RESUMEN
The goal of this study was to investigate the impact of multiple exposomal factors (genetics, lifestyle factors, environmental/occupational exposures) on pulmonary inflammation and corresponding alterations in local/systemic immune parameters. Accordingly, male Sprague-Dawley (SD) and Brown Norway (BN) rats were maintained on either regular (Reg) or high fat (HF) diets for 24wk. Welding fume (WF) exposure (inhalation) occurred between 7 and 12wk. Rats were euthanized at 7, 12, and 24wk to evaluate local and systemic immune markers corresponding to the baseline, exposure, and recovery phases of the study, respectively. At 7wk, HF-fed animals exhibited several immune alterations (blood leukocyte/neutrophil number, lymph node B-cell proportionality)-effects which were more pronounced in SD rats. Indices of lung injury/inflammation were elevated in all WF-exposed animals at 12wk; however, diet appeared to preferentially impact SD rats at this time point, as several inflammatory markers (lymph node cellularity, lung neutrophils) were further elevated in HF over Reg animals. Overall, SD rats exhibited the greatest capacity for recovery by 24wk. In BN rats, resolution of immune alterations was further compromised by HF diet, as many exposure-induced alterations in local/systemic immune markers were still evident in HF/WF animals at 24wk. Collectively, HF diet appeared to have a greater impact on global immune status and exposure-induced lung injury in SD rats, but a more pronounced effect on inflammation resolution in BN rats. These results illustrate the combined impact of genetic, lifestyle, and environmental factors in modulating immunological responsivity and emphasize the importance of the exposome in shaping biological responses.
Asunto(s)
Contaminantes Ocupacionales del Aire , Exposoma , Lesión Pulmonar , Exposición Profesional , Neumonía , Soldadura , Ratas , Masculino , Animales , Ratas Sprague-Dawley , Ratas Endogámicas BN , Lesión Pulmonar/inducido químicamente , Dieta Alta en Grasa/efectos adversos , Exposición por Inhalación/efectos adversos , Exposición por Inhalación/análisis , Neumonía/inducido químicamente , Inflamación , Biomarcadores , Contaminantes Ocupacionales del Aire/toxicidadRESUMEN
Welding fumes were reclassified as a Group 1 carcinogen by the International Agency for Research on Cancer in 2017. Gas metal arc welding (GMAW) is a process widely used in industry. Fume generated from GMAW-mild steel (MS) is abundant in iron with some manganese, while GMAW-stainless steel (SS) fume also contains significant amounts of chromium and nickel, known carcinogenic metals. It has been shown that exposure to GMAW-SS fume in A/J mice promotes lung tumors. The objective was to determine if GMAW-MS fume, which lacks known carcinogenic metals, also promotes lung tumors in mice. Male A/J mice received a single intraperitoneal injection of corn oil or the initiator 3-methylcholanthrene (MCA; 10 µg/g) and, one week later, were exposed by whole-body inhalation to GMAW-MS aerosols for 4 hours/day x 4 days/week x 8 weeks at a mean concentration of 34.5 mg/m3. Lung nodules were enumerated by gross examination at 30 weeks post-initiation. GMAW-MS fume significantly increased lung tumor multiplicity in mice initiated with MCA (21.86 ± 1.50) compared to MCA/air-exposed mice (8.34 ± 0.59). Histopathological analysis confirmed these findings and also revealed an absence of inflammation. Bronchoalveolar lavage analysis also indicated a lack of lung inflammation and toxicity after short-term inhalation exposure to GMAW-MS fume. In conclusion, this study demonstrates that inhalation of GMAW-MS fume promotes lung tumors in vivo and aligns with epidemiologic evidence that shows MS welders, despite less exposure to carcinogenic metals, are at an increased risk for lung cancer.
Asunto(s)
Contaminantes Ocupacionales del Aire/toxicidad , Carcinógenos/toxicidad , Hierro/toxicidad , Neoplasias Pulmonares/inducido químicamente , Acero , Soldadura , Administración por Inhalación , Animales , Neoplasias Pulmonares/patología , Masculino , RatonesRESUMEN
UNLABELLED: One of the factors that can result in musculoskeletal injuries, and time off work, is exposure to repetitive motion. The goal of this study was to determine if skeletal muscle injury induced by exposure to injurious stretch-shortening cycles (iSSCs), resulted in hyperalgesia in the hind limb and changes in calcitonin-gene related peptide (CGRP) immunolabeling in the dorsal root ganglia (DRG) in young and old male rats. METHODS: Young (3months) and old (30months) male Fisher 344×BN F1 rats were anesthetized with isoflurane and the left hind limbs were exposed to 15 sets of 10 SSCs. Control animals were exposed to a single bout of SSCs of equal intensity. Sensitivity to mechanical stimulation was assessed using von Frey filaments prior to beginning the experiment, and on days 2 and 9 following exposure to iSSCs. Rats were euthanized one, 3 or 10days after the exposure. The ipsilateral DRG were dissected from the L4-5 region of the spine, along with the left tibialis anterior (LTA) muscle. RESULTS: Rats exposed to iSSCs were more sensitive to mechanical stimulation than control rats 2days after the exposure, and showed a reduction in peak force 3days after exposure. Changes in sensitivity to pressure were not associated with increases in CGRP labeling in the DRG at 3days. However, 9days after exposure to iSSCs, old rats still displayed an increased sensitivity to mechanical stimulation, and this hyperalgesia was associated with an increase in CGRP immunolabeling in the DRG. Young rats exposed to iSSC did not display a change in CGRP immunolabeling and sensitivity to mechanical stimulation returned to control levels at 10days. CONCLUSIONS: These findings suggest that hyperalgesia seen shortly after exposure to iSSC is not influenced by CGRP levels. However, in cases where recovery from injury may be slower, as it is in older rats, CGRP may contribute to the maintenance of hyperalgesia.
