[Simultaneous Determination of Pesticide Residues in Brown Rice Using GC-MS/MS and LC-MS/MS].

This study developed a method that simultaneously detected 283 pesticide residues in brown rice using GC-MS/MS and LC-MS/MS. In this method, we examined the desirable amount of sodium chloride required for salting out and the SPE cartridge required for clean-up. Pesticide residues from the sample were extracted with acetonitrile using a homogenizer and mixed with salts including anhydrous magnesium, two types of citrate, and sodium chloride. The sample solution of the acetonitrile layer was cleaned up using the GCB/NH2 (200 mg/200 mg, 6 mL) SPE cartridge. The determination method was validated using two concentrations (0.01 and 0.1 µg/g) of 283 pesticides based on the validation guideline of the Ministry of Health, Labor and Welfare in Japan. Of the 283 pesticides, 250 were detected satisfactorily. In addition, 59 brown rice samples sold in Tokyo were surveyed using the same method. Out of 44 samples, 12 pesticide residues below MRLs were detected. Therefore, this developed method is useful for the simultaneous determination of pesticide residues in brown rice.

Simultaneous determination of five carbapenems, highly polar antibiotics, in milk by LC-MS/MS.

The simultaneous determination of five carbapenems (biapenem, doripenem, ertapenem, imipenem, and meropenem) in raw and pasteurised bovine milk samples using LC-MS/MS was achieved and validated. Chromatographic separation was conducted on an InertSustain® AQ-C18 column using 0.1% formic acid in water and acetonitrile as the mobile phase. Target compounds were extracted using acetonitrile/water (20:80, v/v). After the removal of lipids with acetonitrile-saturated hexane, the dissolved protein was denatured with acetic acid. A portion of the supernatant was passed through an Oasis® PRiME HLB cartridge to remove the matrix. This novel method was validated in accordance with the Japanese validation guidelines and exhibited good trueness, ranging from 86.3% to 96.2%, using matrix-matched calibration curves. The relative standard deviation of repeatability ranged from 1.0% to 6.3%, and that of within-laboratory reproducibility ranged from 1.6% to 7.1%. The limit of quantification was 1.0 µg kg-1 for all analytes. None of the 60 milk samples commercially available in Tokyo contained any analytes. This novel method exhibited high-quality performance and can easily be implemented for the routine monitoring of carbapenems, which are highly polar antibiotics in milk.

Development of a simple screening method for analyzing cereulide toxin in fried rice using liquid chromatography-tandem mass spectrometry.

PURPOSE: The presence of cereulide, an emetic toxin produced by Bacillus cereus, in fried rice samples is critical evidence of food poisoning even in situations where B. cereus could not be detected. This study aims to develop a screening method for analyzing cereulide in fried rice using the QuEChERS procedure and liquid chromatography-tandem mass spectrometry (LC-MS/MS). METHODS: Cereulide was identified and quantified in fried rice samples using the QuEChERS extraction method and LC-MS/MS. The accuracies of the methods were determined by analyzing fortified blank samples at two concentrations (10 and 50 µg/kg) conducted on three samples daily for five days. RESULTS: The QuEChERS procedure removed matrix compounds from fried rice. Characteristic MS/MS spectra enabled the identification of cereulide. As the matrix effects in seven fried rice samples were within ± 6%, an external solvent calibration curve could be used for quantification. This method exhibited good accuracy ranging from 88 to 89%. The relative standard deviations for both repeatability and intra-laboratory reproducibility were < 4%. These standard deviations satisfied the criteria of the Japanese validation guidelines for residues (MHLW 2010, Director Notice, Syoku-An No. 1224-1). The limit of quantification was 2 µg/kg. The applicability of this method was confirmed using the analysis of cereulide in fried rice samples incubated with emetic Bacillus cereus. CONCLUSIONS: The QuEChERS extraction procedure described herein showed substantial promise as a reliable screening tool for cereulide in fried rice sample.

