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Large amounts of rice husk ash (RHA) are discharged from rice husk power plants, and the development of an effective system for recycling this RHA waste would be desirable. In the current study, the silica component of RHA obtained from a rice husk power plant in Myanmar was successfully used as raw material for the synthesis of the aluminosilicate nanotube (ASNT) material. The RHA used contained over 90 wt% of silica. The synthesized ASNT had a web-like structure formed of fi brous bundles, each containing many individual ASNT.

Nanosized Mg-Al layered double hydroxides (LDHs) with Cl−, NO3− or SO42− as the interlayer anion have been synthesized by a modified coprecipitation method. The obtained LDHs were confirmed to be composed of a single phase and to be highly substituted by Al (Mg/Al ratio ∼1.9). The abilities of the LDHs to adsorb several harmful anions (F−, CrO42−, HAsO42− and HSeO3−) in aqueous solution were studied. The LDHs exhibit high adsorption abilities. The amount of adsorption onto the LDHs differed between the starting interlayer anions, and decreased in the following order of the interlayer anions: NO3− > Cl− > SO42−. The NO3-formed Mg-Al LDH reached a CrO42− adsorption equilibrium state within only 30 min, much faster than those in previous reports. Thus, the nanocrystallized, highly Al substituted phase of the NO3-formed Mg-Al LDH is found to markedly enhance the anion adsorption ability.

We demonstrate the fabrication of complicated three-dimensional (3D) microstructures embedded in a photosensitive glass by a high-order multiphoton process using a femtosecond (fs) laser. Direct writing of the fs laser followed by a post baking process and preferential etching in a dilute hydrofluoric (HF) acid solution results in a microplate that can freely move in hollow structures embedded in the glass. The fabricated structure functions as a microvalve that can control the flow direction of fluids in the microreactor.

Hebeloma radicosoides sp. nov. is described, illustrated and characterised. It superficially resembles H. radicosum in having a long-rooting stipe with a well-formed, membranous annulus and in having preference for animal wastes as its resources. Its pileus is yellower than that of H. radicosum, and it fruits after addition of urea to soil, whereas H. radicosum does not. It also fruits on various animal wastes whereas H. radicosum specifically fruits on mole or mouse middens. Unlike H. radicosum, H. radicosoides has no distinct odour. Hebeloma radicosoides also resembles Hebeloma luchuense and H. spoliatum in having a long-rooting stipe and in fruiting on urea-treated soil, but differs from them by having a yellowish scaly pileus and distinct, membranous annulus.

This randomised study was performed to assess the anti-emetic efficacy and tolerability of two-dose regimens of granisetron in children with leukaemia. 49 children with leukaemia were treated with three consecutive courses of high-dose methotrexate or cytarabine regimen. During the first course, patients were evaluated regarding the emetogenicity of each regimen. They were randomised in a crossover manner to receive 20 or 40 micrograms/kg of granisetron before the second and third course of chemotherapy. Neither emesis nor severe appetite loss were observed in over 80% of patients within the first 24 h in all treatment groups. There was no significant difference in the anti-emetic efficacy between the two-dose regimens of granisetron. However, complete protection was achieved less frequently on days 2 and 3. Older children and girls appeared to be less well protected. No adverse events attributable to granisetron were observed. Granisetron dose regimens of 20 and 40 micrograms/kg are, comparably, well tolerated and effective in controlling chemotherapy-induced emesis in the first 24 h, though this protection fails thereafter, particularly in older patients and girls.

A new technology for rapid prototyping of lab-on-chip devices is described. Direct write of a near-infrared femtosecond laser forms three-dimensional (3D) latent images inside photostructurable glass. Modified regions are developed by a post-annealing and then preferentially etched away in dilute hydrofluoric acid solution with an etching selectivity of 40–50 times, resulting in the formation of true 3D hollow microstructures inside the glass. Microfluidic structures with microcells and microchannels embedded in the glass are fabricated by this technique.

Several studies have shown that HIV and nutrition operate in tandem. Moreover, it has been shown that the two greatly affect agricultural production due to reduced energy to work, inability to purchase agricultural inputs, low labor, and eventual death. The link between agricultural productivity, malnutrition, and HIV can therefore not be overlooked. People who are inadequately nourished are more susceptible to diseases and poor health. In an attempt to achieve optimal nutrition and good health among vulnerable groups, various intervention programs have used food supplementation and especially the plant-based food products to achieve this. Such programs have proved to be effective in restoring the nutrition and health status of the people. However, much more value would be achieved if such programs were complemented with basic health services such as deworming, water, sanitation, malaria control, hygiene. This chapter explores the benefits of research on nutrition as the basis for improving threatened rural communities’ nutria-health and potential economic performance. The premise is that good nutrition and preventive measures will reverse some of the human health problems associated with HIV, hunger, and/or malnutrition. Emphasis is placed on food preservation, processing, nutrition intervention, and education. Micronutrients through agronomic fortification/fertilization is recommended as an intervention with the benefit of improving the nutritive quality of food and thereby providing essential elements needed by the human body to combat malnutrition and poor health. The conclusion is that better nutrition will contribute to better health and increase productivity and production on the farm.