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Péptido Relacionado con Gen de Calcitonina/biosíntesis , Trastornos de Traumas Acumulados/complicaciones , Contracción Muscular/fisiología , Músculo Esquelético/lesiones , Envejecimiento/metabolismo , Envejecimiento/fisiología , Animales , Péptido Relacionado con Gen de Calcitonina/fisiología , Trastornos de Traumas Acumulados/metabolismo , Trastornos de Traumas Acumulados/patología , Ganglios Espinales/metabolismo , Hiperalgesia/etiología , Hiperalgesia/metabolismo , Hiperalgesia/patología , Masculino , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Estimulación Física/métodos , Ratas Endogámicas F344 , Resistencia a la Tracción/fisiologíaRESUMEN
BACKGROUND: Epidemiological surveys indicate that occupants of mold contaminated environments are at increased risk of respiratory symptoms. The immunological mechanisms associated with these responses require further characterization. OBJECTIVE: The aim of this study was to characterize the immunotoxicological outcomes following repeated inhalation of dry Aspergillus fumigatus spores aerosolized at concentrations potentially encountered in contaminated indoor environments. METHODS: Aspergillus fumigatus spores were delivered to the lungs of naïve BALB/cJ mice housed in a multi-animal nose-only chamber twice a week for a period of 13 weeks. Mice were evaluated at 24 and 48 h post-exposure for histopathological changes in lung architecture, recruitment of specific immune cells to the airways, and serum antibody responses. RESULT: Germinating A. fumigatus spores were observed in lungs along with persistent fungal debris in the perivascular regions of the lungs. Repeated exposures promoted pleocellular infiltration with concomitant epithelial mucus hypersecretion, goblet cell metaplasia, subepithelial fibrosis and enhanced airway hyperreactivity. Cellular infiltration in airways was predominated by CD4(+) T cells expressing the pro-allergic cytokine IL-13. Furthermore, our studies show that antifungal T cell responses (IFN-γ(+) or IL-17A(+) ) co-expressed IL-13, revealing a novel mechanism for the dysregulated immune response to inhaled fungi. Total IgE production was augmented in animals repeatedly exposed to A. fumigatus. CONCLUSIONS & CLINICAL RELEVANCE: Repeated inhalation of fungal aerosols resulted in significant pulmonary pathology mediated by dynamic shifts in specific immune populations and their cytokines. These studies provide novel insights into the immunological mechanisms and targets that govern the health outcomes that result from repeated inhalation of fungal bioaerosols in contaminated environments.
Asunto(s)
Hongos/inmunología , Hipersensibilidad/etiología , Exposición por Inhalación/efectos adversos , Neumonía/etiología , Animales , Anticuerpos Antifúngicos/inmunología , Aspergillus fumigatus/inmunología , Citocinas/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Perfilación de la Expresión Génica , Hipersensibilidad/metabolismo , Hipersensibilidad/patología , Ratones , Fenotipo , Neumonía/metabolismo , Neumonía/patología , Esporas Fúngicas/inmunología , Linfocitos T/inmunología , Linfocitos T/metabolismoRESUMEN
BACKGROUND: Heart rate and cardiovascular function are regulated by the autonomic nervous system. Heart rate variability (HRV) as a marker reflects the activity of autonomic nervous system. The prognostic significance of HRV in cardiovascular disease has been reported in clinical and epidemiological studies. The present study focused on the influence of inhaled multi-walled carbon nanotubes (MWCNTs) on autonomic nervous system by HRV analysis. METHODS: Male Sprague-Dawley rats were pre-implanted with a telemetry device and kept in the individual cages for recovery. At week four after device implantation, rats were exposed to MWCNTs for 5 h at a concentration of 5 mg/m(3). The real-time EKGs were recorded by a telemetry system at pre-exposure, during exposure, 1 day and 7 days post-exposure. HRV was measured by root mean square of successive differences (RMSSD); the standard deviation of inter-beat (RR) interval (SDNN); the percentage of successive RR interval differences greater than 5 ms (pNN5) and 10 ms (pNN10); low frequency (LF) and high frequency (HF). RESULTS: Exposure to MWCNTs increased the percentage of differences between adjacent R-R intervals over 10 ms (pNN10) (p < 0.01), RMSSD (p < 0.01), LF (p < 0.05) and HF (p < 0.01). CONCLUSIONS: Inhalation of MWCNTs significantly alters the balance between sympathetic and parasympathetic nervous system. Whether such transient alterations in autonomic nervous performance would alter cardiovascular function and raise the risk of cardiovascular events in people with pre-existing cardiovascular conditions warrants further study.
