Vitamin E Supplementation Reduces Cellular Loss in the Brain of a Premature Aging Mouse Model.

BACKGROUND: Aging is a highly complex biological process driven by multiple factors. Its progression can partially be influenced by nutritional interventions. Vitamin E is a lipid-soluble anti-oxidant that is investigated as nutritional supplement for its ability to prevent or delay the onset of specific aging pathologies, including neurodegenerative disorders. PURPOSE: We aimed here to investigate the effect of vitamin E during aging progression in a well characterized mouse model for premature aging. METHOD: Xpg-/- animals received diets with low (~2.5 mg/kg feed), medium (75 mg/kg feed) or high (375 mg/kg feed) vitamin E concentration and their phenotype was monitored during aging progression. Vitamin E content was analyzed in the feed, for stability reasons, and in mouse plasma, brain, and liver, for effectiveness of the treatment. Subsequent age-related changes were monitored for improvement by increased vitamin E or worsening by depletion in both liver and nervous system, organs sensitive to oxidative stress. RESULTS: Mice supplemented with high levels of vitamin E showed a delayed onset of age-related body weight decline and appearance of tremors when compared to mice with a low dietary vitamin E intake. DNA damage resulting in liver abnormalities such as changes in polyploidy, was considerably prevented by elevated amounts of vitamin E. Additionally, immunohistochemical analyses revealed that high intake of vitamin E, when compared with low and medium levels of vitamin E in the diet, reduces the number of p53-positive cells throughout the brain, indicative of a lower number of cells dying due to DNA damage accumulated over time. CONCLUSIONS: Our data underline a neuroprotective role of vitamin E in the premature aging animal model used in this study, likely via a reduction of oxidative stress, and implies the importance of improved nutrition to sustain health.

Use of neutral protamine lispro insulin (NPL) in patients with hyperglycemia receiving parenteral nutrition.

AIMS: Nutritional support with parenteral nutrition (PN), a key component in the care of critically ill patients, usually requires insulin therapy in patients with diabetes or may require insulin treatment in patients not known to be diabetic. We wanted verify whether it is possible to use neutral protamine lispro (NPL) in double administration monotherapy in patients receiving artificial nutrition (AN) and if the same NPL is capable of obtaining and maintaining acceptable glycemic control without inducing hypoglycemia. PATIENTS AND METHODS: We studied 18 consecutive patients, who were not taking insulin, they needed to start artificial nutrition, and presenting at least two consecutive blood glucose > 120 mg/dL. Each patient was given at least 1 U of insulin for every 10 grams of glucose infused. RESULTS: Eighteen consecutive patients, not stratified in any way, were judged eligible in the last 24 months, with a mean age of 71 years (range 54-85 yrs). All patients were evaluated after 2, 3 and 5 days of treatment; only 1 patient has not been evaluated to 5 days. Mean glycemic values on days 2, 3, 5 were in range between 145 and 180 mg/dL. Any adjustments in NPL dose were carried out by the team of nutrition and there was no hypoglycemia that required medical intervention in emergency. CONCLUSIONS: Our impression is that also lispro protamine insulin (NPL) in double subcutaneous administration may contribute to improving the glycemic values in patients receiving parenteral nutrition with hyperglycemia.

How to approach to the therapy of diabetes in the elderly.

In gerontology diabetes mellitus (DM) is clinically the most frequent and extremely serious metabolic disorder. DM is an important health issue across the globe. With increasing life expectancy physicians are called upon to manage diabetes in the elderly more often. Senior patients suffer predominantly from the type 2 DM--T2DM (70+ up to 90-95%). Apart from genetic predisposition and an environmental influence, nutritional habits, modern lifestyle, stress and minor physical activity are of particular importance. Treatment options for T2DM in the elderly are diet, physical activity, various oral anti-diabetic drugs and insulin. At the start of treatment should primarily take into account: patient's age; self-sufficiency; late micro- and macro-vascular complications; social status; nutritional assessment (incl. dental status); other handicaps--psychic, motoric, visual and aural. Especially in frail, elderly patients, there should be less emphasis on strict glycaemic control than on avoiding malnutrition and hypoglycemia and achieving the best possible quality of life. Therapy of DM in this population is tightly connected with significant risks of micro- and macrovascular complications on one hand, and possible problems of the treatment (e.g. hypoglycemia) with intensive control on the other hand. To realize a comprehensive approach to therapy of diabetes in the old age a holistic approach with main aim improving quality of life is necessary.

