Stress Protein Levels Research Articles

Impact of Different Rearing Systems and Age on Bovans White Layer’s Performance, Egg Quality Traits and Synthesis of Heat Shock Protein 70 kDa

Abstract This study was conducted to investigate the performance of laying hens and to determine possible age-related changes on external and internal quality traits of their eggs and synthesis of heat shock protein 70 kDa (HSP70) in the conventional cage and organic rearing systems. For this purpose, 4 different Bovans White hybrid flocks of the same age were monitored in each of these two systems for 52 weeks. While a total of 360 eggs were examined to determine the effects of rearing system and age (30 and 60 weeks) on egg quality traits, 48 liver tissue samples were examined for the analysis of HSP70. Egg production (hen-day) and dirty egg ratio were higher in the organic rearing system (P<0.05). The age at 50% yield, the age at peak of lay, and peak production rate were calculated as 156, 218.75 days and 95.98% in the conventional system, and 155.75, 201.50 days, and 96.56% in the organic system, respectively (P>0.05). While egg weight, albumen weight, yolk weight, shell weight, shape index, and yolk colour were higher in the organic system, the crude ash ratio of eggshell and shell ratio were higher in the conventional system (P<0.05). It was also found that the egg weight, albumen weight, yolk weight, shell weight, and the percentage of yolk were higher at 60 weeks of age (P<0.001). The percentage of shell and albumen, shape index, and yolk colour were higher at 30 weeks of age (P<0.001). The interactions between rearing system and age were statistically significant in terms of shell thickness, shape index, crude ash ratio, and yolk colour (P<0.001). Stress protein (HSP70) level was affected only by the rearing system and was higher in the organic system (P<0.001). As a result, the organic system can be considered as advantageous in terms of egg production and quality traits and the eggs of young hens exhibited better properties. The high level of HSP70 in the organic system could indicate that hens were affected by the environmental conditions at higher rates and/or the hens reared in the organic system had a stronger antioxidant defence system.

Antiageing properties of Damaurone D in Caenorhabditis elegans.

This study was conducted to evaluate the longevity potential of damaurone D (DaD), a component of the damask rose, in the animal model Caenorhabditis elegans. To investigate the effect of DaD on the longevity, lifespan assay was carried out. Fluorescence intensity of transgenic mutants was quantified to test the expression levels of stress proteins. A genetic study using single gene knockout mutants was designed to determine the target genes of DaD. DaD prolonged the mean lifespan of wild-type nematodes by 16.7% under normal conditions and also improved their stress endurance under thermal, osmotic, and oxidative stress conditions. This longevity-promoting effect could be attributed to in vivo antioxidant capacity and its up-regulating effects on the expressions of stress-response proteins such as SOD-3 and HSP-16.2. In addition, DaD treatment attenuated food intake, body length, lipofuscin accumulation and age-dependent decline of motor ability. Gene-specific mutant studies showed the involvement of genes such as daf-2, age-1, and daf-16. These results suggest that DaD has beneficial effects on the longevity, and thus it can be a valuable plant origin lead compound for the development of nutraceutical preparations targeting ageing and ageing-related diseases.

Effect of oxytocin on the levels of malondialdehyde, nitric oxide, and s100b protein in newborns

To assess the effects of the administration of oxytocin during labor management on the levels of malondialdehyde (MDA), nitric oxide (NO), and S100B protein in newborns. We selected 80 term pregnant women without obstetric and fetal pathology, forming two groups: pregnant women with normal delivery and pregnant women conducted with oxytocin. Blood was collected immediately after delivery from the umbilical cord vein to measure MDA, ON and from the artery for protein S100B. The concentration of MDA and ON was quantified by spectroscopic methods and the protein S100B by ELISA. Values of 3.4 uMol/L and 3.6 uMol/L of MDA and 1.4 uMol/L and 1.8 uMol/L of NO were obtained in the oxytocin and control group, respectively, without significant difference; S100B levels were higher in the oxytocin managed group, with a median of 1.36 μg/L compared to the normal delivery group 1.11 μg/L (p=0.03). There was no relationship between the dose of oxytocin administered and the levels of MDA, ON, and S100B. There is no difference between MDA and NO levels between pregnant women undergoing a normal or managed birth. There is a significant difference in S100B protein levels in newborns born via an oxytocin-managed delivery. There is no relationship between oxytocin dose and levels of oxidative stress and S100B protein.