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Arritmias Cardíacas/inducido químicamente , Sistema Nervioso Autónomo/efectos de los fármacos , Sistema Cardiovascular/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Nanotubos de Carbono/toxicidad , Administración por Inhalación , Animales , Arritmias Cardíacas/fisiopatología , Sistema Nervioso Autónomo/fisiopatología , Sistema Cardiovascular/inervación , Electrocardiografía Ambulatoria , Masculino , Ratas Sprague-Dawley , Medición de Riesgo , Telemetría , Factores de TiempoRESUMEN
Carbon nanotubes were among the earliest products of nanotechnology and have many potential applications in medicine, electronics, and manufacturing. The low density, small size, and biological persistence of carbon nanotubes create challenges for exposure control and monitoring and make respiratory exposures to workers likely. We have previously shown mitotic spindle aberrations in cultured primary and immortalized human airway epithelial cells exposed to 24, 48 and 96 µg/cm(2) single-walled carbon nanotubes (SWCNT). To investigate mitotic spindle aberrations at concentrations anticipated in exposed workers, primary and immortalized human airway epithelial cells were exposed to SWCNT for 24-72 h at doses equivalent to 20 weeks of exposure at the Permissible Exposure Limit for particulates not otherwise regulated. We have now demonstrated fragmented centrosomes, disrupted mitotic spindles and aneuploid chromosome number at those doses. The data further demonstrated multipolar mitotic spindles comprised 95% of the disrupted mitoses. The increased multipolar mitotic spindles were associated with an increased number of cells in the G2 phase of mitosis, indicating a mitotic checkpoint response. Nanotubes were observed in association with mitotic spindle microtubules, the centrosomes and condensed chromatin in cells exposed to 0.024, 0.24, 2.4 and 24 µg/cm(2) SWCNT. Three-dimensional reconstructions showed carbon nanotubes within the centrosome structure. The lower doses did not cause cytotoxicity or reduction in colony formation after 24h; however, after three days, significant cytotoxicity was observed in the SWCNT-exposed cells. Colony formation assays showed an increased proliferation seven days after exposure. Our results show significant disruption of the mitotic spindle by SWCNT at occupationally relevant doses. The increased proliferation that was observed in carbon nanotube-exposed cells indicates a greater potential to pass the genetic damage to daughter cells. Disruption of the centrosome is common in many solid tumors including lung cancer. The resulting aneuploidy is an early event in the progression of many cancers, suggesting that it may play a role in both tumorigenesis and tumor progression. These results suggest caution should be used in the handling and processing of carbon nanotubes.
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Mitosis/efectos de los fármacos , Nanotubos de Carbono/toxicidad , Mucosa Respiratoria/efectos de los fármacos , Huso Acromático/efectos de los fármacos , Aneuploidia , Ciclo Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Humanos , Mucosa Respiratoria/citologíaRESUMEN
Damage to the CNS results in a complex series of molecular and cellular changes involving the affected targets and the ensuing glial reaction. The initial gene expression events that underlie these cellular responses may serve as early biomarkers of neurotoxicity. Here, we examined gene expression profiles during the initial phase of hippocampal damage resulting from systemic exposure of rats to the organometallic neurotoxicant, trimethyltin (TMT, 8.0 mg/kg, i.p.). Using TMT as a neurodegeneration tool confers several advantages for evaluating molecular events associated with neural damage: 1) regional and cellular targets and time course of damage are known, 2) the blood-brain barrier is not compromised, which limits the contribution of blood-borne factors, e.g. immune, to neural injury responses and 3) known protein and mRNA signatures of TMT-induced neurotoxicity can be used as positive controls to validate novel expression events associated with exposure to this neurotoxicant. Using Affymetrix Gene Chip® to assess gene expression after TMT, combined with Ingenuity Pathway Analysis®, we observed changes consistent for genes known to be affected in hippocampus, while corresponding changes were not detected in cerebellum, a non-target region. In agreement with previous observations, limited changes in expression of inflammation-related genes were observed. Correlated expression profiles were found after exposure to TMT, including changes in gene ontologies associated with neurological disease, cellular assembly and maintenance, as well as signaling pathways associated with cellular stress, energy metabolism and glial activation. Selected gene changes were confirmed from each category by q-RT-PCR and immunoblot analysis. The canonical relationships identified implicate molecular pathways and processes relevant to detection of early stages of hippocampal damage in the TMT model. These observations provide new insight into early events associated with neuronal degeneration and associated glial activation that may serve as the basis for discovery and development of biomarkers of neurotoxicity.