Vitamin D: do we get enough? A discussion between vitamin D experts in order to make a step towards the harmonisation of dietary reference intakes for vitamin D across Europe.

UNLABELLED: On September 29, 2011, acknowledged experts in the field of vitamin D, mainly European, were brought together in order to discuss the recent scientific advances in relation to vitamin D: the current requirements and associations with various health outcomes. In this article, the discussions resulting from the meeting are summarized. INTRODUCTION: Several groups at risk for developing vitamin D insufficiency have been identified. Accordingly, reviews indicate that a significant percentage of the population worldwide have serum 25-hydroxyvitamin D levels below 50 nmol/l. In addition to the role of vitamin D in bone health, recent studies suggest that it may play a pivotal role in other systems, e.g., the cardiovascular system, pancreas, muscle, immune system and brain. Most evidence, however, is obtained from observational studies and yet inconclusive. METHODS: To exchange and broaden knowledge on the requirements for vitamin D and its effect on various health outcomes, a workshop entitled "Vitamin D Expert Meeting: Do we get enough?", was organized. RESULTS: Despite low vitamin D levels worldwide, consensus on the definition of deficiency is not yet reached. In order to define cut-off points for vitamin D whilst taking into account extraskeletal health effects, randomized controlled trials in these fields are warranted. The experts do emphasize that there is evidence to suggest an important role for vitamin D in the maintenance of optimal bone health at all ages and that vitamin D supplementation, in most studies co-administered with calcium, reduces fracture risk in the senior population. CONCLUSION: To reach a serum 25-hydroxyvitamin D level of 50 nmol/l older adults aged ≥65 years are therefore recommended to meet a mean daily vitamin D intake of 20 µg (800 IU), which is best achieved with a supplement.

[Drug consumption and risks of polypharmacotherapy in elderly population]. / Spotreba léciv v seniorské populaci a rizika polyfarmakoterapie ve stárí.

BACKGROUND: One of our previous studies was aimed at the consumption of prescribed drugs by the elderly population. The average per day number of drugs was 4.6 (maximum 13). Existence of freely obtainable drugs with massive advertisements brings a question, how many of those drugs it is necessary to add in order to estimate probability of interaction and undesirable drug effects. In order to achieve valid information, students of the sixth year of General medicine program during their practical course at general practitioners were asked to interview randomly selected senior patients. They asked on the number, type, and price of freely obtainable drugs which they use. Data were evaluated from interviews accomplished during academic years 2001/2002 and 2004/2005. METHODS AND RESULTS: Our cohort included 252 men and 148 women with average age of 78.7 years. Average number of freely obtainable drugs was 2.26 at the beginning and 2.32 at the end of study. Only 34% of questioned did not buy any of those drugs at all or only exceptionally, 66% reported buying once a month or weekly. 44% of seniors buy analgetics, 58% buy vitamins, 37% food supplements, 36% non steroid antirheumatics, 46% cold prevention drugs, 30% anti-constipation drugs. Contrary to our expectation, positive correlation between the sums given for the personal participation on the drug costs and that given for freely obtainable drugs was found. It is not possible to expect, that polymorbidic patient with several prescribed drugs would buy less of freely obtainable drugs even due to the financial requirements. CONCLUSIONS: Freely obtainable drugs, many of them composites, can represent significant source of interactions and undesirable drug effects. They can also significantly modulate compliance of the senior. The high percentage of seniors buying freely obtainable drugs requires aimed questions on the pharmacological history.

Insulin glargine in patients with severe hepato-gastroenterology diseases and hyperglycemia receiving parenteral nutrition.