Ecophysiological determinants of the human skeletal system

This review assessed relationships between external factors and the level of joint pathology (hip, knee, ankle, elbow, shoulder, hands and feet) in humans with dysplastic arthritis, psoriatic arthritis, rheumatoid arthritis, Sjogren’s syndrome, systemic lupus erythematosus, systemic sclerosis, degenerative joint disease, ankylosing spondylitis, osteoporosis, osteomalacia and gout. The accumulation of physiologically different chemical elements in different types of bones in humans of different ages allows for a more accurate assessment of the causal quoted response from parallel biochemical systems. These allow a better understanding of the link between exposure arising from smoking, alcohol, drugs, diseases, heredity, effects of amalgam dental fillings, diet, food preferences, chemical elements, occupational and environmental exposure to toxicants, lipoperoxidation and pro-antioxidant reactions. Physiology of the osteoarticular system determines what is responsible for bone and long-term body accumulation of toxic metals, which may be involved in the pathogenesis of bone diseases. While relationships between stressors, antioxidant system and bone condition predict potential risks in certain abnormalities and change oxidative stress. Elemental instability in the environment combined with salinity, acidity, redox potential and local changes in hydrochemical balance, enhances adverse effects. Processes of remodeling and bone mineralization continue throughout life and therefore may be a determinant of long-term accumulation of toxicants. Environmental issues affecting bioaccumulation of chemical elements in the osteoarticular system in humans is poorly understood. Alloplastic procedures, including the need for prostheses, supplemented by image layouts oxidative enzyme activities, as well as lipoperoxidation and the level of stress proteins, give a complete picture of skeletal response to external stressors. Simultaneously, the analysis of the impact of stressors on bones allows a more accurate tracing of causal quoted responses from parallel reactions. They have a direct relationship with stressors and affect the nature and degree of responses and defense capabilities. Thus they have a role in the diagnosis of diseases of the skeletal system.

Anti-diabetic effect and mechanism of Kursi Wufarikun Ziyabit in L6 rat skeletal muscle cells

Kursi Wufarikun Ziyabit (KWZ) is a traditional prescription that used in folk tea drinking for its health care effect in treatment of type 2 diabetes mellitus (T2DM) in central Asia. However, the underlying mechanism of KWZ in T2DM has not been investigated extensively. This study designed to observe the effect of KWZ on glucose consumption and assess the molecular mechanism on associated proteins in insulin signaling and ER stress pathway in L6 rat skeletal muscle cells. The results showed that, KWZ exhibited proteins of PTP-1B and α-glycosidase inhibitory activity in vitro. No cytotoxicity of KWZ was found on L6 cell line. The best effect of glucose consumption of cells was shown at 6.25 μg/mL after KWZ treatment for 12 h. Expression of PTP-1B protein was inhibited by KWZ in L6 moytubes. PI3K-dependent Akt phosphorylation was found to be activated by KWZ. Moreover, the insulin-mediated induction of IRS-1 and GSK-3 were also activated by KWZ. Western blot results indicated that KWZ significantly improved the levels of ER stress proteins, which reduced the expression of GRP78, enhanced the expression of the PERK, eIF2α and XBP1s. The activation of PERK/eIF2α was likely consequence of GRP78 inhibition, and this might be beneficial for improving the stability of ER and alleviating insulin resistance. These results suggest that KWZ might be serving as the potential drug for the prevention and treatment of T2DM.

