A Narrative Review: The Effect and Importance of Carotenoids on Aging and Aging-Related Diseases.

Aging is generally defined as a time-dependent functional decline that affects most living organisms. The positive increase in life expectancy has brought along aging-related diseases. Oxidative stress caused by the imbalance between pro-oxidants and antioxidants can be given as one of the causes of aging. At the same time, the increase in oxidative stress and reactive oxygen species (ROS) is main reason for the increase in aging-related diseases such as cardiovascular, neurodegenerative, liver, skin, and eye diseases and diabetes. Carotenoids, a natural compound, can be used to change the course of aging and aging-related diseases, thanks to their highly effective oxygen-quenching and ROS-scavenging properties. Therefore, in this narrative review, conducted using the PubMed, ScienceDirect, and Google Scholar databases and complying with the Scale for the Assessment of Narrative Review Articles (SANRA) guidelines, the effects of carotenoids on aging and aging-related diseases were analyzed. Carotenoids are fat-soluble, highly unsaturated pigments that occur naturally in plants, fungi, algae, and photosynthetic bacteria. A large number of works have been conducted on carotenoids in relation to aging and aging-related diseases. Animal and human studies have found that carotenoids can significantly reduce obesity and fatty liver, lower blood sugar, and improve liver fibrosis in cirrhosis, as well as reduce the risk of cardiovascular disease and erythema formation, while also lowering glycated hemoglobin and fasting plasma glucose levels. Carotenoid supplementation may be effective in preventing and delaying aging and aging-related diseases, preventing and treating eye fatigue and dry eye disease, and improving macular function. These pigments can be used to stop, delay, or treat aging-related diseases due to their powerful antioxidant, restorative, anti-proliferative, anti-inflammatory, and anti-aging properties. As an increasingly aging population emerges globally, this review could provide an important prospective contribution to public health.

A Comparison Study of Nutritional Assessment, Diet and Physical Activity Habits, Lifestyle and Socio-Demographic Characteristics in Individuals with and without Dizziness/Vertigo.

Dizziness and vertigo are growing health problems and have become responsible for increases in health expenditures. In this context, a case-control study has been conducted by nutritional assessment, including dietary and physical activity habits, lifestyle, and socio-demographic characteristics in adults with (patient group) and without (control group) dizziness or vertigo, and the outcomes were compared between these groups. The patient (n = 150) and control (n = 150) groups included 300 participants. The 24-h Dietary Recall and the food frequency questionnaire (FFQ-21) were conducted in order to gain detailed information about foods and beverages consumed by the participants. Additionally, a questionnaire was completed, assessing general socio-demographic (age, gender, etc.) and lifestyle (smoking, alcohol consumption, and obesity) characteristics, anthropometric measurements, and dietary and physical activity habits. The results revealed that there is an association between dizziness/vertigo and female gender and increasing age. Smoking status and alcohol consumption did not differ between the groups, whereas differences in body mass index and obesity were significantly higher in the patient group (65%; n = 98) than the control group (46%; n = 69) (p = 0.001). Skipping meals "everyday" was significantly high (p = 0.044), and lunch was the most skipped meal in the patient group. The three most preferred cooking methods were oven baking, boiling, and frying for both groups. Daily water intake in the patient group was lower than in the control group (p = 0.026). Dietary intake for carotene and vitamin K were significantly lower in the patient group than the control group, but the opposite was true for vitamin D intake (p < 0.05). Daily consumption of bread and dairy products were highest in the patient group (p < 0.05). The physical activity rate was 35% (n = 53) in the control group and 28% (n = 42) in the patient group. Regular walking was the most preferred activity in both groups (p = 0.037). Active monitoring of individual diet and hydration along with supporting professional counseling are advisable. In addition, a healthy lifestyle including weight control and regular physical activity can be helpful to reduce symptoms of dizziness/vertigo.

The Integral Role of Diets Including Natural Products to Manage Rheumatoid Arthritis: A Narrative Review.

