CCR2 monocytes as therapeutic targets for acute disc herniation and radiculopathy in mouse models.

OBJECTIVE: Back pain and radiculopathy caused by disc herniation are major health issues worldwide. While macrophages are key players in disc herniation induced inflammation, their roles and origins in disease progression remain unclear. We aim to study the roles of monocytes and derivatives in a mouse model of disc herniation. METHODS: Using a CCR2-CreER; R26R-EGFP (Ai6) transgenic mouse strain, we fate-mapped C-C chemokine receptor type 2 (CCR2) expressing monocytes and derivatives at disc herniation sites, and employed a CCR2RFP/RFP mouse strain and a CCR2-specific antagonist to study the effects of CCR2+ monocytes on local inflammatory responses, pain level, and disc degeneration by immunostaining, flow cytometry, and histology. RESULTS: CCR2+ monocytes (GFP+) increased at the sites of disc hernia over postoperative day 4, 6, and 9 in CCR2-CreER; Ai6 mice. F4/80+ cells increased, and meanwhile, CD11b+ cells trended downward. Co-localization analysis revealed that both GFP+CD11b+ and GFP+F4/80+ constituted the majority of CD11b+ and F4/80+ cells at disc hernia sites. Fluorescence activated cell sorter purified GFP+ cells exhibited higher cytokine expressions than GFP- cells. Inhibition of CCR2 signaling reduced infiltration of monocytes and macrophages, alleviated pain, maintained disc height, and reduced osteoclast activity in adjacent cortical bone for up to 1 month. CONCLUSION: Our findings suggest that circulating CCR2+ monocytes play important roles in initiating and promoting the local inflammatory responses, pain sensitization, and degenerative changes after disc herniation, and thus may serve as therapeutic targets for disc herniation induced back and leg pain.

Recent Advances in CAR-Based Solid Tumor Immunotherapy.

Adoptive cell therapy using chimeric antigen receptor (CAR) technology is one of the most advanced engineering platforms for cancer immunotherapy. CAR-T cells have shown remarkable efficacy in the treatment of hematological malignancies. However, their limitations in solid tumors include an immunosuppressive tumor microenvironment (TME), insufficient tumor infiltration, toxicity, and the absence of tumor-specific antigens. Although recent advances in CAR-T cell design-such as the incorporation of co-stimulatory domains and the development of armored CAR-T cells-have shown promising results in treating solid tumors, there are still challenges that need to be addressed. To overcome these limitations, other immune cells, such as natural killer (NK) cells and macrophages (M), have been developed as attractive options for efficient cancer immunotherapy of solid tumors. CAR-NK cells exhibit substantial clinical improvements with "off-the-shelf" availability and low toxicity. CAR-M cells have promising therapeutic potential because macrophages can infiltrate the TME of solid tumors. Here, we review the recent advances and future perspectives associated with engineered immune cell-based cancer immunotherapies for solid tumors. We also summarize ongoing clinical trials investigating the safety and efficacy of engineered immune cells, such as CAR-T, CAR-NK, and CAR-M, for targeting solid tumors.

Clinical Evaluation of an Antigen Home Test Using Surface-Enhanced Raman Spectroscopy and Stacking Pad for SARS-CoV-2 Screening with Nasal and Salivary Swab Samples.

This prospective study aimed to evaluate the performance of the InstaView COVID-19 (coronavirus diseases 2019) Antigen Home Test (InstaView AHT) which detects severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens. In this test kit, surface-enhanced Raman spectroscopy was used, a stacking pad was inserted, and nasal swab and salivary swab samples were used simultaneously to improve performance. The clinical performance of the InstaView AHT was compared to that of RT-PCR using nasopharyngeal samples. The participants without any prior training were recruited and performed the sample collection, testing, and interpretation of the results by themselves. Of the 91 PCR-positive patients, 85 had positive InstaView AHT results. The sensitivity and specificity of the InstaView AHT were 93.4% (95% confidence interval [CI]: 86.2-97.5) and 99.4% (95% CI: 98.2-99.9). The sensitivity of the InstaView AHT was above 90% for all samples obtained from patients with Ct ≤ 20, 20 < Ct ≤ 25, and 25 < Ct ≤ 30 (100%, 95.1%, and 92.0%, respectively). The InstaView AHT can be used as an alternative to RT-PCR testing because of its relatively high sensitivity and specificity, especially when SARS-CoV-2 prevalence is high, and the availability of RT-PCR testing is limited.