[Clinical course of retinopathy of prematurity and other neonatal complications in Kobe Children's Hospital].

PURPOSE: To report changes in retinopathy of prematurity (ROP) and other neonatal complications for the last 15 years CASES AND METHOD: 638 cases with a birth weight of less than 1000 g were categorized into two groups in oxygen management changes based on reduced oxygen protocol before and after change (Group A: 218 cases from 1997 to 2001, Group B: 420 cases from 2002 to 2011). Prematurity and the clinical course of ROP were reviewed. In addition, the incidence of 5 main neonatal complications before and after oxygen control criterion change was compared. RESULTS: The differences in birth weight and gestational age among the two groups were not significant (A: 765 138 g/26.5 +/- 2.2 weeks, B: 753 +/- 158 g/26.1 +/- 2.3 weeks). The incidence of ROP and the rate of treatment for ROP in Group B were significantly less than in Group A (A: 83.0%/32.6% VS B: 54.8%/ 10.0%; p < 0.01). The rate of forming a cicatricial phase severer than grade 3 was 4.1% in Group A and 1.7% in Group B (p = 0.06). The incidence of neonatal complications before and after oxygen management change was not significantly different. CONCLUSION: Incidence and treatment of ROP were improved after changing to the reduced oxygen protocol. There was no increase in neonatal complications after the management change.

Stepwise Treatment for TAFRO Syndrome.

TAFRO syndrome, a rapidly progressive and fatal disease, is rare, and its etiology remains unknown. It is characterized by thrombocytopenia, anasarca (edema, pleural effusion, and ascites), fever, reticulin fibrosis (or renal insufficiency), and organomegaly with Castleman disease (CD)-like histological features in the lymph nodes. CD is a rare, indolent, lymphoproliferative disorder with no established curative strategies. Most idiopathic multicentric CD cases are controlled with anti-interleukin (IL)-6 therapy (tocilizumab and siltuximab) and/or rituximab. However, it is unclear whether these therapies can be directly applied to treat TAFRO syndrome. Here, we describe stepwise immunotherapy (rituximab induction therapy and cyclosporine maintenance therapy) for two cases of steroid-refractory TAFRO syndrome. A 32-year-old man visited a local hospital with sudden onset of fever and epigastralgia. The diagnosis of TAFRO syndrome was established based on the diagnostic criteria. After rituximab administration, C-reactive protein and IL-6 levels were normalized. However, the ascites persisted, with increased resistance to rituximab. Tocilizumab was also ineffective; therefore, cyclosporine was administered. After the initiation of cyclosporine treatment, the ascites decreased and ultimately disappeared. Twelve months after immunotherapy, the patient remained asymptomatic under cyclosporine maintenance therapy. Similar stepwise immunosuppressive therapy was administered to a 72-year-old man with TAFRO syndrome complicated by renal failure. After rituximab infusion, C-reactive protein was decreased. Although methylprednisolone, rituximab, tocilizumab, and cyclosporine were administered, other laboratory data and clinical symptoms remained unchanged. His level of consciousness subsequently deteriorated due to herpes zoster encephalitis, and he died. We consider the combination of rituximab induction therapy and cyclosporine maintenance therapy to be effective for TAFRO syndrome if initiated at an early stage.

Anti-U1 RNP antibodies in cerebrospinal fluid are associated with central neuropsychiatric manifestations in systemic lupus erythematosus and mixed connective tissue disease.

