IgE-mediated anaphylaxis and allergic reactions to idursulfase in patients with Hunter syndrome.

BACKGROUND: Enzyme replacement therapy (ERT) with recombinant human idursulfase is effective for the treatment of Hunter syndrome, mucopolysaccharidosis (MPS) type II. However, various adverse events can occur by the infusion of idursulfase. The purpose was to evaluate the occurrence of infusion-related allergic reactions, including anaphylaxis, to idursulfase in patients with MPS II receiving ERT and to elucidate its possible mechanism. METHODS: A total of 34 patients with MPS II were enrolled to receive ERT with Elaprase(®) at a dose of 0.5 mg/kg intravenously once a week. Information regarding the symptoms, frequency, and timing of anaphylaxis during treatment was analyzed. Presence of anti-idursulfase IgE antibody was assessed by skin prick test (SPT) and enzyme-linked immunosorbent assay (ELISA). Western blotting was performed to confirm the reaction between idursulfase and specific IgE. RESULTS: Three patients (8.8%) showed anaphylaxis by infusion of idursulfase. No deaths occurred during the study. Anti-idursulfase IgE antibody was detected by SPT and ELISA. Immunoblotting with patients' sera and Elaprase(®) showed a single band of specific IgE binding to the protein around 70 kD, and idursulfase did not display amino acid sequence homology to known allergens. SPT with idursulfase demonstrated positive results in all patients with anaphylaxis. However, we failed to reveal any risk factors for the development of infusion-related immediate-type allergic reactions. CONCLUSIONS: Anaphylaxis related to infusion of idursulfase is mediated by anti-idursulfase IgE antibody, which might be produced by de novo synthesis. SPT is useful in predicting the occurrence of anti-idursulfase IgE-mediated anaphylaxis during infusion.

Melatonin increases striatal dopaminergic function in 6-OHDA-lesioned rats.

The purpose of this study was to assess the in vivo effects of melatonin, as an antioxidant, on striatal dopaminergic function in rats with a unilateral 6-hydroxydopamine (6-OHDA) lesion of the striatum. Compared with sham-operated controls and expressed as a ratio relative to the contralateral side, there was an increase in the lipid peroxidation product malondialdehyde (MDA, 142%) and a significant reduction in tyrosine hydroxylase (TH) enzyme activity (28%) and dopamine (DA, 32%) and its metabolite dihydroxyphenylacetic acid (DOPAC, 50%) 2 weeks after 6-OHDA injection. Melatonin treatment almost completely restored MDA levels to normal, suggesting the in vivo action of melatonin as an antioxidant. In parallel, partial, but statistically significant recovery of striatal dopaminergic function, including TH enzyme activity and DA levels, also occurred following melatonin treatment. Taken together with our previous reports showing behavioral and histochemical effects of melatonin on the nigrostriatal dopaminergic system, the present results strongly support the hypothesis that melatonin, as an antioxidant, may have beneficial effects on therapeutic approaches for the treatment of oxidative stress-induced neurodegenerative disease such as Parkinson's disease (PD).

GT1b ganglioside induces death of dopaminergic neurons in rat mesencephalic cultures.

We examined neurotoxicity of GT1b against dopaminergic neurons in vitro. Cultures of mesencephalic cells deprived of serum underwent the loss of 19% of tyrosine hydroxylase immunopositive (TH-ip) neurons. In cultures deprived of serum, treatment with 10-30 microg/ml GT1b attenuated the number of TH-ip neurons by 26-69%, respectively, compared to non-treated cultures. Intriguingly, cultures deprived of serum were more vulnerable to GT1b-induced neurotoxicity. Application of 60 microg/ml GT1b to cultures grown in serum containing media resulted in the loss of 26% of TH-ip neurons, similar to that (28%) observed in serum-deprived cultures treated with 10 microg/ml GT1b. Moreover, in our cultures, absence of nitric oxide (NO) production after GT1b treatment was obvious. The present results strongly suggest direct neurotoxic actions of GT1b against dopaminergic neurons regardless of NO.

Lung fibrosis in Sprague-Dawley rats, induced by exposure to manual metal arc-stainless steel welding fumes.

