Effects of Dietary Nasunin on the Serum Cholesterol Level in Rats

The effects of nasunin, a major anthocyanin in eggplant, and its aglycone, delphinidin, on the serum cholesterol conpentration were determined in rats fed with a cholesterol-enriched diet. The serum total cholesterol and HDL-cholesterol concentrations tended to be decreased and increased, respectively, by feeding nasunin and delphinidin, while the fecal excretion of both cholesterol and bile acids tended to be increased by feeding with the anthocyanins. There was no difference between the nasunin and delphinidin activity to decrease serum total cholesterol, nor to increase serum HDL-cholesterol or the fecal excretion of cholesterol and bile acids. These results suggest that the slightly lower serum total cholesterol concentration in rats fed with nasunin and delphinidin may in part be due to inhibition of the intestinal absorption of both cholesterol and bile acids by these anthocyanins, and that the delphinidin moiety of nasunin mainly contributes to this activity.     

Erythropoietin Amplifies Stroke-Induced Oligodendrogenesis in the Rat

Background

Erythropoietin (EPO), a hematopoietic cytokine, enhances neurogenesis and angiogenesis during stroke recovery. In the present study, we examined the effect of EPO on oligodendrogenesis in a rat model of embolic focal cerebral ischemia.

Methodology and Principal Findings

Recombinant human EPO (rhEPO) at a dose of 5,000 U/kg (n = 18) or saline (n = 18) was intraperitoneally administered daily for 7 days starting 24 h after stroke onset. Treatment with rhEPO augmented actively proliferating oligodendrocyte progenitor cells (OPCs) measured by NG2 immunoreactive cells within the peri-infarct white matter and the subventricular zone (SVZ), but did not protect against loss of myelinating oligodendrocytes measured by cyclic nucleotide phosphodiesterase (CNPase) positive cells 7 days after stroke. However, 28 and 42 days after stroke, treatment with rhEPO significantly increased myelinating oligodendrocytes and myelinated axons within the peri-infarct white matter. Using lentivirus to label subventricular zone (SVZ) neural progenitor cells, we found that in addition to the OPCs generated in the peri-infarct white matter, SVZ neural progenitor cells contributed to rhEPO-increased OPCs in the peri-infarct area. Using bromodeoxyuridine (BrdU) for birth-dating cells, we demonstrated that myelinating oligodendrocytes observed 28 days after stroke were derived from OPCs. Furthermore, rhEPO significantly improved neurological outcome 6 weeks after stroke. In vitro, rhEPO increased differentiation of adult SVZ neural progenitor cells into oligodendrocytes and enhanced immature oligodendrocyte cell proliferation.

Conclusions

Our in vivo and in vitro data indicate that EPO amplifies stroke-induced oligodendrogenesis that could facilitate axonal re-myelination and lead to functional recovery after stroke.     

Changes of Uric Acid Level in Rat Brain After Focal Ischemia

Changes of uric acid level in rat cerebral hemisphere after left middle cerebral artery (MCA) occlusion were studied by reversed-phase HPLC with electrochemical detection. Uric acid level in the normal group was 2.98 nmol/g tissue. Uric acid concentration of the left hemisphere in the left MCA-occluded group progressively increased after occlusion, and reached a maximum value of 67.26 nmol/g tissue 24 h after ischemia. Uric acid levels in the right hemisphere remained unchanged. Uric acid concentration of the left hemisphere in sham-operated group was 9.29 nmol/g tissue 24 h after the operation.     