Genetic and environmental factors including lifestyle are thought to play a key role in the pathophysiology of rheumatoid arthritis (RA). There is evidence that diet can enhance the inflammatory response in genetically predisposed individuals. On the other hand, certain types of diets can alleviate RA symptoms due to their anti-inflammatory and antioxidant activities. Also, natural compounds with potential effectiveness in RA management belong to different chemical classes such as flavonoids, polyphenols, carotenoids, and alkaloids with their antioxidant characteristics as well as probiotics. The nutritional approaches to prevent or extenuate the disease progress were examined in this narrative review which was conducted using the PubMed, ScienceDirect and Google Scholar databases and conforms to the Scale for the Assessment of Narrative Review Articles (SANRA) guidelines. Mediterranean and vegan diets equally have been shown to exhibit positive effects on RA as the consumption of dietary fiber, antioxidants and anti-inflammatory compounds from fruits, vegetables, grains, nuts, and seeds are high. Whereas Mediterranean diet additionally includes beneficial nutrients of animal origin such as omega-3 polyunsaturated fatty acids from fish and seafood, patients on vegan diet need to be monitored closely for intake of all critical nutrients. Certain calorie restrictions and intermittent fasting diets have been shown to benefit RA patients although there is an obvious need for further studies to establish solid evidence-based recommendations and guidelines. The research data available strongly suggest that dietary approaches with anti-inflammatory properties may help delay the onset of RA and/or improve symptoms and thus nutrition should be routinely addressed to facilitate management of the disease.

Anti-Inflammatory and Antioxidant Effects of Carvacrol on N-Methyl-N'-Nitro-N-Nitrosoguanidine (MNNG) Induced Gastric Carcinogenesis in Wistar Rats.

Carvacrol is a dietary polyphenol from Lamiaceae plants that has been shown to possess a wide range of biological activities including antioxidant and antitumor effects. This study aimed to investigate its anti-inflammatory and antioxidant effects on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced gastric carcinogenesis in Wistar rats. Forty-nine rats were randomly assigned to four treatment and three control groups. Over 60 days, MNNG (200 mg/kg BW) was orally applied to animals of groups 1-5 while the rats in groups 2-5 also received different doses of carvacrol (10, 25, 50, and 100 mg/kg BW, respectively) until the end of the experiment. Group 6 rats were treated with 100 mg/kg BW carvacrol and no MNNG whereas group 7 was the control group without any treatment. After the euthanasia of all rats, the inflammatory cytokines and oxidative stress parameters were assessed in the blood and tissues. The expression of caspase 9, Bax, and Bcl-2 proteins in the stomach tissues were investigated through histopathological examinations. Statistically significant differences were observed in the body weight, oxidative stress, and inflammation parameters of groups 1 to 6 compared to group 7 (p ≤ 0.001). Animals in MNNG groups 2 and 3 treated with the low dose carvacrol (10 and 25 mg/kg BW) showed significantly reduced oxidative stress, inflammation, and apoptotic effect compared to animals of the MNNG groups receiving increased doses of carvacrol (50 and 100 mg/kg BW) or no carvacrol. Rats exposed to MNNG exhibited gastric cancer cells in several areas. In the MNNG group receiving 100 mg/kg BW carvacrol, the inflammatory cell infiltration was observed in gastric mucosal and submucosal areas whereas MNNG rats supplemented with 10 and 25 mg/kg BW carvacrol showed no pathological alterations of the gastric cells. The results of this study indicate that significant antioxidant and anti-inflammatory effects induced by carvacrol at doses of 10 and 25 mg/kg BW interfered with gastric carcinogenesis induced by MNNG in Wistar rats as well as provide hepatoprotection. However, high doses of carvacrol (50 and 100 mg/kg BW) increased oxidative stress, inflammation, and apoptosis.

Preparation and evaluation of novel functional fermented dairy products containing propolis and cinnamon.

Novel functional food products might be an easy accessible and eligible approach to help reduce the risk of severe viral infections including SARS-CoV-2. Hence a product containing probiotics, propolis and cinnamon was developed and interferences of the ingredients were characterized. Yogurts were prepared using starter cultures with propolis (0.03%) and cinnamon in various concentrations (0.3%, 1%, and 2.5%). Bifidobacterium animalis ssp. lactis, Lactobacillus acidophilus, Streptococcus thermophilus, and Lactobacillus delbrueckii subsp. bulgaricus were used as microorganisms for yogurt production. Chemical analysis revealed a decline of fat matter in the presence of propolis and/or cinnamon. Propolis had statistically significant suppressive effects on Bifidobacterium animalis ssp. lactis as well as on Lactobacillus delbrueckii subsp. bulgaricus numbers (p < 0.05). These effects were diminished in the presence of increasing cinnamon concentrations. For Lactobacillus acidophilus a statistically significant reducing effect on the number of colonies was observed in all products investigated. Nevertheless all samples met the standard of recommended level of ≥ 106 viable cells/g of a product. Propolis showed an inverse effect on Streptococcus thermophilus by increasing its colony numbers in yogurts. The probiotic yogurt samples containing propolis (0.03%) and cinnamon (2.5%) gained the highest number of points in the sensory evaluation compared to control.

