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Background:In acute encephalopathy, deterioration of the condition can be rapid, and early intervention is essential to prevent progression of the disease. However, in the acute period, differentiating acute encephalopathy from febrile seizures is difficult. Thus, an early diagnostic marker has been sought to enable early intervention. Proton nuclear magnetic resonance (1H NMR) spectroscopy is used to study the chemical characteristics of biological fluids such as cerebrospinal fluid (CSF). The purpose of this study was to ascertain if pattern recognition of 1H NMR spectra could differentiate CSF obtained from patients with acute encephalopathy and febrile seizures.Methods:CSF was obtained from patients with acute encephalopathy (n = 4), complex febrile seizures (n = 9), and simple febrile seizures (n = 9).Results:NMR spectra of CSF did not visually differ across the three groups. Spectral data were analyzed by partial least squares discriminant analysis and visualized by plotting the partial least squares scores of each sample. The three patient groups clustered separately on the plots.Conclusion:In this preliminary study, we were able to visualize different characteristics of CSF obtained from patients with acute encephalopathy and simple and complex febrile seizures using pattern recognition analysis of 1H NMR data.

OBJECTIVE: Osteonecrosis of the femoral head (ONF) is a necrosis due to disruption of the blood flow. The disease often occurs in association with corticosteroid treatment. The pathology of corticosteroid-induced ONF is unclear, although abnormalities in the coagulation and fibrinolytic systems or in the lipid metabolism have been reported to be involved. We examined the relationships between development of ONF and genetic variations and plasma level of lipoprotein(a) (Lp(a)), which is closely involved in the coagulation and fibrinolytic systems and lipid metabolism. METHODS: The study population consisted of 112 renal transplant patients undergoing corticosteroid treatment. Their apolipoprotein (a) [apo(a)] isoform was determined by Western blotting, and patients were classified into low molecular weight (LMW) or high molecular weight (HMW) groups. The plasma Lp(a) level was measured. Patients were also examined for 3 single-nucleotide polymorphisms (SNP), -773 (G/A), +93 (C/T), and +121 (G/A). Relationships between these 3 genetic factors of Lp(a) and ONF development were examined using statistical methods including multivariate analysis. RESULTS: A strong relationship was observed between the apo(a) molecular weight phenotype and ONF development, with an increased risk of ONF development for the LMW group (adjusted odds ratio 5.75, 95% CI 1.76-18.74, p = 0.0038). No significant relationships were observed between ONF and plasma Lp(a) level and SNP. CONCLUSION: Apo(a) molecular weight phenotype would be a useful predictor of ONF that develops after corticosteroid treatment.

ACEI/D and AT1R1166A/C polymorphisms are considered to comprise individual risk factors for the development of coronary disease. We sought to demonstrate that the ACEI/D and AT1R1166A/C polymorphisms affect coronary artery stenosis in patients with Kawasaki disease (KD). We examined 147 healthy controls and 281 Japanese children with KD. The patients were further divided into group N (n246, no ischemia) and group I (n35, severe coronary artery stenosis with myocardial ischemia), and we studied the genotype of ACEI/D and AT1R1166A/C polymorphisms. We also examined ACE activity in patients with acute KD. We did not detect any prevalent genotypes of the ACEand AT1Rpolymorphisms between controls and KD patients. However, the prevalence of the D allele in the ACEpolymorphism and of the C allele in the AT1Rpolymorphism tended to be higher in group I than in group N (odds ratios, 2.00 and 2.32, respectively). In addition, the presence of the D and/or C alleles significantly increased the relative risk of developing myocardial ischemia (odds ratio, 2.71; p0.038). During the convalescent phase of KD, ACE activity was increased despite significant attenuation during the acute phase. These results suggested that the renin-angiotensin system is associated with the formation of severe coronary artery stenosis and myocardial ischemia.

