Early Histological and Functional Effects of Chronic Copper Exposure in Rat Liver

Cu is an essential trace element capable of producing toxic effects in animals and man when ingested acutely or chronically in excess. Although chronic Cu exposure is increasingly recognized as a public health issue, its early effects remain largely unknown. We approached the significance of a moderate chronic Cu load in young rats to correlate early hepatic histopathological changes with functional alterations of liver cells. For this purpose, supplementation with 1200 ppm of Cu in rat food for 16 weeks was chosen. In these conditions, Cu load elicited a significant decrease in growth curves. There were mild light microscopy alterations in Cu-treated rats, although increasing intracellular Cu storage was correlated with longer Cu exposure both by histological and biochemical measurements. Ultrastructural alterations included lysosomal inclusions as well as mitochondrial and nuclear changes. Liver perfusion studies revealed higher rates of basal O₂ consumption and colloidal carbon-induced O₂ uptake in Cu-treated rats, with enhanced carbon-induced O₂/carbon uptake ratios and NF-κB DNA binding activity. These changes were time-dependent and returned to control values after 12 or16 weeks. It is concluded that subchronic Cu loading in young rats induces early hepatic morphological changes, with enhancement in Küpffer cell-dependent respiratory burst activity and NF-κB DNA binding, cellular responses that may prevent or alleviate the hepatotoxicity of the metal.     

Induction of Apoptosis in Rat Myocardium under Anoxic Conditions

The effect of anoxic incubation of small slices of isolated rat hearts on respiration, internucleosomal DNA fragmentation, and mitochondrial ultrastructure was investigated. Anoxic incubation for 72 h induced apoptosis accompanied by internucleosomal DNA fragmentation and changes in respiration and mitochondrial ultrastructure. The mitochondrial population was characterized by morphological heterogeneity. In a significant part of the mitochondrial population there were signs of mitochondrial swelling and appearance of electron-dense mitochondria. Anoxia also induced the appearance of an atypical (and previously unknown) population of small electron-dense mitochondria. They were characterized by unusual localization inside electron-light mitochondria. Under anoxic conditions the inner mitochondrial membrane formed electron-dense ordered structures. All changes described here reflect two opposing processes occurring in mitochondria: apoptotic destruction and compensatory processes responsible for maintenance of mitochondria.     

生物阻抗测量系统的改良

目的:针对目前现有的生物阻抗测量系统测量速度和精度不够的问题,提出相应的改进方法。方法:在信号发生部分采用AD公司的ADuC847微处理器作为核心微处理器;在信号采集部分采用Atmel公司的AT91RM9200微控制器芯片和TI公司的ADS1256芯片作为核心芯片。结果:生物阻抗测量系统的信号发生精度提高到12位,信号采集精度提高到24位,速度提高到30,000SPS。结论:对信号发生和信号采集方面速度和精度的提高,有效地提高了整个测量系统的速度和精确度。     

Effects of Mild Chronic Intermittent Cold Exposure on Rat Organs

Cold adaptation is a body''s protective response to cold stress. Mild chronic intermittent cold (CIC) exposure has been used to generate animal models for cold adaptation studies. However, the effects of mild CIC exposure on vital organs are not completely characterized. In the present study, we exposed rats to mild CIC for two weeks, and then measured the body weights, the weights of brown adipose tissue (BAT), the levels of ATP and reactive oxygen species (ROS) in the brains, livers, hearts, muscles and BATs. Rats formed cold adaptation after exposure to CIC for two weeks. Compared to rats of the control group that were hosted under ambient temperature, rats exposed to mild CIC showed a lower average body weight, but a higher weight of brown adipose tissue (BAT). Rats exposed to CIC for two weeks also exhibited higher levels of ATP and ROS in all examined organs as compared to those of the control group. In addition, we determined the expression levels of cold-inducible RNA binding protein (Cirbp) and thioredoxin (TRX) in rat tissues after 2 weeks of CIC exposure. Both Cirbp and TRX were increased, suggesting a role of these two proteins for establishment of cold adaptation. Together, this study reveals the effects of mild CIC exposure on vital organs of rats during CIC exposure.     