Outcomes Following Anterior Cervical Discectomy and Fusion in Patients With Ehlers-Danlos Syndrome.

STUDY DESIGN: Retrospective database analysis. OBJECTIVES: To study postoperative complication rates following anterior cervical discectomy and fusion (ACDF) in patients with Ehlers-Danlos syndrome (EDS) compared with patients without EDS. METHODS: The Mariner database was utilized to identify patients with EDS undergoing one or two level anterior cervical discectomy and fusion (ACDF). Postoperative short-term outcomes assessed included medical complications, readmissions, and ED-visits within 90 days of surgery. Additionally, surgical complications including wound complications, surgical site infection, one- and two-year anterior revision along with posterior revision, pseudarthrosis, and hardware failure within 2 years were assessed. Multivariate logistic regression was used to adjust for demographic variables, comorbidities and number of levels operated on. RESULTS: The present study identified 533 patients in the EDS group and 2634 patients in the matched control group. EDS patients undergoing ACDF are at an increased risk for 90-day major medical complications (OR 3.31; P < .001). EDS patients were also found to be associated with surgical complications including wound complications (OR 2.94; P < .001), surgical site infection (OR 8.60; P < .001) within 90 days, pseudarthrosis (OR 2.33; P < .001), instrument failure (OR 4.03; P < .001), anterior revision (OR 22.87; P < .001), and posterior revision (OR 3.17; P < .001) within 2 years. CONCLUSIONS: EDS is associated with higher rates of both medical and surgical complications following ACDF. Spine surgeons should be cognizant of the increased risks in this population to provide appropriate preoperative counseling and enhanced perioperative medical management.

Inhalation exposure to chloramine T induces DNA damage and inflammation in lung of Sprague-Dawley rats.

Chloramine T has been widely used as a disinfectant in many areas such as kitchens, laboratories and hospitals. It has been also used as a biocide in air fresheners and deodorants which are consumer products; however, little is known about its toxic effects by inhalation route. This study was performed to identify the subacute inhalation toxicity of chloramine T under whole-body inhalation exposure conditions. Male and female groups of rats were exposed to chloramine T at concentrations of 0.2, 0.9 and 4.0 mg/m³ for 6 hr/day, 5 days/week during 4 weeks. After 28-day repeated inhalation of chloramine T, there were dose-dependently significant DNA damage in the rat tissues evaluated and inflammation was histopathologically noted around the terminal airways of the lung in both genders. As a result of the expression of three types of antioxidant enzymes (SOD-2, GPx-1, PRX-1) in rat's lung after exposure, there was no significant change of all antioxidant enzymes in the male and female rats. The results showed that no observed adverse effect level (NOAEL) was 0.2 mg/m³ in male rats and 0.9 mg/m³ in female rats under the present experimental condition.

Differential gene expression profiles in spontaneously hypertensive rats induced by administration of enalapril and nifedipine.

