Acute liver damage and interaction of cisplatin with silver nanoparticles depend on particle size.

Nanomaterials are innovative materials that have novel properties that differ from those of macroscale materials in terms of response to stimuli such as heat, light, and voltage. However, the potential unknown effects of nanomaterials on living organisms have raised concerns. There are few reports describing the effects of silver nanoparticles on living organisms and the effects of nanoparticle interactions with chemical substances such as pharmaceuticals. Previously, we investigated the effects of silver nanoparticles on living organisms and their interactions with drugs. In that study, silver nanoparticles with a particle size of 10 nm induced acute liver injury, and silver nanoparticles with a particle size of 10, 50, or 200 nm interacted with drugs when administered to mice via the tail vein. Therefore, to investigate the relationship between the particle size of silver nanoparticles and degree of injury, we examined silver nanoparticles of 5, 10, 20, 30, 40, and 50 nm and the extent of acute liver injury and liver injury due to interactions with drugs. We found that silver nanoparticles ≤30 nm in size induced acute liver injury. Silver nanoparticles with a 5-nm particle size induced the most severe liver injury.

Emergence of resistant variants detected by ultra-deep sequencing after asunaprevir and daclatasvir combination therapy in patients infected with hepatitis C virus genotype 1.

Daclatasvir (DCV) and asunaprevir (ASV) are NS5A and NS3 protease-targeted antivirals respectively, currently under development for the treatment of chronic hepatitis C virus (HCV) infection. We analysed the relationship between pre-existing drug-resistant variants and clinical outcome of the combination treatment with DCV and ASV. Ten patients with HCV genotype 1b were orally treated with a combination of ASV and DCV for 24 weeks. The frequencies of amino acid (aa) variants at NS3 aa positions 155, 156 and 168 and at NS5A aa31 and 93 before and after treatment were analysed by ultra-deep sequencing. We established a minimum variant frequency threshold of 0.3% based on plasmid sequencing. Sustained virological response (SVR) was achieved in 8 out of 10 patients (80%), and relapse of HCV RNA after cessation of the treatment and viral breakthrough occurred in the other two patients. Pre-existing DCV-resistant variants (L31V/M and/or Y93H; 0.9-99.4%) were detected in three out of eight patients who achieved SVR. Pre-existing DCV-resistant variants were detected in a relapsed patient (L31M, Y93H) and in a patient with viral breakthrough (Y93H); however, no ASV-resistant variants were detected. In these patients, HCV RNA rebounded with ASV- and DCV- double resistant variants (NS3 D168A/V plus NS5A L31M and Y93H). While pre-existing DCV-resistant variants might contribute to viral breakthrough in DCV and ASV combination therapy, the effectiveness of prediction of the outcome of therapy based on ultra-deep sequence analysis of pre-existing resistant variants appears limited.

In patients with ulcerative colitis, adsorptive depletion of granulocytes and monocytes impacts mucosal level of neutrophils and clinically is most effective in steroid naïve patients.

BACKGROUND: The aetiology of ulcerative colitis is inadequately understood, and drug therapy has been empirical rather than based on sound understanding of disease aetiology. This has been a major factor for refractoriness and adverse drug effects as additional complications. However, ulcerative colitis by its very nature is exacerbated and perpetuated by inflammatory cytokines, which are released by peripheral granulocytes and monocytes as well. Additionally, active ulcerative colitis is often associated with elevated peripheral granulocytes and monocytes with activation behaviour and are found in vast numbers within the colonic mucosa. Hence, from the clinicopathologic viewpoint, granulocytes and monocytes are appropriate targets for therapy in ulcerative colitis. Based on this thinking, an Adacolumn has been developed for depleting excess granulocytes and monocytes by adsorption. METHODS: By colonoscopy, biopsy and histology, we investigated the impact of granulocyte and monocyte adsorption (GMA) on the mucosal level of granulocytes and monocytes in patients with active ulcerative colitis. Forty-five patients (26 steroid naïve and 19 steroid-dependent), mean age 44.7 yr, were included. Twenty patients had total colitis and 25 had left-sided colitis. Each patient was given up to 11 GMA sessions over 12 weeks. No patient received additional medications within 4 weeks (steroid) to 8 weeks (other immunosuppressants) prior to entry or during the GMA course. Colonoscopy together with biopsy was done at entry and within 2 weeks after the last GMA session. RESULTS: At entry, the mean clinical activity index was 12.6; range 10-16. A total of 400 colonic biopsies were examined, which revealed massive infiltration of the colonic mucosa by granulocytes, and GMA was associated with striking reduction of granulocytes in the mucosa. At week 12, 33 of 45 patients (73.3%, P<0.01) had achieved clinical remission (the mean clinical activity index

A probative approach for noninvasive evaluation of airway hyperresponsiveness and remodeling in adult asthmatics.