Exercise Training Attenuates Non-Alcoholic Fatty Liver Disease in rats with Diabetes via Endoplasmic Reticulum Stress

Non-alcoholic fatty liver disease (NAFLD) is highly prevalent in type 2 diabetes (T2D). Recent evidence supports that endoplasmic reticulum (ER) stress plays a critical role in the development of NAFLD. Although exercise training has been commonly prescribed for the treatment of NAFLD, the effects of different types of exercise on ER stress in NAFLD under type 2 diabetic condition are largely unknown. PURPOSE: The purpose of this study was to determine the effects of aerobic and resistance exercises on hepatic ER stress response and NAFLD in rats with T2D. METHODS: Male Sprague-Dawley rats were randomly assigned to four groups (n=10/group): Control (CON), T2D, T2D with aerobic exercise (T2D+AE; treadmill walking at 30 m/min, 0° incline, 60 mins/day, 5 day/week, for 8 weeks) and T2D with resistance exercise (T2D+RE; climbing a 80° incline vertical ladder with weights progressively increased from 50% to 100% of maximal carrying capacity on the tail, 3 times/day, 5 days/ week, for 8 weeks). Liver tissue samples were collected for histopathological analysis of the density of lipid droplets, and immunoblot analysis of expression levels of ER stress proteins, including glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP), caspase 12, and c-Jun N-terminal kinase (JNK). One-way ANOVAs and Tukey’s -test were used for data analysis. RESULTS: The density of lipid droplets in the liver was significantly higher in the T2D group than in the CON group (p<0.01), but was significantly lower in the T2D+AE and T2D+RE groups when compared to the T2D group (both p<0.01). In addition, the T2D group had significantly higher levels of protein expression of GRP78, CHOP, caspase 12 and JNK when compared to the CON group (all p<0.01). Both exercise groups had significantly lower levels of protein expression of GRP 78, CHOP, Caspase 12 and JNK when compared to the T2D group (p<0.05 to p<0.01). CONCLUSION: Our findings suggest that both aerobic and resistance exercises are protective against NAFLD in rats with T2D by potentially regulating proteins involved in ER stress response.

Changes in salivary chromogranin A levels in adults with atopic dermatitis are correlated with changes in their condition.

Stress-induced scratching is an issue in patients with adult atopic dermatitis (AD). Symptoms of stress-induced AD are common in clinical practise. Salivary chromogranin A (CgA) level has research value as a possible index related to a patient's psychological stress. Using saliva, which is easily collectable, we compared two assessments of the severities of AD and stress with the levels of stress proteins in the saliva of 30 patients with AD in the Department of Dermatology of Shimane University between April 2015 and May 2017. The severities of AD and stress were assessed using the Scoring Atopic Dermatitis (SCORAD) score and State-Trait Anxiety Inventory score, respectively. Additionally, the assessments included those of personality using the Tokyo University Egogram (TEG)-II score and quality of life using the Dermatology Life Quality Index score. Simultaneously, we measured their salivary CgA levels. The change in salivary CgA per protein in patients with AD was correlated with their changes in SCORAD score (correlation coefficient, r = 0.596, P = 0.001) and objective SCORAD (r = 0.608, P < 0.001). The changes in CgA per protein correlated with those in TEG-II A (r = 0.370, P = 0.022), while the changes in SCORAD score correlated with those in DLQI (r = 0.309, P = 0.048). Our results suggest that changes in a patient's condition are reflective of the changes in the patient's stress. The changes in salivary CgA level in patients with AD correlated with the changes in their condition.

Serum Heat Shock Protein Levels and the Relationship of Heat Shock Proteins with Various Parameters in Chronic Obstructive Pulmonary Disease Patients.