Enalapril and nifedipine are used as antihypertensive drugs; however, the therapeutic target molecules regulated by enalapril and nifedipine have yet to be fully identified. The aim of this study was to identify novel target genes that are specifically regulated by enalapril and nifedipine in tissues from spontaneously hypertensive rats (SHR) using DNA microarray analysis. We found that administration of SHR with enalapril and nifedipine differentially regulated 33 genes involved in the pathogenesis of cardiovascular diseases. Furthermore, we identified 16 genes that have not previously been implicated in cardiovascular diseases, including interleukin-24 (IL-24). Among them, exogenous administration of IL-24 attenuated the expression of vascular inflammation and hypertension-related genes induced by H2O2 treatment in mouse vascular smooth muscle (MOVAS) cells. This study provides valuable information for the development of novel antihypertensive drugs. In addition, the genes identified may be of use as biomarkers and therapeutic targets for cardiovascular diseases, including hypertension.

Korea National Survey for Environmental Pollutants in the human body 2008: 1-hydroxypyrene, 2-naphthol, and cotinine in urine of the Korean population.

The Korea National Survey for Environmental Pollutants in the human body conducts representative Korean population studies, which were first initiated in 2005 in Korea. This study was conducted from 2008 to 2009 to determine the exposure levels of polycyclic aromatic hydrocarbons and nicotine in the Korean general population. The study population consisted of 4702 adult subjects from 196 sampling locations including coastal, rural, and urban areas. The urinary levels of 1-hydroxypyrene, 2-naphthol, and cotinine were measured for exposure of polycyclic aromatic hydrocarbons and nicotine. The geometric means of the urinary 1-hydroxypyrene, 2-naphthol and cotinine concentrations in the Korean general population were 0.15 µg/L (95% confidence interval (CI): 0.13-0.17), 3.84 µg/L (95% CI: 3.57-4.11) and 47.42 µg/L (95% CI: 40.52-54.32) respectively. When these values were compared with reference ranges for the United States and Germany, the levels of 1-hydroxypyrene, 2-naphthol, and cotinine were very similar for Korea and Germany, however, these levels were slightly lower in the United States. This study is the first nationwide survey of exposure to polycyclic aromatic hydrocarbons and nicotine in Korea and provides a background reference range for exposure to polycyclic aromatic hydrocarbons and nicotine in the Korean general population.

Subacute inhalation toxicity assessment of fly ash from industrial waste incinerators.

Fly ash from industrial waste incinerators has been a significant concern because of their constituent toxic heavy metals and organic compounds. The objective of this study was to identify the subacute inhalation toxicity of fly ash from industrial waste incinerators, using whole body inhalation exposure chambers. Male and female groups of Sprague-Dawley rats were exposed to fly ash by inhalation of concentrations of 0, 50, 100, 200 mg/m(3), for 6 h/day, 5 days/week for 4 weeks. There was no significant difference in body weight, and relative organ weight to body weight, between the exposure groups and the control group. Hematological examinations revealed a significant increase of monocyte counts in fly ash exposed rats and brown pigment laden macrophage was found in the lungs of rats exposed to high concentration of fly ash. A decrease of blood glucose levels and an increase in glutamate oxaloacetate transaminase activity were observed in fly ash treated rats. There was also a significant increase of lactate dehydrogenase levels in rat blood exposed fly ash. A significant dose-dependent increase of DNA damage was found in lymphocytes, spleen, bronchoalveolar lavage, liver, lung, and thymus of rats exposed to fly ash. In addition, the level of lipid peroxidation was increased in the plasma of rats exposed to a high concentration of fly ash. These results suggest that inhalation of fly ash from industrial waste incinerators can induce histopathologic, hematological, and serum biochemical changes and oxidative damage.

Korea National Survey for Environmental Pollutants in the Human Body 2008: heavy metals in the blood or urine of the Korean population.