To investigate the disease process of pneumoconiosis induced by welding-fume exposure, a lung fibrosis model was established by building a stainless steel arc welding fume generation system and exposing male Sprague-Dawley rats for 90 days. The rats were exposed to welding fumes with concentrations of 57-67 mg/m3 (low dose) and 105-118 mg/m3 (high dose) total suspended particulates for 2 h per day in an inhalation chamber for 90 days. The concentrations of the main metals, Fe, Mn, Cr, and Ni, were measured in the welding fumes, plus the gaseous compounds, including nitrous gases and ozone, were monitored. During the exposure period, the animals were sacrificed after the initial 2-h exposure and after 15, 30, 60, and 90 days. Histopathological examinations were conducted on the animals' upper respiratory tract, including the nasal pathway and conducting airway, plus the gas exchange region, including the alveolar ducts, alveolar sacs, and alveoli. When compared to the control group, the lung weights did not increase significantly in the low-dose group, yet in the high-dose group there was a significant increase from day 15 to day 90. The histopathological examination combined with fibrosis-specific staining (Masson's trichrome) indicated that the lungs in the low-dose group did not exhibit any progressive fibrotic changes. Whereas, the lungs in the high-dose group exhibited early delicate fibrosis from day 15, which progressed into the perivascular and peribronchiolar regions by day 30. Interstitial fibrosis appeared at day 60 and became prominent by day 90, along with the additional appearance of pleural fibrosis. Accordingly, it would appear that a significant dose of welding-fume exposure was required to induce lung fibrosis.

Pattern of deposition of stainless steel welding fume particles inhaled into the respiratory systems of Sprague-Dawley rats exposed to a novel welding fume generating system.

In order to investigate occupational diseases related to welding fume exposure, such as nasal septum perforation, pneumoconiosis and manganese intoxication, we built a welding fume exposure system that included a welding fume generator, exposure chamber and fume collector. The fume concentrations in the exposure chamber were monitored every 15 min during a 2-h exposure. Fume (mg/m(3)) concentrations of major metals, including Fe, Mn, Cr, and Ni were found to be consistently maintained. An acute inhalation toxicity study was conducted by exposing male Sprague-Dawley rats to the welding fumes generated in this apparatus by stainless steel arc welding. The rats were exposed in the inhalation chamber to a welding fume with a concentration of 62 mg/m(3) total suspended particulates for 4 h. Animals were sacrificed at 4 h and at 1, 3, 7, 10, and 14 days after exposure. Histopathological examinations were conducted on the animals' upper respiratory tracts, including the nasal pathway and the conducting airway, and on the gas exchange region including the alveolar ducts, alveolar sacs, and alveoli. Diameters of fume particles varied from 0.02 to 0.81 microm and were distributed log normally, with a mean diameter of 0.1 microm and geometric standard deviation of 1.42. Rats exposed to the welding fume for 4 h did not show any significant respiratory system toxicity. The mean particle diameter of 0.1 microm resulted in little adsorption of the welding fume particles in the upper respiratory tract. Particle adsorption took place principally in the lower respiratory tracts, including bronchioles, alveolar ducts, alveolar sacs, and alveoli.

Reproductive toxicity of 2-bromopropane in Sprague Dawley rats.

OBJECTIVES: In an effort to clarify the mass intoxication of workers at an electronic company in Korea, the possible causative chemical for reproductive toxicity, 2-bromopropane (2BP), was investigated. METHODS: 2BP was tested through the use of repeated dose experiments among male Sprague Dawley rats. Ten rats were assigned to each treatment group. Vehicle control olive oil and 2BP concentrations of 125 mg/kg, 250 mg/kg, and 500 mg/kg were injected into the intraperitoneum on 28 consecutive days. RESULTS: The rats showed significant decreases in body weight depending on the 2BP dose. The right and left testes showed typical weight loss depending on the dose of 2BP. The red blood cells, hemoglobin, and hematocrit showed some degree of decline with the high dose. The amount of hemoglobin, the mean platelet volume, the number of white blood cells, and the number of lymphocytes decreased significantly with the high dose, while the number of granulocytes and monocytes had a tendency to decrease depending on the dose of 2BP. The histopathology of the testes treated with the middle and high 2BP dose showed a typical patch appearance with severely depleted atrophic tubules, exhibiting germ cell necrosis of spermatogonia and spermatocytes in the seminiferous tubules. Leydig cell hyperplasia or hypertrophy in the interstitial tissue was also noticeable. The epididymis showed some degree of atrophy with vacuolization of the epididymal epithelium. CONCLUSIONS: The testes are the main target organs tested for 2BP toxicity. 2BP also affect the hematopoietic system and thus induces leukopenia and normocytic anemia. Besides the reproductive organs and the hematopoietic system, no significant toxicity has been found.