OBJECTIVE: To determine the significance of anti-U1 RNP antibodies in the cerebrospinal fluid (CSF) of patients with systemic lupus erythematosus (SLE) or mixed connective tissue disease (MCTD) who have central neuropsychiatric SLE (NPSLE). METHODS: The frequency of antinuclear antibodies including anti-U1 RNP antibodies in the sera and CSF of 24 patients with SLE and 4 patients with MCTD, all of whom had neuropsychiatric syndromes, was determined using an RNA immunoprecipitation assay and an enzyme-linked immunosorbent assay. The frequency of anti-U1 RNP antibodies in the CSF of patients with central NPSLE was examined, and the anti-U1 RNP index ([CSF anti-U1 RNP antibodies/serum anti-U1 RNP antibodies]/[CSF IgG/serum IgG]) was compared with CSF interleukin-6 (IL-6) levels and the albumin quotient (Qalb, an indicator of blood-brain barrier damage). CSF and serum antibodies against U1-70K, U1-A, and U1-C, including autoantigenic regions, were examined, and the U1-70K, U1-A, and U1-C indices as well as the anti-U1 RNP index were calculated. RESULTS: CSF anti-U1 RNP antibodies with an increased anti-U1 RNP index showed 64.3% sensitivity and 92.9% specificity for central NPSLE. The anti-U1 RNP index did not correlate with CSF IL-6 levels or the Qalb. The anti-U1-70K index was higher than the anti-U1-A and anti-U1-C indices in the CSF of anti-U1 RNP antibody-positive patients with central NPSLE. The major autoantigenic region for CSF anti-U1-70K antibodies appeared to be localized in U1-70K amino acid 141-164 residue within the RNA-binding domain. CONCLUSION: The frequency of anti-U1 RNP antibodies in the CSF and the anti-U1 RNP index are useful indicators of central NPSLE in anti-U1 RNP antibody-positive patients. The predominance of anti-U1-70K antibodies in CSF suggests intrathecal anti-U1 RNP antibody production.

Two cases of late-onset drug-induced lupus erythematosus caused by ticlopidine in elderly men.

We have experienced two cases of drug-induced lupus erythematosus caused by ticlopidine in the last three years. Both were late-onset cases (1 and 4 years) that occurred in elderly men (76 and 81 years old). The common features were fever, arthralgia, myalgia, serositis, and the presence of anti-histone autoantibodies. Because ticlopidine is widely used in elderly people with ischemic vascular disease, ticlopidine-induced lupus should be considered when patients taking ticlopidine present lupus-like symptoms.

Proteomic study of DBA/2J mice retina: Down-regulation of Integrin beta7 correlated with retinal ganglion cell death.

To identify and determine the function of the proteins associated with the death of retinal ganglion cells (RGCs) in DBA/2J mice, an animal model of glaucoma, retinas of DBA/2J mice, were analyzed by proteomics at 5-, 7-, and 11-months-of-age. The proteins showing significant alterations were selected for identification by MS and 18 proteins were differentially expressed and the identified proteins included cell membrane receptors and proteins associated with intracellular signaling pathways. Among of identified proteins, the expression of Integrin beta7 at 7-months-of-age was decreased by about 89% of that at 5-months-of-age. Integrin beta7 was expressed in the RGCs. The effect of glutamate toxicity on the expression pattern of Integrin beta7 in a RGC line was also investigated and the glutamate-induced death of RGC was inhibited by the RNA knockdown of Integrin beta7. Our data showed also that the expression of 18 proteins in the DBA/2J was significantly altered in DBA2 mice and down-regulation of Integrin beta7 may have a protective effect on glutamate-induced death of RGCs.

(-)-Epigallocatechin-3-gallate potentiates the cytotoxicity induced by benzyl isothiocyanate and hydrogen peroxide in human Jurkat T lymphocytes.

(-)-Epigallocatechin-3-gallate (EGCG)-induced apoptosis was along both the extracellular signal-regulated protein kinase (ERK) and c-jun N-terminal kinase (JNK) pathways in Jurkat cells. Co-treatment with EGCG potentiated the cytotoxicity induced by benzyl isothiocyanate (BITC) and H(2)O(2), both being inhibited by ERK and JNK inhibitors. These results suggest the significant role of mitogen-activated protein kinase (MAPK) signaling in the apoptosis induction regulated by EGCG alone and in combination.

Survey of Nitrate Ion Concentrations in Vegetables Cultivated in Plant Factories: Comparison with Open-Culture Vegetables.

