Oregano (Origanum vulgare) Essential Oil and Its Constituents Prevent Rat Kidney Tissue Injury and Inflammation Induced by a High Dose of L-Arginine.

This study aimed to evaluate the protective action of oregano (Origanum vulgare) essential oil and its monoterpene constituents (thymol and carvacrol) in L-arginine-induced kidney damage by studying inflammatory and tissue damage parameters. The determination of biochemical markers that reflect kidney function, i.e., serum levels of urea and creatinine, tissue levels of neutrophil-gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1), as well as a panel of oxidative-stress-related and inflammatory biomarkers, was performed. Furthermore, histopathological and immunohistochemical analyses of kidneys obtained from different experimental groups were conducted. Pre-treatment with the investigated compounds prevented an L-arginine-induced increase in serum and tissue kidney damage markers and, additionally, decreased the levels of inflammation-related parameters (TNF-α and nitric oxide concentrations and myeloperoxidase activity). Micromorphological kidney tissue changes correlate with the alterations observed in the biochemical parameters, as well as the expression of CD95 in tubule cells and CD68 in inflammatory infiltrate cells. The present results revealed that oregano essential oil, thymol, and carvacrol exert nephroprotective activity, which could be, to a great extent, associated with their anti-inflammatory, antiradical scavenging, and antiapoptotic action and, above all, due to their ability to lessen the disturbances arising from acute pancreatic damage. Further in-depth studies are needed in order to provide more detailed explanations of the observed activities.

Acutely applied melatonin prevents CCl4-induced testicular lesions in rats: the involvement of the oxidative capacity and arginine metabolism

Abstract Carbon tetrachloride (CCl4) represents an organic chemical that causes reactive oxygen species derived organ disturbances including male infertility. Melatonin (MLT) is a neurohormone with strong antioxidant capacity, involved in numerous physiological processes. In this study we evaluated the capability of MLT, administered in a single dose of 50 mg/kg, to preserve the testicular tissue function after an acute administration of CCl4 to rats. The disturbance in testicular tissue and the effects of MLT after CCl4 exposure were estimated using biochemical parameters that enabled us to determine the tissue (anti)oxidant status and the intensity of arginine/nitric oxide metabolism. Also, the serum levels of testosterone and the histopathological analysis of tissue gave us a better insight into the occurring changes. A significant diminution in tissue antioxidant defences, arginase activity and serum testosterone levels, followed by the increased production of nitric oxide and extensive lipid and protein oxidative damage, was observed in the CCl4-treated group. The application of MLT after the CCl4 caused changes, clearly visible at both biochemical and histological level, which could be interpreted mainly as a consequence of general antioxidant system stimulation and a radical scavenger. On the other hand, the application of MLT exerted a limited action on the nitric oxide signalling pathway.

Protective effects of anthocyanins from bilberry extract in rats exposed to nephrotoxic effects of carbon tetrachloride.

This study examined the nephroprotective effects of 15 different anthocyanins from the bilberry extract on the acute kidney injury caused by CCl4. The acute nephrotoxicity in rats was induced 24 h after the treatment with a single dose of CCl4 (3 mL/kg, i.p.).The nephroprotective effects of the anthocyanins were examined in the animals that had been given the bilberry extract in a single dose of 200 mg of anthocyanins/kg daily, 7 days orally, while on the seventh day, 3 h after the last dose of anthocyanins, the animals received a single dose of CCl4 (3 mL/kg, i.p.) and were sacrificed 24 h later. When the nephrotoxicant alone was administered, it resulted in a substantial increase of the pro-oxidative (TBARS, CD, H2O2, XO, and GSSG) and pro-inflammatory markers (TNF-α, NO, and MPO), as well as a noticeable reduction of the antioxidant enzymes (CAT, SOD, POD, GPx, GST, GR) and GSH when compared to the results of the control group. Moreover, the application of CCl4 significantly influenced a reduction of the renal function, as well as an increase in the sensitive and specific injury indicators of the kidney epithelial cells (ß2-microglobulin, NGAL, KIM1/TIM1) in the serum and urine of rats. The pretreatment of the animals poisoned with CCl4 with the anthocyanins from the bilberry extract led to a noticeable reduction in the pro-oxidative and pro-inflammatory markers with reduced consumption of the antioxidant defence kidney capacity, compared to the animals exposed to CCl4 alone. Anthocyanins have been protective for the kidney parenchyma, with an apparent absence of the tubular and periglomerular necrosis, severe degenerative changes, inflammatory mononuclear infiltrates and dilatation of proximal and distal tubules, in contrast to the CCl4-intoxicated animals. The nephroprotective effects of anthocyanins can be explained by strong antioxidant and anti-inflammatory effects achieved through the stabilization and neutralization of highly reactive and unstable toxic CCl4 metabolites.