ACE I/D and AT1R 1166A/C polymorphisms are considered to comprise individual risk factors for the development of coronary disease. We sought to demonstrate that the ACE I/D and AT1R 1166A/C polymorphisms affect coronary artery stenosis in patients with Kawasaki disease (KD). We examined 147 healthy controls and 281 Japanese children with KD. The patients were further divided into group N (n = 246, no ischemia) and group I (n = 35, severe coronary artery stenosis with myocardial ischemia), and we studied the genotype of ACE I/D and AT1R 1166A/C polymorphisms. We also examined ACE activity in patients with acute KD. We did not detect any prevalent genotypes of the ACE and AT1R polymorphisms between controls and KD patients. However, the prevalence of the D allele in the ACE polymorphism and of the C allele in the AT1R polymorphism tended to be higher in group I than in group N (odds ratios, 2.00 and 2.32, respectively). In addition, the presence of the D and/or C alleles significantly increased the relative risk of developing myocardial ischemia (odds ratio, 2.71; p = 0.038). During the convalescent phase of KD, ACE activity was increased despite significant attenuation during the acute phase. These results suggested that the renin-angiotensin system is associated with the formation of severe coronary artery stenosis and myocardial ischemia.

Nontraumatic osteonecrosis of the femoral head (ONFH) is one of the complications that may occur after renal transplantation. We investigated the relationship between the incidence of ONFH and polymorphisms in the genes for plasminogen activator inhibitor (PAI)-1, which is one of the major regulatory proteins of the fibrinolytic system, and 5,10-methylenetetrahydrofolate reductase (MTHFR), which is associated with the plasma levels of homocysteine in Japanese subjects.

We investigated risk factors for osteonecrosis of the femoral head (ONF) in renal transplant recipients, who are susceptible to the disease. Among 287 renal transplant recipients, 18 ONF patients with enough data were included, and 18 age- and sex-matched recipients without ONF were nominated as reference cases. Risk factors were analyzed using a conditional logistic regression method. There were no differences between the ONF patients and the reference cases regarding the types of immunosuppressant or the donor (living or cadaveric, father or mother, matching blood type and human leukocyte antigens). The daily oral steroid dosage (prednisolone 25.0 mg/day or more) and blood urea nitrogen level 2 months after transplantation were the only factors with relevance to the occurrence of ONF. We propose that oral steroid dosages should be low or reduced after renal transplantation, and acute rejection should be controlled with pulsed therapy.

We investigated risk factors for osteonecrosis of the femoral head (ONF) in renal transplant recipients, who are susceptible to the disease. Among 287 renal transplant recipients, 18 ONF patients with enough data were included, and 18 age- and sex-matched recipients without ONF were nominated as reference cases. Risk factors were analyzed using a conditional logistic regression method. There were no differences between the ONF patients and the reference cases regarding the types of immunosuppressant or the donor (living or cadaveric, father or mother, matching blood type and human leukocyte antigens). The daily oral steroid dosage (prednisolone 25.0?mg/day or more) and blood urea nitrogen level 2 months after transplantation were the only factors with relevance to the occurrence of ONF. We propose that oral steroid dosages should be low or reduced after renal transplantation, and acute rejection should be controlled with pulsed therapy.

Advances in transplantation technology have brought about great benefits to patients suffering from organ failure, but the problem still remains of complications induced by steroids used for post-transplant immunosuppression. Among the side-effects caused by steroids, non-traumatic osteonecrosis of the femoral head (ONF) constitutes a serious problem. The same protocol for steroid administration induces ONF in some patients, but not in others, indicating the presence of individual difference in steroid sensitivity. We hypothesized that this difference might be mediated by the drug-transport protein, P-glycoprotein (P-gp), and investigated the relationship between single nucleotide polymorphisms in the multidrug resistance gene 1 (ABCB1, MDR1) encoding P-gp and ONF. Subjects comprised 136 patients receiving kidney transplantation. Thirty patients developed post-transplant ONF. Genomic DNA was extracted from peripheral blood, and genotypes of ABCB1C3435T (exon 26) and G2677T/A (exon 21) were determined by direct sequencing. Multivariate analyses based on clinical information were performed to determine the relationship between ABCB1genotypes and ONF. The dose/concentration (D/C) ratios of tacrolimus were also determined to estimate the activity of P-gp in patients with different genotypes of ABCB1C3435T (CC, CT, TT), and in those who did and did not develop ONF. The ABCB13435TT genotype showed a significantly lower incidence of ONF (adjusted odds ratio = 0.10, P= 0.034). The D/C ratio in the 3435TT genotype was significantly higher than that in the 3435CC genotype. The D/C ratio in patients developing ONF was significantly higher than in those patients who did not develop ONF. The results suggest increased activity of P-gp in patients with the 3435TT genotype and in those who did not develop ONF. The ABCB12677 homozygous variant type also showed a lower incidence of ONF (adjusted odds ratio = 0.26, P= 0.056). The 3435T and 3435C alleles were in linkage disequilibrium with the 2677T and the 2677G alleles, respectively, in the study population. An assessment of C3435T and G2677T/A polymorphisms preceding steroid treatment could be useful for predicting the resistance to ONF development.