城市人群血清营养水平指标与膳食习惯关系的分析

目的:分析血清营养状态指标与膳食习惯的关联关系与意义。方法:采用横断面调查的数据,用标准问卷询问1694人相关信息,内容包括人口学资料、自报疾病现患史、膳食习惯、健康自评、认知功能信息;同时采集被调查人群血样本;测定血红蛋白、白蛋白、叶酸和维生素B12水平。采用R 3.2.5和SPSS 20.0软件进行相关和显著性差异分析。结果:人体血红蛋白、白蛋白、叶酸和维生素B12水平与被调查个体进食水果和鸡蛋习惯有密切关联;天天进食水果、鸡蛋的个体上述指标水平明显高于不进食此类食物的个体,且血红蛋白、白蛋白水平随增龄下降程度减低;天天进食水果习惯的个体同时具有天天进食蛋、奶、菜习惯;有进食水果习惯的人群表现为受教育程度高;自评健康状态好和相对自报患病率低等特点。结论:合理膳食习惯可保持较高的血营养状态指标水平;且延缓相关指标水平随增龄下降的程度。天天进食水果习惯可被视为合理膳食的指标,为膳食评估提供简易、可行、有效的指标。     

High Resolution Mapping of Modafinil Induced Changes in Glutamate Level in Rat Brain

Modafinil is marketed in the United States for the treatment of narcolepsy and daytime somnolence due to shift-work or sleep apnea. Investigations of this drug in the treatment of cocaine and nicotine dependence in addition to disorders of executive function are also underway. Modafinil has been known to increase glutamate levels in rat brain models. Proton magnetic resonance spectroscopy (¹HMRS) has been commonly used to detect the glutamate (Glu) changes in vivo. In this study, we used a recently described glutamate chemical exchange saturation transfer (GluCEST) imaging technique to measure Modafinil induced regional Glu changes in rat brain and compared the results with Glu concentration measured by single voxel ¹HMRS. No increases in either GluCEST maps or ¹HMRS were observed after Modafinil injection over a period of 5 hours. However, a significant increase in GluCEST (19±4.4%) was observed 24 hours post Modafinil administration, which is consistent with results from previous biochemical studies. This change was not consistently seen with ¹HMRS. GluCEST mapping allows regional cerebral Glu changes to be measured and may provide a useful clinical biomarker of Modafinil effects for the management of patients with sleep disorders and addiction.     

Changes in Serum Zinc Levels Associated with Giardiasis and Dietary Zinc Intake in Mice

The association of giardiasis with the malabsorption of zinc remains controversial. This study investigated changes in serum zinc levels in Giardia-infected mice subjected to different dietary zinc regimens. Thirty-five mice (strain C₃H/H_eJ) were randomly categorized into two groups. The first group was inoculated with 5 × 10⁶ Giardia trophozoites (n = 18), and the second group remained Giardia free (n = 17). Each group (Giardia infected and Giardia free) was randomly classified into three subgroups and given low (9 mg Zn/kg), normal (33 mg Zn/kg), and high levels (288 mg Zn/kg) of dietary zinc over a 2-week period for acclimation. Fourteen days post-Giardia infection, all of the mice were euthanized and blood samples were collected. The number of trophozoites was quantified (hematocytometer), and serum zinc levels were determined via atomic absorption spectrophotometry. Significant increases in the median weights were only found in the Giardia-free mice (p < 0.05). A higher final median weight was found in the Giardia-free group when compared with that of the Giardia-infected group given low dietary zinc (p = 0.013). In the Giardia-infected group with low dietary zinc, the geometric mean of trophozoites was 3,498 ± 101 (SE) per milliliter. The Giardia-infected group had lower serum zinc levels than did the Giardia-free group with the high dietary zinc regimens (p < 0.05). Our results are consistent with studies among human populations, but further studies are required to elucidate the actual mechanism governing the zinc–giardiasis interaction.     

Regional Changes in the Cellular Level of Adenine Nucleotides in Ischemic Rat Brain Subjected to Single Embolization

Regional changes in adenine nucleotides in the rat brain were studied after 1 h of ischemia produced by the embolization method. The animals were divided into three groups according to neurological symptoms: sham-operation group, group A (hemiparesis only), and group B (hemiparesis with unconsciousness). Marked ATP depletion was detected in the hippocampus on the embolized side and extended to the other regions on the same side in group B. The results suggest that this damage in regional energy metabolism arises from regional reduction in blood flow and/or tissue vulnerability. ATP levels in the hypothalamus, hippocampus, and striatum on the opposite side of embolization decreased markedly in group B, and may be caused by extension of brain edema or diaschisis.     