Mutagenicity of 2-bromopropane.

2-Bromopropane (2BP, isopropyl bromide), a substitute for freon, has recently been suspected to be the causative chemical for the outbreak of some reproductive dysfunctions such as amenorrhea and oligospermia in workers who has been exposed to this solvent in an electronic factory. Bacterial mutation assays, chromosome aberration analysis in vitro, and micronucleus tests in vivo, were carried out to clarify the mutagenicity of 2BP. 2BP induced mutagenicity in Salmonella typhimurium TA100 with metabolic activation in a dose-dependant manner. 2BP induced mutagenicity in TA1535 as well, with or without metabolic activation. These observations indicated that 2BP induced the base-pair substitution type mutations in Salmonella strains. The chromosome aberration analysis showed negative results in Chinese hamster lung cells treated with different concentrations, ranging from 0.077 to 2.46 mg/ml for 6 h with metabolic activation and for 24 h without metabolic activation. The micronucleus frequencies were recorded by examining polychromatic erythrocytes in the bone marrows of rats which were intraperitoneally injected with 2BP for 28 days. There was no significant increase in the micronucleus frequencies at any of the different doses of 2BP (125 mg/ kg b.w./day, 250 mg/kg b.w./day, and 500 mg/kg b.w./day). However, in comparison to controls, there was a significant decrease in the percentage of polychromatic erythrocytes in the total number of erythrocytes. This suggests that there may be bone marrow depression in hematopoiesis at these dose levels of 2BP. Despite the dose levels which showed hematopoietic inhibition in the bone marrow, no micronucleus formation was induced.

Effects of 2-bromopropane on the female reproductive function in Sprague-Dawley rats.

2-Bromopropane (2BP) has been implicated to be the reason for the mass intoxication of workers at an electronic company in Korea. A case series study indicated that 2BP was the possible causative chemical for reproductive toxicity, causing severe anemia accompanied by amenorrhea among female workers, and azoospermia or oligospermia among male workers. To clarify the effect of 2BP on the female reproductive function, repeated doses of 2BP were tested on female Sprague-Dawley rats for 21 days. Ten rats were assigned to each treatment group. The rats were maintained in a 12 hr: 12 hr light-dark cycle and vaginal smears were monitored daily for 3 cycles. After the rats had completed 3 estrous cycles, vehicle control olive oil, 300 mg, 600 mg, and 900 mg/kg of 2BP were injected into intraperitoneum for 14 days. The female rats were then mated with male rats on a 1:1 ratio basis. The treatment continued for an additional 7 days. Rats treated with 2BP experienced a significant decrease in body weight gain depending on the dose of 2BP. The estrous cycles of the rats continued at a normal duration of time before the initiation of treatment, showing 4.32 days for the control group, 4.79 days for the low dose, 4.63 days for the middle dose, and 4.75 days of estrous cycle for the high dose group. 2BP treatment, however, induced a significant delay of the estrous cycle in the high dose treated group, showing 11.1 +/- 3.82 days of the estrous cycle. 2BP decreased the fertility and tended to decrease in the number of pups born, depending on the dose. A 900 mg/kg treatment of 2BP decreased ovarian weight, but 2BP did not affect the length of gestation. Our results indicated that 2BP seemed to be the causative agent for amenorrhea observed in female workers.

Clinical characterization and molecular classification of 12 Korean patients with pseudohypoparathyroidism and pseudopseudohypoparathyroidism.

CONTEXT: Pseudohypoparathyroidism (PHP) is defined as resistance toward parathyroid hormones. PHP and pseudopseudohypoparathyroidism (PPHP) are rare disorders resulting from genetic and epigenetic aberrations within or upstream of the GNAS locus. This study investigated the clinical characteristics and performed a molecular analysis of PHP and PPHP. METHODS: A total of 12 patients with (P)PHP from 11 unrelated families (4 with PHP-Ia, 6 with PHP-Ib, and 2 with PPHP) were characterized using both clinical and molecular methods. Clinical features included the presenting symptoms, Albright hereditary osteodystrophy features, and resistance to hormones. Comprehensive analysis of the GNAS and STX16 loci was undertaken to investigate the molecular defects underlying (P)PHP. RESULTS: All PHP-Ib patients displayed hypocalcemic symptoms. All PHP-Ia patients showed resistance toward TSH, in addition to PTH. In most patients with PHP, when the diagnosis of PHP was first established, hypocalcemia and hyperphosphatemia were associated with a significant increase in serum PTH levels. One patient with PHP-Ia was diagnosed with growth hormone deficiency and showed a good response to human recombinant growth hormone therapy. 6 patients with PHP-Ia and PPHP showed 5 different mutations in the GNAS gene. 5 patients with PHP-Ib displayed a loss of differentially methylated region (DMR) imprints of the maternal GNAS. One PHP-Ib patient showed a de novo microdeletion in STX16 and a loss of methylation of exon A/B on the maternal allele. No patients revealed paternal disomy among 4 patients with PHP-Ib. CONCLUSIONS: Identification of the molecular causes of PHP and PPHP explains their distinctive clinical features and enables confirmation of the diagnosis and exact genetic counseling.

