[Determination of Chlorothalonil Metabolite I in Livestock Products by LC-MS/MS].

An analytical method based on LC-MS/MS was developed for the determination of chlorothalonil metabolite I in livestock products. Chlorothalonil metabolite I in livestock products was extracted with acetone. The crude extracts were defatted by acetonitrile and n-hexane partitioning. Cleanup was carried out using a combination of ethylene diamine-N-propyl silylation silica gel (PSA) and silica gel (SI) mini columns with acidic condition. The sample solution was subjected to LC-MS/MS using an external solvent calibration curve. The average recovery (n=5) of chlorothalonil metabolite I from five types of livestock products (cattle muscle, cattle fat, cattle liver, milk and egg) spike at the maximum residue limits (MRLs) or at a uniform limit of 0.01 mg/kg was 97.1-102.9%, with a relative standard deviation of 1.4-6.8%. The limit of quantitation of the developed method was calculated to be 0.01 mg/kg.

Collagen Q is a key player for developing rational therapy for congenital myasthenia and for dissecting the mechanisms of anti-MuSK myasthenia gravis.

Acetylcholinesterase (AChE) at the neuromuscular junction (NMJ) is anchored to the synaptic basal lamina via a triple helical collagen Q (ColQ) in the form of asymmetric AChE (AChE/ColQ). We exploited the proprietary NMJ-targeting signals of ColQ to treat congenital myasthenia and to explore the mechanisms of autoimmune myasthenia gravis (MG). Mutations in COLQ cause congenital endplate AChE deficiency (CEAD). First, a single intravenous administration of adeno-associated virus serotype 8 (AAV8)-COLQ to Colq−/− mice normalized motor functions, synaptic transmission, and partly the NMJ ultrastructure. Additionally, injection of purified recombinant AChE/ColQ protein complex into gluteus maximus accumulated AChE in non-injected forelimbs. Second, MuSK antibody-positive MG accounts for 5-15 % of MG. In vitro overlay of AChE/ColQ to muscle sections of Colq−/− mice, as well as in vitro plate-binding of MuSK to ColQ, revealed thatMuSK-IgG blocks binding of ColQ to MuSK in a dose-dependent manner. Passive transfer of MuSK-IgG to wild-type mice markedly reduced the size and intensity of ColQ signals at NMJs. MuSK-IgG thus interferes with binding of ColQ to MuSK. Elucidation of molecular mechanisms of specific binding of ColQ to NMJ enabled us to ameliorate devastating myasthenic symptoms of Colq−/− mice and also to reveal underlying mechanisms of anti-MuSK-MG.

[Survey of pesticide residues in imported spices and herbs (1997.4-2011.3)].

A survey of pesticide residues in 313 samples of imported spices and herbs on the Tokyo market from April 1997 to March 2011 was carried out. Thirty-seven kinds of pesticides, including organophosphorus, organochlorine, pyrethroid, carbamate and others, were detected between levels of trace (below 0.01 ppm) and 3.3 ppm from 64 samples. The rate of detection was highest in peel (100%) followed by stem (66.7%), fruit (34.5%), bark (33.3%), flower (31.3%) and leaf (14.7%). No residues were detected in root, seed or whole grass. Organochlorine pesticides were detected in all plant parts. The insecticides were detected in products from all production areas, suggesting that their use is common. Residue levels of these pesticides were calculated as less than 1% of their ADI values, based on the daily intake of spices and herbs. Therefore, these spices and herbs should be safe when consumed in customary amounts.

[Survey of pesticide residues in imported tea (1992.4-2010.3)].