s S39 on immune system function in relationship to INTERMACS class prior to MCSD-implantation is virtually non-existent. We hypothesized that preMCSD peripheral blood mononuclear cell (PBMC) gene expression profiling (GEP) by next-generation sequencing (NGS) identifies a subgroup of patients with higher risk of post MCSD MODS. Methods: PBMC samples from 8 AdHF patients (53±25 years old) undergoing MCSD implantation included in the prospective MultiOrgan Dysfunction Expression (MOD-E) profiling study between August 2012 2013. Obtained 1 day before surgery, and at days 1, 3, 5 and 8 postoperatively were collected for 3 INTERMACS level 3 patients (n= 3) and 5 patients (n= 5) INTERMACS level 1-2. Purified mRNA was subjected to whole-genome NGS analysis. Differentially expressed genes where identified by Two-way ANOVA and Benjamini-Hochberg correction. Only those with fold change of at least 1.5 were included in the analysis. Hierarchically clustering was used for visualization. Biological significance was assessed by gene ontology and pathway analysis. Results: Compared to INTERMACS level 3, patients in the higher risk categories had dramatic differences in their gene expression profiles before and across all 5 time points (Figure 1). Over 3100 different genes were identified and mechanistically linked to the systemic inflammatory response by their biological significance. Gene ontology and pathway analysis revealed significant enrichment of immune related categories and pathways. Conclusion: In AdhF patient facing MCSD-implantation, NGS-based PBMC GEP correlates with clinical severity as assessed by INTERMACS class prior to as well as early after MCSD-implantation and may be further develop to assist in improved patient selection.

We describe the prevalence of community-acquired M. pneumoniae pneumonia diagnosed by culture methods in a single institute in Japan from January 1979 to December 1999. Cultures were performed in 2971 pneumonia cases and yielded M. Pneumoniae in 508 cases. The epidemic peaks recurred regularly at 4-year intervals (1980, 84, 88 and 91–2). Although a large epidemic has not occurred since 1992, traces of epidemic periodicity have still persisted from 1992 to 1999 at 3-year intervals.

Methods HIV-specific proliferative CD4+ T cell responses were assessed by CFSE assays performed after stimulation with recombinant HIV p24 in the presence of a blocking antiIL10R antibody or an isotype control, with or without depletion of CD25+ cells. IL-10 production was measured in cell subsets by ICS or by luminex of culture supernatants from PBMCs depleted of CD14+, CD19+ or CD25+ cells. Finally, monocytes and regulatory T cells were sorted and cultured alone or together in transwell plates and secreted IL-10 was measured.

1. The spatial patterns of reflexes elicited by localized pressure stimulation of the foot skin were analysed by recording electromyographic activities of various hindlimb muscles or muscle nerve discharges in cats anaesthetized with sodium pentobarbitone. 2. Reflex discharges evoked by stimulation of the central pad occurred mainly in physiological toe extensors located in the foot. Stimulus-response relationships of single motor units revealed characteristically wide ranges of graded response and recruitment. 3. Within the central pad, the strongest excitation was evoked from the central lobe and was distributed to extensors of all four toes. Excitation from the medial and the lateral lobes was usually asymmetrical and weaker in toe muscles of the stimulated side. It is suggested that the weakness was in part due to concomitant inhibition. 4. Stimulation of a toe pad caused marked suppression of central pad-evoked activity of toe extensors with a highly specific spatial pattern. The inhibition was strongest in extensors of its own toe, and gradually weaker in the more distant toes. Weak excitation was occasionally evoked in extensors of the most medial or lateral toes, when the most lateral or the most medial toe-pad, respectively, was stimulated. 5. A similar pattern of relfex to that from the toe pad was evoked from the claw base and the hairy toe dorsum of each digit. 6. Reflex effects, both inhibitory and excitatory, from the central and toe pads, claw bases and toe dorsum were maintained during prolonged stimuli, indicating that slowly adapting receptors contributed to these reflexes. 7. It is concluded that stimulation of localized skin areas of the foot, particularly the pads, evokes highly specialized reflexes, which may be important in controlling movements of individual digits.