BACKGROUND: Recently, there have been several nationwide episodes involving imported toys contaminated with toxic metals and environmental hormones. In addition, cadmium intoxication has occurred due to soil contamination with cadmium from abandoned metal mines. OBJECTIVES: To investigate the distribution, extent and factors influencing the levels of toxic metals in the blood or urine of the Korean general population over twenty years of age, we studied the blood or urine concentrations of heavy metals in a representative sample of 5087 Koreans in 2008. METHODS: Multiple biological substrates were collected from each participant to determine the most suitable samples for an environmental health survey system. Information regarding exposure conditions of all subjects was collected by questionnaire-based interviews. RESULTS: The geometric means of the blood lead, mercury and manganese levels were 19.1, 3.23 and 10.8 µg/L, respectively. The geometric means of urinary arsenic and cadmium concentrations were 43.5 and 0.65 µg/L, respectively. Blood mercury and urinary arsenic levels in the Korean general population were significantly higher than in European and American populations. CONCLUSIONS: The higher levels of blood mercury and urinary arsenic could be explained by the greater seafood consumption among the Korean population. This biomonitoring study of blood or urine heavy metals in the Korean general population provides important reference data stratified by demographic and lifestyle factors that will be useful for the ongoing surveillance of environmental exposure of Koreans to toxic metals.

Ethanol-induced DNA damage and repair-related molecules in human intestinal epithelial Caco-2 cells.

The acute administration of ethanol to intestinal epithelial cells causes increased intestinal permeability and the translocation of endotoxins. The changes caused by ethanol in intestinal cells may be related to oxidative stress and DNA damage. However, DNA damage and repair-related molecules which act against stresses, including ethanol, have not been fully investigated in intestinal cells. Heat shock proteins (Hsps) are involved in the recovery and protection from cell damage and may be associated with DNA repair. Therefore, the aim of our study was to investigate cytotoxicity, DNA damage and the expression of DNA repair-related molecules, antioxidant proteins and Hsps in intestinal cells exposed to ethanol. Human intestinal Caco-2 cells were incubated with 1-8% ethanol for 1 h. Cell viability and DNA damage were determined using the MTT and comet assays, respectively. We measured DNA repair-related molecules, including DNA polymerase ß, apurinic/apyrimidinic endonuclease/redox factor-1 (APE/Ref-1), growth arrest and DNA damage 45α (GADD45α) and proliferating cell nuclear antigen (PCNA), in Caco-2 cells using western blot analysis. We also measured glutathione peroxidase-1 (GPx-1), peroxiredoxin-1 (PRX-1), superoxide dismutase-2 (SOD-2), Hsp10, Hsp27, Hsp60, heat shock cognate (Hsc)70, Hsp70 and Hsp90. The viability of the Caco-2 cells exposed to ethanol decreased at concentrations ≥ 7% (P<0.05). The Olive tail moment, indicating DNA damage, increased dose dependently in ≥ 3% ethanol (P<0.05). Among the DNA repair proteins, the expression of PCNA and APE/Ref-1 increased significantly at 1% ethanol. Antioxidant enzymes, including GPx-1, PRX-1 and SOD-2, had an increased expression at 1% ethanol. Hsp10, Hsp27 and Hsp70 expression also increased significantly at 1% ethanol. In conclusion, the expression of DNA repair molecules, antioxidants and Hsps increased in intestinal Caco-2 cells exposed to low concentrations of ethanol. In particular, PCNA, APE/Ref-1, Hsp10, Hsp27 and Hsp70 were sensitive to low ethanol concentrations, indicating that they may be useful in evaluating the DNA repair and cytoprotective effects of the drug against stress in intestinal cells.

Comet assay analysis of DNA damage in T- and B-lymphocytes separated by MACS for human biomonitoring studies.

In this study, we developed and tested a method for human biomonitoring using Comet assays with human T- and B-lymphocytes obtained by magnetic cell sorting (MACS). We evaluated DNA damage induced by treatment with hydrogen peroxide (H(2)O(2); 5, 25 and 50µM) and methyl methane sulfonate (MMS; 5, 25 and 50µM) in both human B- and T-lymphocytes obtained by MACS, and compared their DNA damage levels. Significant, dose-dependent levels of DNA damage were found in T-lymphocytes and B-lymphocytes. Furthermore, the level of DNA damage was significantly greater in B-lymphocytes than in T-lymphocytes, suggesting that human B-lymphocytes may be a more sensitive target than T-lymphocytes for the evaluation of DNA damage. In addition, we compared these in vitro exposure data with previous studies that showed DNA damage in B- and T-lymphocyte and granulocytes of control subjects and industrial workers exposed in vivo to environmental toxicants. The use of single types of human peripheral blood mononuclear cells obtained by MACS, for Comet assays gave sensitive and reliable data for human biomonitoring for environmental toxicants.