Oral supplementation with melatonin reduces oxidative damage and concentrations of inducible nitric oxide synthase, VEGF and matrix metalloproteinase 9 in the retina of rats with streptozotocin/nicotinamide induced pre-diabetes.

Hyperglycemia mediated oxidative stress and pro-angiogenic molecules such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP9) are considered important for diabetic retinopathy onset and progression. Melatonin is a pineal hormone that regulates circadian and seasonal rhythms and most likely is involved in regulating glucose metabolism. We aimed to evaluate the potential benefit of melatonin supplementation to the pre-diabetic retina by assessing melatonin effects on lipid peroxidation (thiobarbituric acid reactive substances, TBARS), protein oxidation (advanced oxidation protein products, AOPP) and concentrations of inducible nitric oxide synthase (iNOS), VEGF and MMP9 in the retina of rats with pre-diabetes. Pre-diabetes was induced by streptozotocin (45 mg/kg, i.p.) following nicotinamide injection (110 mg/kg, i.p.). Beside mild hyperglycemia, lower serum insulin, increased fructosamine and lower HDL cholesterol, the present study demonstrated decreased serum melatonin in pre-diabetic rats, as well as, increased concentration of retinal TBARS, AOPP, iNOS, VEGF, and MMP9. Oral supplementation with melatonin (85 µg/animal/day) caused melatonin and HDL cholesterol levels to rise in treated rats and reduced levels of fasting serum glucose and fructosamine. It also affected serum insulin and quantitative insulin sensitivity check index (QUICKI) in treated groups but had no significant effect on non-fasting glucose. Finally, supplementation with melatonin reduced concentrations of TBARS, AOPP, iNOS, VEGF, and MMP9 in significant level, thereby exerting an overall positive effect on oxidative stress and pro-angiogenic signaling in the pre-diabetic retina. Thus, oral melatonin might be considered in an early treatment or in the prevention of retinal changes associated with pre-diabetes.

Influence of different wild-garlic (Allium ursinum) extracts on the gastrointestinal system: spasmolytic, antimicrobial and antioxidant properties.

OBJECTIVES: As there are no previous studies of the European wild-garlic (Allium ursinum) effects on the gastrointestinal system, despite its traditional applications in gastrointestinal disorders' treatment and regular use in the human diet, we have quantified and compared spasmolytic, antimicrobial and antioxidant activities of its different leaf extracts. METHODS: Wild-garlic extracts were tested for spasmolytic activity on isolated rat ileum, antimicrobial activity on selected Gram-positive and Gram-negative bacteria and fungi by microdilution method and antioxidant capacity by DPPH radical-scavenging assay. KEY FINDINGS: Wild-garlic extracts were found to decrease ileal basal tone. As the relaxation of K+ -induced contractions was similar to one caused by papaverin, the observed spasmolytic effect was most likely mediated through Ca2+ -channel inhibition. Ethanolic extract (with the highest phenolic and high alk(en)yl cysteine sulphoxides' levels) produced the strongest spasmolytic activity. In case of acetylcholine-induced contractions, only hydromethanolic extract showed no statistical difference in comparison with positive control. All samples exhibited certain antioxidant potential and strong antimicrobial activity against tested enteropathogenic strains (Salmonella enteritidis was the most sensitive, followed by Escherichia coli, Proteus mirabilis and Enterococcus faecalis). CONCLUSION: Besides other already established health-promoting effects, wild garlic could be useful in treatment of mild gastrointestinal disturbances.

Bilberry: Chemical Profiling, in Vitro and in Vivo Antioxidant Activity and Nephroprotective Effect against Gentamicin Toxicity in Rats.