[A survey among breeders of South American camelids concerning breeding and reproduction management]. / Umfrage unter Neuweltkameliden-Züchtern zum Zucht- und Fortpflanzungsmanagement.

OBJECTIVE: South American camelids have become popular with an increasing number of breeders in Austria, Germany and Switzerland. The aim of the present online survey was to gain information concerning housing and management of South American camelids, particularly during pregnancy and parturition. MATERIAL AND METHODS: A link to the online survey was sent via email to 8 breeder associations in Austria, Germany and Switzerland. The questionnaire included a total of 24 closed or open questions. The period for participation was May to October 2019. RESULTS: A total of 92 breeders participated and 64 answered all questions. The survey revealed that South American camelids were mainly used for breeding (87 %) and production of fibre (74 %). The median flock size was 27 animals, mainly females. Most breeders (74 %) acquired their knowledge on South American camelids via personal contacts and advanced training courses. The majority (57 %) mated their females with an own sire that is kept separately. Using teaser males was the most popular pregnancy test (91 %), often combined with other techniques, especially ultrasonography. In most mares (88 %) duration of pregnancy was in the normal range of 335-360 days. Repeated problems with loss of pregnancy were reported by 76 % of breeders for less than 5 % of their mares. Postpartum problems occurred sporadically, e. g. lack of milk or mastitis (each < 5 %). Problems with the cria at parturition within the last 3 years were reported by > 50 % of the breeders, but reported reasons for dystocia showed no clear focus. The majority of the participants (69 %) were satisfied with the veterinary care in general and in the fields of pregnancy and obstetrics. CONCLUSION: The results of this survey provide new insights into the management practices of South American camelids breeders in Austria, Germany and Switzerland. Further studies could compare these practices with those in countries where breeding of South American camelids possesses a longer tradition and different commercial backgrounds. CLINICAL RELEVANCE: The results provide information to practitioners in preparation for problems and challenges in the veterinary care of South American camelids.

In Vitro Hormetic Effect Investigation of Thymol on Human Fibroblast and Gastric Adenocarcinoma Cells.

The concept of hormesis includes a biphasic cellular dose-response to a xenobiotic stimulus defined by low dose beneficial and high dose inhibitory or toxic effects. In the present study, an attempt has been made to help elucidate the beneficial and detrimental effects of thymol on different cell types by evaluating and comparing the impact of various thymol doses on cancerous (AGS) and healthy (WS-1) cells. Cytotoxic, genotoxic, and apoptotic effects, as well as levels of reactive oxygen species and glutathione were studied in both cell lines exposed to thymol (0-600 µM) for 24 h. The results showed significant differences in cell viability of AGS compared to WS-1 cells exposed to thymol. The differences observed were statistically significant at all doses applied (P ≤ 0.001) and revealed hormetic thymol effects on WS-1 cells, whereas toxic effects on AGS cells were detectable at all thymol concentrations. Thymol at low concentrations provides antioxidative protection to WS-1 cells in vitro while already inducing toxic effects in AGS cells. In that sense, the findings of the present study suggest that thymol exerts a dose-dependent hormetic impact on different cell types, thereby providing crucial information for future in vivo studies investigating the therapeutic potential of thymol.

Intracranial abscess formation in an adult alpaca: a case report.

BACKGROUND: Intracranial abscess formation is an extremely rare and sporadically documented disease in South American Camelids (SACs). Herein we report the first case of otogenic brain abscess formation in this species. CASE PRESENTATION: A 4 years old female alpaca was presented to our veterinary hospital with a 6 month history of neurologic disorder symptoms, mainly head tilt to the right and emaciation. A comprehensive workup (ultrasound and computed tomography) revealed irreversible cranial nerve abnormalities, extensive lesions in the region of external, middle and internal right ear including destruction of bony structures (tympanic bulla, parts of temporal bone) and severe brain deformation caused by an intracranial abscess. The lesion was up to 6x7x4 cm and occupying almost 40% of the cranial cavity. No pathological findings were evident in other organs or structures. The late referral of the alpaca at this advanced stage of destructive disease precluded surgical intervention. CONCLUSIONS: This case report describes the clinical signs, diagnostic procedures and pathological findings in an adult female alpaca suffering from cranial nerve abnormalities caused by a massive otogenic brain abscess. Camelids suffering from otitis may not present with clinical signs until the pathology is severe. The importance of considering intracranial abscess formation as differential diagnosis in SACs showing the merest hint of nerve deficits cannot be emphasized enough in order to diagnose such pathological processes at an early and treatable stage.