Chronic Obstructive Pulmonary Disease (COPD) is accompanied by increased cellular stress and inflammation. Most of the Heat Shock Proteins (HSPs) have strong cytoprotective effects. The role of HSPs in COPD pathogenesis has not determined completely. We investigated the serum level of HSPs in COPD patients, smokers without COPD and healthy non-smoking controls. Also, we evaluated the relationship of HSPs with various parameters (inflammatory, oxidative, functional status, quality of life) in COPD patients. The levels of stress protein (HSP27, HSP70, HSP60, HSP90, CyPA), interleukin-6, C-reactive protein and malondialdehyde were measured in 16 healthy non-smoker, 14 smokers without COPD and 50 patients with stable COPD. Pulmonary function tests (PFT) and arterial blood gases parameters were measured. Health Related Quality of Life was evaluated and exercise capacity was measured with 6 minute walking test. Only HSP27 levels was significantly higher in COPD patients when compared with both healthy non-smoker and smokers without COPD (for both, p< 0.001). There was a weak-moderate negative correlation between serum levels of HSP27 and PFT parameters and between HSP27 levels and PaO2. Serum levels of HSP27 showed a weak-moderate positive correlation with symptom, activity and total scores. Subjects evaluated only smokers without COPD and patients with COPD; HSP27 had an area under the curve (AUC) in the receiver operating characteristic (ROC) curve of 0.819 (0.702-0.935; 95% CI; p= 0.000). Increased serum levels of HSP27 was found in COPD patients and our results showed sensitivity and specificity of serum HSP27 as diagnostic markers for COPD.

Extracellular cell stress (heat shock) proteins-immune responses and disease: an overview.

Extracellular cell stress proteins are highly conserved phylogenetically and have been shown to act as powerful signalling agonists and receptors for selected ligands in several different settings. They also act as immunostimulatory 'danger signals' for the innate and adaptive immune systems. Other studies have shown that cell stress proteins and the induction of immune reactivity to self-cell stress proteins can attenuate disease processes. Some proteins (e.g. Hsp60, Hsp70, gp96) exhibit both inflammatory and anti-inflammatory properties, depending on the context in which they encounter responding immune cells. The burgeoning literature reporting the presence of stress proteins in a range of biological fluids in healthy individuals/non-diseased settings, the association of extracellular stress protein levels with a plethora of clinical and pathological conditions and the selective expression of a membrane form of Hsp70 on cancer cells now supports the concept that extracellular cell stress proteins are involved in maintaining/regulating organismal homeostasis and in disease processes and phenotype. Cell stress proteins, therefore, form a biologically complex extracellular cell stress protein network having diverse biological, homeostatic and immunomodulatory properties, the understanding of which offers exciting opportunities for delivering novel approaches to predict, identify, diagnose, manage and treat disease.This article is part of the theme issue 'Heat shock proteins as modulators and therapeutic targets of chronic disease: an integrated perspective'.

Is Cr(III) toxic to bacteria: toxicity studies using Bacillus subtilis and Escherichia coli as model organism.

The objective of this work is to detect the mode of damage caused by Cr(III), one of the widely used industrial pollutant on Bacillus subtilis-industrial strain 168 and Escherichia coli MTCC 40. Bioassays are very sensitive, precise, economical and rapid for detecting early stages of pollution. The detrimental effect of trivalent chromium becomes clear from the growth profile and growth inhibition studies. Mode of action of damage by trivalent chromium in bacterial model was found to be oxidative, as chromium is one of the redox active metals. The generation of reactive oxygen species (ROS) resulted in membrane damage which in turn had a detrimental effect on the membrane proteins as well as the DNA. The structural changes in the SEM and AFM images clearly reveals the damage caused by Cr(III) to the test bacterial models. Trivalent chromium causes greater DNA, protein and membrane damage in case of E. coli than B. subtilis. Membrane damage caused by ROS becomes evident from the production of Thiobarbituric acid reactive substances (TBARs) as the mechanism of killing followed by DNA damage and the production of elevated levels of stress proteins known as extracellular cellular proteins.

An investigation of methyl tert‑butyl ether‑induced cytotoxicity and protein profile in Chinese hamster ovary cells.