Exposure to Cobalt Causes Transcriptomic and Proteomic Changes in Two Rat Liver Derived Cell Lines

Cobalt is a transition group metal present in trace amounts in the human diet, but in larger doses it can be acutely toxic or cause adverse health effects in chronic exposures. Its use in many industrial processes and alloys worldwide presents opportunities for occupational exposures, including military personnel. While the toxic effects of cobalt have been widely studied, the exact mechanisms of toxicity remain unclear. In order to further elucidate these mechanisms and identify potential biomarkers of exposure or effect, we exposed two rat liver-derived cell lines, H4-II-E-C3 and MH1C1, to two concentrations of cobalt chloride. We examined changes in gene expression using DNA microarrays in both cell lines and examined changes in cytoplasmic protein abundance in MH1C1 cells using mass spectrometry. We chose to closely examine differentially expressed genes and proteins changing in abundance in both cell lines in order to remove cell line specific effects. We identified enriched pathways, networks, and biological functions using commercial bioinformatic tools and manual annotation. Many of the genes, proteins, and pathways modulated by exposure to cobalt appear to be due to an induction of a hypoxic-like response and oxidative stress. Genes that may be differentially expressed due to a hypoxic-like response are involved in Hif-1α signaling, glycolysis, gluconeogenesis, and other energy metabolism related processes. Gene expression changes linked to oxidative stress are also known to be involved in the NRF2-mediated response, protein degradation, and glutathione production. Using microarray and mass spectrometry analysis, we were able to identify modulated genes and proteins, further elucidate the mechanisms of toxicity of cobalt, and identify biomarkers of exposure and effect in vitro, thus providing targets for focused in vivo studies.     

Decreased Nitric Oxide Production in the Rat Brain after Chronic Arsenic Exposure

Chronic arsenic exposure is associated with nervous system damage, vascular disease, hepatic and renal damage as well as different types of cancer. Alterations of nitric oxide (NO) in the periphery have been detected after arsenic exposure, and we explored here NO production in the brain. Female Wistar rats were exposed to arsenite in drinking water (4–5 mg/kg/day) from gestation, lactation and until 4 months of age. NOS activity, NO metabolites content, reactive oxygen species production (ROS) and lipid peroxidation (LPx) were determined in vitro in the striatum, and NO production was estimated in vivo measuring citrulline by microdialysis. Exposed animals showed a significantly lower response to NMDA receptor stimulation, reduction of NOS activity and decreased levels of nitrites and nitrates in striatum. These markers of NO function were accompanied by significantly higher levels of LPx and ROS production. These results provide evidence of NO dysfunction in the rat brain associated with arsenic exposure.     

Effect of Chronic Ethanol Feeding on Oxysterols in Rat Liver

It was our hypothesis that, as a consequence of increased oxidative stress, cholesterol-derived hydroperoxides and oxysterols are increased in livers of rats exposed to ethanol. To test this we dosed Wistar rats (approximately 0.1 kg initial body weight) with ethanol chronically (rats fed a nutritionally complete liquid diet containing ethanol as 35% of total calories; sampled liver at approximately 6-7 weeks). We measured concentrations of 7 &#102 - and 7 &#103 -hydroperoxycholest-5-en-3 &#103 -ol (7 &#102 -OOH and 7 &#103 -OOH) as well as 7 &#102 - and 7 &#103 -hydroxycholesterol (7 &#102 -OH and 7 &#103 -OH), and 3 &#103 -hydroxycholest-5-en-7-one (also termed 7-ketocholesterol; 7-keto). In response to chronic alcohol feeding, there were significant elevations in the concentrations of 7 &#102 -OOH (+169%, P =0.005 ) and 7 &#103 -OOH (+199%, P =0.011 ). Increases in the concentrations of hepatic 7-keto (+74%, P =0.01 ) and decreases in cholesterol ( &#109 43%; P =0.03 ) also occurred. In contrast, the concentrations of both 7 &#102 -OH and 7 &#103 -OH were not significant (NS). However, when oxysterols in chronic ethanol-fed rats were expressed relative to cholesterol there were significant increases in 7-keto/cholesterol ( P =0.0006), 7 &#102 -OH/cholesterol ( P =0.0018) and 7 &#103 -OH/cholesterol ( P =0.0047). In conclusion, this is the first report of increased 7 &#102 -OOH, 7 &#103 -OOH, and 7-keto in liver of rats and their elevation in chronic experimental alcoholism represent evidence of increased oxidative stress.     