We radiographically investigated 36 hips in 34 patients with osteoarthritis of the hip who had undergone total hip arthroplasty. Their mean age was 59.2 years (range 36–79 years), and the mean follow-up period was 11.2 years (range 10–14 years). The long-term outcome and the chronological changes in the bulk autograft were examined. The acetabular component of the prosthesis was a Lord-type threaded cup with a smooth surface. At follow-up, bone absorption was minor in 17 joints, moderate in 11, and major in 8. The hips with graft coverage of ≥20% (group A) had a significantly higher loosening rate than hips with coverage of <20% (group B) (P< 0.05). The cup position changed markedly in group A. Our findings indicate that graft coverage should be less than 20% when a bulk graft is used together with a smooth-surfaced cementless cup.

We radiographically investigated 36 hips in 34 patients with osteoarthritis of the hip who had undergone total hip arthroplasty. Their mean age was 59.2 years (range 36–79 years), and the mean follow-up period was 11.2 years (range 10–14 years). The long-term outcome and the chronological changes in the bulk autograft were examined. The acetabular component of the prosthesis was a Lord-type threaded cup with a smooth surface. At follow-up, bone absorption was minor in 17 joints, moderate in 11, and major in 8. The hips with graft coverage of ?20% (group A) had a significantly higher loosening rate than hips with coverage of «20% (group B) ( P « 0.05). The cup position changed markedly in group A. Our findings indicate that graft coverage should be less than 20% when a bulk graft is used together with a smooth-surfaced cementless cup.

With advanced organ transplantation technology, steroid-induced osteonecrosis of the femoral head (ONF) is one of the most troublesome complications. Steroid sensitivity varies among individuals, and the involvement of polymorphism of various genes relating to steroid metabolism is suggested. The present study investigated the relation between single nucleotide polymorphism (SNP) on the DNA sequence of cytochrome P450and ONF development. The subjects were 80 renal transplant patients. Genome DNA was obtained from the peripheral blood, and SNP analyses for CYP3A4, CYP2D6, and CYP2C19 were conducted using various methods: direct sequencing, polymerase chain reaction restriction fragment length polymorphism, and DNA Chip. The relation between ONF development and SNP was statistically analyzed. It is useful if the generating risk judged according to SNPs to prevention of steroid-induced ONF is possible. This time, SNPs, which are clearly related to ONF, were not accepted, although the possibility that SNP related to steroid metabolism is involved in ONF development is important. We think that it is necessary to examine this area more closely.

Abstract:  With advanced organ transplantation technology, steroid-induced osteonecrosis of the femoral head (ONF) is one of the most troublesome complications. Steroid sensitivity varies among individuals, and the involvement of polymorphism of various genes relating to steroid metabolism is suggested. The present study investigated the relation between single nucleotide polymorphism (SNP) on the DNA sequence of cytochrome P450 and ONF development. The subjects were 80 renal transplant patients. Genome DNA was obtained from the peripheral blood, and SNP analyses for CYP3A4, CYP2D6, and CYP2C19 were conducted using various methods: direct sequencing, polymerase chain reaction restriction fragment length polymorphism, and DNA Chip. The relation between ONF development and SNP was statistically analyzed. It is useful if the generating risk judged according to SNPs to prevention of steroid-induced ONF is possible. This time, SNPs, which are clearly related to ONF, were not accepted, although the possibility that SNP related to steroid metabolism is involved in ONF development is important. We think that it is necessary to examine this area more closely.

Bacillus circulansIAM1165 produces isoforms of β-1,3-glucan-hydrolases. Of these enzymes, the 42-kDa enzyme BglM degrades Aspergillus oryzaecell walls the most actively. A gene coding for a BglM precursor consisting of 411 amino acid residues was cloned. The 27 N-terminal amino acid sequence of the precursor is a signal peptide. The 141 C-terminal amino acid sequence showed a motif of carbohydrate-binding module family 13. This domain bound to pachyman, lichenan, and A. oryzaecell walls. The central domain showed a bacterial β-1,3-glucan-hydrolase motif belonging to glycosyl hydrolase family 16. By removal of the C-terminal domain in the IAM1165 culture, mature BglM was processed to several 27-kDa fragments that hydrolyze a soluble β-1,3-glucan.