We propose a probative approach for noninvasive evaluation of airway hyperresponsiveness (AHR) and remodeling to investigate their outcome in adult asthmatics treated according to the Global Initiative for Asthma (GINA) guideline. Pulmonary function and AHR to methacholine were measured twice with an interval of 24.3 +/- 3.4 months in 18 adult asthmatics during the ongoing treatments. Mathematical formulas previously used in an animal model were applied in human asthmatics to eliminate the effect of airway wall thickening on respiratory resistance (Rrs), calculating indices for the proportional changes with time in airway wall thickness (PW(1)/PW(0)) and airway smooth muscle shortening (PMS(1)/PMS(0)), respectively. The minimum cumulative dose of methacholine (Dmin), an ordinary index of AHR measured with the oscillometry Asthograph, correlated with the asthma severity. The disease periods significantly correlated with the indices of airflow limitation. While there was no change in PW(1)/PW(0) (1.00 +/- 0.07) during the assessment periods, methacholine-induced airway smooth muscle shortening was attenuated by 46% (PMS(1)/PMS(0)=0.54 +/- 0.16). Less improvement in PMS(1)/PMS(0) was seen with a correlation to the disease periods, but PMS(1)/PMS(0) improved correlating to the relative length of the assessment period with ongoing treatments in the disease period. In conclusion, this probative approach may be useful to investigate the outcome of AHR and remodeling in human asthmatics, and shows that remodeling may get worse with time or may halt and AHR may improve with a stepwise, early intervention and prolonged treatment given according to the GINA guideline.

Y-chromosomal STR haplotype in the Japanese population.

Allele and haplotype frequencies of seven Y-chromosome STR loci from samples of 108 unrelated Japanese males living in Aichi Prefecture.

Application of nitric oxide for a case of veno-occlusive disease after peripheral blood stem cell transplantation.

A 5-year-old girl at high risk for acute lymphoblastic leukemia was treated with high-dose chemotherapy and autologous peripheral blood stem cell transplantation (PBSCT). However, her condition was complicated by veno-occlusive disease of the liver (VOD) after PBSCT. For treatment of VOD, transdermal isosorbide tape was applied as a nitric oxide (NO) donor. The signs of VOD improved immediately after NO treatment was initiated, and the patient showed no side effects from the transdermal isosorbide tape.

NK-1 receptor desensitization and neutral endopeptidase terminate SP-induced pancreatic plasma extravasation.

Substance P (SP) induces plasma extravasation and neutrophil infiltration by activating the neurokinin-1 receptor (NK1-R). We characterized the mechanisms regulating this response in the rat pancreas. Anesthetized rats were continuously infused with SP, and plasma extravasation was quantified using Evans blue (EB) dye. Continuous infusion of SP (8 nmol. kg(-1). h(-1)) resulted in a threshold increase in EB at 15 min, a peak effect at 30 min (150% increase), and a return to baseline by 60 min. The NK1-R antagonist CP-96,345 blocked SP-induced plasma extravasation. After 60 min, the NK1-R was desensitized to agonist challenge. Resensitization was first detected at 20 min and increased until full recovery was seen at 30 min. Inhibition of the cell-surface protease neutral endopeptidase (NEP) by phosphoramidon potentiated the effect of exogenous SP; therefore endogenous NEP attenuates SP-induced plasma extravasation. Thus the continuous infusion of SP stimulates plasma extravasation in the rat pancreas via activation of the NK1-R, and these effects are terminated by both desensitization of the NK1-R and the cell-surface protease NEP.

Substance P is a determinant of lethality in diet-induced hemorrhagic pancreatitis in mice.

BACKGROUND: The neuropeptide substance P (SP) induces plasma extravasation and neutrophil infiltration by activating the neurokinin 1-receptor (NK1-R). SP-induced neurogenic inflammation is terminated by the cell surface enzyme neutral endopeptidase (NEP), which degrades SP. We determined whether genetic deletion of the NK1-R reduces mortality and, conversely, whether genetic deletion of NEP increases mortality in a lethal model of hemorrhagic pancreatitis. METHODS: Necrotizing pancreatitis was induced by feeding mice a diet deficient in choline and supplemented with ethionine. We determined the length of survival, the severity of pancreatitis (by measuring the neutrophil enzyme myeloperoxidase [MPO] and by histologic evaluation), and the severity of pancreatitis-associated lung injury (lung MPO and histology) in NK1-R (+/+)/(-/-) and NEP (+/+)/(-/-) mice. RESULTS: Genetic deletion of the NK1-R significantly improved survival (100% vs 8% at 120 hours, P <.001) and reduced pancreatic MPO and acinar cell necrosis. Conversely, genetic deletion of NEP significantly worsened survival (0% vs 90% at 120 hours, P <.001) and exacerbated pancreatic MPO and pancreatitis-associated lung injury. CONCLUSIONS: Substance P is an important determinant of lethality in this model of necrotizing pancreatitis. Defects in NEP expression could lead to uncontrolled inflammation.