A survey of nitrate-ion concentrations in plant-factory-cultured leafy vegetables was conducted. 344 samples of twenty-one varieties of raw leafy vegetables were examined using HPLC. The nitrate-ion concentrations in plant-factory-cultured leafy vegetables were found to be LOD-6,800 mg/kg. Furthermore, the average concentration values varied among different leafy vegetables. The average values for plant-factory-cultured leafy vegetables were higher than those of open-cultured leafy vegetables reported in previous studies, such as the values listed in the Standard Tables of Food Composition in Japan- 2015 - (Seventh revised edition). For some plant-factory-cultured leafy vegetables, such as salad spinach, the average values were above the maximum permissible levels of nitrate concentration in EC No 1258/2011; however, even when these plant-factory-cultured vegetables were routinely eaten, the intake of nitrate ions in humans did not exceed the ADI.

High frequency of open-angle glaucoma in Japanese patients with Alzheimer's disease.

The clinical and genetic relationships between Alzheimer's disease (AD) and glaucoma remain obscure. The aim of this study was to determine the prevalence of open-angle glaucoma (OAG) in patients with AD and whether the apolipoprotein E (APOE) 4 allele is associated with AD, with or without OAG, in Japanese. The groups consisted of 172 patients with the diagnostic criteria of AD and 176 age-matched controls. Ophthalmic examinations were conducted, and genomic analysis was performed by PCR and digestion of products with an enzyme. OAG was found in 41 (23.8%) of the AD patients, which was a significantly (p = 0.0002) higher prevalence than that in the controls (9.9%). Furthermore, there was no significant difference between intraocular pressures (IOPs) in AD patients with OAG and without OAG. The percentage of AD patients who carried an APOE epsilon4 allele (29.5%) was significantly (p = 0.0007) higher than that of the controls (9.1%). However, the percentage of AD patients with OAG who carried an APOE epsilon4 allele (35.7%) was not significantly different than that of AD patients without OAG (27.7%, p = 0.42). In summary, the prevalence of OAG is high in Japanese patients with AD, suggesting that common factors other than APOE may contribute to the two diseases.

Traffic injuries of the pregnant women and fetal or neonatal outcomes.

OBJECTIVE: To investigate the relationship between pregnancy outcome and injury severity of pregnant woman in traffic accidents. METHOD: We reviewed insurance reports of traffic accidents and collected data on injuries of pregnant women and outcomes of their pregnancies. RESULT: A total of 135 pregnant women, with a mean injury severity score of 1.8+/-4.0, were involved in traffic accidents from 1994 through 2003. Injury severity score, abdominal abbreviated injury scale score were significantly higher in women whose neonates died than in women with healthy newborns. However, neither the likelihood of having been subjected to direct external forces during the accident nor injury severity differed between women with spontaneous abortions and woman with healthy newborns. CONCLUSION: Predicting abortion on the basis of maternal injury severity is difficult. Because unknown variables may contribute to fetal loss, further studies of traffic injuries are needed.

Accidental choking in a patient with Treacher Collins syndrome.

A 3-year-old boy with Treacher Collins syndrome had undergone tracheostomy and placement of a secured cannula at the age of 4 months. When he was 3 years old, he manually extracted the secured cannula by himself and choked to death. Autopsy revealed upper airway obstruction with posterior deviation and mucosal hyperplasia of the radix linguae, mandibular hyperplasia, and occlusion of the artifical airway owing to intratracheal granuloma due to the long-standing tracheotomy. For safe, long-term use of a tracheostomy to maintain the airway, children with craniofacial abnormalities should be carefully supervised by their families to prevent accidental decannulation.

Prevalence of glaucoma in adults with Down's syndrome.