Temporal Changes in Rat Liver Gene Expression after Acute Cadmium and Chromium Exposure

U.S. Service Members and civilians are at risk of exposure to a variety of environmental health hazards throughout their normal duty activities and in industrial occupations. Metals are widely used in large quantities in a number of industrial processes and are a common environmental toxicant, which increases the possibility of being exposed at toxic levels. While metal toxicity has been widely studied, the exact mechanisms of toxicity remain unclear. In order to further elucidate these mechanisms and identify candidate biomarkers, rats were exposed via a single intraperitoneal injection to three concentrations of CdCl₂ and Na₂Cr₂O₇, with livers harvested at 1, 3, or 7 days after exposure. Cd and Cr accumulated in the liver at 1 day post exposure. Cd levels remained elevated over the length of the experiment, while Cr levels declined. Metal exposures induced ROS, including hydroxyl radical (•OH), resulting in DNA strand breaks and lipid peroxidation. Interestingly, ROS and cellular damage appeared to increase with time post-exposure in both metals, despite declines in Cr levels. Differentially expressed genes were identified via microarray analysis. Both metals perturbed gene expression in pathways related to oxidative stress, metabolism, DNA damage, cell cycle, and inflammatory response. This work provides insight into the temporal effects and mechanistic pathways involved in acute metal intoxication, leading to the identification of candidate biomarkers.     

Validation of Bioelectrical Impedance Analysis in Adolescents Across Different Ethnic Groups

Adolescent obesity is difficult to assess in multi‐ethnic populations using BMI, due to variability in the BMI–fatness relationship. We aimed to describe body composition (BC), and to validate leg–leg bioelectrical impedance analysis (BIA), in adolescents from different ethnic groups using deuterium (D₂O) as the reference method. Measurements were made of weight, height, total body water (TBW), and BIA (TANITA TBF‐300) in 110 white, 170 Asian, and 102 black adolescents aged 11–15 years. TBW was converted to lean mass (LM) using assumed hydration of lean tissue. General linear models were used to compare BC by D₂O between the ethnic groups. BC values from D₂O were compared with TANITA values, and used to generate ethnic‐specific prediction equations in the whole sample, and also in equation‐generation (group 1) and cross‐validation (group 2) subsamples. Ethnic variability in BMI did not reflect variability in adiposity. Asians had less LM than white and black adolescents, and less fat mass (FM) than white girls. TANITA in‐built equations did not predict BC accurately across ethnic groups, with significant bias in white and Asian males, and Asian and black females. The new equation generated from the entire sample removes ethnic‐specific mean biases. The group 1 equation showed no significant bias in any ethnic group when tested in group 2. We found significant variability in BC between ethnic groups that was not reflected by BMI. Manufacturers' equations are unsuitable for predicting BC in multi‐ethnic populations, and our new equations are recommended.     