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Perfilación de la Expresión Génica/métodos , Gliosis/inducido químicamente , Hipocampo/efectos de los fármacos , Enfermedades Neurodegenerativas/inducido químicamente , Neurotoxinas/toxicidad , Compuestos de Trimetilestaño/toxicidad , Animales , Enfermedad Crónica , Diagnóstico Precoz , Femenino , Gliosis/diagnóstico , Gliosis/genética , Hipocampo/patología , Hipocampo/fisiopatología , Immunoblotting/métodos , Masculino , Enfermedades Neurodegenerativas/diagnóstico , Enfermedades Neurodegenerativas/genética , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Ratas , Ratas Long-Evans , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Factores de TiempoRESUMEN
Engineered carbon nanotubes are newly emerging manufactured particles with potential applications in electronics, computers, aerospace, and medicine. The low density and small size of these biologically persistent particles makes respiratory exposures to workers likely during the production or use of commercial products. The narrow diameter and great length of single-walled carbon nanotubes (SWCNT) suggest the potential to interact with critical biological structures. To examine the potential of nanotubes to induce genetic damage in normal lung cells, cultured primary and immortalized human airway epithelial cells were exposed to SWCNT or a positive control, vanadium pentoxide. After 24 hr of exposure to either SWCNT or vanadium pentoxide, fragmented centrosomes, multiple mitotic spindle poles, anaphase bridges, and aneuploid chromosome number were observed. Confocal microscopy demonstrated nanotubes within the nucleus that were in association with cellular and mitotic tubulin as well as the chromatin. Our results are the first to report disruption of the mitotic spindle by SWCNT. The nanotube bundles are similar to the size of microtubules that form the mitotic spindle and may be incorporated into the mitotic spindle apparatus.
Asunto(s)
Aneuploidia , Nanotubos de Carbono , Línea Celular Transformada , Humanos , Hibridación Fluorescente in Situ , Tamaño de la PartículaRESUMEN
The objectives of this study are to examine the fundamental characteristics of the biodynamic responses of a rat tail to vibration and to compare them with those of human fingers. Vibration transmission through tails exposed to three vibration magnitudes (1 g, 5 g, and 10 g r.m.s.) at six frequencies (32 Hz, 63 Hz, 125 Hz, 160 Hz, 250 Hz, and 500 Hz) was measured using a laser vibrometer. A mechanical-equivalent model of the tail was established on the basis of the transmissibility data, which was used to estimate the biodynamic deformation and vibration power absorption at several representative locations on the tail. They were compared with those derived from a mechanical-equivalent model of human fingers reported in the literature. This study found that, similar to human fingers, the biodynamic responses of the rat tail depends on the vibration magnitude, frequency, and measurement location. With the restraint method used in this study, the natural frequency of the rat tail is in the range 161-368 Hz, which is mostly within the general range of human finger resonant frequencies (100-350 Hz). However, the damping ratios of the rat tail at the unconstrained locations are from 0.094 to 0.394, which are lower than those of human fingers (0.708-0.725). Whereas the biodynamic responses of human fingers at frequencies lower than 100 Hz could be significantly influenced by the biodynamics of the entire hand-arm system, the rat tail biodynamic responses can be considered independent of the rat body in the frequency range used in this study. Based on these findings it is concluded that, although there are some differences between the frequency dependences of the biodynamic responses of the rat tail and human fingers, the rat tail model can provide a practical and reasonable approach to examine the relationships between the biodynamic and biological responses at midrange to high frequencies, and to understand the mechanisms underlying vibration-induced finger disorders.
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Aceleración , Modelos Biológicos , Movimiento/fisiología , Oscilometría/métodos , Estimulación Física/métodos , Animales , Simulación por Computador , Masculino , Ratas , Ratas Sprague-Dawley , VibraciónRESUMEN
Progressive massive fibrosis (PMF) is a chronic interstitial lung disease with a complex aetiology that can occur after cumulative dust exposure. A case-control study was conducted to test the hypothesis that single nucleotide polymorphisms (SNPs) within genes involved in inflammatory and fibrotic processes modulate the risk of PMF development. The study population consisted of 648 underground coal miners participating in the National Coal Workers Autopsy Study, of which 304 were diagnosed with PMF. SNPs that influence the regulation of interleukin (IL)-1, IL-6, tumour necrosis factor-alpha, transforming growth factor-beta1, vascular endothelial growth factor (VEGF), epidermal growth factor intercellular cell adhesion molecule (ICAM)-1 and matrix metalloproteinase-2 genes were determined using a 5'-nuclease real-time PCR assay. There were no significant differences in the distribution of any individual SNP or haplotype between the PMF and control groups. However, the polygenotype of VEGF +405/ICAM-1 +241/IL-6 -174 (C-A-G) conferred an increased risk for PMF (odds ratio 3.4, 95% confidence interval 1.3-8.8). The present study suggests that the examined genetic variations that help regulate inflammatory and fibrotic processes are unlikely to strongly influence susceptibility to this interstitial lung disease, although the role of vascular endothelial growth factor, intercellular cell adhesion molecule-1 and interleukin-6 polymorphisms in the development of progressive massive fibrosis may require further investigation.