A survey of pesticide residues in 116 imported tea samples on the Tokyo market from April 1992 to March 2010 was carried out. Twenty-two kinds of pesticides, including organophosphorus, organochlorine, pyrethroid and others, were detected at levels between trace (below 0.01 ppm) and 4.0 ppm in 76 samples. The rate of detection was 90% in non-fermented tea, 89% in semi-fermented tea, and 49% in fermented tea. Organophosphorus pesticide was not detected in puer tea, which is fermented with bacteria. The pesticide residues tended to be decreased by a fermentation process. However, organochlorine and pyrethroid pesticides appeared to be chemically stable, so they were still detected in fermented teas. Residue levels of these pesticides were calculated as less than 1% of ADI, except for ethion (45% of ADI) based on the daily intake of tea. Therefore, these teas should be safe when drunk in customary amounts.

[Survey of pesticide residues in imported frozen vegetables and fruits (1989.4~2008.3)].

A survey of pesticide residues in 595 imported frozen products on the Tokyo market from April 1989 to March 2008 was carried out. Forty three kinds of pesticides, including organophosphorus, organochlorine, carbamate, pyrethroid and others, were detected between levels of trace (below 0.01 ppm) and 4.6 ppm from 162 samples. Chlorpyrifos, cypermethrin and omethoate were frequently detected in green vegetables (komatsuna leaf and spinach), cypermethrin and methamidophos were detected in pods and seeds (green soybean and string pea), chlorpropham (CIPC) was detected in potato, and captan and carbaryl were detected in berries (blueberry, raspberry and strawberry). The hydrophilic pesticide methamidophos was detected in flesh of lychee. Residue levels of these pesticides were calculated as between less than 0.5% and 30% of their ADI values according to the daily intake of frozen products. Therefore, these frozen products should be safe when they were eaten in customary amounts.

[Transfer of pesticide residues to crops via cardboard boxes].

Cardboard boxes used to transport crops are often reused in the distribution process, and therefore transfer of pesticides between crops might occur. So, we designed model experiments to investigate whether or not transfer of pesticide residues from crops to other crops via cardboard boxes occurs. Under severe experimental conditions, 6.2% of the pesticide residues of grapefruit was found to be transferred to spinach via cardboard boxes. In the case of the mandarin orange, 0.57% was transferred. The actual amount of transferred pesticides in the market may be less than that in these model experiments, but it is clear that transfer of pesticide residues to other crops via cardboard boxes can occur. Therefore more attention must be given to reuse of cardboard boxes in the distribution process.

[Survey of pesticide residues in imported fruit products (1994.4-2006.3)].

A survey of pesticide residues in 600 imported fruit products on the Tokyo market from April 1994 to March 2006 was carried out. Thirty kinds of pesticides, including organophosphorus, organochlorine, carbamate, and pyrethroid, were detected between levels of Tr (below 0.01 ppm) to 0.37 ppm from 75 samples. Residual pesticides were detected from fruit products, dried fruits and pressed juice. A high frequency of pesticide residues was observed in peels, whole body of fruits and products which contained them. Residue levels of these pesticides were calculated as between less than 0.1 and 3.9% of their ADI values according to the daily intake of fruit products. Therefore, these fruit products should be safe when eaten in customary amounts.

Acute annular outer retinopathy.

Acute annular outer retinopathy is an uncommon disorder of uncertain aetiology characterized by a unilateral visual field defect with an irregular annular band of grey-white, deep retinal opacification in a peripapillary location. Acute annular outer retinopathy is considered to be a variant of acute zonal occult outer retinopathy. The case is reported of acute annular outer retinopathy with spontaneous good visual recovery and no ophthalmoscopic sequelae.

A case of posterior ischemic optic neuropathy in a posterior-draining dural cavernous sinus fistula.

A 79-year-old woman presented with sudden unilateral visual loss after an ocular motor disturbance and pulsatile tinnitus. Neuro-ophthalmologic examination showed a presumed right posterior ischemic optic neuropathy (PION), oculosympathetic, and third, sensory fifth, and sixth cranial nerve pareses. Selective angiography of the right internal and external carotid arteries confirmed a posterior-draining dural carotid cavernous sinus fistula (CCF) fed by the right meningohypophyseal trunk and right middle meningeal artery. Angiography also showed an ophthalmic-middle meningeal arterial anastomosis. We postulate that the PION was caused by an arterial steal, because blood was drawn into the fistula and away from the intraorbital optic nerve.