We assessed possible protective effect of bilberry diet in rat model of nephrotoxicity. In vivo and in vitro antioxidant activity and chemical profiling of this functional food was performed. With aid of HPLC-DAD and spectrophotometric method, 15 individual anthocyanins were quantified alongside total tannin, phenylpropanoid, and anthocyanin content. The study was conducted on four groups of rats: control, treated with only gentamicin, treated with only bilberry, and treated with both gentamicin and bilberry. Kidney function was evaluated by tracking urea and creatinine. Morphology of renal tissue and its changes were recorded pathohistologically and quantified morphometrically. Bilberry (100 mg/kg daily) showed strong nephroprotective effect against gentamicin toxicity in rats (as shown through MDA, AOPP, and catalase levels). In conclusion, the demonstrated protective activity of bilberry extract matched well with the assessed in vivo and in vitro antioxidant activity as well as with its polyphenolic content, particularly with high anthocyanin levels. Copyright © 2016 John Wiley & Sons, Ltd.

The effect of bilberries on diabetes-related alterations of interstitial cells of Cajal in the lower oesophageal sphincter in rats.

Diabetic gastroenteropathy involves not only the parasympathetic and sympathetic autonomic nerves, but also enteric neurons, smooth muscle cells and interstitial cells of Cajal (ICC). ICC are the cells of mesenchymal origin that occur within and around the muscle layers in the gastrointestinal tract. The objective of the present study was to investigate the alterations of ICC in the lower oesophageal sphincter (LOS) of streptozotocin-nicotinamide non-insulin-dependent diabetes rats. Moreover, we investigated possible ICC in rats with the same type of diabetes, treated with bilberry fruit extract, bearing in mind that its hypoglycemic effect had been already proven. Male Wistar rats (10 weeks old) were used, and diabetes was induced by an intraperitoneal injection of streptozotocin, immediately after intraperitoneal application of nicotinamide. The specimens were exposed to anti-c-kit antibodies to investigate the distribution of ICC, and the smooth muscle cells were immunohistochemically labelled using anti-desmin antibodies. Intramuscular ICC were very abundant in the LOS of rats. They were spindle-shaped, with two long processes connecting them into long linear sequences. In the LOS of diabetic rats, intramuscular ICC were rarely present and linear cell-cell connections between these cells were completely missing. In groups treated with bilberry, the number and distribution of ICC were exactly the same as in the above described rats with induced diabetes. In summary, a decrease of intramuscular ICC, discontinuities and breakdown of contacts between ICC were observed in streptozotocin-nicotinamide induced diabetes rats and in groups treated with bilberry. Bilberry fruit extract was shown to have hypoglycemic activity, but without any protective effects on ICC in the LOS of diabetic rats.

Antioxidant and proapoptotic effects of anthocyanins from bilberry extract in rats exposed to hepatotoxic effects of carbon tetrachloride.

AIMS: The aim of this research was to determine the hepatoprotective effects of anthocyanins from bilberry extract in rats exposed to carbon tetrachloride (CCl4) by monitoring the parameters of oxidative stress and apoptosis, and by performing the histopathological and morphometric analyses. MAIN METHODS: Animals were divided into four groups: Group I (0.9% NaCl-10days), Group II (bilberry extract, 75mg/kg-10days), Group III (0,9% NaCl-9days, and on the tenth day CCl4-2ml/kg), Group IV (bilberry extract, 75mg/kg-10days and on the tenth day CCl4-2ml/kg). KEY FINDINGS: Bilberry extract led to a significant decrease in the activity of biochemical parameters in serum (AST, GGT, LDH, and ALT), the activity of pro-oxidative enzyme xanthine oxidase, as well as the level of lipid peroxidation in the liver in Group IV compared to Group III (p<0.01). Bilberry extract resulted in a significant increase in the activity of the antioxidant markers-catalase (p<0.05), superoxide dismutase, glutathione S-transferase and glutathione peroxidase (p<0.01), and the concentration of reduced glutathione (p<0.05) in Group IV in relation to Group III. The application of bilberry extract resulted in an increase in the number of apoptotic hepatocytes and the activity of caspase-3 in the liver tissue (p<0.01). The reduction of coagulation necrotic areas was proved (p<0.001) as well as the number of macrovesicular hepatocytes (p<0.01), along with an increased mitotic activity (p<0.01) in Group IV compared to Group III. SIGNIFICANCE: Anthocyanins from bilberry extract have strong antioxidant properties and therefore can be considered as powerful hepatoprotectives in natural products.

Impact of folic acid supplementation on single- and double-stranded RNA degradation in human colostrum and mature milk.