Adiponectin Protects Rat Myocardium against Chronic Intermittent Hypoxia-Induced Injury via Inhibition of Endoplasmic Reticulum Stress

Obstructive sleep apnea syndrome (OSAS) is associated with many cardiovascular disorders such as heart failure, hypertension, atherosclerosis, and arrhythmia and so on. Of the many associated factors, chronic intermittent hypoxia (CIH) in particular is the primary player in OSAS. To assess the effects of CIH on cardiac function secondary to OSAS, we established a model to study the effects of CIH on Wistar rats. Specifically, we examined the possible underlying cellular mechanisms of hypoxic tissue damage and the possible protective role of adiponectin against hypoxic insults. In the first treatment group, rats were exposed to CIH conditions (nadir O2, 5–6%) for 8 hours/day, for 5 weeks. Subsequent CIH-induced cardiac dysfunction was measured by echocardiograph. Compared with the normal control (NC) group, rats in the CIH-exposed group experienced elevated levels of left ventricular end-systolic dimension and left ventricular end-systolic volume and depressed levels of left ventricular ejection fraction and left ventricular fractional shortening (p<0.05). However, when adiponectin (Ad) was added in CIH + Ad group, we saw a rescue in the elevations of the aforementioned left ventricular function (p<0.05). To assess critical cardiac injury, we detected myocardial apoptosis by Terminal deoxynucleotidyl transfer-mediated dUTP nick end-labeling (TUNEL) analysis. It was showed that the apoptosis percentage in CIH group (2.948%) was significantly higher than that in NC group (0.4167%) and CIH + Ad group (1.219%) (p<0.05). Protein expressions of cleaved caspase-3, cleaved caspase-9, and cleaved-caspase-12 validated our TUNEL results (p<0.05). Mechanistically, our results demonstrated that the proteins expressed with endoplasmic reticulum stress and the expression of reactive oxygen species (ROS) were significantly elevated under CIH conditions, whereas Ad supplementation partially decreased them. Overall, our results suggested that Ad augmentation could improve CIH-induced left ventricular dysfunction and associated myocardial apoptosis by inhibition of ROS-dependent ER stress.     

Reliability of Multifrequency Bioelectrical Impedance Analysis to Quantify Body Composition in Patients After Musculoskeletal Trauma

BackgroundChanges in body composition, especially loss of lean mass, commonly occur in the orthopedic trauma population due to physical inactivity and inadequate nutrition. The purpose of this study was to assess inter-rater and intra-rater reliability of a portable bioelectrical impedance analysis (BIA) device to measure body composition in an orthopedic trauma population after operative fracture fixation. BIA uses a weak electric current to measure impedance (resistance) in the body and uses this to calculate the components of body composition using extensively studied formulas.MethodsTwenty subjects were enrolled, up to 72 hours after operative fixation of musculoskeletal injuries and underwent body composition measurements by two independent raters. One measurement was obtained by each rater at the time of enrollment and again between 1-4 hours after the initial measurement. Reliability was assessed using intraclass correlation coefficients (ICC) and minimum detectable change (MDC) values were calculated from these results.ResultsInter-rater reliability was excellent with ICC values for body fat mass (BFM), lean body mass (LBM), skeletal muscle mass (SMM), dry lean mass (DLM), and percent body fat (PBF) of 0.993, 0.984, 0.984, 0.979, and 0.986 respectively. Intra-rater reliability was also high for BFM, LBM, SMM, DLM, and PBF, at 0.994, 0.989, 0.990, 0.983, 0.987 (rater 1) and 0.994, 0.988, 0.989, 0.985, 0.989 (rater 2). MDC values were calculated to be 4.05 kg for BFM, 4.10 kg for LBM, 2.45 kg for SMM, 1.21 kg for DLM, and 4.83% for PBF.ConclusionPortable BIA devices are a versatile and attractive option that can reliably be used to assess body composition and changes in lean body mass in the orthopedic trauma population for both research and clinical endeavors. Level of Evidence: III     