Relationship of dissociated optic nerve fiber layer appearance to internal limiting membrane peeling.

PURPOSE: The dissociated optic nerve fiber layer (DONFL) appearance features numerous arcuate striae after vitrectomy for idiopathic epiretinal membrane or macular hole. We studied the relation between the DONFL appearance and internal limiting membrane (ILM) peeling and performed static microperimetry to explore the possible mechanisms underlying the DONFL appearance. DESIGN: Prospective cohort study. PARTICIPANTS: Ninety-five eyes of 88 consecutive patients undergoing vitrectomy for idiopathic macular hole. METHODS: Vitrectomy and gas tamponade were performed with or without ILM peeling. All eyes were examined postoperatively using fundus photography with blue filters. Static microperimetry was performed in 31 eyes with a clearly visible DONFL appearance. MAIN OUTCOME MEASURE: The postoperative incidence of the DONFL appearance on blue-filtered fundus photographs. The secondary outcome measure included the microperimetry threshold value in the arcuate striae and the surrounding normal retina within the papillomacular bundle. RESULTS: On blue-filtered photographs, 46 (62.2%) of 74 eyes with ILM peeling exhibited the DONFL appearance, compared with 0 (0%) of 21 eyes without ILM peeling (P = 0.0002). The area of the DONFL appearance was limited to the area of ILM peeling. There was no difference in the microperimetry threshold value between the arcuate striae and the surrounding normal retina (P = 0.9312). CONCLUSION: The present study indicates that the DONFL appearance might be related to ILM peeling and is not associated with the loss of optic nerve fibers. These results are consistent with the suggestion that the DONFL appearance might be caused by cleavage of the optic nerve fiber bundles due to damage to the Müller cells, which are in close contact with the ILM and act to maintain the close proximity of the nerve fiber bundles.

An unusual case of congenital convergent convergent strabismus fixus.

BACKGROUND: Convergent strabismus fixus is classified into congenital and acquired types; however, there are only a few detailed case reports of congenital convergent strabismus fixus. Here, we report a rare case of this condition in which the eyeball was shown by imaging diagnosis to be fixed with a hard cord-like material in the orbit. To our knowledge, there have been no previous reports of congenital convergent strabismus fixus caused by an abnormal cord-like material that was clearly identifiable by imaging. CASE REPORT: The patient was a 16 year old female who visited our department for consultation regarding restoration of normal appearance of her strabismus. She had undergone strabismus surgery at the age of 3 years, but a forced duction test was strongly positive in all directions, and the eyeball was not mobile at the time. The initial surgery performed was a tenotomy of the medial rectus muscle and inferior rectus muscle and resection of the lateral rectus muscle of the right eye. The eye position and eye movement remained unchanged after surgery. At this time, MRI of the orbital region was performed, and an abnormal cord-like material connecting the posterior eyeball to the orbital wall was detected in the orbit. The cord-like material was pulling strongly on the posterior eyeball, congenitally fixing the eye position internally. CONCLUSION: We experienced a rare case in which a hard cord-like material in the orbit congenitally fixed the position of the eyeball. On MRI, the intensity of the cord-like material was clearly different from those of the muscle and nerve tissues. Considering the linearity of the cord-like material and the results of a forced duction test, the cord-like material probably consisted of fairly hard tissue, similar to bone.

Transition from upbeat to downbeat nystagmus observed in a patient with Wernicke's encephalopathy.