Effects of carvacrol on human fibroblast (WS-1) and gastric adenocarcinoma (AGS) cells in vitro and on Wistar rats in vivo.

Carvacrol is a natural phenolic compound found in essential oils of Lamiaceae species. In the present study, an attempt has been made to elucidate the mechanism behind the anti-cancer potential of carvacrol on human gastric adenocarcinomas (AGS) by comparing its effects on cancer cells AGS to those on normal human fibroblast (WS-1) cells, in vitro. Cytotoxicity, reactive oxygen species (ROS) generation, glutathione (GSH) levels, genotoxicity, and apoptotic effects of carvacrol (0-600 µM) were studied in both cell lines. Additionally, the effect of high dose carvacrol (100 mg/kg BW) on the oxidative status was investigated in vivo. For this purpose, carvacrol was administered orally to male Wistar rats over a period of 60 days. Rats were weighed regularly. At the end of the experiment, rats were euthanized. Blood and stomach tissues were collected for biochemical and pathological examinations. The in vitro results showed significant differences in cell viability of AGS compared to WS-1 cells exposed to carvacrol. Also the extent of ROS generation, GSH reduction and DNA damage differed significantly between the cell lines studied (P ≤ 0.001). The differences observed were statistically significant at all concentrations applied (P ≤ 0.001). The results found in AGS cells were mirrored in the pathohistological findings obtained from animals of the in vivo experimental group. Changes in body weight, and oxidative stress index for plasma and stomach tissues of animals in this group were found to differ statistically significant from those found in the control group of Wistar rats (P ≤ 0.001). The data obtained from our present study uncovered that carvacrol has the potential to cause toxic effects in both, AGS and WS-1 cells but more effectively in cancer cells than in normal cells. The carvacrol-mediated responses observed in the in vitro and in vivo experiments presented suggest a double-edged pro-oxidative effect. Via this mechanism carvacrol induced cytotoxicity, apoptosis, and DNA damage in a dose-dependent manner in both cancer and normal cells and these activities were higher in cancer cells than those of normal cells.

Llamas and alpacas in Europe: Endoparasites of the digestive tract and their pharmacotherapeutic control.

There are distinctive specifications for veterinary medical care of South American camelids (SACs), namely, llamas, alpacas, vicunas and guanacos. Camelids are classified as food-producing animals, but as veterinary medicinal products are often only licensed for domestic food-producing species such as horses, goats, sheep and cattle, treatment of SACs generally requires off-label use of drugs. Endoparasitism is a major health concern in camelids and can result in severe clinical diseases and economic losses. There is still a lack of work on the pharmacokinetics, safety and efficacy for most antiparasitic drugs used in SACs. Even when choosing an appropriate route of administration, several aspects must be considered such as the fact that pour-on formulations are largely ineffective in camelids due to the unique features of llama and alpaca skin and hair that result in extremely low drug bioavailability. This review focuses on the main endoparasites of the digestive tract in llamas and alpacas in Europe and pharmacotherapeutic options based on current knowledge.

Investigations into cytotoxic effects of the herbal preparation Abnormal Savda Munziq.

OBJECTIVE: To evaluate the effects of Abnormal Savda Munziq (ASMq), a traditional herbal medicine, for the prevention and treatment of human diseases, e.g. bowel cancer. METHODS: The parameters total polyphenol content, cell proliferation and DNA-damage as well as RNA and protein-oxidation were analysed in vitro. Besides, the expressions of miRNA and tumor suppressor genes as well as cellular senescence were evaluated. RESULTS: ASMq had a high polyphenol content and induced damage to proteins, RNA as well as to DNA, which is correlated with its cytotoxicity. Furthermore ASMq up-regulated the tumor suppressor genes p21, p53 and p16 and down-regulated the micro-RNAs hsa-mir-17 and hsa-mir-106b. In addition cellular growth arrest and SA-ß-gal-staining were induced. CONCLUSION: ASMq has the ability to induce DNA damage and cellular senescence, which are double-edged mechanisms in fighting cancer, as they might also have harmful side effects.