亚慢性砷中毒对大鼠肝脏自噬的抑制作用

目的:在建立亚慢性砷暴露模型的基础上,探讨砷暴露对SD大鼠肝脏自噬水平的影响.方法:出生21天断乳雄性SD大鼠分为不同水平砷暴露组,通过饮水给与NaAsO2的方式染毒;全自动生化分析仪检测血清肝功能项目的变化;HE染色检测肝脏组织病理学改变;透射电镜法超微结构的改变.Western blot方法检测自噬相关蛋白Beclin1、LC3表达水平变化.结果:砷暴露组肝脏出现显著的异常,同时伴随肝组织形态学改变.于是此同时,LC3Ⅱ/LC3Ⅰ比值以及beclin1的表达水平均随着砷作用浓度的升高而降低.结论:亚慢性砷暴露在诱导肝脏损伤的同时可伴随自噬水平的抑制,这种水平的改变可能参与了砷的肝脏毒性.     

慢性乙醇摄入大鼠肝脏醇脱氢酶和谷胱甘肽硫转移酶活性影响

为探讨长期摄入乙醇大鼠肝脏ＡＤＨ和ＧＳＴ活性的动态变化,旨在为酒精中毒的发病机理提供实验依据,进行了下述实验研究。选用体重１００～１２０ｇＳＤ雄性大鼠４８只,随机分成两组：（１）对照组：２４只喂普通饲料,饮用自来水,（２）乙醇组：２４只白天饮用１０％蔗糖水,夜间饮用１０５乙醇水溶液,平均每只实验动物的乙醇摄入量为８．０ｇ／ｋｇ体重／ｄ,饲料同对照组。实验开始后的３０ｄ、６０ｄ时分批处死动物,第９０天时处死全部动物,除肉眼观察肝脏形态学改变外,同时留取病理标本和进行组织中ＡＤＨ和ＧＳＴ活性测定,结果显示：（１）大鼠肝细胞胞浆中ＡＤＨ活性,在服用乙醇３０ｄ时即可看到明显减低,较对照组有显著差别（Ｐ＜０．０５）,至实验９０ｄ结束时,其减低的更明显（Ｐ＜０．０１）．（２）大鼠肝细胞胞浆中ＧＳＴ活性　在６０ｄ和９０ｄ时乙醇组较对照组均明显减低,经统计学处理均有显著差别（Ｐ＜０．０１）。上述结果提示肝脏ＡＤＨ和ＧＳＴ活性表达在酒精性肝病发病机制中起着重要作用。     

The Interactions between the Chronic Exposure to Aluminum and Liver Regeneration on Bile Flow and Organic Anion Transport in Rats

The chronic exposure to Aluminum (Al) may compromise different liver functions, mainly during the hepatic regeneration. The aim of this study is to investigate the interactions between the chronic i.p. exposure to Al and hepatic regeneration (HR) on bile flow and organic anion transport in experimental animals. For this purpose, we studied bile flow and fractional transfer rates for the transport of hepatic organic anions (hepatic uptake, sinusoidal efflux, and canalicular excretion), as well as parameters related with the oxidative stress (OS), on rats chronically treated with Al at 0 and 2 days of HR. The Al treatment and time of HR caused a decrease in the biliary flow and in the hepatic uptake and canalicular excretion constants. In addition, Al and HR increased the lipoperoxidation associated with a reduction of the glutathione content and glutathione peroxidase and catalase enzyme’s activities. Since the effects of Al and HR on biliary flow and transport systems were additive, but not on the oxidative status, different mechanisms might be involved on these alterations. Even though the OS may play a key role on the hepatic deleterious effects, there is no unique cause–effect relationship between OS and liver dysfunction in this experimental animal model.     