A cDNA encoding for human monocyte chemotactic and activating factor (MCAF) was ligated into the retroviral vector pLXSN. These pMCAF‐LXSN and antisense p‐antiMCAF‐LXSN vectors were transfected into HBT20 and HBT28 human brain tumor cells. HBT28 cells constitutively express high amounts of MCAF, whereas HBT20 cells express much less MCAF.HBT20 cells transfected with pMCAF‐LXSN (HBT20‐MCAF) showed significantly higher MCAFmRNA expression and MCAF protein production than the HBT20‐parent or HBT20 cells transfected with control vector (HBT20‐LXSN). In contrast, supernatant from HBT28 cells transfected with p‐antiMCAF‐LXSN (HBT28‐antiMCAF) contained less MCAFthan HBT28‐parent, HBT28‐LXSN, and HBT28‐MCAFcells. Activated human monocytes killed HBT20‐MCAFcells more efficiently compared with HBT20‐parent, HBT20‐LXSN, and HBT20‐antiMCAFcells (P< 0.02), whereas HBT28‐antiMCAFcells were killed more efficiently by activated monocytes compared with HBT28‐parent, HBT28‐LXSN, and HBT28‐MCAFcells (P< 0.05). Cultured supernatants from activated monocytes plus HBT20‐MCAFcells or from activated monocytes plus HBT28‐antiMCAFcells inhibited the growth of HBT20 and HBT28 cells, respectively. Altered MCAFexpression can therefore enhance the ability of activated monocytes to kill brain tumor cells. This increased cytotoxicity is partially dependent upon the basal state of MCAFin the individual tumor cells.

Liposome-encapsulated muramyl tripeptide phosphatidylethanolamine (L-MTP-PE) is a novel immune modulator that is now under investigation against metastatic melanoma and osteosarcoma. We have already reported that L-MTP-PE induced monocyte-mediated tumoricidal activity and up-regulation of the tumor necrosis factor and interleukin-1 (IL-1) in vivo and in vitro. We now demonstrate that L-MTP-PE also induces monocyte chemotactic and activating factor (MCAF) mRNA expression at both the transcriptional and post-transcriptional levels. Monocyte chemotactic activity was also present in the supernatants of L-MTP-PE-stimulated cells. In monocytes, the increased expression of MCAF was induced rapidly (by 2 h) but was short-lived. By 4 h, MCAF mRNA had decreased to background level. We found no change in MCAF mRNA levels in lymphocytes exposed to L-MTP-PE. We therefore conclude that L-MTP-PE selectively up-regulates MCAF expression in monocytes and that MCAF may play a role in the tumoricidal and immune-stimulating activity of L-MTP-PE.

The purpose of this study was to examine the mechanisms by which liposome-encapsulated muramyl tripeptide phosphatidylethanolamine (L-MTP-PE) stimulates monocytes to produce tumor necrosis factor (TNF) and interleukin-1 (IL-1). We have previously shown that secretion of TNF protein occurred 2–4 h following incubation of monocytes with L-MTP-PE and that this stimulation of TNF production was associated with an increase in TNF mRNA. Increased intracellular interleukin-1a (IL-1a) and IL-1ß were not detected until 8 h after exposure to L-MTP-PE. To determine whether TNF played a role in the stimulation of IL-1 production by L-MTP-PE, normal human monocytes were incubated with L-MTP-PE or medium in the presence or absence of anti-TNF or anti IL-1a plus anti IL-1ß. Enhanced expression of IL-1a and IL-1ß mRNA was inhibited at 4 h but not 24 h when monocytes were incubated with L-MTP-PE plus anti-TNF compared with L-MTP-PE alone. By contrast, enhanced expression of TNF mRNA wasnot inhibited at any time when monocytes were incubated with L-MTP-PE and anti-IL-1a plus anti-IL-1ß. These data indicate that the up-regulation of IL-1 seen in monocytes following L-MTP-PE exposure may be due in part to the production of TNF. The up-regulation of TNF, however, appears to be independent of IL-1 production.

