%) surgical specimens and 11 of the 16 (68.8%) xenografts of NSCLC showed GM-CSF gene expression. Forty-one of the 44 (93.2%) GM-CSF-positive NSCLC specimens showed IL-10 expression, and GM-CSF expression was significantly correlated with IL-10 expression (p0.0171, Mann-Whitney's U-test). These results suggested that IL-10 is one of the cytokines up-regulating the GM-CSF expression in NSCLC.

A new reagent for tert-butoxycarbonylation: 2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile intermediate: 2-hydroxyimino-2-phenylacetonitrile product: 2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile C6H5CH2 4-CH3OC6H4CH2 Cl3CCH2 Keywords: phosgene

We report here a case of primary pulmonary sarcoma complicated with neurofibromatosis type 1 (NF1). A 60-year-old man was urgently admitted to Tokai University Hospital, Japan, due to severe right chest pain. Skin inspection showed subcutaneous tumor lesions and café-au-lait spots, and therefore a diagnosis of neurofibromatosis type 1 (NF1) was made. Chest X-ray showed marked fluid accumulation in the right pleural cavity, and he was diagnosed as having empyema by thoracic drainage. Chest X-ray showed a large round mass lesion (80 mm in diameter) in the right lower lung field. Percutaneous needle biopsy of the tumor revealed that the empyema was caused by actinomycotic infection, and then he underwent right middle and lower lobectomy. Histopathological examination of the surgical specimens revealed features of spindle cell sarcoma. Several postoperative systemic examinations showed no other lesions except in the right lung. We concluded that this was a complicated case of primary pulmonary sarcoma with NF1.

Background: The type 2 cytokine interleukin (IL)-10 has been reported to inhibit the antitumour activity of the regional immunity against various neoplasms. Certain lung cancers produce IL-10, but the clinical significance of IL-10 expression is not well understood. Patients and methods: We examined IL-10 and IL-10 receptor (IL-10R) mRNA expression in 82 non-small-cell lung cancers (NSCLC) by reverse transcription-polymerase chain reaction (RT-PCR) assay. Immunohistochemistry (IHC) and enzyme immunoassay ( EIA) were applied to evaluate the cellular localisation and the serum levels of IL-10. Results: RT-PCR assay revealed IL-10 mRNA expression in 68 (83%) of 82 NSCLC surgical specimens (40 of 50 adenocarcinomas, 22 of 26 squamous cell carcinomas, 5 of 5 large-cell carcinomas, 1 of 1 adenosquamous-cell carcinoma). RT-PCR assay also revealed IL-10R mRNA expression in 79 cases of NSCLC (96.1%). IL-10 expression was confirmed within tumour cells by 1HC. EIA showed no significant serum IL-10 elevation in the 12 NSCLC positive for IL-10 mRNA expression (0-2.99 pg/ml). The NSCLC patients with IL-10 production showed significantly poorer prognosis than those without IL-10 production (P < 0.05, Kaplan-Meier, log-rank test). Conclusions: These results suggested that the cytoplasmic IL-10 correlated to clinical prognosis, and that IL-10 expression is a prognostic factor for NSCLC.

This paper proposes the new display system by windmill blade. The proposed method is a system by the visual afterimage phenomenon, using windmill blade in which the light emitter has been embedded. This system shows new effectiveness of the wind power generator system.

Heterogametic sex chromosomes have evolved independently in various lineages of vertebrates. Such sex chromosome pairs often contain nonrecombining regions, with one of the chromosomes harboring a master sex-determining (SD) gene. It is hypothesized that these sex chromosomes evolved from a pair of autosomes that diverged after acquiring the SD gene. By linkage and association mapping of the SD locus in fugu (Takifugu rubripes), we show that a SNP (C/G) in the anti-Müllerian hormone receptor type II (Amhr2) gene is the only polymorphism associated with phenotypic sex. This SNP changes an amino acid (His/Asp384) in the kinase domain. While females are homozygous (His/His384), males are heterozygous. Sex in fugu is most likely determined by a combination of the two alleles of Amhr2. Consistent with this model, the medaka hotei mutant carrying a substitution in the kinase domain of Amhr2 causes a female phenotype. The association of the Amhr2 SNP with phenotypic sex is conserved in two other species of Takifugu but not in Tetraodon. The fugu SD locus shows no sign of recombination suppression between X and Y chromosomes. Thus, fugu sex chromosomes represent an unusual example of proto–sex chromosomes. Such undifferentiated X-Y chromosomes may be more common in vertebrates than previously thought., Author Summary Diverse systems of sex determination have evolved independently in the animal and plant kingdoms. In vertebrates, so far four master sex-determining (SD) genes, Sry, Dmrt1, Dmy, and Dm-W, have been identified. These genes code for transcription factors and are located on only one of the sex chromosomes surrounded by nonrecombining regions. It is hypothesized that these sex chromosomes evolved from a pair of homologous chromosomes that diverged after acquiring the SD gene. We investigated the SD locus in fugu by high-resolution genetic mapping and association mapping. We found that a SNP that changes an amino acid (His/Asp384) in the kinase domain of anti-Müllerian hormone receptor type II (Amhr2) is perfectly associated with phenotypic sex. A combination of the two alleles of the SNP (homozygous females and heterozygous males) is likely to be responsible for sex determination in fugu. While these alleles are conserved in two other species of Takifugu, they are absent in the freshwater pufferfish, Tetraodon. Furthermore, Fugu Amhr2 lies in a region that shows no evidence for recombination suppression between X and Y chromosomes. Thus, fugu sex chromosomes represent an unusual example of a pre-differentiated phase of sex chromosomes in vertebrates.