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Minas de Carbón , Predisposición Genética a la Enfermedad , Polimorfismo de Nucleótido Simple , Fibrosis Pulmonar/genética , Anciano , Estudios de Casos y Controles , Humanos , Molécula 1 de Adhesión Intercelular/genética , Interleucina-6/genética , Interleucina-6/inmunología , Masculino , Fibrosis Pulmonar/inmunología , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
BACKGROUND: In certain occupations, including farm work, workers are exposed to hazardous substances, some of which are known to be toxic to the nervous system and may adversely affect muscle strength. Measurement of hand-grip strength may be useful for detecting neurotoxic exposure. METHODS: The authors studied 3522 participants of the Honolulu Heart Program and the Honolulu-Asia Aging Study to determine whether occupational exposures to pesticides, solvents, and metals assessed at exam I (1965-68) are associated with hand-grip strength at exam IV (1991-93) and change in hand-grip strength over 25 years. Correlation, analysis of variance and covariance, and linear regression were used to evaluate the associations. RESULTS: At exam IV, participants ranged in age from 71-93 years; mean hand-grip strength was 39.6 kg at exam I and 30.3 kg at exam IV. Over 25 years, the decline in hand-grip strength was an average of 8-9 kg for all exposures. Hand-grip strength was inversely associated with age and glucose but directly associated with cognitive function, BMI, and haemoglobin level. No other exposures were associated with hand-grip strength. CONCLUSION: This study did not provide evidence that occupational exposure to pesticides, solvents, and metals adversely affected hand-grip strength in this population, but confirmed other important associations with hand-grip strength.
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Fuerza de la Mano , Sustancias Peligrosas/toxicidad , Exposición Profesional/efectos adversos , Anciano , Anciano de 80 o más Años , Envejecimiento/fisiología , Monitoreo del Ambiente/métodos , Estudios de Seguimiento , Humanos , Modelos Lineales , Masculino , Metales/toxicidad , Exposición Profesional/análisis , Plaguicidas/toxicidad , Estudios Prospectivos , Factores de Riesgo , Solventes/toxicidadRESUMEN
BACKGROUND: Oxidative stress plays a major role in the pathogenesis of interstitial lung diseases. The antioxidant enzymes glutathione S-transferases (GST) and manganese superoxide dismutase (MnSOD) are important components of lung defence against oxidative stress, and polymorphisms in the genes which regulate their expression may represent important disease modifiers. METHODS: A matched case-control study was conducted to determine the influence of the GSTP1, GSTT1 and MnSOD polymorphisms on susceptibility to progressive massive fibrosis (PMF). Seven hundred ex-coal miners were included in the study; 350 were classified as PMF cases while 350 with a similar underground mining tenure but no clinical or histological evidence of lung disease served as controls. Genotype analysis was performed on genomic DNA, using a 5' nuclease PCR assay. RESULTS: None of the individual investigated polymorphisms and two-way gene-gene interactions had a statistically significant association with PMF. CONCLUSION: The results of this study suggest that polymorphic genotypes within the GST gene cluster and MnSOD do not affect individual susceptibility to PMF.
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Minas de Carbón , Glutatión Transferasa/genética , Isoenzimas/genética , Neumoconiosis/genética , Polimorfismo Genético/genética , Superóxido Dismutasa/genética , Anciano , Antioxidantes , Estudios de Casos y Controles , Frecuencia de los Genes , Predisposición Genética a la Enfermedad/genética , Gutatión-S-Transferasa pi , Humanos , Mutación/genética , Reacción en Cadena de la Polimerasa/métodosRESUMEN
The accuracy of extended histopathology to detect immunotoxic chemicals in female B6C3F1 mice was evaluated under the auspices of the National Toxicology Program (NTP). A workgroup was formed consisting of four pathologists who conducted extended histopathological evaluation of lymphoid tissues obtained from a subset of NTP toxicology studies, in which previously detailed immunotoxicity assessment was performed. In addition, a positive control data set of three known immunosuppressive agents, one negative control data set, and an additional negative control group composed of the vehicle only treated groups were included. Data obtained from extended histopathology evaluations were compared to more traditional immune test results (both functional and nonfunctional) from previously conducted immunotoxicity assessments. Analyses of the data indicated that the ability to identify immunotoxic chemicals using histological endpoints decreased linearly as the level of stringency used to determine significant histopathological changes increased. A relatively high (80%) accuracy level was achieved when histological changes were considered in toto (i.e., any histological abnormality in the three tissues examined), using minimal or mild criteria for scoring. When minimal or mild histological changes were considered significant for a specific tissue, a 60% level of accuracy in identifying immunotoxic chemicals was obtained as compared to a 90% accuracy level that was achieved with this data set using the antibody plaque forming cell response, considered to represent the most predictive functional test. A minimal classification was obtained in the analyses of the negative control groups, suggesting that use of the minimal classification for hazard identification is inappropriate as it will likely result in a high incidence of false positives. This was not the case when mild classifications were used as an indicator of significance, which in