Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is the only known intracellular antioxidant enzyme that can directly reduce lipid hydroperoxide in membrane. Mitochondrial and ...non-mitochondrial PHGPx and sperm nuclei GPx are transcribed from one gene by alternative transcription using different first exons Ia and Ib, respectively. To examine the role of PHGPx in development, we generated mice deficient in PHGPx by a targeted disruption of all exons of the PHGPx gene. Heterozygotes are viable, fertile, and appear normal, despite having decreased levels of three types of PHGPx mRNA and protein. Embryos homozygous for PHGPx-null die between 7.5 and 8.5 days post coitum (dpc), probably developing distal apoptosis. We examined the expression of PHGPx in mouse embryos using immunohistochemical analysis with anti-PHGPx mAb. The expression of PHGPx was detected in the embryonic ectoderm and the yolk sac membrane at 7.5
dpc. The results demonstrated that PHGPx is expressed in early gastrulation stage at 7.5
dpc and that the expression of PHGPx was essential for normal mouse development.
Cigarette smoke (CS) is a complex mixture of chemicals and interacts with various physiological processes. We previously reported that nuclear factor erythroid 2‐related factor 2 (NRF2) was the most ...sensitive transcription factor to aqueous CS extract (AqCSE) exposure in monolayer cultured human bronchial epithelial cell lines. Recently, in vitro three‐dimensional (3D) culture models have been used to supplement pharmacological and toxicological assessments. Bronchial epithelium models in particular are useful for the evaluation of substances that directly contact the respiratory tract, such as CS. In the present study, we used 3D‐cultured human bronchial epithelial cells (HBECs) to assess activation of transcription factors and relevant gene expression in response to AqCSE, primarily focusing on NRF2 and nuclear factor‐kappa B (NF‐κB) pathways. The 3D‐cultured HBECs exposed to AqCSE showed expression of NRF2 and its nuclear translocation in addition to upregulation of genes related to oxidative stress. Our results suggest that the NRF2 pathway was the dominant pathway when 3D‐cultured HBECs were exposed to AqCSE at a low dose, supporting our previous findings that NRF2 was the most sensitive transcription factor in response to AqCSE. Expression and nuclear translocation of NF‐κB were not increased, although proinflammatory genes were upregulated. However, another inflammation‐related transcription factor, activation protein 1, was induced by AqCSE. Gene classification analysis suggested that induction of the inflammatory response by AqCSE was dependent on NRF2 and activation protein 1 rather than NF‐κB.
We assessed activation of transcription factors and relevant signaling pathways when human bronchial epithelial cells in three‐dimensional culture were exposed to aqueous cigarette smoke extract (AqCSE). Our results suggest that the nuclear factor erythroid 2‐related factor 2 (NRF2) pathway is the dominant pathway at low‐dose exposure, supporting our previous findings that NRF2 is the most sensitive transcription factor to AqCSE. Moreover, the AqCSE‐induced inflammatory response is dependent on NRF2 and activation protein 1 rather than nuclear factor‐kappa B.
Purposes:
(a) To measure the urinary excretion of antineoplastic drugs of three patients during 48 h after the administration of cyclophosphamide (two patients) and 5-fluorouracil (one patient). (b) ...To evaluate environmental contamination with antineoplastic drugs via excreta of patients in the home setting. (c) To evaluate exposure of family members to antineoplastic drugs by measuring the drugs in their urine during the 48 h after completion of the chemotherapy by the patients.
Methods:
Two patients were administered cyclophosphamide by i.v. bolus injection. One patient was administered 5-fluorouracil by i.v. bolus injection and thereafter immediately administered the same drug by continuous infusion for 46 h. Urine samples from the patients administered cyclophosphamide and their family members, and wipe samples from their home environment, were analysed for the unchanged form of cyclophosphamide. For 5-fluorouracil, the urine samples from the patient and the family member were analysed for the 5-fluorouracil metabolite α-fluoro-β-alanine. Wipe samples were analysed for 5-fluorouracil. Drugs were detected and quantified with gas chromatography in tandem with mass spectroscopy-mass spectroscopy or by high-performance liquid chromatography with ultraviolet-light detection.
Results:
A total of 35 and 16 urine samples were collected from the three patients and their family members, respectively. The drugs were detected in all samples.
Cyclophosphamide was detected at levels of 0.03–7.34 ng/cm2 in 8 of the 12 wipe samples obtained from the homes of the patients administered cyclophosphamide. For the patient administered 5-fluorouracil, drug levels in his home environment were below the limit of detection.
Conclusion:
We demonstrated contamination of the home setting and exposure of family members to cyclophosphamide via the excreta of outpatient receiving chemotherapy. Exposure of the family member of the patient administered 5-fluorouracil was also demonstrated. These findings indicate the importance of strict precautions by the members of treated cancer patients as well as healthcare workers, to reduce the risk of exposure to antineoplastic drugs.
Flow cytometric detection of anti-HLA antibodies has yet to be optimally standardized. There is no consensus on whether serum or plasma specimens should be used. We studied interspecimen differences ...in anti-HLA antibody levels between serum and plasma using the flow cytometric method FlowPRA®. We simultaneously examined the serum and plasma of a total of 55 cases that were positive or negative for anti-HLA antibodies. The percentage of class I and class II beads that reacted with plasma was significantly correlated with those that reacted with serum (class I: r = 0.8570 p < 0.0001, class II: r = 0.7529 p < 0.0001). Plasma and serum, samples that were positive for anti-HLA antibodies were identical in all 55 cases for class I and in 54 cases for class II. In one case, class II anti-HLA antibodies were detected in plasma but not in serum. Mean fluorescence intensity in plasma samples was significantly higher than that in serum. Non-specific reaction due to high background intensity was seen in 2 plasma specimens out of 55 cases. Therefore, both serum and plasma specimens are useful for detecting anti-HLA antibodies using Flow PRA®, although the features of each specimen should be considered.
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