SummaryA 7-year-old boy with dysmorphic features was found to have a recombinant chromosome 18, rec(18), resulting from meiotic recombination of a maternal pericentric inversion, inv(18) (p11.2q21.3), as defined by high-resolution banding. He was trisomic for the long arm (q21.3-qter) and monosomic for the short arm (p11.2-pter) of chromosome 18. His clinical features were compared with those in other rec(18) cases, and also those in monosomy 18p, trisomy 18qter and full trisomy 18 syndromes. The risk of recombinant formation for inv(18) carriers was also discussed.

We have investigated the possibility of overcoming the resistance of human brain tumour cells (HBT20) to etoposide by transferring the normal human topoisomerase IIα (H-topo II) gene into these cells. H-topo II in a mammalian expression vector containing a glucocorticoid-inducible mouse mammary tumour virus (MMTV) promoter was transfected into etoposide-resistant HBT20 cells (HBT20-hTOP2MAM). HBT20 cells transfected with pMAMneo vector alone served as control cells (HBT20-MAM). These were stable transfections. Following a 2 h dexamethasone treatment, H-topo II mRNA expression, protein production, etoposide-induced DNA-protein complex formation and sensitivity to etoposide were increased in HBT20-hTOP2MAM cells compared with control HBT20-MAM cells and with HBT20-hTOP2MAM cells not treated with dexamethasone. However, mRNA and protein levels and cell sensitivity returned to baseline when incubation with dexamethasone was continued for 24 h. This decrease from the 2 h values could not be explained by a loss of the MMTV promoter response to dexamethasone. (H-topo IIα promoter)-(chloramphenicol acetyltransferase) constructs containing regions -559–0 and -2400–0 were significantly down-regulated in HBT20-hTOP2MAM cells treated for 24 h with dexamethasone compared with dexamethasone-treated control cells. H-topo II mRNA stability after 24 h of dexamethasone treatment was not altered compared with that in control cells. Our data indicate that the exogenously produced H-topo II may have a negative-feedback effect on the endogenous topoisomerase II promoter, causing down-regulation of the endogenous gene.

Liposomal muramyl tripeptide phosphatidylethanolamine (L-MTP-PE) is a biological agent in phase I and II trials for osteosarcoma and melanoma. Its mechanism of action has been linked to its ability to activate monocyte tumoricidal function and to stimulate monocyte production of tumor necrosis factor (TNF) and interleukins(IL)-1, -6, and -8. Our ultimate goal is to combine L-MTP-PE with chemotherapy. The purpose of this study was to determine whether doxorubicin (Adriamycin) interfered with the ability of L-MTP-PE to activate monocyte cytokine production. Human monocytes were cultured with or without 5–500 ng/ml of Adriamycin for 3 h and washed before being exposed to 2 µg/ml L-MTP-PE for 16 h. Cultured supernatants were collected and assayed for TNF, IL-1, IL-6, and IL-8. The messenger RNA expression of IL-1a, IL-1ß, TNFa, IL-6, and IL-8 was quantified with northern blot analysis. Adriamycin did not suppress the upregulation of any of these cytokines. We concluded that combination therapy with L-MTP-PE and Adriamycin is feasible and that this combination warrents further investigation in a clinical setting.

We report a case of infantile acute monocytic leukemia associated with solid tumors in the skin. Light microscopy showed that the tumor cells were mainly spindle-shaped and arranged in tandem. Immunohistochemical staining showed that fibronectin was detected in the extracellular matrix (and partially on the surface of tumor cells), integrin α5β1 on the cell surface, and vinculin and actin were detected in the cytoplasm of the tumor cells. Electron microscopy showed that the tumor cells had perpendicular transmembranous fibrils and closely situated collections of cytoplasmic microfilaments beneath the cell membrane suggesting fibronexus-like structures. We conclude that the expression of fibronectin, integrin α5β1, vinculin, cytoplasmic actin and fibronexus-like structure in leukemic cells indicate these cells possess an adhesive function. The mechanism of formation of the extramedullar solid tumors in this patients involved these adhesion molecules of the tumor cells.

No abstract

Ink-jet printable thin-film transistors (TFTs) on flexible plastic substrates are an important focus of research because present silicon-based electronics cannot realize such devices. In the present study, we fabricated single-walled carbon nanotube (SWCNT) TFTs on plastic substrates using the ink-jet printing method, and realized high-on/off current ratio (?104) and flexibility, respectively. The present study therefore represents a major step towards "flexible SWCNT electronics".

For Abstract see ChemInform Abstract in Full Text.