Inter- and intra-individual variations of urinary endogenous metabolites in healthy male college students using (1)H NMR spectroscopy.

BACKGROUND: Most human metabolomics studies have shown that spectral outputs of (1)H nuclear magnetic resonance fingerprinting are strongly influenced by inter- and intra-individual variations; however, few studies have been performed to evaluate the inter- and intra-individual variations in urinary endogenous metabolites. METHODS: We recruited 30 male college students to evaluate the factors affecting intra- and inter-individual variations in urinary endogenous metabolites. Statistical analysis for variations in urinary metabolites was performed after eliminating outliers found in principal component analysis (PCA) plots. RESULTS: Inter-individual variations were relatively low for 2-oxoglutarate, succinate, citrate, dimethylglycine, and taurine, but high for trimethylaminoxide (TMAO), hippurate, and lactate. Intra-individual variations for 2-oxoglutarate, citrate, dimethylglycine, and taurine were relatively low, but high for TMAO and hippurate. The factors affecting inter-individual variation of lactate were age, body mass index, beverages, and alcohol, whereas the factors affecting intra-individual variation of lactate were age and fish. Kim Chi intake affected the inter-individual variation of succinate, citrate, TMAO, and hippurate; however, it did not affect the intra-individual variation of endogenous metabolites. CONCLUSIONS: Our results showed that inter- and intra-individual variations in urinary endogenous metabolites were very large, and significant factors affecting inter- and intra-individual variation were diverse, even after eliminating outliers in PCA analysis.

Characterization of biological effect of 1763 MHz radiofrequency exposure on auditory hair cells.

PURPOSE: Radiofrequency (RF) exposure at the frequency of mobile phones has been reported not to induce cellular damage in in vitro and in vivo models. We chose HEI-OC1 immortalized mouse auditory hair cells to characterize the cellular response to 1763 MHz RF exposure, because auditory cells could be exposed to mobile phone frequencies. MATERIALS AND METHODS: Cells were exposed to 1763 MHz RF at a 20 W/kg specific absorption rate (SAR) in a code division multiple access (CDMA) exposure chamber for 24 and 48 h to check for changes in cell cycle, DNA damage, stress response, and gene expression. RESULTS: Neither of cell cycle changes nor DNA damage was detected in RF-exposed cells. The expression of heat shock proteins (HSP) and the phosphorylation of mitogen-activated protein kinases (MAPK) did not change, either. We tried to identify any alteration in gene expression using microarrays. Using the Applied Biosystems 1700 full genome expression mouse microarray, we found that only 29 genes (0.09% of total genes examined) were changed by more than 1.5-fold on RF exposure. CONCLUSION: From these results, we could not find any evidence of the induction of cellular responses, including cell cycle distribution, DNA damage, stress response and gene expression, after 1763 MHz RF exposure at an SAR of 20 W/kg in HEI-OC1 auditory hair cells.

Molecular responses of Jurkat T-cells to 1763 MHz radiofrequency radiation.