Single and combined effects of deoxynivalenol mycotoxin and a microbial feed additive on lymphocyte DNA damage and oxidative stress in broiler chickens.

The immune and intestinal epithelial cells are particularly sensitive to the toxic effects of deoxynivalenol (DON). The aim of this experiment was to study the effects of DON and/or a microbial feed additive on the DNA damage of blood lymphocytes and on the level of thiobarbituric acid reactive substance (TBARS) as an indicator of lipid peroxidation and oxidative stress in broilers. A total of forty 1-d-old broiler chicks were randomly assigned to 1 of 4 dietary treatments (10 birds per group) for 5 wk. The dietary treatments were 1) basal diet; 2) basal diet contaminated with 10 mg DON/kg feed; 3) basal diet contaminated with 10 mg DON/kg feed and supplemented with 2.5 kg/ton of feed of Mycofix Select; 4) basal diet supplemented with Mycofix Select (2.5 kg/ton of feed). At the end of the feeding trial, blood were collected for measuring the level of lymphocyte DNA damage of blood and the TBARS level was measured in plasma, heart, kidney, duodenum and jejunum. The dietary exposure of DON caused a significant increase (P = 0.001) of DNA damage in blood lymphocytes (31.99 ± 0.89%) as indicated in the tail of comet assay. Interestingly addition of Mycofix Select to DON contaminated diet decreased (P = 0.001) the DNA damage (19.82 ± 1.75%) induced by DON. In order to clarify the involvement of lipid peroxidation in the DNA damage of DON, TBARS levels was measured. A significant increase (P = 0.001) in the level of TBARS (23 ± 2 nmol/mg) was observed in the jejunal tissue suggesting that the lipid peroxidation might be involved in the DNA damage. The results indicate that DON is cytotoxic and genotoxic to the chicken intestinal and immune cells and the feed additive have potential ability to prevent DNA damage induced by DON.

Efficacy and safety of oral praziquantel against Dicrocoelium dendriticum in llamas.

Dicrocoelium dendriticum can cause severe pathological changes of the liver and bile system in camelids, and therapeutic options for treatment are limited. To address this problem, the efficacy of two different dose rates of praziquantel was investigated in llamas suffering from natural D. dendriticum infections. 53 llamas were examined under field conditions on two occasions: before and two weeks after treatment. At the beginning of the study, the animals were weighed, randomly allocated to one of the treatment groups (n=21 each) or the control group (n=11) and dosed orally using a praziquantel-containing paste (250 mg/ml) at a dose of either 25 mg (group 1) or 50 mg (group 2) per kg of body weight. Criteria for efficacy were faecal egg count reduction (FECR) and extensity effect. Animals treated with 25 mg/kg of body weight showed a FECR of 85%. Therapy with 50 mg/kg led to a FECR of 91%. Almost twice the number of animals of group 1 (33%) still shed eggs two weeks after treatment compared with group 2. The results of this study indicate that 50 mg/kg oral praziquantel is required for efficacious dosing and that this dose rate is safe in llamas and thus is recommended for the treatment of camelids naturally infected with D. dendriticum.

Metabolic changes associated with anaesthesia and cherry poisoning in a pony.

OBSERVATIONS: A case of a pony with severe cyanide intoxication as a result of cherry ingestion is presented. General anaesthesia was performed for colic surgery. Severe metabolic lactate acidosis in combination with a high arterial oxygen partial pressure and clinically good peripheral perfusion parameters were the remarkable signs during anaesthesia. Severe hypothermia was obvious during recovery. Ten hours post-surgery the pony was euthanized as a result of neurological signs. The diagnosis of cyanide intoxication was made post-mortem. CONCLUSION: Cherry ingestion can lead to lethal cyanide intoxication in horses indicated by severe nonhypoxic lactic acidosis during anaesthesia.

Use of endoscopy for examination of the sacral epidural space in standing cattle.