We have fabricated 3-D microreactor structures embedded in photosensitive glass by femtosecond laser direct writing followed by heat treatment and chemical etching in HF solution. The etching condition has been optimized for getting higher resolution. This technique was applied for manufacturing microfluidic structures containing microvalves which controlled the direction of reagent flow. This technique would be very attractive for rapid prototyping of micro-total-analysis systems.

Of the cytochrome P450 (CYP) family of carcinogen-activating enzymes, CYP3A is the major form found in human livers. The purpose of this study was to investigate the cytotoxic effects of dental resin monomers after being metabolized by CYP3A4 and CYP3A7, using a colony formation assay and a neutral red assay. Specimen wells were plated with transfected cells derived from the Chinese hamster lung at 100 cells well(-1). The experimental group consisted of CYP-producing 3A4-10 and 3A7-40 cells, while the control group consisted of non-CYP-producing CR-119 cells. Bisphenol A (BPA) and bisphenol A glycidyl methacrylate (Bis-GMA) and a positive control (Aflatoxine Bl) were added separately to each well and cultured for 7 days. After cultivation, the number of the colonies was counted and IC50 values were determined. The data were statistically analysed by a Student's t-test. The resultant of IC50 values indicated that the monomers were not metabolically activated by CYP3A4 or CYP3A7 as compared with the control (P0.05). We also confirmed that these monomers act neither as activators nor as inhibitors of CYP3A4 and CYP3A7.

Four cultures of monkey kidney cell line, JTC-12, were flown on the Spacelab-J (SL-J) mission during 8 days. The results of the present study showed that the space flight gave no essential effect on morphology, cell cycle, glucose consumption and urokinase production of the mammalian culture cell. However, the cell proliferation slightly decreased under microgravity. Moreover, the lack of gravity induced the trypsin-treated dissociated cells to keep floating in the culture medium. Therefore, the attachment of the cells onto the substratum was delayed, and that caused difficulties in subculturing the cells. The present research also offered some important information on techniques for establishment of cell cultures in space laboratories.

New antitumor agents must be added to the current neuroblastoma treatment regimens to improve the clinical results. We investigated whether recombinant human endostatin (rhEndostatin), an antiangiogenic agent, is effective against human neuroblastoma in the human neuroblastoma xenograft model designated TNB9. When tumors on the back of nude mice grew to a weight of 90-95 mg, rhEndostatin 10 mg/kg/day was administered subcutaneously every day for 10 consecutive days. Mean relative tumor weight in mice administered rhEndostatin (n=5) was significantly less than that in controls (n=12) on days 2, 4, and 6 after the start of administration (p

The expression of Fas, a cell surface receptor directly responsible for triggering cell death by apoptosis, and its ligand (FasL) was investigated on both human colonic intraepithelial T lymphocytes (IELs) and peripheral blood mononuclear lymphocytes (PBMLs). FACS analysis indicated that IELs have increased expression of Fas compared with PBMLs, together with the progress activation marker, CD45RO. A discrete fraction of freshly isolated IELs also constitutively expressed FasL, perhaps as a result of recent in vivo activation. Using monoclonal antibody APO2.7, which detects mitochondrial 7A6 antigen specifically expressed by cells undergoing apoptosis, we further investigated the apoptosis-inducing effect of anti-Fas monoclonal antibody (CH11) on both IELs and PBMLs. FACS analysis revealed that CH11 increased the percentage of apoptotic cells, in IELs but not in PBMLs. Culture with anti-FasL monoclonal antibody (4H9) significantly recovered cell viability in IELs, but not in PBMLs. These results indicate that IELs constitutively express both Fas and FasL and that Fas crosslinking generates signals resulting in apoptosis, outlining a potential mechanism involved in intestinal tolerance.

The surveillance study was conducted to determine the antimicrobial activity of fluoroquinolones (ofloxacin, levofloxacin, ciprofloxacin, tosufloxacin) and other 20 antimicrobial agents against 5,180 clinical isolates obtained from 26 medical institutions during 1998 in Japan. The resistance to fluoroquinolones was remarkable in Enterococci, methicillin-resistant staphylococci and Pseudomonas aeruginosa from UTI. However, many of the common pathogens such as Streptococcus pneumoniae including penicillin-resistant isolates, methicillin-susceptible Stahylococcus aureus, Moraxella catarrhalis, the family of Enterobacteriaceae, Haemophilus influenzae including ampicillin-resistant isolates have been kept to be susceptible to fluoroquinolones. About 90% of P. aeruginosa isolates from RTI were susceptible to fluoroquinolones. In conclusion, the results from this surveillance study suggest that fluoroquinolones are useful in the treatment of various bacterial infections including respiratory infections.