PURPOSE: The biological effects of exposure to mobile phone emitted radiofrequency (RF) radiation are the subject of intense study, yet the hypothesis that RF exposure is a potential health hazard remains controversial. In this paper, we monitored cellular and molecular changes in Jurkat human T lymphoma cells after irradiating with 1763 MHz RF radiation to understand the effect on RF radiation in immune cells. MATERIALS AND METHODS: Jurkat T-cells were exposed to RF radiation to assess the effects on cell proliferation, cell cycle progression, DNA damage and gene expression. Jurkat cells were exposed to 1763 MHz RF radiation at 10 W/kg specific absorption rate (SAR) and compared to sham exposed cells. RESULTS: RF exposure did not produce significant changes in cell numbers, cell cycle distributions, or levels of DNA damage. In genome-wide analysis of gene expressions, there were no genes changed more than two-fold upon RF-radiation while ten genes change to 1.3 approximately 1.8-fold. Among ten genes, two cytokine receptor genes such as chemokine (C-X-C motif) receptor 3 (CXCR3) and interleukin 1 receptor, type II (IL1R2) were down-regulated upon RF radiation, but they were not directly related to cell proliferation or DNA damage responses. CONCLUSION: These results indicate that the alterations in cell proliferation, cell cycle progression, DNA integrity or global gene expression was not detected upon 1763 MHz RF radiation under 10 W/kg SAR for 24 h to Jurkat T cells.

Panax ginseng effects on DNA damage, CYP1A1 expression and histopathological changes in testes of rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

The effects of Panax ginseng extracts on DNA damage, expression of cytochrome P450 (CYP) 1A1 and reproductive toxicity were evaluated in the testis of rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxinthe (TCDD). Fifty rats were divided into five groups according to treatment with 2,3,7,8-TCDD and P. ginseng extracts. Single cell gel electrophoresis assays were performed to evaluate DNA damage that occurred in the lymphocytes of rats. Histological changes in the seminiferous tubules of the testis were determined using Johnsen's scoring system and Real Time-PCR was performed to evaluate the mRNA expression of CYP1A1. Significant pathological effects were observed in the 2,3,7,8-TCDD treated rats including a reduced seminiferous tubular diameter, an increased number of damaged tubules (maturation arrest, eosinophilic degeneration and spermatid giant cells) and increased Johnsen's score. DNA damage and the expression of CYP1A1 mRNA were significantly increased in rat testes. There were no significant differences between the control and animals treated with P. ginseng extracts. However, a significantly decreased level of DNA damage, decreased CYP1A1 expression and reduced pathological effects were observed in the 2,3,7,8-TCDD with P. ginseng extracts treated groups when compared with the TCDD treated group. In summary, our study demonstrates that 2,3,7,8-TCDD induces the pathological and genotoxical damage in rat testes, while P. ginseng extract treatment exhibits a therapeutic capacity to reduce these effects via reduction of CYP1A1 mRNA.

[Increased DNA damage of lymphocytes in Korean male smokers].

OBJECTIVE: The purpose of this study was to evaluate the levels of DNA damage in human lymphocytes caused by smoking and other lifestyle factors. METHODS: The study population consisted of 173 normal healthy male adults from 21 to 59 years old. The demographic and lifestyle variables were obtained from administered questionnaires. The level of lymphocytic DNA damage in the peripheral blood was evaluated by the Comet assay. Statistical analyses were done by general linear model analysis and Dunnett's multiple comparison. RESULTS: The difference in DNA damage between smokers and non-smokers was statistically significant. The means for the Tail%DNA were found to be 10.48 in the current smokers and 9.60 in the non-smokers (p < 0.05). The tail moment means were 1.58 and 1.45 (p < 0.05) for the current smokers and non-smokers, respectively. The number of cigarettes smoked per day did not result in a significant difference in the level of DNA damage among the smokers. Other lifestyle factors such as age, and drinking and exercise habits were not related to DNA damage. CONCLUSIONS: The DNA damage in the lymphocytes of smokers was found to be significantly higher than that for non-smokers. However, the number of cigarettes smoked per day was not related to DNA damage. Further study is needed to evaluate the relationship between the amount of smoking and level of damage to DNA. In addition, the status of DNA repair activities should be assessed.