OBJECTIVE: The aim of this nonrandomized observational study is to verify and confirm whether it is possible to use insulin glargine (Lantus) subcutaneously in patients receiving parenteral nutrition (PN) and whether the analogue is capable of obtaining and maintaining good glycemic control without inducing hypoglycemia. MATERIALS AND METHODS: The sample is made up of 25 patients with severe hepato-gastroenterology diseases receiving parenteral nutrition, diagnosed diabetics and non-diabetics, who had previously been prescribed traditional insulin therapy. All were to be given subcutaneous insulin glargine at a dosage equal to the average of insulin/day administered in the preceding days spent receiving PN. RESULTS: Twenty-five consecutive patients, not stratified in any way, were judged eligible in the last six months of 2004 and first eight months of 2005. Four out of these 25 cannot be evaluated, either because (2/4) they did not begin or complete the treatment with Lantus or because the proper number of glycemic tests were not done (2/4); 21/25 patients, 84% of the sample with a mean age of 65.9 years (range 46-93 yr), finished the study and can be evaluated. The mean glycemic values after treatment with glargine were already better on the second day, and on the seventh day the difference was statistically significant. No hypoglycemias occurred which required medical intervention. CONCLUSIONS: This study confirms the possibility of using insulin glargine in patients receiving parenteral nutrition with hyperglycemia diagnosed diabetics or not diabetics.

Epigallocatechin gallate (EGCG) (TEAVIGO) does not impair nonhaem-iron absorption in man.

A number of studies have shown that tea catechins can inhibit intestinal iron absorption, mostly iron in the nonhaem form. This randomized, double-blind, placebo-controlled, 3-periods cross-over study examined the degree of inhibition of nonhaem iron absorption by pure crystalline epigallocatechin gallate (EGCG). The study was designed to show the maximum inhibitory action of EGCG by selecting 30 healthy women with low iron stores. Treatments were 150 mg, 300 mg EGCG and placebo each for 8 consecutive study days with a wash-out period of 14 days between treatments. Iron incorporation was assessed by supplying 57Fe orally and 58Fe intravenously. Differences in fractional nonhaem iron absorption between the treatments were evaluated by using two-sided ANOVA. Results showed a relative nonhaem iron absorption reduction of 14% with 150mg EGCG and 27% for 300mg EGCG treatment compared to placebo. Differences were statistically significant (p < or = 0.05) between the placebo and the 300mg EGCG treatments and between the 150 and 300 mg EGCG treatments. The inverse relation between EGCG dose and fractional nonhaem iron absorption was linear (p = 0.0002). In this study the magnitude of the inhibitory action of EGCG on nonhaem iron absorption was found to be much lower than that reported in the literature for black tea and similar compounds. The doses of EGCG in supplements, which will be lower than those used in this study, are not expected to have any health relevant effects on iron absorption in subjects with normal iron stores.

[Evaluation of levofloxacin susceptibility against strains isolated from lower urinary tract infections in the community]. / Evaluation de la sensibilité à la lévofloxacine des souches isolées d'infections urinaires basses communautaires.

The aim of this study was to evaluate the in vitro activity of levofloxacin (LVX) in comparison to nalidixic acid (NAL), ofloxacin (OFX), norfloxacin (NOR), amoxicillin (AMX), cefixime (CFM), cotrimoxazole (SXT) and nitrofurantoin (FT), against 402 strains recently isolated from urine specimens in outpatient women suffering from lower urinary tract infections for which short-term treatment was not indicated. MICs were determined by the agar dilution method on Mueller-Hinton medium (Bio-Rad) according to the recommendations of the Comite de l'Antibiogramme de la Societe Francaise de Microbiologie (CA-SFM). Strains were classified as susceptible (S), intermediate (I) or resistant (R) according to the CA-SFM recommended breakpoints. Quality control was carried out using three reference strains: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853. For E. coli, the most prevalent species (345 isolates: 85.3%), susceptibilities were as follows: AMX: 60.6%, CFM: 99.1%, NAL: 94.8%, NOR: 97.4%, OFX: 97.4%, LVX: 97.4%, SXT: 84.5%, FT: 98%. This study confirms the good in vitro activity of LVX, OFX, and CFM against strains isolated from urinary tract infections in the community and particularly against E. coli, which is by far the most prevalent pathogen, 90% of strains, with more than 97% of strains being susceptible.