During October and December of each year of from 1994 to 1996, 3,849 strains of 10 species of bacteria were isolated from clinical materials in 21 institutions nationwide. The minimum inhibitory concentrations (MICs) for these bacteria of four carbapenems (imipenem [IPM], panipenem [PAPM], meropenem [MEPM], and biapenem [BIPM]) and other representative antibacterial agents were measured to investigate annual changes in antibacterial activity. Carbapenems showed potent activity against methicillin-sensitive S. aureus (MSSA), S. pneumoniae, E. faecalis, H. influenzae, E. coli, K. pneumoniae, E. cloacae, S. marcescens, and the B. fragilis group, with the activity being stable. However, these drugs showed weak activity against methicillin-resistant S. aureus (MRSA) and P. aeruginosa. The antibacterial activity (MIC90) against the tested organisms generally remained stable. Particularly, there was annual improvement of the MIC90 values of IPM and BIPM for S. pneumoniae, as well as the values of IPM and PAPM for H. influenzae, and those of IPM, PAPM, and BIPM for S. marcescens. On the other hand, the activity of carbapenems (including IPM) against MRSA was not necessarily strong, but there was annual improvement of MIC90 values.

The purpose of this study was to investigate the biological characteristics and prognostic value of in vitro three-drug resistance to prednisolone, L-asparaginase, and vincristine in childhood acute lymphoblastic leukemia (ALL). We carried out in vitro tests with a 4-day culture and a methyl-thiazol-tetrazolium assay on bone marrow samples from 209 children newly diagnosed with ALL. After testing the resistance of leukemic cells to 14 drugs, we classified the patients into two groups according to their sensitivity to three drugs (prednisolone, L-asparaginase, and vincristine) used in remission induction therapy. The three-drug resistant group (RR: sensitive to no drugs or to one drug) correlated with both short-term and long-term treatment failure. Three-year event-free survival (95% confidence interval) for the sensitive group (SS: sensitive to two or three drugs) was 0.813 (0.773-0.853) and that of the RR group was 0.616 (0.569-0.669) (P = 0.0001). Univariate analysis showed that Philadelphia-chromosome (Ph1) positivity and immunophenotype of mixed lineage were also prognostic factors in the 209 patients. The prognosis of the SS/RR drug resistance profile within 14 Ph1 patients was marginally significant (P = 0.062). Multivariate Cox regression analysis showed that Ph1 was an overwhelmingly adverse factor in event-free survival, with a relative hazard of 5.37 (2.57-11.21, P0.0001), followed by RR, with a relative hazard of 2.98 (1.69-5.25, P = 0.0001). Furthermore, we clarified the characteristics of the RR group by examination of the pattern of drug resistance to other drugs in comparison with the SS group. The leukemic cells of RR patients were more resistant than those of SS patients (P0.0001) to all the drugs tested, with resistance ratios of 1.6 to 13.1 (mean 3.4). In conclusion, in vitro three-drug resistance at the initial stage is an important independent predictor of treatment failure for both induction response and long-term outcome in childhood ALL.

In response to 1-methyladenine, a maturation-inducing substance, starfish oocytes undergo reinitiation of meiosis with germinal vesicle breakdown through activation of p34cdc2-cyclin B, which results in the dispersal of the nucleolus. Little information has been elucidated thus far on nucleolar proteins that are phosphorylated by p34cdc2-cyclin B during meiotic maturation. Here, we describe a novel nucleolar protein of the starfish Asterina pectinifera oocyte, which is designated ANO39 and which is phosphorylated during meiotic maturation. A full-length ANO39 cDNA of 2106 base pairs encodes a polypeptide of 346 amino acids having a calculated Mr of 39 005. The amount of ANO39 is kept nearly constant during oocyte maturation and embryogenesis up to the midgastrula stage. The transcript encoding ANO39 was present in growing oocytes but not in full-grown ones, as evidenced by Northern blot hybridization. Ser145 is specifically phosphorylated when ANO39 is incubated in vitro with purified starfish p34cdc2-cyclin B. This phosphorylation site corresponds to that is phosphorylated during meiotic maturation in vivo. Immunoblot analysis using phosphoserine145-specific antibody as a probe revealed that some populations of ANO39 of the immature oocytes at the G2 stage have been already phosphorylated on Ser145 and Ser145 is maximally phosphorylated during meiotic maturation.