PURPOSE: To compare the prevalence of glaucoma in adults with Down's syndrome (DS) to that in non-DS control adults. METHODS: Twenty-six patients (14 men and 12 women) with DS and 188 control subjects (105 men and 83 women) were studied. The mean age was 35.1 +/- 6.9 (+/- SD) years in the DS group and 36.9 +/- 5.2 years in the control group. There were no significant differences in age or sex distribution between the two groups. Glaucoma was diagnosed by two glaucoma specialists based on the optic disc findings obtained through dilated pupils. RESULTS: The prevalence of patients with glaucoma in the DS group was 11.5%, significantly higher (P = 0.014) than that in the control group, 1.1%. There was no significant difference in intraocular pressure between glaucomatous eyes (12.2 +/- 3.2 mmHg) and nonglaucomatous eyes (11.1 +/- 4.1 mmHg) in the DS group (P = 0.465). CONCLUSIONS: The prevalence of glaucoma in adult patients with DS was significantly higher than that in age-matched control subjects.

Accidental strangulation of a mentally retarded patient by a clothing collar: a case report.

A 40-year-old mentally retarded woman died of accidental strangulation in a nursing home. She was found in a kneeling position with her hands on her knees and the collar of her clothing compressing the front and sides of the neck. Before the accident, a nurse had dressed the patient in one-piece overall-style pyjamas put on back to front so that she could not remove the garment herself. The post-mortem findings and reconstruction of the scene of death suggested that the patient had been strangled by the collar of her backward-facing clothing while in a kneeling position. Because patients with psychiatric illnesses may have a limited ability to recognize or communicate symptoms of physical danger, they must be closely monitored by knowledgeable medical and nursing staff. This case highlights the importance of preventing the accidental deaths of mentally retarded patients in nursing homes.

3. Sudden death due to pulmonary thromboembolism after car driving: a case report.

An autopsy case of sudden death due to pulmonary thromboembolism after a car trip is reported. A 56-year-old Japanese man with hypertension and atrial fibrillation suddenly died immediately after having driven for two and a half hours. At autopsy, the pulmonary arteries were found to be filled with dark-red, coiling thrombi consisting of fibrin and red blood cells. In the dilated right popliteal vein, a thrombus consisting of fibroblasts, a large number of collagen fibres, and newly formed capillaries was attached to the vessel wall. The cause of death was determined to be pulmonary embolism due to the thrombus of the right popliteal vein. Two and a half hours of prolonged sitting while driving a car encouraged thrombus formation. We believe that this case, the first autopsy case of fatal pulmonary thromboembolism after a car trip to be reported, highlights the importance of predicting venous thrombosis of the leg or fatal venous thromboembolism after a car trip.

A five-year review of patients admitted with the diagnosis of bacterial endophthalmitis.

We conducted a retrospective, hospital-based study of patients who were diagnosed with bacterial endophthalmitis on admission to the Department of Ophthalmology, Hiroshima University Hospital, between January 1999 and December 2003. Thirty eyes of 30 patients were identified. Of these patients, 19 eyes had postoperative endophthalmitis, 8 eyes had penetrating trauma and 3 eyes were infected from an endogenous source. All of the patients underwent immediate three-port pars plana vitrectomy. Vitreous specimens of diabetic patients demonstrated a significantly higher incidence of positive bacterial culture. Diabetic vitreous appeared to be a good medium for culture. Visual acuity of hand motion or less at the latest follow-up visit was associated with the presence of diabetes. Prompt treatment with vitrectomy and intra-vitreal antibiotics is crucial for patients with bacterial endophthalmitis, especially if they are diabetic.

[Sudden death while bathing].

We describe the epidemiologic and pathophysiologic features of sudden death while bathing. The leading cause of death is cardiovascular disease (60.4%), followed by cerebrovascular disease (18.7%) and drowning (11.2%). Because hydrostatic pressure, autonomic nervous reflexes or endocrine system reflect on hemodynamic changes while bathing, the elderly persons or patients with cardiovascular disease are often suffered from fatal cardiac events. To find the detail pathophysiologic changes while bathing, exact cause of death should be examined by autopsies. Furthermore, elderly or handicapped persons should be supervised by their families for preventing drowning.