Plasma-kinetic characteristics of purified and isolated green tea catechin epigallocatechin gallate (EGCG) after 10 days repeated dosing in healthy volunteers.

This randomized, double-blind, placebo-controlled study assessed the safety, tolerability, and plasma-kinetic behavior of 94% pure crystalline epigallocatechin gallate (EGCG) after ten days' repeated dosing in 36 healthy male volunteers. Each of the three treatment groups consisted of 12 subjects; nine of them received oral EGCG in one dose of 200, 400, or 800 mg daily, and three received a placebo. Blood samples for plasma-kinetic EGCG characterization were taken on day 1 and day 10. Kinetic parameters for rate and extent, elimination half-lives, and accumulation factor (R) were determined and compared between day 1 and day 10 for each dosage group. Orally administered EGCG is rapidly absorbed from the gut. Dose linearity was applied for single-dose application (day 1). After repeated dosing (day 10) dose linearity was applied between the 200 mg and 400 mg group. Dose escalation to 800 mg was more than dose-proportional in rate and extent, and statistically different from the 200 mg and 400 mg group. An increase in elimination half-life (t1/2.z) and in the accumulation factor (R) in the 800 mg dosage group indicates dose-dependent saturation of capacity-limited excretion routes or an increase of hepato-duodenal re-circulation. Ten days' repeated administration of oral doses of EGCG of up to 800 mg per day were found to be safe and very well tolerated.

Polyunsaturated fatty acids and cardiovascular risk: interference at the level of gene expression.

Long-chain polyunsaturated fatty acids (PUFA) are important structural components of the cell membrane as well as precursors of highly active lipid mediators, the eicosanoids. In addition, they have been identified as novel intracellular signaling molecules which either directly or indirectly through the modulation of other signaling pathways activate transcription factors and gene expression. With regard to cardiovascular risk and diseases, effects of PUFA have been observed on genes whose products regulate either metabolic processes, especially fatty acid metabolism, or are involved in inflammatory and mitogenic processes of vascular cells. Often, the effects of n-3 and n-6 PUFA on the expression of both types of genes are antagonistic indicating a balancing role of n-3 and n-6 PUFA in cell metabolism and function already at the level of gene expression.

Effect of either gamma-tocotrienol or a tocotrienol mixture on the plasma lipid profile in hamsters.

BACKGROUND/AIMS: Tocotrienols has been shown to inhibit the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity; however, the published animal and human studies yield conflicting results. We investigated the effects of a 4-week dietary supplement of either gamma-tocotrienol (86% gamma-T3) or a mixture of tocotrienols (29.5% alpha-T3, 3.3% beta-T3, 41.4% gamma-T3, 0.1% delta-T3: mix-T3) on the plasma lipid profile in hamsters receiving a high fat diet. METHODS: The hamsters were randomized into 7 groups: no treatment, 16 mg/day/kg BW simvastatin, 23, 58, 263 mg/day/kg BW gamma-tocotrienol, and 39 or 263 mg/day/kg BW for the mixture of tocotrienols. Plasma lipid levels were measured after 2 and 4 weeks of treatment. RESULTS: In all groups treated with tocotrienol total cholesterol levels were decreased, ranging from 7 to 23% after 2 weeks of treatment and from 7 to 15% after 4 weeks. Low-density lipoprotein plasma levels changed accordingly: a decline of 6-37% after 2 weeks and of 12-32% at the end of the study was observed. After 4 weeks of treatment, total cholesterol and low-density lipoprotein plasma levels were significantly reduced in the 263 mg/day/kg BW mixed tocotrienols and the 58 mg/day/kg BW and 263 mg/day/kg BW gamma-tocotrienol groups when compared to the no treatment group. Plasma triglycerides and high-density lipoprotein levels did not change significantly. CONCLUSION: This study provides further evidence that tocotrienols lower total cholesterol and low density lipoprotein plasma levels in hamsters and that gamma-tocotrienol is a more potent agent than a mixture of tocotrienols.