SUMMARYIntestinal intraepithelial lymphocytes (IEL) constitute the first lymphoid compartment to encounter dietary antigens and intestinal pathogens. IEL are proposed to be involved in the defence against bacterial and viral invasion and to play an important role in mucosal immunity. Fas (CD95/APO-1) is a surface receptor that induces apoptotic cell death upon ligation with Fas ligand (FasL). The aim of this study was to examine the expression and function of Fas and FasL on freshly isolated normal human colonic IEL. The expression and function of Fas and FasL on IEL isolated from 40 normal colonic specimens were examined by flow cytometry, reverse transcriptase-polymerase chain reaction, immunohistochemistry, and DNA-release cytotoxicity assay. Virtually all CD3+ IEL (95.2 ± 4.3%) expressed Fas and were sensitive to agonistic anti-Fas antibody, whereas only 56.6 ± 8.4% of peripheral T lymphocytes expressed Fas and were resistant to the antibody. We also detected FasL mRNA and protein (40.1 ± 4.2%) on IEL, and found that IEL exerted FasL-mediated cytotoxicity against Fas-expressing target cells. These findings suggest that human IEL are activated in situ but are tightly regulated by the constitutive expression of functional Fas and FasL to maintain homeostasis of the mucosal immune system.

Fas ligand (FasL) belongs to the TNF superfamily. It is induced in activated lymphocytes and eliminates Fas-positive lymphocytes, resulting in the down-regulation of immune responses. FasL has also been detected in tissues other than lymphoid cells. We investigated the expression and function of FasL on human colon cancer cells. FasL mRNA was detected by RT-PCR in all six colon cancer cell lines tested and was not found on fibroblasts. FasL protein was detected in DLD-1, LoVo, HCT-116 and RPMI 4788 cells by immunohistochemical staining. DLD-1, LoVo and WiDr were cytotoxic against mouse T lymphoma cells which were transfected with human Fas receptor cDNA. The cytotoxicity was significantly enhanced by phorbol 12-myristate 13-acetate (PMA) and ionomycin. Our data suggest that the FasL expressed in human colon cancer cells may be regulated by endogenous factors in the microenvironment of the host and facilitates the escape from the host immune system.

Research groups were formed in 21 institutions nationwide to investigate carbapenem resistance. The activities of various antibacterial agents, principally carbapenems were tested against clinical isolates collected from these institutions. The broth microdilution method was used to determine the minimum inhibitory concentrations (MIC) of 17 antibacterial agents for 1,282 strains of 11 bacterial species isolated at all institutions between October and December 1995. The results were as follows: 1. Carbapenems exhibited strong antibacterial activities against MSSA and Streptococcus pneumoniae. Their activities against Enterococcus faecalis were comparable to that of ABPC. Carbapenems showed low activities against MRSA. 2. OFLX exhibited the greatest antibacterial activity against Haemophilus influenzae, followed by MEPM. The antibacterial activities of the other carbapenems were comparable to those of FMOX and CTM. 3. The carbapenems showed high activities against Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Bacteroides fragilis group. Their activities were greater than that exhibited by other beta-lactam antibacterial agents. The carbapenems also exhibited greater antibacterial activities against Serratia marcescens than the other beta-lactam antibacterial agents, but some resistant strains were detected. 4. The antibacterial activities of carbapenems against Pseudomonas aeruginosa were comparable to those of CAZ, AZT, AMK.

Correlations between alterations of the p53 gene and clinical features were examined in childhood acute lymphoblastic leukemia (ALL). We analyzed 147 patients and 38 cell lines for p53 mutations within exons 5 to 9 (2 to 11 in some of them) by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis and direct sequencing. p53 gene mutations were found in 3 of 62 (5%) patients at diagnosis, 1 of 14 (7%) patients at relapse, and 13 of 20 (65%) cell lines in T-ALL, 2 of 20 (10%) patients at diagnosis, 4 of 4 (100%) patients at relapse, and 4 of 5 (80%) cell lines in t(1;19)-ALL, 1 of 23 (4%) patients at diagnosis, 2 of 22 (9%) patients at relapse, and 5 of 12 (42%) cell lines in common ALL other than t(1;19) or t(9;22)-ALL and 3 of 3 (100%) patients at diagnosis in B-ALL. In t(1;19)-ALL, p53 gene alterations were associated with a poor prognosis. The patients with p53 mutations had a trend towards poor prognosis in childhood ALL without B-ALL. p53 gene mutation is not always associated with the current prognostic factors. This alteration may become one of the important prognostic factors, if the detection of a small number of the leukemic cells with the p53 gene mutation would be possible.