BACKGROUND: We report an unusual case of Wernicke's encephalopathy presenting with transient upbeat nystagmus that changed to a persistent downbeat nystagmus. CASE: A 27-year-old man presented with upbeat nystagmus. Three months earlier, he had been diagnosed with Wernicke's encephalopathy after fasting for a month. OBSERVATIONS: This diagnosis was supported by his symptoms (ataxia, a confused state). Clinical recovery followed thiamine therapy. His upbeat nystagmus had linear slow phases with average amplitude and frequency (+/-SD) during fixation straight ahead of 2.8 +/- 0.7 degrees and 4.6 +/- 2.2 Hz, respectively. Two months later, the primary position upbeat nystagmus had diminished and downbeat nystagmus (0.9 +/- 0.5 degrees and 3.2 +/- 0.7 Hz on average) for a 20 degrees downward gaze had developed. Then, 8 months later, he showed only downbeat nystagmus, which obeyed Alexander's law. His primary position downbeat nystagmus was completely suppressed by clonazepam, a gamma-aminobutyric acid (GABA) agonist. CONCLUSIONS: Owing to an underlying central vestibular imbalance, even after the recovery of acute neurological symptoms, Wernicke's encephalopathy can be complicated by persistent downbeat nystagmus, which can be treated by a GABA agonist.

Glyceraldehyde-derived advanced glycation end-products preferentially induce VEGF expression and reduce GDNF expression in human astrocytes.

The blood-brain barrier (BBB) is a biological unit composed of capillary endothelial cells and astrocytes. Here we examined the effects of various types of advanced glycation end-products (AGEs) on astrocytes and BBB-forming endothelial cells. While no type of AGE we examined changed the permeability of endothelial sheets, glyceraldehyde-derived AGE induced VEGF expression most significantly in astrocytes. The expression of glial cell line-derived neurotrophic factor (GDNF), which reduces the vascular permeability, was decreased in the astrocytes by treatment with glyceraldehyde-derived AGE. These results indicate that glyceraldehyde-derived AGE is the biologically active substance for astrocytes by regulating the VEGF and GDNF expression, which is causally contributing to an increase in the permeability of the BBB.

A review of 244 orbital tumors in Japanese patients during a 21-year period: origins and locations.

PURPOSE: To review 244 cases of orbital tumors to determine the distribution of patient age, pathology, and tumor origin and location. METHODS: We reviewed cases of histopathologically or radiologically verified orbital tumors at our institution from 1981 through 2002, and studied the distribution of patient age, pathology, origin of tumors, and tumor location in the orbit in 244 consecutive patients (aged from 0 to 90 years, mean, 48.7 years; 114 men, 130 women). RESULTS: Among the 244 cases, 213 (89%) were primary orbital tumors, 23 (9%) were secondary tumors originating in contiguous spaces, and 8 (2%) were metastatic tumors. Regarding the location in the orbit, 122 tumors (50%) were extraconal, 36 (15%) were intraconal, and 86 (35%) were in the lacrimal gland area. The common tumors were reactive lymphoid hyperplasia (22%) and malignant lymphoma (20%) in the extraconal area; cavernous hemangioma (25%), optic nerve glioma (14%), and optic nerve sheath meningioma (14%) in the intraconal area; and malignant lymphoma (40%) and pleomorphic adenoma (24%) in the lacrimal gland area. The age distribution of the 244 patients exhibited two peaks, at 0 to 9 years and 60 to 69 years. In the 0- to 9-year-old patients, the most common tumors were dermoid cyst (26%), optic nerve glioma (11%), capillary hemangioma (11%), and hemorrhagic lymphangioma (11%). On the other hand, in patients over 40 years of age, the common tumors were malignant lymphoma (31%), orbital pseudotumor (24%), pleomorphic adenoma (10%), and cavernous hemangioma (9%). CONCLUSIONS: The pathologic profiles of orbital tumors were characterized according to patient age and the location of the tumor in the orbit. The age of onset, tumor location, and radiological findings provide us with important information for the diagnosis of a tumor prior to biopsy or tumor resection and for the determination of the treatment strategy.