Antibody repertoire development in fetal and neonatal piglets. IV. Switch recombination, primarily in fetal thymus, occurs independent of environmental antigen and is only weakly associated with repertoire diversification.

The epitheliochorial placenta of swine is considered a barrier to Ag and selective transport of IgG, so this species should be an excellent model with which to determine whether switch recombination is Ag dependent. Analysis of Ig levels and Ig isotype profiles in >150 normal and virus-infected fetuses from 38-110 days of gestation (DG) suggested that IgG, IgA, and IgM were most likely the result of de novo fetal synthesis. Although transcripts for IgM could be recovered at DG 50 (114 DG is full gestation) in all major fetal lymphoid tissues, those for IgG and IgA first became prominent at 60 DG in thymus, and transcription and spontaneous secretion became especially pronounced in this organ in older fetuses. Data on transcription, secretion, and serum isotype profiles suggest that although all fetal IgA and IgM may result from de novo synthesis, some IgG may result from low-level selective transport. The complementarity-determining region 3 spectratypes of thymic IgA and IgG transcripts at 70 and 90 days, respectively, were as polyclonal as that of IgM, indicating a broad repertoire of switched B cells although the VDJs transcribed with these switched isotypes in normal fetuses were not diversified in comparison to those from animals exposed to environmental Ags such as age-matched, virus-infected fetuses, colonized isolator piglets, and conventional adults. However, VDJs expressed with switched isotypes were more diversified than those expressed with IgM. Thus, switch recombination in fetal life does not appear to be driven by environmental Ag and is only weakly coupled to VDJ diversification. These findings, and the fact that the oligoclonal IgA and IgM repertoires in a noninductive site of the mucosal immune system (parotid gland) become polyclonal in piglets reared germfree, suggest that initial expansion of the switched cells in the B cell compartment of fetal and neonatal piglets is not driven by environmental Ag.

A colorimetric assay for high-throughput screening of inhibitors of herpes simplex virus type 1 alkaline nuclease.

Herpes simplex virus type 1 (HSV-1) encodes a deoxyribonuclease that is frequently referred to as alkaline nuclease (AN) because of its elevated pH optimum. Studies with recombinant viruses which contain deletions in the HSV-1 gene encoding AN have indicated that this enzyme is required for efficient virus replication and therefore represents a potential target for novel antiviral therapies. A simple colorimetric assay for deoxyribonuclease activity employing a DNA-methyl green substrate was adapted for use in a high-throughput screen to identify small molecule inhibitors of this enzyme. This screen identified 1,2-benzoisothiazolin-3-one as a specific inhibitor of AN, since it exhibited activity against AN but was completely inactive against bovine pancreatic DNaseI. Subsequent studies revealed that this compound most likely inhibited AN by forming disulfide linkages with one or more exposed cysteine residues on the surface of the enzyme and that AN was sensitive to sulfhydryl-group-modifying reagents in general. These results demonstrated the utility of this DNA-methyl green substrate-based assay in both the rapid identification and the characterization of novel small molecule inhibitors of the AN encoded by HSV-1 and other herpesviruses.

Triglyceride-lowering effect of omega-3 LC-polyunsaturated fatty acids--a review.

There is increasing evidence that serum triglycerides are a significant and independent risk factor for CVD. The aim of this report is to review recent literature pertinent to the triglyceride-lowering effect of omega-3 long chain polyunsaturated fatty acids (LC-PUFA). Animal data are not considered because they are difficult to extrapolate to the human situation. A large body of evidence derived from epidemiological studies and clinical trials has consistently demonstrated that this effect is dose-dependent and can be achieved by diet. The smallest amount of omega-3 LC-PUFA needed to significantly lower serum triglycerides appears to be approximately 1 g/day as provided by a fish diet. Use of fish oil administering as little as 0.21 g EPA and 0.12 g DHA per day significantly lowered serum triglycerides in hyperlipidemics. In normolipidemics, a daily intake of 0.17 g EPA and 0.11 g DHA, given as a fish oil supplement, induced a non-significant reduction of 22%. These findings must be considered as preliminary and warrant further research. Intake of omega-3 LC-PUFA is frequently reported to modestly increase LDL cholesterol. However, in normo- or slightly hyperlipidemic individuals who received omega-3 LC-PUFA for 4 months or longer, changes of LDL cholesterol were not significantly different from a placebo group. Both EPA and DHA lower serum triglycerides, but they may have a differential effect on lipoproteins. Intake of omega-3 LC-PUFA in the amount mentioned above is safe.