Validity of a New Abdominal Bioelectrical Impedance Device to Measure Abdominal and Visceral Fat: Comparison With MRI

Abdominal fat, and in particular, visceral adipose tissue (VAT), is the critical fat depot associated with metabolic aberrations. At present, VAT can only be accurately measured by computed tomography or magnetic resonance imaging (MRI). This study was designed to compare a new abdominal bioelectrical impedance (BIA) device against total abdominal adipose tissue (TAAT) and VAT area measurements made from an abdominal MRI scan, and to assess its reliability and accuracy. One‐hundred twenty participants were recruited, stratified by gender and BMI. Participants had triplicate measures of abdominal fat and waist circumference (WC) with the AB‐140 (Tanita, Tokyo, Japan) and WC measurements using a manual tape measure. A single abdominal MRI scan was performed as the reference method. Triplicate measures with the AB‐140 showed excellent precision for “visceral fat level,” trunk fat %, and WC. AB‐140 “visceral fat level” showed significantly stronger correlations with MRI TAAT area than with MRI VAT area (r = 0.94 vs. 0.65 in men and 0.92 vs. 0.64 in women). AB‐140 WC showed good correlation with manual WC measurements (r = 0.95 in men and 0.90 in women). AB‐140 and manual WCs showed comparable correlations with MRI TAAT area (r = 0.92 and 0.96 in men and 0.88 and 0.88 in women). AB‐140 is a simple, quick, and precise technique to measure abdominal fat and WC in healthy adults. It provides a useful proxy for TAAT measured by MRI, comparable to the correlation obtained with manual WC measurements. Neither the AB‐140 abdominal fat measures nor WC measurement appear to provide a useful proxy measure of VAT.     

Effect of Adaptation to Graduated Physical Exercises on the Function of the Rat Myocardium

On isolated hearts obtained from control rats and rats subjected to regular physical exercises (forced swimming) during 6 weeks, we studied the contractile activity of the heart, resistance of the myocardium to ischemia/reperfusion-induced injuries, as well as the dependence of the developed and end-diastolic pressures in the aortic ventricle (AV) on the strain of the myocardium (by means of a dosed increase in the volume of a polyethylene reservoir inserted into the ventricle). It was demonstrated that adaptation to regular graduated physical exercises exerts a positive effect on the functional state of the AV myocardium and its contractile function. This was manifested in intensification of the contractile activity of the myocardium, a decrease in its oxygen “job cost,” and an increase in the resistance to injuries induced by ischemia-reperfusion. In addition, regular physical trainings led to an increase in the resistance of the AV myocardium to the strain. In trained rats, the plateau of the Frank–Starling plot was significantly greater than that in control animals, while the rigidity of the AV myocardium was significantly lower. Neirofiziologiya/Neurophysiology, Vol. 41, No. 1, pp. 41–47, January–February, 2009.     

Morphometric Indices of Scots Pine Needle under Chronic Radiation Exposure

We have studied the morphometric indices of needles in Scots pine populations that grow in Bryansk oblast in sites with radioactive contamination long after the Chernobyl accident. The variability in needle weight and length, as well as the fluctuating asymmetry indices and occurrence of necroses and morphoses, were studied in four contaminated and two reference populations of Scots pine in 2011, 2013, and 2014. The exposure of needles in radioactively contaminated sites varied from 7 to 130 mGy/yr. We found brachyblasts with three needles in contaminated Scots pine populations; this morphosis was absent in the reference populations. A dependence of the occurrence of needles damaged by necrosis on the levels of radiation exposure in 2011 was found. However, it was statistically insignificant in other years. The length and weight of needles in contaminated populations differed from the control values; however, the dependence of these indices on the level of radiation exposure was not revealed in the studied range of doses. In 2011 and 2013, the index of fluctuating asymmetry in needle length exceeded the control levels in sites where the absorbed doses were 90 and 130 mGy; this index also tended to grow (statistically significantly in 2011) with an increase in the characteristics of radioactive contamination of the studied plots: the exposure dose rate and the specific activity of ⁹⁰Sr and ¹³⁷Cs in cones and of ¹³⁷Cs in the soil. Therefore, one can observe the consequences of chronic radiation exposure at the organismic level in populations of Scots pine (one of the most radiosensitive plant species) even 25 years after the accident. The data that we obtained in natural habitats confirm the international estimates, according to which the annual chronic radiation exposure of 100 mGy can be considered the limit of safe radiation exposure of natural populations according to morphological and ontogenetic parameters.