Gene expression profiling in lung tissues from rats exposed to formaldehyde.

Formaldehyde is a ubiquitous toxic organic compound recently classified as a carcinogen by the International Agency for Research on Cancer and one of the major factors causing sick building syndrome. In this study, we have investigated the effects of formaldehyde on mRNA expression in rat lung tissues by applying genomics. Rats were exposed to ambient air and two different concentrations of formaldehyde (0, 5, 10 ppm) for 2 weeks at 6 h/day and 5 days/week in an inhalation chamber. Malondialdehyde (MDA) assay and carbonyl spectrometric assay were conducted to determine lipid peroxidation and protein oxidation levels and Comet assays were used for genotoxicity evaluation. Level of MDA, carbonyl insertion and DNA damage in the lungs of rats exposed to FA were found to be dose dependently increased. Gene expression was evaluated by using a bio-array hybridization analysis. A total of 21 (2 up- and 19 down-regulated) genes were identified as biomarkers for formaldehyde effects. Several differentiated gene groups were found. Genes involved in apoptosis, immunity, metabolism, signal transduction, transportation, coagulation and oncogenesis were found to be up- and down-regulated. Among these genes, the mRNA expressions of cytochrome P450, hydroxymethylbilane synthase, glutathione reductase, carbonic anhydrase 2, natriuretic peptide receptor 3, lysosomal associated protein transmembrane 5, regulator of G-protein signaling 3, olfactomedin related ER localized protein, and poly (ADP-ribose) polymerase-1 were confirmed by quantitative RT-PCR analysis. In summary, the MDA lipid peroxidation and the carbonyl protein oxidation assays showed that cytotoxic effects increased with increasing formaldehyde levels. Genomic analysis showed that 21 genes were up- or down-regulated. Of these genes, nine were confirmed by quantitative RT-PCR and could be potential biomarkers for human diseases associated with formaldehyde exposure.

Evaluation of toxicological monitoring markers using proteomic analysis in rats exposed to formaldehyde.

Formaldehyde (FA) is known as a low molecule weight organic compound and one of major components that causes sick building syndrome (SBS), and it has been reported that FA has cytotoxic, hemotoxic, immunotoxic, and genotoxic properties. The International Agency for Research on Cancer (IARC) has characterized FA as a carcinogen. In this study, we investigated the effects of FA on rat plasma proteins by using proteomic approach. Rats were exposed to three different concentrations of FA (0, 5, 10 ppm) for 2 weeks at 6 hours/day and 5 days/week in an inhalation chamber. Malondialdehyde (MDA) assay and carbonyl spectrometric assay were conducted to determine lipid peroxidation and protein oxidation levels and Comet assays were used for genotoxicity evaluation. Level of MDA, carbonyl insertion and DNA damage in plasma, livers, and in the lymphocytes of rats exposed to FA were found to be dose dependently increased. Proteomic analysis using three different pI ranges (3.5-5.6, 5.3-6.9, 6-9) and large size two-dimensional gel electrophoresis (2-DE) showed the presence of 3491 protein spots. A total of 32 (19 up- and 13 down-regulated) proteins were identified as biomarkers of FA, all showed dose dependent expressions in the plasma of rats exposed to FA and of these, 27 protein spots were identified by MALDI-TOF/MS. Several differentiated protein groups were found. Proteins involved in apoptosis, transportation, signaling, energy metabolism, and cell structure and motility were found to be up- or down-regulated. Among these, the identities of SNAP 23, apolipoprotein A-1 and E, clusterin, kinesin, and fibrinogen gamma were confirmed by Western blot assay, and apo E was further analyzed by using 2-DE immunoblot assays to determine isoform patterns. Two cytokine including IL4 and INF-gamma were measured in plasma with respect to fibrinogen gamma changes. In summary, cytotoxicity, and genotoxicity assays, namely MDA lipid peroxidation assay, the carbonyl protein oxidation assay, and Comet genotoxic assay showed that these effects increased on increasing FA levels. Proteomic analysis with three different pI ranges and long size 2-DE gel electrophoresis showed that 32 protein spots were up-or down-regulated. Of these 32 proteins, 7 proteins were confirmed by western blot assay. They could be potential biomarkers for human diseases associated with FA exposure.