Methyl tert-butyl ether (MTBE) is widely used as an oxygenating agent in gasoline to reduce harmful emissions. However, previous studies have demonstrated that MTBE is a cytotoxic substance that has harmful effects in vivo and in vitro. Although remarkable progress has been made in elucidating the mechanisms underlying the MTBE-induced reproductive toxicological effect in different cell lines, the precise mechanisms remain far from understood. The present study aimed to evaluate whether mammalian ovary cells were sensitive to MTBE exposure in vitro by assessing cell viability, lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA) content and antioxidant enzyme activities. In addition, the effect of MTBE exposure on differential protein expression profiles was examined by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. MTBE exposure induced significant effects on cell viability, LDH leakage, plasma membrane damage and the activity of antioxidant enzymes. In the proteomic analysis, 24 proteins were demonstrated to be significantly affected by MTBE exposure. Functional analysis indicated that these proteins were involved in catalytic activity, binding, structural molecule activity, metabolic processes, cellular processes and localization, highlighting the fact that the cytotoxic mechanisms resulting from MTBE exposure are complex and diverse. The altered expression levels of two representative proteins, heat shock protein family A (Hsp70) members 8 and 9, were further confirmed by western blot analysis. The results revealed that MTBE exposure affects protein expression in Chinese hamster ovary cells and that oxidative stress and altered protein levels constitute the mechanisms underlying MTBE-induced cytotoxicity. These findings provided novel insights into the biochemical mechanisms involved in MTBE-induced cytotoxicity in the reproductive system.

Amelioration of bleomycin-induced pulmonary fibrosis by chlorogenic acid through endoplasmic reticulum stress inhibition

To investigate the inhibitory effects of chlorogenic acid on pulmonary fibrosis and the internal mechanisms in vivo and in vitro. 30 male BALB/C mice were randomized into 5 groups: control group, pulmonary fibrosis model group, low, middle and high dose of chlorogenic acid groups. Mice in pulmonary fibrosis model group were administered 5.0mg/kg bleomycin with intracheal instillation and mice in 3 chlorogenic acid groups were treated with chlorogenic acid every day for 28 days after bleomycin administration. Lung tissue histology was observed using HE staining. Primary pulmonary fibroblasts were isolated and cultured. The expressions of fibrosis related factors (α-SMA and collagen I), as well as ER stress markers (CHOP and GRP78) were determined by both real-time PCR assay and Western blotting, while the expressions of other ER stress signaling pathway factors PERK, IRE-1, ATF-6 and protein levels of caspase-12, caspase-9, caspase-3, PARP were determined by Western blotting. RLE-6TN cell line induced by TGF-β1 was also used to verify the amelioration effects in vitro study. In both in vivo and in vitro studies, TUNEL staining was used to evaluate cell apoptosis. Expressions of collagen I, α-SMA, GRP78, and CHOP were significantly inhibited by chlorogenic acid in dose-dependent manner. Similarly, decreasing levels of cleaved caspase-12, caspase-9, caspase-3 and increasing level of uncleaved PARP were observed in chlorogenic acid groups compared with those in the fibrosis group both in vivo and in vitro. Chlorogenic acid could also significantly down-regulate the level of phosphorylation of PERK and cleaved ATF-6 in vivo study. Moreover, MTT assay demonstrated chlorogenic acid could enhance proliferation of RLE-6TN cells induced by TGFβ1 in vitro. And the apoptosis assays indicated that chlorogenic acid could significantly inhibit cell apoptosis both in vivo and in vitro studies. Chlorogenic acid could inhibit the pulmonary fibrosis through endoplasmic reticulum stress inhibition in vivo and in vitro.

Reduction of oxidative-nitrosative stress underlies anticataract effect of topically applied tocotrienol in streptozotocin-induced diabetic rats.