We analyzed 60 B precursor acute lymphoblastic leukemia (ALL) primary samples and 15 cell lines for homozygous deletions of p16 and p15 genes and mutations of p16 gene. These included five cell lines and 13 primary samples with the t(1;19)(q23;pl3), and eight primary samples with the t(9;22)(q34;qll). Of 10 cell lines without t(1;19), homozygous deletion of both p16 and p15 genes was found in eight cell lines (80%), and a rearrangement of p16 in one cell line (10%). In contrast, only one (20%) of the five cell lines with t(1;19) showed homozygous deletion or rearrangement of p16/p15 gene. Thirteen of 60 (22%) primary samples demonstrated p16 gene homozygous deletion. No case with t(1;19) showed homozygous deletion of p16 gene (0/13, 0%), while cases without t(1;19) showed considerable incidence of p16 gene homozygous deletion (13/47, 28%). These results suggest that the incidence of deletions of p16 gene differs according to the subtypes of B precursor ALL. We also compared the frequency of p16 gene homozygous deletion between the patients at diagnosis and at relapse. Nine of 45 (20%) samples at diagnosis and four of 22 (18%) samples at relapse showed p16 homozygous deletions. The similarity of the rate in these two groups raises the question of the role of p16 gene in progression of B precursor ALL. Mutations were found in three of the primary cases (5%); the mutations included two nonsense mutations at codon 72 and one missense mutation at codon 98. All the mutations found in this study were heterozygous, and the clinical relevance of p16 gene mutation is yet to be determined in these case

We have investigated the mutation of the TP53 gene in hepatoblastomas (HBLs) by using polymerase chain reaction-single strand conformation polymorphism and direct sequencing in 38 HBL tumor samples and in two HBL cell lines. We detected the TP53 gene mutation in an anaplastic hepatoblastoma cell line, but no aberration of the TP53 gene (exons 5-9) was found in tumor samples and in the other HBL cell line. The mutation of the cell line was a missense mutation from GAC (asparagine) to CAC (histidine) at codon 281, which was different from the G-to-T transversion of codon 249 that is frequently found in adult hepatocellular carcinomas (HCCs). In addition, we performed Southern blot analysis of the MDM2 gene, but we did not find MDM2 gene amplification in 19 cases tested. Our results suggest that, in contrast to the findings in HCCs in adults, TP53 gene aberrations are not involved in the development or progression of HBLs in children.

Forty-one patients were treated with epidural neurolysis using 50 % ethyl alcohol 2 ml. Thirty eight patients were suffering from cancer pain and three patients were complaining of chronic benign pain. Alcohol block was repeated 2.3 times (mean) in the same patient. Thirty patients were followed after the treatment. Forty-seven percent of the patients reported 70 % or greater pain relief and 20 % of the patients reported about 50 % pain relief. Duration of pain relief was from 9 days to 203 days with a mean duration of 54 days. Adverse effects were reported 43 % of the patients. There is no miserable adverse effect. Adverse effect reported most was pain with epidural injection of drugs after the alcohol block had been performed.

Antibody titres against Plasmodium falciparum and P. vivax were examined using the indirect fluorescent antibody test (IFAT) for 183 Gidra-speaking adults and adolescents in four ecologically different villages of lowland Papua New Guinea. The findings highlight that 1) in Gidraland P. falciparum was more prevalent than P. vivax, 2) the proportion of antibody titres of 1:64 or higher markedly differed among the villages, ranging from 35.3% to 100% for males and from 31.6% to 100% for females, and 3) in the two villages with high prevalences, these were higher among males than females. The inter-village and sex differences can be largely explained by microenvironmental conditions and behavioural patterns of the population. The population-based analyses of this study intend to contribute to a better understanding of the prevalence of malaria in human-environment settings and thus to the planning of malaria prevention.

We have investigated the frequency of p53 gene mutations in Ewing's sarcoma (ES) and neuroblastoma (NB) by using polymerase chain reaction-single strand conformation polymorphism analysis for genomic DNA or complementary DNA generated from total RNA. Mutations of the p53 gene were found in six of seven ES cell lines: a missense mutation of TGC (Cys)--TAC (Try) at codon 141 in one, a missense mutation of CGT (Arg)--TGT (Cys) at codon 273 in one, a missense mutation of TGC (Cys)--TTC (Phe) at codon 176 in three, and one base deletion of CGC--CG at codon 283 in one. Further analysis of 14 ES and related primary tumors showed mutations of the p53 gene in only two: one base insertion of CCG--CCCG at codon 152 in one and a missense mutation of GGC (Gly)--GTC (Val) at codon 154 in the other. Both of the two tumors were obtained from patients with an advanced stage disease. Three of the eight ESs with mutations of the p53 gene showed the same missense mutation at codon 176, suggesting the mutational hot spot of the p53 gene in ESs. In contrast to ES, none of 6 NB cell lines or 48 NB tumors including advanced-stage ones with or without N-myc amplification showed any aberration of the p53 gene. Our findings suggest that mutations of the p53 gene in ES might represent late genetic events related to tumor progression, and that aberrations of the p53 gene might not be involved in the development or the progression of NB.