most instances allowed the successful identification of negatives. When moderate to marked histopathological changes were used to identify immunotoxic chemicals, the level of accuracy that could be achieved was poor. A considerably higher level of accuracy was obtained for the positive control data set than the test chemical data set suggesting that the ability to detect an immunotoxic agent histologically is proportional to the potency of the immunotoxic agent. Comparison of immune function test results and histopathological results obtained from the high-dose treatment groups and the lower-dose treatment group did not reveal any significant differences between the two endpoints to predict immunotoxicity as a function of dose. Of the three lymphoid organs examined, (i.e., lymph node, thymus, and spleen), the most consistent and discernible histological lesions were observed in the thymus cortical region. These lesions correlated with thymus: body weight ratios and to a slightly lesser extent, the antibody plaque forming cell response. Addition of general toxicological endpoints such as body weight and leukocyte counts did not significantly improve the sensitivity of extended histopathology for this data set. Taken together, these data suggest that, while not as sensitive as functional analyses, extended histopathology may provide a reasonable level of accuracy as a screening test to identify immunotoxic chemicals, provided the level of stringency used to score histological lesions is carefully considered to allow for detection of immunotoxic agents while limiting false positives.
Asunto(s)
Sistema Inmunológico/efectos de los fármacos , Ganglios Linfáticos/patología , Bazo/patología , Timo/patología , Pruebas de Toxicidad/métodos , Animales , Peso Corporal/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Determinación de Punto Final , Reacciones Falso Positivas , Técnica de Placa Hemolítica , Inmunidad Celular/efectos de los fármacos , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Recuento de Linfocitos , Ratones , Ratones Endogámicos , Tamaño de los Órganos/efectos de los fármacos , Especificidad de Órganos , Vehículos Farmacéuticos , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Bazo/inmunología , Timo/inmunologíaRESUMEN
There has been considerable interest in the use of expanded histopathology as a primary screen for immunotoxicity assessment. To determine the utility of a semiquantitative histopathology approach for examining specific structural and architectural changes in lymphoid tissues, a validation effort was initiated. This study addresses the interlaboratory reproducibility of extended histopathology, using tissues from studies of ten test chemicals and both negative and positive controls from the National Toxicology Program's immunotoxicology testing program. We examined the consistency between experienced toxicologic pathologists, who had varied expertise in immunohistopathology in identifying lesions in immune tissues, and in the sensitivity of the individual and combined histopathological endpoints to detect chemical effects and dose response. Factor analysis was used to estimate the association of each pathologist with a so-called "common factor" and analysis-of-variance methods were used to evaluate biases. Agreement between pathologists was highest in the thymus, in particular, when evaluating cortical cellularity of the thymus; good in spleen follicular cellularity and in spleen and lymph node-germinal center development; and poorest in spleen red-pulp changes. In addition, the ability to identify histopathological change in lymphoid tissues was dependent upon the experience/training that the individual pathologist possessed in examining lymphoid tissue and the apparent severity of the specific lesion.
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Alergia e Inmunología/normas , Sistema Inmunológico/efectos de los fármacos , Laboratorios/normas , Toxicología/normas , Animales , Biología Computacional , Interpretación Estadística de Datos , Relación Dosis-Respuesta a Droga , Histología/normas , Modelos Estadísticos , Reproducibilidad de los Resultados , Terminología como AsuntoRESUMEN
The objective of the present study was to investigate the impact of muscle length during stretch-shortening cycles on static and dynamic muscle performance. Animals were randomly assigned to an isometric (control, Con, n = 12), a short-muscle-length (S-Inj, 1.22-2.09 rad, n = 12), or a long-muscle-length (L-Inj, 1.57-2.44 rad, n = 12) group. The dorsiflexor muscles were exposed in vivo to 7 sets of 10 stretch-shortening contractions (conducted at 8.72 rad/s) or 7 sets of isometric contractions of the same stimulation duration by using a custom-designed dynamometer. Performance was characterized by multipositional isometric exertions and positive, negative, and net work before exposure, 6 h after exposure, and 48 h after exposure to contractions. Real-time muscle performance during the stretch-shortening cycles was characterized by stretch-shortening parameters and negative, positive, and net work. The S-Inj group recovery (force difference) was similar to the Con group force difference at 48 h, whereas the L-Inj group force difference was statistically greater at 1.39, 1.57, and 1.74 rad than the Con group force difference (P < 0.05). Negative work (P < 0.05) and net work (P < 0.05) were statistically lower in the S-Inj and L-Inj groups than in the Con group 48 h after exposure to contractions. Of the real-time parameters, there was a difference in cyclic force with treatment during the stretch-shortening cycles (P < 0.0001), with the L-Inj group being the most affected. Thus longer ranges of motion result in a more profound isometric force decrement 48 h after exposure to contractions and in real-time changes in eccentric forces.