Of 29 hematopoietic cell lines tested for susceptibility to human immunodeficiency virus (HIV)-1HTLV-IIIB infection, all CD4+ cell lines became infected. Continuous culturing of infected cell lines resulted in nine HIV-1 carrier cell lines, including, for the first time, an HIV-1 carrier megakaryoblastic cell line, MEG-01/HIV. The immunophenotypic profiles of a total of 17 HIV-1 carrier cell lines (nine newly and eight previously established cell lines) were compared with their respective parental noninfected cell lines. Except for total absence of CD4 expression, the expression of other antigens was variable among the 17 HIV-1 carrier cell lines. Persistent and consistent replication of infectious HIV-1 was detected in all of them in varying quantities. The great variability observed in both the altered marker expression, with respect to that of the noninfected parental cell lines, and in the quantities of persistently produced infectious HIV-1 was, nevertheless, specific to the individual cell lines. Furthermore, the present study demonstrates that there is no apparent correlation in the quantity of HIV-1 produced to either T cell, myelomonocytic cell, or megakaryocytic cell types. Instead, the results suggest that a particular interaction between HIV-1 and individual clonal cell lines may provide insight into the extremely complex immune dysregulation associated with the pathogenesis of acquired immune deficiency syndrome. Thus, the 17 HIV-1 carrier cell lines of diverse origin presented here provide valuable and unique models for further understanding acquired immune deficiency syndrome pathogenesis at the cellular and molecular levels.

The murine interleukin 5 receptor (IL-5R) was identified by utilizing an immobilized IL-5 and an immobilized monoclonal antibody against the murein IL-5R (designated H7 mAb). The H7 mAb immunoaffinity-purified materials from the extract of cell-surface radioiodinated T88-M cells (an IL-5-dependent early B cell line) using 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) were reacted with an immobilized IL-5 matrix. SDS-PAGE of the adsorbed fraction revealed a single band at approximately 60 kDa. The binding of the 60 kDa protein to the immobilized IL-5 matrix was inhibited by the excess IL-5. The CHAPS-extract depleted of the 60 kDa protein by the absorption with H7 mAb did not contain any IL-5 binding proteins. Immunoaffinity procedure provided a final 7400-fold purification, based on an estimation of the content of the 60 kDa protein (approximate purity: 20%) from the silver-stained pattern of SDS-PAGE. Actin was copurified with the 60 kDa protein at an approximate ratio of 1:1, suggesting that the intracytoplasmic domain of the IL-5R may interact with actin. Furthermore, soluble IL-5R (molecular mass: 50 kDa) was purified by the H7 mAb-immunoaffinity chromatography. The purified soluble IL-5R was capable of inhibiting the binding of IL-5 to T88-M cells. Preparative SDS-PAGE followed by electroblotting onto a membrane permitted the determination of the N-terminal sequence of the IL-5R. The determined N-terminal sequence of the IL-5R and the deduced primary sequence from recently isolated cDNA were compared.

The structures of nocardicins A and B, novel monocyclic beta-lactam antibiotics produced by a strain of Nocardia, have been established as 1 and 2, respectively, on the basis of spectroscopic and chemical evidence. They are the first examples of monocyclic beta-lactam antibiotics possessing relatively high antimicrobial activity, and are stereochemically and biologically related to penicillins and cephalosporins.