Nowadays the use of combination chemotherapy in children with acute lymphoblastic leukemia (ALL) results in a complete remission rate of more than 95%. With the best currently available treatment, about two-thirds of these children will remain in continuous complete remission and can therefore be considered cured. Patients suffering from a relapse, however, have a cure rate which is much lower. One of the main causes of this poor prognosis is probably a resistance of the leukemic cells to a number of drugs used for treatment. At present the knowledge of drug resistance in childhood ALL is very limited. It is unknown for instance for which drugs resistance occurs and which mechanisms are of clinical importance. Currently, much attention is given to the multidrug resistance (MDR) phenomenon: a resistance to vincra alkaloids and anthracyclines, mediated by the drug efflux pump p-glycoprotein (-gp), which can at least be partially overcome by so-called resistance modifiers. Recently, we and others adapted and improved assays to detect drug resistance of leukemic cells obtained from patients showing good correlations between in vitro results and clinical response to chemotherapy [1–8]. Because of the development of these short-term assays it has recently become possible to study drug resistance of patients with ALL. In the present study we assessed the resistance profiles of children with relapsed ALL and the clinical relevance of the MDR model in these patients.

Parathyroid hormone-related protein (PTHrP), which is responsible for producing hypercalcemia in patients with humoral hypercalcemia of malignancy, has recently been identified in several normal tissues. Because PTHrP, like parathyroid hormone (PTH), is known to exhibit vasodilatory properties, we investigated the expression and regulation of PTHrP mRNA in cultured rat aortic smooth muscle cells (SMC). We report here that PTHrP mRNA is expressed in SMC and is markedly induced by serum in a time- and concentration-dependent fashion. Addition of 10% fetal calf serum to serum-deprived, confluent cells, resulted in a marked induction of PTHrP mRNA by 2 h with a peak at 4-6 h. PTHrP was detected in SMC by immunocytochemistry and radioimmunoassay of conditioned medium, and was shown to be up-regulated within 24 h after the addition of serum. The serum induction of PTHrP mRNA was blocked by actinomycin D and by cycloheximide indicating the need for protein synthesis to evoke the serum effect on PTHrP gene transcription. In addition, treatment with dexamethasone, which has been previously shown to reduce the constitutive expression of PTHrP in human cancer cells, also blunted the serum induction of PTHrP mRNA in SMC. Treatment of quiescent cells with the serum mitogens platelet-derived growth factor or insulin-like growth factor-I had no effect on PTHrP, whereas the vasoactive peptides endothelin, norepinephrine and thrombin stimulated PTHrP expression. Exogenous addition of recombinant PTHrP-(1-141) had no significant effect on SMC DNA synthesis as measured by [3H]thymidine incorporation. In summary, the abundance of PTHrP mRNA and the characteristics of its regulation in SMC suggest a major role for PTHrP as a local modulator in vascular smooth muscle.

This paper described the morphological changes in the trabecular bone orientation of the jaw bone due to reduction in the applied functional pressure. All teeth on the right side of the upper and lower jaws were extracted from canine specimens. Experimental periods were set at 0.5, 1, 1.5, 2, 2.5, 3, 5, 7, 9, 11, and 13 months after tooth extraction. The trabecular bone orientation was measured by an image analyzer as morphometrical parameters. The loss of teeth changed the support system for functional pressure, resulting in the marked disordering of the trabecular bone orientation.

Lymphoblasts from 21 previously untreated patients with acute lymphoblastic leukemia (ALL) and 31 patients in relapse were tested for chemosensitivity. Blast cells were cultured with 22 anticancer drugs for 4 days and assayed by MTT dye using a scanning microplate photometer. The percent cytotoxicity index (%CI) and LD50 (micrograms/ml) were calculated for each drug. The mean absorbances (+/- S.D.) of 1 x 10(5) cells in the untreated group and relapsed groups in control wells were 0.219 (+/- 0.126) and 0.385 (+/- 0.147), respectively (p less than 0.01). Cells in the untreated group were more sensitive in vitro to vincristine, prednisolone, L-asparaginase (L-ASP), vinblastine, 5-fluorouracil, epirubicin, bleomycin (BLM), and etoposide (VP16) with respect to the %CI value and to L-ASP, VP16, BLM, and mitoxantrone with respect to the LD50 value than those in the relapsed groups. In contrast, no significant differences were observed for the other 13 drugs. There was also a significant difference in sensitivity within the relapsed group--13 having good clinical response and 15 showing no response to chemotherapy--with regard to four drugs, mitomycin C, neocarzinostatin, L-ASP, and teniposide. Cells in the relapsed group had more heterogeneous chemosensitivity than those in the untreated group, and divided into sensitive and resistant types, but large interindividual differences existed. The MTT assay and LD50-drug resistance percentile curves are useful for the selection of effective drugs in both untreated and relapsed patients with acute leukemia.