Asunto(s)
Contracción Isométrica/fisiología , Músculo Esquelético/lesiones , Músculo Esquelético/fisiología , Esguinces y Distensiones/fisiopatología , Animales , Estimulación Eléctrica , Masculino , Rango del Movimiento Articular , Ratas , Ratas Sprague-DawleyRESUMEN
Muscle damage due to stretch-shortening cycles (i.e., cyclic eccentric/concentric muscle actions) is one of the major concerns in sports and occupational related activities. Mechanical responses of whole muscle have been associated with damage in neural motor units, in connective tissues, and the force generation mechanism. The objective of this study was to introduce a new method to quantify the real-time changes in skeletal muscle forces of rats during injurious stretch-shortening cycles. Male Sprague Dawley rats ( n=24) were selected for use in this study. The dorsi flexor muscle group was exposed to either 150 stretch-shortening cycles ( n=12) or 15 isometric contractions ( n=12) in vivo using a dynamometer and electrical stimulation. Muscle damage after exposure to stretch-shortening cycles was verified by the non-recoverable force deficit at 48 h and the presence of myofiber necrosis. Variations of the dynamic forces during stretch-shortening cycles were analyzed by decomposing the dynamic force signature into peak force ( F(peak)), minimum force ( F(min)), average force ( F(mean)), and cyclic force ( F(a)). After the 15th set of stretch-shortening cycles, the decrease in the stretch-shortening parameters, F(peak), F(min), F(mean), and F(a), was 50% ( P<0.0001), 26% ( P=0.0055), 68% ( P<0.0001), and 50% ( P<0.0001), respectively. Our results showed that both isometric contractions and stretch-shortening cycles induce a reduction in the isometric force. However, the force reduction induced by isometric contractions fully recovered after a break of 48 h while that induced by stretch-shortening cycles did not. Histopathologic assessment of the tibialis anterior exposed to stretch-shortening cycles showed significant myofiber degeneration and necrosis with associated inflammation, while muscles exposed to isometric contractions showed no myofiber degeneration and necrosis, and limited inflammation. Our results suggest that muscle damage can be identified by the non-recoverable isometric force decrement and also by the variations in the dynamic force signature during stretch-shortening cycles.
Asunto(s)
Trastornos de Traumas Acumulados/patología , Trastornos de Traumas Acumulados/fisiopatología , Contracción Isométrica , Músculo Esquelético/patología , Músculo Esquelético/fisiopatología , Enfermedades Musculares/patología , Enfermedades Musculares/fisiopatología , Animales , Estimulación Eléctrica , Masculino , Músculo Esquelético/lesiones , Músculo Esquelético/inervación , Periodicidad , Ratas , Ratas Sprague-Dawley , Estrés MecánicoRESUMEN
OBJECTIVES: International standard ISO 10819 was established in order to quantify the vibration attenuation characteristics of anti-vibration gloves. One problem that exists with the standard is possible misalignment of the palm adaptor that is placed underneath the test glove. If the adaptor becomes misaligned, the measured glove transmissibility will be lower than the actual value. A tri-axial accelerometer was installed in the adaptor and was used as the basis for providing visual feedback of the adaptor alignment to the test subjects. The objective of this study was to test the hypothesis that adaptor misalignment could be reduced by providing feedback to the test subjects. METHODS: Eight male volunteers (mean age 24.8 yr) were used in the study. Each subject performed two sets of tests: the standard ISO 10819 glove test and the modified version. Three different anti-vibration gloves were tested. Glove transmissibility and adaptor misalignment were calculated for each glove. A three-way analysis of variance was used to analyze the results. RESULTS: A comparison of the two testing methods showed that the modified glove testing method did reduce misalignment significantly, which, in turn, resulted in an increase in the measured glove transmissibility. CONCLUSIONS: The proposed method greatly improved the standard deviation of transmissibility and made the test results more consistent.
Asunto(s)
Ergonomía , Guantes Protectores/normas , Vibración/efectos adversos , Adulto , Traumatismos del Brazo/prevención & control , Diseño de Equipo , Traumatismos de la Mano/prevención & control , Humanos , Masculino , Ensayo de Materiales , Valores de Referencia , SíndromeRESUMEN
This study tests the hypothesis that the bronchial hyperreactivity induced by chronic cigarette smoke (CS) exposure involves the increased expression and release of tachykinins and calcitonin gene-related peptide (CGRP) from afferent nerve fibers innervating the airways. In guinea pigs chronically exposed to CS (20 min twice daily for 14-17 d), peak response in total lung resistance to capsaicin (1.68 microg/kg, intravenously) was significantly greater than that evoked by the same dose of capsaicin in control (air-exposed) animals. This augmented response in CS-exposed animals was abolished after treatment with CP-99994 and SR-48968, the neurokinin (NK)-1 and NK-2 receptor antagonists, suggesting the involvement of tachykinins in chronic CS-induced airway hyperresponsiveness (AHR). Further, substance P (SP)-like immunoreactivity (LI) and CGRP-LI in the airway tissue were significantly greater in the CS animals than in the control animals. Finally, beta-preprotachykinin (PPT, a splice variant from the PPT A gene encoding tachykinins including SP and NKA) messenger RNA levels as measured by in situ hybridization histochemistry displayed a significant increase in jugular ganglion neurons but not in dorsal root or nodose ganglion neurons. These data suggest that chronic CS-induced AHR is related to an increase in SP synthesis and release in jugular ganglion neurons innervating the lungs and airways.