Cataract, a leading cause of blindness, is of special concern in diabetics as it occurs at earlier onset. Polyol accumulation and increased oxidative-nitrosative stress in cataractogenesis are associated with NFκB activation, iNOS expression, ATP depletion, loss of ATPase functions, calpain activation and proteolysis of soluble to insoluble proteins. Tocotrienol was previously shown to reduce lens oxidative stress and inhibit cataractogenesis in galactose-fed rats. In current study, we investigated anticataract effects of topical tocotrienol and possible mechanisms involved in streptozotocin-induced diabetic rats. Diabetes was induced in Sprague Dawley rats by intraperitoneal injection of streptozotocin. Diabetic rats were treated with vehicle (DV) or tocotrienol (DT). A third group consists of normal, non-diabetic rats were treated with vehicle (NV). All treatments were given topically, bilaterally, twice daily for 8 weeks with weekly slit lamp monitoring. Subsequently, rats were euthanized and lenses were subjected to estimation of polyol accumulation, oxidative-nitrosative stress, NFκB activation, iNOS expression, ATP levels, ATPase activities, calpain activity and total protein levels. Cataract progression was delayed from the fifth week onwards in DT with lower mean of cataract stages compared to DV group (p<0.01) despite persistent hyperglycemia. Reduced cataractogenesis in DT group was accompanied with lower aldose reductase activity and sorbitol level compared to DV group (p<0.01). DT group also showed reduced NFκB activation, lower iNOS expression and reduced oxidative-nitrosative stress compared to DV group. Lenticular ATP and ATPase and calpain 2 activities in DT group were restored to normal. Consequently, soluble to insoluble protein ratio in DT group was higher compared to DV (p<0.05). In conclusion, preventive effect of topical tocotrienol on development of cataract in STZ-induced diabetic rats could be attributed to reduced lens aldose reductase activity, polyol levels and oxidative-nitrosative stress. These effects of tocotrienol invlove reduced NFκB activation, lower iNOS expression, restoration of ATP level, ATPase activities, calpain activity and lens protein levels.

Chemical Chaperone 4-Phenylbutyric Acid Reduces Cardiac Ischemia/Reperfusion Injury by Alleviating Endoplasmic Reticulum Stress and Oxidative Stress.

BackgroundCardiovascular diseases are the leading cause of death in many countries and myocardial ischemia-reperfusion (I/R) injury is the cause of many serious heart diseases. Recent reports suggested that endoplasmic reticulum (ER) stress is associated with the progress of ischemia/reperfusion (I/R) injury. In a previous study, we illustrated that 4-phenylbutyric acid (4-PBA) reduces I/R-induced cell death in vitro through inhibiting the ER stress-initiated cell apoptosis. In the present study we investigated whether 4-PBA improves heart function in isolated rat hearts subjected to I/R and elucidated the potential mechanisms involved in 4-PBA-induced cardioprotective effects.Material/MethodsThe isolated rat hearts were subjected to global ischemia and reperfusion in the absence or presence of 4-PBA. Hemodynamic parameters (LVSP, LVEDP, ±dP/dtmax, and HR) were monitored and histopathological examination was applied. The biomarkers related to oxidative stress were detected by LDH, ROS, MDA, CK, SOD, and GSH-Px kits. A TUNEL apoptosis assay kit was used to detect apoptosis. The expression levels of ER stress and apoptosis proteins were evaluated by Western blotting.ResultsWe found that 4-PBA (5 mM, 10 mM) pretreatment significantly attenuated cardiac dysfunction and depressed oxidative stress induced by I/R. Moreover, I/R activated the ER stress proteins Grp78 and PERK, which are all decreased by 4-PBA. 4-PBA pretreatment also inhibited the expression of CHOP, Caspase-12, and Bax, reduced the phosphorylation of JNK, and enhanced the expression of anti-apoptotic protein Bcl-2.ConclusionsWe elucidated the significant protective effects of 4-PBA against I/R injuries by inhibition of ER stress, oxidative stress, and their associated apoptosis.

Weight loss-induced cellular stress in subcutaneous adipose tissue and the risk for weight regain in overweight and obese adults.

Weight loss is often followed by weight regain after the dietary intervention (DI). Cellular stress is increased in adipose tissue of obese individuals. However, the relation between cellular stress and weight regain is unclear. Previously, we observed increased adipose tissue cellular stress of participants regaining weight compared with participants maintaining weight loss. In the current study, we further investigated the relation between weight regain and changes in the expression of stress-related genes and stress protein levels to determine possible predictors of weight regain. In this randomized controlled trial, sixty-one healthy overweight/obese participants followed a DI of either a 5-week very-low-calorie diet (500 kcal per day) or a 12-week low-calorie diet (1250 kcal per day; WL period) with a subsequent 4-week weight stable diet (WS period), and a 9-month follow-up. The WL and WS period taken together was named the DI. Abdominal subcutaneous adipose tissue biopsies were collected in 53 participants for microarray and liquid chromatography-mass spectrometry analysis. RNA and protein levels for a broad set of stress-related genes were correlated to the weight regain percentage. Different gene sets correlated to weight regain percentage during WS and DI. Bioinformatics clustering suggests that during the WS phase-defined genes for actin filament dynamics, glucose handling and nutrient sensing are related to weight regain. HIF-1 (hypoxia-inducible factor-1) is indicated as an important regulator. With regard to DI, clustering of correlated genes indicate that LGALS1, ENO1 and ATF2 are important nodes for conferring risk for weight regain. Our present findings indicate that the risk for weight regain is related to expression changes of distinct sets of stress-related genes during the first 4 weeks after returning to energy balance, and during the DI. Further research is required to investigate the mechanistic significance of these findings and find targets for preventing weight regain.