Designation of a GLP facility by the Korean Ministry of Environment GLP authority: the case of the Center for Occupational Toxicology.

In this report, the process of designating a GLP facility by the Korean Ministry of Environment (MOE) is described in detail using the case of the Center of Occupational Toxicology (COT). The COT, which had been prepared as a GLP facility, filed an application to the National Institute of Environmental Research (NIER) of the MOE. The GLP system of the COT was evaluated by a harmonized evaluation team that consisted of several authorities including the NIER, the National Institute of Agricultural Science and Technology (NIAST), and the National Institute of Toxicological Research (NITR). The evaluation was arranged for mutual acceptance of data among GLP authorities. The designation process, additional documents necessary for applying GLP facility, the process of test facility evaluation including reviewing the application and site inspection, and inspection results and submission of correction plans are explained by using the instance of the inspection process of the COT. COT was evaluated as a suitable GLP facility for acute oral and inhalation toxicity tests and the Ames test.

Chromosome aberration and lipid peroxidation in chromium-exposed workers.

Chromosome aberration frequency and lipid peroxidation levels were analyzed to investigate their efficacy as biological markers for monitoring the genotoxicity and oxidative damage in Korean chromium (Cr)-exposed workers. Fifty-one Cr-exposed workers and 31 age-matched controls in ten chrome-plating plants were sampled. The Cr level was measured in the workers' blood and urine, and in the ambient air at the workplaces. The conventional Giemsa staining method and fluorescence in situ hybridization (FISH) technique were used for chromosome aberration analysis. Spectrum green whole chromosome paint specific for chromosome 4 was used in the FISH procedure. As for lipid peroxidation, malondialdehyde (MDA) was measured in the blood plasma as thiobarbituric acid-reactive substances (TBARS). The blood Cr concentration was statistically correlated with both the frequency of chromatid exchange and the total frequency of chromosome/chromatid breaks and exchanges, as detected by the Giemsa staining. Meanwhile, the frequency of translocation, as detected by the FISH technique, was significantly higher in the Cr-exposed workers than in the controls and it correlated with the blood Cr concentration. Although the concentration of MDA, the metabolite of lipid peroxidation, in the exposed workers was higher than that of the controls, no statistically significant correlation between the MDA level and the blood or urine Cr levels was observed. Accordingly, the genotoxicity and oxidative damage (plasma lipid peroxidation) in the Korean Cr-exposed workers were consequential at quite low exposure levels, plus chromosome rearrangement, especially translocation, was clearly evident as a biological response marker for Cr exposure based on a significant positive correlation between the translocations detected by FISH and the Cr in the blood.

History of the Korean GLPs and the activities and perspectives of the Korean Society of GLP.

The Korean Society of Good Laboratory Practice (KSGLP) was established Dec. 10, 1998. The objectives of the KSGLP are to enhance the quality of domestic studies and the level of GLP compliance, in safety testing, and to promote information exchange among its members. The activities of KSGLP include: offering workshops and symposiums, linking with related governmental organizations, collecting GLP related information and providing the information to the related organizations, building international networks to collect information and to establish relationship, developing training materials and publishing periodicals, and other business necessary to achieve the objectives of the KSGLP. The KSGLP achieved its goals within a short period of time by offering workshops and symposia, and by providing important GLP related information in newspapers or via the KSGLP's internet homepage (www.ksglp.or.kr). The main role of the KSGLP will be to disseminate GLP technology nationwide. The KSGLP would like to help many labs that are preparing their facilities for GLP compliance. Further, the KSGLP is hoping to share GLP experiences with other members.