OBJECTIVE: To develop an epiduroscopic technique for use in standing cattle and describe the endoscopically visible anatomic structures of the epidural space in the sacrococcygeal area. ANIMALS: 6 healthy nonlactating, nonpregnant cows (mean +/- SD age, 60 +/- 18.5 months; mean weight, 599.7 +/- 63.87 kg) and 3 bovine cadavers. PROCEDURES: Cadavers were used to allow familiarization with the equipment and refinement of the technique. Following these experiences, procedures were performed in live animals. Each cow was restrained in a stock. After sedation with xylazine (0.03 mg/kg, IV), 2% lidocaine hydrochloride (0.25 mg/kg) was injected epidurally in the first intercoccygeal or the sacrococcygeal intervertebral space. By use of an introducer set (guidewire and dilation trocar and shaft), a flexible endoscope (length, 75 cm; diameter, 2.3 mm) was inserted through the dilation shaft into the epidural space. To obtain an optimal view, small amounts of air were insufflated into the epidural space through the working channel of the endoscope via a syringe with special filter. RESULTS: Anatomic structures of the epidural space that were viewed by means of the endoscopic procedure included blood vessels, connective tissue, fat, nerves, and the spinal dura mater. No adverse events were detected during epiduroscopy, and it was tolerated well by all 6 cows. CONCLUSIONS AND CLINICAL RELEVANCE: In ruminants, epidural structures can be viewed via endoscopy. Such epiduroscopic procedures may be useful in anatomic studies as well as for the diagnosis of disease or therapeutic interventions in ruminants.

Laparoscopic-assisted implantation of a urinary catheter in male sheep.

OBJECTIVE-To evaluate a laparoscopic technique for implantation of a urinary catheter in the right paramedian area in male sheep and to determine feasibility, benefits, and risks for this technique. DESIGN-Evaluation study. ANIMALS-6 Healthy male sheep (mean +/- SD body weight, 42.16 +/- 11.95 kg [92.75 +/- 26.29 lb]). PROCEDURES-Each sheep was anesthetized and positioned in dorsal recumbency. A 10-mm laparoscope was inserted in the right paramedian area between the xiphoid and preputial orifice. After creation of capnoperitoneum, grasping forceps were inserted in the left paramedian area at the level of the teats and used to immobilize the urinary bladder. A pigtail balloon catheter was implanted transcutaneously in the right paramedian area between the preputial orifice and teats and directed into the urinary bladder by use of laparoscopic guidance. The catheter was removed 10 days after implantation. Fourteen days after initial surgery, a second laparoscopy was performed to evaluate pathologic changes. RESULTS-Inadvertent insertion of the first trocar into the rumen of 1 sheep was the only intraoperative complication encountered. Laparoscopic-assisted implantation of the urinary catheter was successfully performed in all sheep. No postoperative complications were detected. CONCLUSIONS AND CLINICAL RELEVANCE-Laparoscopic-assisted implantation of a urinary catheter in the right paramedian area was successfully performed and may be a feasible method for use in sheep. This method can be considered as an alternative to tube cystotomy performed by laparotomy.

Low dose magnetic fields do not cause oxidative DNA damage in human placental cotyledons in vitro.

The biological impact of low dose magnetic fields generated by electric appliances present in the human environment is still uncertain. In this study, human placentas served as a model tissue for the evaluation of the potential effect of oscillating low intensity magnetic fields on the concentration of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in cellular DNA. Cotyledons were dissected from placentas obtained immediately after physiological labours and exposed to magnetic fields (groups MF A, 2 mT, 50 Hz and MF B, 5 mT, 50 Hz) or sham exposed (group C) during an in vitro perfusion of 3 h. Cellular DNA was isolated, hydrolyzed and analyzed by HPLC. Native nucleosides were monitored at 254 nm and 8-OH-dG by electrochemical detection. Results were expressed as mumol 8-OH-dG/mol deoxyguanosine (dG). The concentrations of 8-OH-dG in group C, MF A and MF B were 28.45+/-15.27 micromol/mol dG, 62.80+/-31.91 mumol/mol dG, and 27.49+/-14.23 micromol/mol dG, respectively, demonstrating no significant difference between the groups. The results suggest that placental tissues possess a capacity to protect DNA against oxidative alterations by magnetic field of intensities previously shown to produce radical mediated DNA damage in rat brain cells in vivo and imbalances in electrolyte release of cotyledons under in vitro conditions.

Cardiac myocyte-specific HIF-1alpha deletion alters vascularization, energy availability, calcium flux, and contractility in the normoxic heart.