Dose-dependent effects of cisplatin on the severity of testicular injury in Sprague Dawley rats: reactive oxygen species and endoplasmic reticulum stress.

Cisplatin (CIS) is used in the treatment of cancer, but its nonspecific systemic actions lead to toxic effects on other parts of the body. This study investigated the severity of CIS toxicity by increasing its dose over a constant time period. Sprague Dawley rats were divided into five treatment groups and control group with CIS (2, 4, 6, 8, and 10 mg/kg) administered intraperitoneally for 5 days. The body and organs were weighed, epididymal sperm was counted, and sperm motility and sperm apoptosis were evaluated. Blood samples were evaluated for complete blood count, reactive oxygen and nitrogen species, malondialdehyde levels, and total testosterone. The testicular tissue was examined for steroidogenic acute regulatory protein and endoplasmic reticulum stress protein. Epididymal sperm was collected for CatSper Western blot. The toxic effects of different doses of CIS on the testis and kidney were compared histologically. The weights of body, testis, epididymis, prostate, seminal vesicle, and kidney; sperm count; sperm motility; steroidogenic acute regulatory protein level; and epididymal sperm count were significantly lower in the CIS-treated groups than in the control group. In contrast, sperm apoptosis, plasma reactive oxygen and nitrogen species, and malondialdehyde, testosterone, red blood cell, hematocrit, hemoglobin, and endoplasmic reticulum stress protein levels all increased. Though CIS effectively treats cancer, at an increased dose it is toxic and life-threatening to the genitourinary system and other parts of the body.

Bonny light crude oil-induced alteration in levels of testicular stress proteins is accompanied by apoptosis in rats after treatment withdrawal.

The folkloric use of Bonny light crude oil (BLCO) in the treatment of gastrointestinal disorders and as an anti-poison is a generally acceptable practice in the Niger Delta area of Nigeria. The testicular dysfunction induced by BLCO exposure is of public concern with a view to its folkloric usage. The present study investigated the effects of BLCO exposure and withdrawal on the levels of testicular stress proteins and apoptosis-related proteins in rats. Adult male Wistar rats were exposed to 800 mg/kg body weight of BLCO for 7 days. One-half of the rats in each group were sacrificed on day 8, while the remaining one-half stayed an additional 45 days without treatment. Western blot analysis showed that administration of BLCO resulted in a significant increase in the levels of stress proteins and apoptosis-related proteins by 50% and above relative to control, except cytosolic nuclear factor-κB (NF-κB), which decreased significantly relative to control. This was followed by a concomitant increase in the expression of caspase-3, FasL, and NF-κB by immunofluorescence staining within the testicular germ cells. Apoptosis showed a significant increase in TUNEL-positive cells. Following withdrawal of treatment, BLCO-mediated alteration in stress proteins and induction of apoptosis persisted relative to control. Collectively, BLCO induced irreversible alteration in testicular stress proteins and apoptosis in rats within the time course of investigation. These findings highlight the potential long-term adverse effects of BLCO on individuals unduly exposed to BLCO.