Sufficient intake of folic acid is necessary for normal embryogenesis, fetal, and neonatal development. Folic acid facilitates nucleic acid internalization, and protects cellular DNA from nuclease degradation. Human milk contains enzymes, antimicrobial proteins, and antibodies, along with macrophages, that protect against infections and allergies. However, little to no information is available on the effects of folic acid supplementation on degradation of nucleic acids in human milk. In the present study, we aimed to determine the RNase activity (free and inhibitor-bound) in colostrum and mature milk, following folic acid supplementation. The study design included a total of 59 women, 27 of whom received 400 µg of folic acid daily periconceptionally and after. Folic acid supplementation increased the free RNase and polyadenylase activity following lactation. However, the increased RNase activity was not due to de novo enzyme synthesis, as the inhibitor-bound (latent) RNase activity was significantly lower and disappeared after one month. Folic acid reduced RNase activity by using double-stranded RNA as substrate. Data suggests that folic acid supplementation may improve viral RNAs degradation and mRNA degradation, but not dsRNA degradation, preserving in this way the antiviral defense.

Predicting the biological effects of mobile phone radiation absorbed energy linked to the MRI-obtained structure.

The nature of an electromagnetic field is not the same outside and inside a biological subject. Numerical bioelectromagnetic simulation methods for penetrating electromagnetic fields facilitate the calculation of field components in biological entities. Calculating energy absorbed from known sources, such as mobile phones when placed near the head, is a prerequisite for studying the biological influence of an electromagnetic field. Such research requires approximate anatomical models which are used to calculate the field components and absorbed energy. In order to explore the biological effects in organs and tissues, it is necessary to establish a relationship between an analogous anatomical model and the real structure. We propose a new approach in exploring biological effects through combining two different techniques: 1) numerical electromagnetic simulation, which is used to calculate the field components in a similar anatomical model and 2) Magnetic Resonance Imaging (MRI), which is used to accurately locate sites with increased absorption. By overlapping images obtained by both methods, we can precisely locate the spots with maximum absorption effects. This way, we can detect the site where the most pronounced biological effects are to be expected. This novel approach successfully overcomes the standard limitations of working with analogous anatomical models.

Skin ageing: natural weapons and strategies.

The fact that the skin is the most visible organ makes us aware of the ageing process every minute. The use of plant extracts and herbs has its origins in ancient times. Chronological and photo-ageing can be easily distinguished clinically, but they share important molecular features. We tried to gather the most interesting evidence based on facts about plants and plant extracts used in antiaging products. Our main idea was to emphasize action mechanisms of these plant/herbal products, that is, their "strategies" in fighting skin ageing. Some of the plant extracts have the ability to scavenge free radicals, to protect the skin matrix through the inhibition of enzymatic degradation, or to promote collagen synthesis in the skin. There are some plants that can affect skin elasticity and tightness. Certainly, there is a place for herbal principles in antiaging cosmetics. On the other hand, there is a constant need for more evaluation and more clinical studies in vivo with emphasis on the ingredient concentration of the plant/herbal products, its formulation, safety, and duration of the antiaging effect.

Alkaline phosphatase activity in human colostrum as a valuable predictive biomarker for lactational mastitis in nursing mothers.

AIMS: Biochemical investigations have shown that an indigenous milk enzyme - alkaline phosphatase (ALP) - which is detectable in the lactocytes, plays a very important diagnostic role in clinical medicine, since its activity varies in different tissues and serves as a specific indicator of disease states. The purpose of this study was to evaluate ALP activity in human colostrum as a possible early predictive biomarker for lactational mastitis in nursing mothers. PATIENTS & METHODS: During a period from May to July 2010, a total of 60 healthy nursing mothers were recruited for this study. RESULTS: The mean level of colostrum ALP activity from the affected breasts was significantly higher when compared with ALP activity from the contralateral asymptomatic as well as 'healthy' breasts (p < 0.01). CONCLUSION: Determining ALP activity in colostrum could be a valuable biochemical marker for an early prediction of mastitis in nursing mothers.

Correlation of nitric oxide levels in the cerebellum and spinal cord of experimental autoimmune encephalomyelitis rats with clinical symptoms.