Oxidative damages in the DNA, lipids, and proteins of rats exposed to isofluranes and alcohols.

DNA damage, lipid peroxidation and protein oxidation were evaluated in rats exposed to a 1% isoflurane atmosphere with or without alcohol administration (administrated by gastric intubation at 4 g/kg body weight as a 50% solution). Single cell gel electrophoresis assays were performed in order to evaluate DNA damage occurring in the lymphocytes, spleen, bone marrow, brain, livers and lung of rats exposed to 1% isoflurane for 30 or 60 min with/without ethanol. Levels of malondialdehydes (MDA), a metabolite of lipid peroxidation, were determined in plasma and tissues. Carbonyl contents were also analyzed to determine levels of protein oxidation in plasma and tissues. Levels of DNA damage in lymphocytes, bone marrow, and the organ tissues of rats exposed to isoflurane were found to increase time dependently, and alcohol increased DNA damage. Lipid peroxidation and protein oxidation results showed patterns that differed from those of DNA damage. Levels of MDA in plasma, bone marrow, spleen, and the livers of rats exposed to isoflurane with/without ethanol were found to be time dependently increased, but this was not observed in the brain or lung. However, protein oxidation levels were significantly increased in the plasma, brains, and lungs of rats exposed to isoflurane, and exposure to isoflurane and alcohol, significantly increased these levels in plasma and brain. The present study demonstrates that isoflurane exposure results in significant DNA damage in rat lymphocytes, bone marrow, spleen, brain, livers, and lung. Moreover, alcohol was found to be as a strong inducer of DNA damage, lipid peroxidation and protein oxidation. However, no evidence in association between DNA damage, lipid peroxidation and protein oxidation was found. Regarding the effects of isoflurane and alcohol on oxidative damages, single strand DNA damages may be a useful biomarkers and blood cells and plasma appear to be more sensitive targets to oxidative damage than other tissues.

Comparison of immunnological and genotoxicological parameters in automobile emission inspectors exposed to polycyclic aromatic hydrocarbons.

In this study, we investigated the immunotoxicities of polycyclic aromatic hydrocarbons in 54 automobile emission inspectors and in 84 control subjects, and evaluated associations between immunological and genotoxicological parameters. Specific surface antigens of peripheral lymphocytes, namely, CD3, CD4, CD8, CD19, and CD69 were subjected to measure immune status in automobile emission inspectors and control subjects. T-and B-cells showed no significant differences between automobile emission inspectors and control subjects (p=0.740 and 0.395). In addition, the ratio of T helper cells to T cytotoxic cells was not deferent (p=0.144). However, T-cell activation was found to be significantly higher in automobile emission inspectors (p=0.041), but not B-cell activation. The levels of two cytokines (IL-4 an INF-γ) and four immunoglobulins (IgA, IgE, IgG, and IgM) were also determined in automobile emission inspectors and control subjects. All immunoglobulin types were lower in automobile emission inspectors, but this was significant only for IgG (0.047). In addition, the levels of two cytokines, IL-4 and INF-γ, were also higher in automobile emission inspectors, though this was not significant. DNA damage in mononuclear and polynuclear lymphocytes and in the level of urinary metabolites, 1-OHP and 2-naphthol, were evaluated in automobile emission inspectors and in control subjects and significant differences were found between the two groups. Examinations of urinary metabolites, DNA damage, and immunological parameters, including leukocyte subpopulations, immunoglobulins, and cytokines, showed that the cytokines levels were associated with the levels of two urinary metabolites, 1-OHP and 2-naphthol.