We have studied the effects of urokinase (UK) on concentration changes of α2-antiplasmin (α2-AP) and on fibrinogenolysis. Medium dose (480.000u) or large dose (960, 000u) of UK was given to each of seven normal volunteers by intravenous drip infusion within six hours, and then blood and urine analyses were carried out. Total α2-AP, which includes free α2-AP and α2-AP-plasmin complex, decreased about 50 per cent of the original value with large dose of UK. α2-AP-plasmin complex appeared in the plasma one hour after UK infusion and increased up to 50 per cent of total α2-AP at the end of UK infusion. Bβ1-42, which liberated from fibrinogen at the very early stage of fibrinogenolysis, increased significantly with UK infusion, and was 65 times as much as the normal range at the end of UK infusion. Urinary Bβ1-42 increased as well as plasma Bβ1-41. On the other hand, fibrinogen degradation products (FDP) measured with enzyme immunoassay (ETA) increased only slightly, and moreover, urinary FDP was not detectable at any time. Plasma fibrinogen level did not decrease and it changed within the normal range in both groups. We then gave 960, 000u of UK to three patients with deep vein thrombosis and blood analysis was carried out as with normal volunteers. The most significant observation different from that of normal volunteers was shown in FDP levels. Serum FDP levels of three patients increased significantly in comparison with normal volunteers. Urinary FDP increased as significantly as plasma FDP.In conclusion, we consider that infusion of 960, 000u of UK is enough to neutralyze α2-AP and enough to express thrombolytic activity. It causes only very early stage of fibrinogenolysis without advanced fibrinogenolysis which may result in bleeding tendency in normal volunteers. On the other hand, in thrombotic patients, advanced fibrinolysis was observed with UK infusion.

The syntheses of FR-33289 (3), originally isolated from a microorganism, and FR-32863 (6), first designed on the basis of a biogenetic consideration and recently found to be also present in nature, are described.

Several t-butoxycarbonyl derivatives of oximes were prepared through the corresponding oxime chloroformates. Of these, diethyl (t-butoxycarbonyloxyimino)malonate and 2-(t-butoxycarbonyloxyimino)-2-phenylacetonitrile were utilized for the preparation of t-butoxycarbonylamino acids under various conditions. The results are summarized in a table.

Monoclonal antibodies (MAbs) were obtained from hybridoma clones established by cell fusion between P3X63Ag8.653 mouse myeloma cells and spleen cells of mice or rats hyperimmunized against human bladder cancer tissue or BC47 rat bladder cancer cells. RBS-31 and RBS-85 mouse MAbs and RBA-1 rat MAb were raised against BC47 cells and HBP-1 MAb was raised against human bladder cancer tissues. Urinary antigens detected by these MAbs were quantitatively assayed by means of ELISA using 50 microliters of 1:2 diluted urine samples. The cut-off value of the assay was set up as the mean + 4 X SD of the mean using data from the healthy individual urine samples. The reactivity of all healthy control urine samples were under the cut-off value (negative). By contrast, urine from bladder cancer patients reacted positively with the RBS-31 MAb at 72%, with the RBS-85 MAb at 63%, with the RBA-1 MAb at 51% and with the HBP-1 MAb at 35%. The urine samples from some patients with renal calculi, acute cystitis or complicated urinary tract infections showed only a weak reactivity with our MAbs. As for extra-bladder cancers, some patients with renal, renal pelvis, prostate or ureter cancer, but no patients with esophageal, gastric, colon or liver cancer or leukemia, had reactive urinary antigens.

The synthesis of analogs of 3-(N-acetyl-N-hydroxyamino) propylphosphonic acid (FR-900098, Ia) and an analysis of their structure-activity relationship are described. Among them, compounds Ib (FR-31564) and XIIb (FR-32863) have recently been found to be produced naturally by Streptomyces lavendulae. The antibacterial activity of Ib against Pseudomonas species is especially significant.

Monocyclic β-lactams related to nocardicins were enantioselectively synthesized from phthal-imidoacetyl chloride and hexahydro-1, 3, 5-triazines (4) via a [2+2] cycloaddition reaction. The preparation and biological activity of some typical acyl derivatives are also described.

The total synthesis of monocyclic β-lactam antibiotics, nocardicins A ( 1a ) and D ( 1d ), is described. 3-Aminonocardicinic acid (3-ANA, 2) was synthesized from p -hydroxyphenylglycine via an acid chloride-imine cyloaddition reaction. The side chain amino acid 13 was prepared via a key step of condensation of p -hydroxyacetophenone and α-phthalimidobutyrolactone. Acylation of 3-ANA with 13 gave nocardicin D, from which nocardicin A was obtained by oximation.

Synthesis and effect on antibacterial activity of 2- and/or 3-methyl group(s) in the cephalosporins (4a-c) with (Z)-2-(2-amino-4-thiazolyl)-2-(methoxyimino)acetyl side chain were described.