The capacity of the v-myc-transformed, chicken myelomonocytic cell line HD-11 to metabolize 25-hydroxyvitamin D3 (25-OHD3) was examined. HD-11 cells produced and secreted a metabolite of 25-OHD3 that was bound with high affinity by receptor for 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. On normal-phase HPLC, this metabolite cochromatographed with authentic 1,25-(OH)2D3 in both hexane- and methylene chloride-based solvent systems. The 25-OHD3 1-hydroxylation reaction was substrate saturable with a Km of 73 nM 25-OHD3 and a maximal velocity of 167 fmol per 10(6) cells per h. This reaction was inhibited by ketoconazole, a recognized inhibitor of cytochrome P450 mixed-function oxidases including the authentic, renal 25-OHD3 1-hydroxylase. On the other hand, HD-11 cell 1,25-(OH)2D3 production was not affected by the antioxidant DPPD, a known inhibitor of free radical-generated 1,25-(OH)2D3. In addition to synthesizing 1,25-(OH)2D3, this monocyte-macrophage cell line also has the potential to be a target for the hormone; HD-11 cells express high-affinity receptor for 1,25-(OH)2D3 (Kin = 0.06 nM).

We applied the MTT dye reduction assay to the anti-cancer drug sensitivity test using short-term microplate cultures. Blast cells were cultured with approximately 25 anti-cancer drugs for 4 days. After cultivation, MTT dye was placed in each well, and the formazans generated by living cells were dissolved in acidified isopropyl alcohol. The absorbance of each well was measured at a scanning microplate photometer. When we made the table of the cytotoxicity index (CI) that was classified into anti-cancer drugs and concentrations for each leukemic sample, it was possible to compare efficacy with different drugs and to select the effective ones. Retrospectively, the in vitro results were compared with the clinical responses of the 34 patients (26 of acute lymphocytic leukemia [ALL] and eight of acute nonlymphoblastic leukemia [ANLL]) who were treated by combination chemotherapy. The following results were obtained: true-positive rate, 78.1%; true-negative rate, 57.1%; and predictive accuracy, 74.4%. Therefore, the MTT assay-CI table might serve as a reliable tool for the selection of effective chemotherapy in patients with acute leukemia.

A (H+ + K+)-ATPase-enriched membrane fraction derived from the fundic portion of hog gastric mucosa was obtained by a combination of differential and repeated 7% Ficoll gradient centrifugation. The microsomal membrane fraction isolated by repeated 7% Ficoll gradient centrifugation was free of ouabain-sensitive (Na+ + K+)-ATPase, 5'-nucleotidase and succinate dehydrogenase; and it was highly enriched in (H+ + K+)-ATPase and K(+)-stimulated p-nitrophenylphosphatase (p-NPPase). The (H+ + K+)-ATPase had a pH optimum of 7.4 and was stimulated by Tl+, K+, Rb+ and NH4+ with Ka values of 0.0667, 0.526, 0.667 and 3.03 mM, respectively, at this pH. On the other hand, monovalent cations such as Na+, Li+ and (CH3)4N+ as well as divalent cations such as Cu2+, Ca2+, Ba2+, Sr2+ and Cd2+ inhibited this enzyme activity concentration-dependently. Ouabain and oligomycin had no effect, whereas omeprazole, a specific (H+ + K+)-ATPase inhibitor, inhibited this enzyme activity in a pH-dependent manner. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis showed a major band (greater than or equal to 90% of protein) at 97,400 daltons, which was phosphorylated in the presence of Mg2+ and [gamma-32P]-ATP and dephosphorylated in the presence of K+. The present method was very simple, and the (H+ + K+)-ATPase activity of the microsomal fraction obtained by this method was much higher compared with those obtained by other methods such as free-flow electrophoresis.

External measurements of the zygomatic bone have been reported, but there are no reports on the internal structure. We studied the internal structure of the zygomatic bone and determined the changes in cortical bone width, cortical bone ratio to total cross-sectional area, and trabecular bone width through six periods of tooth growth and development. Zygomatic bone were fixed in resin and sliced, then the cross-sectional specimens were photographed with soft X-rays. The images were analyzed with an image processor. The specimens included the surrounding cortical bone and its internal spongy substance. The cortical bone width, cortical bone ratio, and trabecular bone width increased significantly from the period of deciduous dentition to the first early period of mixed dentition, but leveled off after that. This indicates that in addition to the growth and development of the zygomatic bone itself, its structure is also affected by the forces generated in the maxilla by mastication.