At a resting pulse rate the heart consumes almost twice-as much oxygen per gram tissue as the brain and more than 43 times more than resting skeletal muscle (1). Unlike skeletal muscle, cardiac muscle cannot sustain anaerobic metabolism. Balancing oxygen demand with availability is crucial to cardiac function and survival, and regulated gene expression is a critical element of maintaining this balance. We investigated the role of the hypoxia-inducible transcription factor HIF-1alpha in maintaining this balance under normoxic conditions. Cardiac myocyte-specific HIF-1alpha gene deletion in the hearts of genetically engineered mice caused reductions in contractility, vascularization, high-energy phosphate content, and lactate production. This was accompanied by altered calcium flux and altered expression of genes involved in calcium handling, angiogenesis, and glucose metabolism. These findings support a central role for HIF-1alpha in coordinating energy availability and utilization in the heart and have implications for disease states in which cardiac oxygen delivery is impaired. Heart muscle requires a constant supply of oxygen. When oxygen supply does not match myocardial demand cardiac contractile dysfunction occurs, and prolongation of this mismatch leads to apoptosis and necrosis. Coordination of oxygen supply and myocardial demand involves immediate adaptations, such as coronary vasodilatation, and longer-term adaptations that include altered patterns of gene expression (2-4). How the expression of multiple genes is coordinated with oxygen availability in the heart and the impact of oxygen-dependent gene expression on cardiac function are insufficiently understood. Further elucidating these relationships may help clarify the molecular pathology of various cardiovascular disease states, including ischemic cardiomyopathy and myocardial hibernation (5, 6).

Oxidative DNA damage in placentas from normal and pre-eclamptic pregnancies.

Placental oxidative stress was suggested to play a role in the pathogenesis of pre-eclampsia (PE). In this study, levels of 8-hydroxy-2'-deoxyguanosine (8-OH-dG), a well-established marker of oxidative DNA damage, were analysed in placental cellular DNA from normal (group NP) and pre-eclamptic (group PE) pregnancies as well as from PE pregnancies complicated by intrauterine growth restriction (group PE-IUGR). Placental samples obtained immediately after delivery were frozen at -80 degrees C until analysis. Cellular DNA was isolated, hydrolysed and analysed using high-performance liquid chromatography. Native nucleosides were monitored at 254 nm and 8-OH-dG using electrochemical detection. Concentrations of 8-OH-dG were expressed as micro mol/mol 2'-deoxyguanosine. In group NP, mean concentration of 8-OH-dG reached 179.97+/-80.58 (+/-SEM; micro mol/mol dG). 8-OH-dG levels were higher in group PE (273.44+/-110.14 micro mol/mol), but the difference was not significant in comparison with group NP. Highest concentrations of 8-OH-dG were found in group PE-IUGR (428.97+/-141.40 micro mol/mol), with levels significantly higher than in group NP, but not group PE. The results indicate a positive correlation between the severity of PE and the degree of oxidative stress and corroborate previous studies suggesting reactive oxygen species to be involved in the pathophysiology of PE.

Hypoxia-induced gene expression occurs solely through the action of hypoxia-inducible factor 1alpha (HIF-1alpha): role of cytoplasmic trapping of HIF-2alpha.

The hypoxia-inducible factors 1alpha (HIF-1alpha) and 2alpha (HIF-2alpha) have extensive structural homology and have been identified as key transcription factors responsible for gene expression in response to hypoxia. They play critical roles not only in normal development, but also in tumor progression. Here we report on the differential regulation of protein expression and transcriptional activity of HIF-1alpha and -2alpha by hypoxia in immortalized mouse embryo fibroblasts (MEFs). We show that oxygen-dependent protein degradation is restricted to HIF-1alpha, as HIF-2alpha protein is detected in MEFs regardless of oxygenation and is localized primarily to the cytoplasm. Endogenous HIF-2alpha remained transcriptionally inactive under hypoxic conditions; however, ectopically overexpressed HIF-2alpha translocated into the nucleus and could stimulate expression of hypoxia-inducible genes. We show that the factor inhibiting HIF-1 can selectively inhibit the transcriptional activity of HIF-1alpha but has no effect on HIF-2alpha-mediated transcription in MEFs. We propose that HIF-2alpha is not a redundant transcription factor of HIF-1alpha for hypoxia-induced gene expression and show evidence that there is a cell type-specific modulator(s) that enables selective activation of HIF-1alpha but not HIF-2alpha in response to low-oxygen stress.