Nutritional assessment of peripheral blood stem cell transplantation in children.

Peripheral blood stem cell transplantation (PBSCT) has many advantages for patients because hematopoiesis and general condition return to normal more rapidly than they do following bone marrow transplantation. Thus, the authors hypothesize that the nutritional condition of patients also returns to normal more rapidly after PBSCT. The duration of insufficient nutrition was investigated in children undergoing PBSCT. The subjects of this study were 8 patients with malignant diseases. The factors measured were body weight, body fat, cholesterol, albumin, pre-albumin, and retinol-binding protein. These parameters were measured a day before transplantation, and then once a week for 4 weeks after transplantation. All parameters were recovered until day 28 from the lowest level in transplantation. In this study, all parameters returned to normal comparatively early. PBSCT causes little damage to patients' nutrition.

Substance P mediates inflammatory oedema in acute pancreatitis via activation of the neurokinin-1 receptor in rats and mice.

Pancreatic oedema occurs early in the development of acute pancreatitis, and the overall extent of fluid loss correlates with disease severity. The tachykinin substance P (SP) is released from sensory nerves, binds to the neurokinin-1 receptor (NK1-R) on endothelial cells and induces plasma extravasation, oedema, and neutrophil infiltration, a process termed neurogenic inflammation. We sought to determine the importance of neurogenic mechanisms in acute pancreatitis. Pancreatic plasma extravasation was measured using the intravascular tracers Evans blue and Monastral blue after administration of specific NK1-R agonists/antagonists in rats and NK1-R(+/+)/(-/-) mice. The effects of NK1-R genetic deletion/antagonism on pancreatic plasma extravasation, amylase, myeloperoxidase (MPO), and histology in cerulein-induced pancreatitis were characterized. In rats, both SP and the NK1-R selective agonist [Sar(9) Met(O(2))(11)]SP stimulated pancreatic plasma extravasation, and this response was blocked by the NK1-R antagonist CP 96,345. Selective agonists of the NK-2 or NK-3 receptors had no effect. In rats, cerulein stimulated pancreatic plasma extravasation and serum amylase. These responses were blocked by the NK1-R antagonist CP 96,345. In wildtype mice, SP induced plasma extravasation while SP had no effect in NK1-R knockout mice. In NK1-R knockout mice, the effects of cerulein on pancreatic plasma extravasation and hyperamylasemia were reduced by 60%, and pancreatic MPO by 75%, as compared to wildtype animals. Neurogenic mechanisms of inflammation are important in the development of inflammatory oedema in acute interstitial pancreatitis.

Protection of hematopoietic progenitor cells by nitric oxide.

The free radical gas nitric oxide (NO) has been reported to trigger apoptosis of a variety of cell types, but there have also been contradictory reports of an inhibitory effect of NO on this form of programmed cell death. We hypothesized that NO may inhibit apoptosis of hematopoietic cells. In this study, therefore, hematopoietic cells obtained from umbilical cord blood (CB) were incubated with NO, and NO inhibited apoptosis of hematopoietic cells in umbilical CB, although it did not affect the number of mononuclear cells (MNC). Inhibition of apoptosis of hematopoietic cells is important for their use in transplantation. Our results suggest that it might be possible to prevent the loss of hematopoietic cells using NO.

A case of malignant lymphoma in a patient with high levels of CA125 and CA19-9.

The authors encountered a case of malignant lymphoma in a patient who had high levels of CA125 and CA19-9. These tumor markers showed almost identical changes during the clinical course of the disease. To date, there has been no investigation of these markers as they relate to malignant lymphoma. Unfortunately, the normal values of CA125 and CA19-9 in children were not known and thus this case could not be compared with disease-free children. This study shows CA125 and CA19-9 levels of the children that do not have the malignant diseases, and reports on one case of malignant lymphoma in a patient who presented with high levels of these markers.

A case of hemolytic uremic syndrome improved with nitric oxide.

Hemolytic uremic syndrome (HUS) after transplantation is difficult to treat, and there is no consensus regarding optimal mode of treatment. We attached transdermal isosorbide tape as a nitric oxide (NO) donor to patients with HUS after bone marrow transplantation (BMT). This was very effective in ameliorating the hemolysis and increasing platelet numbers. We report here the successful use of an isosorbide in a patient with HUS after transplantation. Bone Marrow Transplantation (2000) 25, 109-110.