Experimental autoimmune encephalomyelitis (EAE) is a well-established cell-mediated autoimmune inflammatory disease of the CNS, which has been used as a model of the human demyelinating disease. EAE is characterized by infiltration of the CNS by lymphocytes and mononuclear cells, microglial and astrocytic hypertrophy, and demyelination which cumulatively contribute to clinical expression of the disease. EAE was induced in female Sprague-Dawley rats, 3 months old (300 g ± 20 g), by immunization with myelin basic protein (MBP) in combination with Complete Freund's adjuvant (CFA). The animals were divided into 7 groups: control, EAE, CFA, EAE + aminoguanidine (AG), AG, EAE + N-acetyl-L-cysteine (NAC) and NAC. The animals were sacrificed 15 days after EAE induction, and the level of nitric oxide (NO(·)) production was determined by measuring nitrite and nitrate concentrations in 10% homogenate of cerebellum and spinal cord. Obtained results showed that the level of NO(·) was significantly increased in all examined tissues of the EAE rats compared to the control and CFA groups. Also, AG and NAC treatment decreased the level of NO(·) in all tissues compared to the EAE group. The level of NO(·) is increased significantly in the spinal cord compared to the cerebellum. The clinical course of the EAE was significantly decreased in the EAE groups treated with AG and NAC during the development of the disease compared to EAE group and its correlates with the NO(·) level in cerebellum and spinal cord. The findings of our work suggest that NO(·) and its derivatives play an important role in multiple sclerosis (MS). It may be the best target for new therapies in human demyelinating disease and recommend the new therapeutic approaches based on a decreased level of NO(·) during the course of MS.

Protective effect of selenium on gentamicin-induced oxidative stress and nephrotoxicity in rats.

Gentamicin (GM) is a widely used antibiotic against serious, life-threatening infections, but its usefulness is limited by the development of nephrotoxicity. The present study was designed to determine the protective effect of selenium (Se) in GM-induced nephrotoxicity in rats. Experiments were done on 32 adult Wistar rats divided into four groups of 8 animals each. The GM group received gentamicin (100 mg/kg), whereas the GM+Se group received the same dose of GM and selenium (1 mg/kg) by intraperitoneal (i.p.) injections on a daily basis. Animals in the Se group, serving as a positive control, received only selenium (1 mg/kg) and the control group received saline (1 mL/day), both given i.p. All groups were treated during 8 consecutive days. Quantitative evaluation of GM-induced structural alterations and degree of functional alterations in the kidneys were performed by histopathological and biochemical analyses in order to determine potential beneficial effects of selenium coadministration with GM. GM was observed to cause a severe nephrotoxicity, which was evidenced by an elevation of serum urea and creatinine levels. The significant increases in malondialdehyde levels and protein carbonyl groups indicated that GM-induced tissue injury was mediated through oxidative reactions. On the other hand, simultaneous selenium administration protected kidney tissue against oxidative damage and the nephrotoxic effect caused by GM treatment. Exposure to GM caused necrosis of tubular epithelial cells. Necrosis of tubules was found to be prevented by selenium pretreatment. The results from our study indicate that selenium supplementation attenuates oxidative-stress-associated renal injury by reducing oxygen free radicals and lipid peroxidation in GM-treated rats.

Aminoguanidine and N-acetyl-cysteine supress oxidative and nitrosative stress in EAE rat brains.

Experimental autoimmune encephalomyelitis (EAE) is a well-established animal model of human multiple sclerosis (MS). We have evaluated the role of oxidative and nitrosative stress, as the causal factors in the development of EAE, responsible for the damage of cardinal cellular components, such as lipids, proteins and nucleic acids, resulting in demyelination, axonal damage, and neuronal death. EAE was induced in female Sprague-Dawley rats, 3 months old (300±20 g), by immunization with myelin basic protein in combination with Complete Freund's adjuvant (CFA). The animals were divided into seven groups: control, EAE, CFA, EAE+aminoguanidine (AG), AG, EAE+N-acetyl-L-cysteine (NAC) and NAC. The animals were sacrificed 15 days after EAE induction, and the levels of nitrosative and oxidative stress were determined in 10% homogenate of the whole encephalitic mass. In EAE rats, brain NO production and MDA level were significantly increased (P<0.001) compared to the control values, whereas AG and NAC treatment decreased both parameters in EAE rats compared to EAE group (P<0.001). Glutathione (GSH) was reduced (P<0.001) in EAE rats in comparison with the control and CFA groups, but increased in EAE+AG and EAE+NAC group compared to the EAE group (P<0.01). Superoxide dismutase (SOD) activity was significantly decreased (P<0.001) in the EAE group compared to all other experimental groups. The clinical expression of EAE was significantly decreased (P<0.05) in the EAE groups treated with AG and NAC compared to EAE rats, during disease development. The obtained results prove an important role of oxidative and nitrosative stress in the pathogenesis of EAE, whereas AG and NAC protective effects offer new possibilities for a modified combined approach in MS therapy.