Lipids in vascular function.

Physiological and pathological vascular responses depend on the action of numerous intercellular mediators, ranging from hormones to gases like nitric oxide, proteins, and lipids. The last group consists not only of the different types of lipoproteins, but also includes a broad array of other lipophilic signaling molecules such as fatty acids, eicosanoids, phospholipids and their derivatives, sphingolipids and isoprenoids. Due to space limitations, it is impossible to discuss all the vascular effects of lipophilic mediators or compounds. Therefore, we will focus on one of the most important lipid-mediated diseases, atherosclerosis. Lipoproteins and especially their native or oxidized lipid compounds affect vascular function in many different ways, and these effects do not only modulate atherogenesis but are of paramount physiological and pathophysiological importance in other diseases, such as inflammation, tumor metastasis, or normal wound healing.

TP5 triggers signal transduction involving mitogen activated protein kinases in monocytes.

The pentapetide thymopentin (TP5) corresponding to the aminoacids RKDVY represents the residues 32-36 of thymopoietin (TP), which was originally isolated from bovine thymus. Both were observed to induce T-cell differentiation and maturation. Recently however it was shown, that TP represents the N-terminal 49 aa of the human thymopoietin (TMPO) isoforms TMPO alpha, beta and gamma, which are localized in the nucleus. TP5 was investigated in a variety of diseases and showed efficacy by improving the immune balance, whereby different cells increased in cell number or activity. Findings which support the assumption of multifunctional efficacy and a description of TP and TP5 modulating T cells lack any interpretation on molecular level. In the present study we investigated the binding of TP5 on white blood cells. We identified monocytes and neutrophils as TP5-binding cells by displacing fluorescein-labelled TP5 with an excess of unlabelled TP5 in competition assays. Binding of TP5 on cell surface proteins resulted in cellular signalling and we report here that TP5 triggers signal transduction involving mitogen activated protein kinases p42/p44 (MAPKs) in monocytes.

Alumina gel injections into the temporal lobe of rhesus monkeys cause complex partial seizures and morphological changes found in human temporal lobe epilepsy.

The goal of the present study was to determine whether alumina gel injections into temporal lobe structures cause complex partial seizures (CPS) and pathological changes observed in human temporal lobe epilepsy. Rhesus monkeys with alumina gel injections in the amygdala, perirhinal and entorhinal cortices, or Ammon's horn and dentate gyrus all initially displayed focal pathological electroencephalographic (EEG) slowing limited to the site of injection. After clinical seizures developed, they also displayed widespread pathological EEG slowing over both hemispheres, interictal and ictal epileptiform EEG abnormalities limited to the mesial-inferior temporal lobe on the side of injection, and different degrees of spread to other ipsilateral and contralateral structures. Noninjected control and nonepileptic monkeys with injections into the middle and inferior temporal gyri displayed no hippocampal neuronal loss or mossy fiber sprouting. When alumina gel was injected into the amygdala, CPS began within 3-6 weeks and degeneration of neurons and gliosis occurred in the perirhinal cortex or the hippocampus, with consequent sprouting of mossy fibers in the dentate gyrus. Dispersion of the granule cell layer was also observed. Other monkeys with alumina gel in the perirhinal and entorhinal cortices developed CPS within 2-3 weeks after the injections and displayed mossy fiber sprouting only after 4 weeks after the injections. Alumina gel in Ammon's horn and the dentate gyrus also induced CPS, but mossy fiber sprouting was limited to sites immediately adjacent to the injection, probably because none survived more than 4 weeks after the injections. This nonhuman primate model of CPS displayed similar anatomical, behavioral, and EEG features as observed in human temporal lobe epilepsy and provides opportunities to analyze the chronological sequence of epileptogenesis and to test potential therapies.