Substance P inhibits pancreatic exocrine secretion via a neural mechanism.

We investigated the effects of the sensory neuropeptide substance P (SP) on amylase and fluid secretion in the isolated vascularly perfused rat pancreas. SP inhibited CCK-induced amylase release and secretin-induced juice flow via the pancreatic duct in a dose-related fashion. Threshold inhibition occurred following addition of 10(-10) M SP to the perfusate, and maximal inhibition was seen with 10(-8) M SP. The effects of SP were partially blocked by both the neurokinin-1 (NK1) and neurokinin-2 (NK2) receptor antagonists. Atropine and TTX blocked SP-induced effects on both amylase secretion (26 and 63% blockade, respectively) and pancreatic juice flow (21 and 79% blockade, respectively). Excitation of pancreatic sensory nerves using capsaicin (in the absence of SP) inhibited both amylase and pancreatic juice flow via activation of the NK1 receptor. We conclude that SP inhibits exocrine secretion via an indirect neural mechanism.

A case of myelodysplastic syndrome complicated by pulmonary alveolar proteinosis with a high serum KL-6 level.

Serious hematological diseases often cause respiratory disorders. Because these are related to the prognoses of patients with hematological diseases, their early diagnosis is necessary. This study describes a 6-year-old girl with myelodysplastic syndrome complicated by pulmonary alveolar proteinosis who showed a remarkable increase in her serum KL-6 level. Three years and 2 months after the end of therapy for neonatal melanoma, a diagnosis of myelodysplastic syndrome with leukemic change was made. Ten months after the onset of leukemia, she had respiratory distress with an increased serum KL-6 level of 75,000 U/mL (reference range; < 500 U/mL). Despite various treatments for pulmonary complications, she died 3 months after developing respiratory distress. A diagnosis of pulmonary alveolar proteinosis was made at autopsy. Earlier treatment of respiratory distress could be achieved if serum KL-6 levels were examined earlier.

The relation between the number of PBSC and number of leukaphereses in children.

Although PBSC transplantation has an advantage over BM transplantation in that fewer burdens are placed on the patient at the time of stem cell collection, the number of collected cells decreases when leukapheresis is done repeatedly. We examined the relation between the number of times leukapheresis is performed and the number of mononuclear cells (MNC), CD34+ cells, and colony-forming unit-granulocyte-macrophages (CFU-GM) collected. The percentage of CD34+ cells was measured by flow cytometry and the number of CFU-GM was measured by a progenitor assay. The number of cells collected was significantly decreased by the third collection. Therefore, to secure enough cells for transplantation, leukapheresis ideally should be performed no more than twice if PBSC collection is to be efficient.

[Ventricular outflow tract obstruction by cardiac tumors during ACTH therapy of an infant with tuberous sclerosis].

In an infant with tuberous sclerosis and West syndrome, ACTH treatment was interrupted because of augmentation of the biventricular outflow tract obstruction. A 5-month-old boy, who had been diagnosed to have multiple cardiac tumors since in utero, manifested West syndrome with typical hypsarhythmia in EEG. Several days after starting low dose ACTH treatment, a systolic murmur due to biventricular outflow tract obstruction intensified with ventricular arrhythmia. A tumor in the left ventricular outflow tract was enlarged. Discontinuation of daily administration of ACTH and administration of a beta-blocking agent improved the cardiac problems. Acceleration of blood velocity in the outflow tracts disappeared two months later. Intracardiac morphology and hemodynamic performance should be monitored to avoid critical complications during ACTH treatment in patients with tuberous sclerosis.

Pancreatitis with normal serum amylase associated with sodium valproate: a case report.

The patient was a 1-year-old infant with severe postencephalitis syndrome. Diarrhea and elevation of the pancreatic enzymes, except for serum amylase (elastase 1 > 1, 5000 ng/dl (100-400); lipase, 885 IU/I/37 degrees C (10-48); trypsin, > 900 ng/ml (110-460)), were observed starting 70 days after starting valproate (dose, 70 mg/kg; serum level, 83.8 micrograms/ml). These findings as well as those obtained by abdominal ultrasonography suggested a diagnosis of pancreatitis, which was thought to be caused by sodium valproate. Important signs of valproate-induced pancreatitis may be easily overlooked in patients with neurological impairment, such as in ours. Because the blood half-life of amylase is short, not only amylase but some other pancreatic enzymes should be promptly investigated when valproate-related pancreatitis is clinically suspected in physically or mentally handicapped children.