Exposure to extremely low frequency electromagnetic fields alters the behaviour, physiology and stress protein levels of desert locusts

Electromagnetic fields (EMFs) are present throughout the modern world and are derived from many man-made sources including overhead transmission lines. The risks of extremely-low frequency (ELF) electromagnetic fields are particularly poorly understood especially at high field strengths as they are rarely encountered at ground level. Flying insects, however, can approach close to high field strength transmission lines prompting the question as to how these high levels of exposure affect behaviour and physiology. Here we utilise the accessible nervous system of the locust to ask how exposure to high levels of ELF EMF impact at multiple levels. We show that exposure to ELF EMFs above 4 mT leads to reduced walking. Moreover, intracellular recordings from an identified motor neuron, the fast extensor tibiae motor neuron, show increased spike latency and a broadening of its spike in exposed animals. In addition, hind leg kick force, produced by stimulating the extensor tibiae muscle, was reduced following exposure, while stress-protein levels (Hsp70) increased. Together these results suggest that ELF EMF exposure has the capacity to cause dramatic effects from behaviour to physiology and protein expression, and this study lays the foundation to explore the ecological significance of these effects in other flying insects.

Autophagy Activation by Transcription Factor EB (TFEB) in Striatum of HDQ175/Q7 Mice.

Background: Mutant huntingtin (mHTT) is encoded by the Huntington’s disease (HD) gene and its accumulation in the brain contributes to HD pathogenesis. Reducing mHTT levels through activation of the autophagosome-lysosomal pathway may have therapeutic benefit. Transcription factor EB (TFEB) regulates lysosome biogenesis and autophagy.Objective: To examine if increasing TFEB protein levels in HD mouse striatum induces autophagy and influences mHTT levels.Methods: We introduced cDNA encoding TFEB with an HA tag (TFEB-HA) under the control of neuron specific synapsin 1 promoter into the striatum of 3 month old HDQ175/Q7 mice using adeno-associated virus AAV2/9. The levels of exogenous TFEB were analyzed using qPCR and Western blot. Proteins involved in autophagy, levels of huntingtin, and striatal-enriched proteins were examined using biochemical and/or immunohistochemical methods.Results: In HD mice expressing TFEB-HA, HA immunoreactivity distributed throughout the striatum in neuronal cell bodies and processes and preferentially in neuronal nuclei and overlapped with a loss of DARPP32 immunoreactivity. TFEB-HA mRNA and protein were detected in striatal lysates. There were increased levels of proteins involved with autophagosome/lysosome activity including LAMP-2A, LC3II, and cathepsin D and reduced levels of mutant HTT and the striatal enriched proteins DARPP32 and PDE10A. Compared to WT mice, HDQ175/Q7 mice had elevated levels of the ER stress protein GRP78/BiP and with TFEB-HA expression, increased levels of the astrocyte marker GFAP and pro-caspase 3.Conclusion: These results suggest that TFEB expression in the striatum of HDQ175/Q7 mice stimulates autophagy and lysosome activity, and lowers mHTT, but may also increase a neuronal stress response.

Effects of short-term exposure of Acheta domesticus to nanodiamonds in food: DNA damage but no histological alteration in tissues

The aim of this work is to evaluate toxicity of nanodiamonds (ND, 20 and 200 mg g−1 food) to Acheta domesticus after short-term exposure (10 days) in food. Catalase (CAT), total antioxidant capacity (TAC), stress protein level (HSP70) and DNA damage to hemocytes were monitored every two days. On day 2 and 10, the levels of reactive oxygen species (ROS), apoptosis (annexin V and multicaspase tests), cell cycle and DNA damage to the gut were examined. The ultrastructure of the gut epithelium and testis was assessed on day 10 of the experiment. CAT, TAC, HSP70 in hemolymph, and histological morphology of gut and testis did not indicate ND toxicity. In the gut we observed a transient increase in CAT activity on day 2,4 and 6; and an increase in HSP70 level on day 6, 8 and 10. TAC level in the gut did not increase until day 10. ND did not influence ROS level in cells and the cell cycle. Yet, we observed a moderate increase in apoptosis, particularly at the higher ND concentration. ND induced dose-dependent DNA damage. We can conclude that short-term, occasional exposure to trace amounts of ND, like in medical applications, is probably safe for organisms.