Asunto(s)
Bronquios/inervación , Hiperreactividad Bronquial/fisiopatología , Neuronas Aferentes/fisiología , Fumar , Taquicininas/biosíntesis , Análisis de Varianza , Animales , Benzamidas/farmacología , Bronquios/efectos de los fármacos , Bronquios/fisiología , Péptido Relacionado con Gen de Calcitonina/biosíntesis , Capsaicina/farmacología , Cobayas , Hibridación in Situ , Masculino , Piperidinas/farmacología , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Radioinmunoensayo , Distribución Aleatoria , Receptores de Neuroquinina-2/antagonistas & inhibidores , Taquicininas/genética , Taquicininas/metabolismoRESUMEN
Because of the potential of arsenic for causing cancer in humans, and of the fact of widespread environmental and occupational exposure, deriving acceptable human-limit values has been of major concern to industry as well as to regulatory agencies. Based upon epidemiological evidence and mechanistic studies, it has been argued that a non-linear dose-response model at low-level exposures is more appropriate for calculating risk than the more commonly employed linear-response models. In the present studies, dose-response relationships and recovery studies employing a cancer precursor marker, i.e., activating protein (AP)-1 DNA-binding activity, were examined in bladders of mice exposed to arsenic in drinking water and compared to histopathological changes and arsenic tissue levels in the same tissue. While AP-1 is a functionally pleomorphic transcription factor regulating diverse gene activities, numerous studies have indicated that activation of the MAP kinase pathway and subsequently increased AP-1 binding activities, is a precursor for arsenic-induced cancers of internal organs as well as the skin. We observed previously that within 8 weeks of exposure AP-1 activation occurs in urinary bladder tissue of mice exposed to arsenic in the drinking water. In the present studies, C57BL/6 mice were exposed to sodium arsenite at various concentrations in the drinking water for 8 consecutive weeks. Minimal but observable AP-1 activity occurred in bladder tissue at exposure levels below which histopathological changes or arsenic tissue accumulation was detected. Marked AP-1 DNA-binding activity only occurred at exposure levels of sodium arsenite above 20 microg/ml, where histopathological changes and accumulation of arsenic in the urinary bladder epithelium occurred. Although the experimental design did not allow statistical modeling of the entire dose-response curve, the general shape of the dose-response curve is not inconsistent with the previously proposed hypothesis that arsenic-induced cancer follows a non-linear dose-response model.
Asunto(s)
Arsénico/toxicidad , Carcinógenos Ambientales/toxicidad , Epitelio/metabolismo , Factor de Transcripción AP-1/biosíntesis , Vejiga Urinaria/efectos de los fármacos , Vejiga Urinaria/metabolismo , Animales , Arsénico/farmacocinética , Carcinógenos Ambientales/farmacocinética , Daño del ADN , Proteínas de Unión al ADN , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Ingestión de Líquidos , Epitelio/efectos de los fármacos , Epitelio/patología , Femenino , Ratones , Ratones Endogámicos C57BL , Medición de Riesgo , Distribución Tisular , Vejiga Urinaria/patologíaRESUMEN
Clinical studies demonstrate that estrogen replacement therapy in postmenopausal women may enhance cognitive function and reduce neurodegeneration associated with Alzheimer's disease and stroke. This study assesses whether physiologic levels of estradiol prevent brain injury in an in vivo model of permanent focal ischemia. Sprague-Dawley rats were ovariectomized; they then were implanted, immediately or at the onset of ischemia, with capsules that produced physiologically low or physiologically high 17beta-estradiol levels in serum (10 or 60 pg/mL, respectively). One week after ovariectomy, ischemia was induced. Estradiol pretreatment significantly reduced overall infarct volume compared with oil-pretreated controls (mean+/-SD: oil = 241+/-88; low = 139+/-91; high = 132+/-88 mm3); this protective effect was regionally specific to the cortex, since no protection was observed in the striatum. Baseline and ischemic regional CBF did not differ between oil and estradiol pretreated rats, as measured by laser Doppler flowmetry. Acute estradiol treatment did not protect against ischemic injury. Our finding that estradiol pretreatment reduces injury demonstrates that physiologic levels of estradiol can protect against neurodegeneration.