Changes in the Serum Urate Level Can Predict the Development of Parkinsonism in the 6-Hydroxydopamine Animal Model

Epidemiological studies indicate that a higher plasma level of uric acid (UA) associates with the reduced risk of Parkinson’s disease (PD). To confirm the role of UA as a biomarker for PD, we evaluated changes in the serum UA level in the 6-hydroxydopamine (6-OHDA)-induced hemiparkinsonism in rat. For this purpose, 6-OHDA was administered in the medial forebrain bundle by stereotaxic surgery. According to the apomorphine-induced rotational test, the increased intensity of behavioral symptoms as a function of time was associated with the further reduction of UA level. On the other hand, the level of UA increased in the midbrain of the injured hemisphere. The level of reduction in the serum UA level of rats with severe and moderate symptoms was significantly higher than that of rats with mild symptoms. The immunohistofluorescence and biochemical analyses showed that the serum UA level was also correlated with the death of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra pars compacta (SNc), reduced level of striatal dopamine, and severity of oxidative stress in the midbrain. The rats with mild symptoms also showed a significant decrease in TH-positive neurons and striatal dopamine level. These findings suggest a positive correlation between the level of reduction in the serum urate level and severity of 6-OHDA-induced Parkinsonism. In addition, our findings indicated that UA had no marked neuroprotective effects, at least at concentrations obtained in this study. On the other hand, UA was introduced as a biomarker for PD, as a significant decline was observed in the serum UA level of rats with mild behavioral symptoms but with significant dopaminergic cell death in the SNc.     

Effects of Dietary Polychlorinated Biphenyls and Protein Level on Liver and Serum Lipid Metabolism of Rats

The effects of polychlorinated biphenyls (PCB) and dietary protein level on the liver and serum lipid metabolism of rats were studied. Rats were fed an experimental diet containing 7 or 30% casein with or without 0.1 % PCB for 24 days. Dietary PCB increased the level of triglyceride, phospholipid and cholesterol in the liver. The accumulation of triglyceride and cholesterol in liver was markedly increased with a low protein diet. The incorporation of injected ³H₂O into liver cholesterol was increased by PCB, but not affected by the dietary level of protein. The incorporation of the tracer into liver fatty acids was not increased by PCB intake. Dietary PCB also raised serum cholesterol and phospholipid, while PCB decreased triglyceride level, especially in rats on low protein diet. In addition, PCB intake clearly raised serum high density lipoprotein and diminished very low density lipoprotein. In the low protein group, PCB markedly repressed the incorporation of ³H₂O into serum lipids. The results suggest that the hepatic lipids accumulation by the addition of 0.1 % PCB to a low protein diet might be mainly ascribed to a repression in the transport of triglyceride from liver to blood. KEY WORDS: PCB, dietary protein, liver lipids, serum lipoprotein.     

Serum Proteomic Changes after Randomized Prolonged Erythropoietin Treatment and/or Endurance Training: Detection of Novel Biomarkers

IntroductionDespite implementation of the biological passport to detect erythropoietin abuse, a need for additional biomarkers remains. We used a proteomic approach to identify novel serum biomarkers of prolonged erythropoiesis-stimulating agent (ESA) exposure (Darbepoietin-α) and/or aerobic training.MethodsSerum proteins were separated according to charge and molecular mass (2D-gel electrophoresis). The identity of proteins from spots exhibiting altered intensity was determined by mass spectrometry.ResultsSix protein spots changed in response to Darbepoietin-α treatment. Comparing all 4 experimental groups, two protein spots (serotransferrin and haptoglobin/haptoglobin related protein) showed a significant response to Darbepoietin-α treatment. The haptoglobin/haptoglobin related protein spot showed a significantly lower intensity in all subjects in the training-ESA group during the treatment period and increased during the washout period.ConclusionAn isoform of haptoglobin/haptoglobin related protein could be a new anti-doping marker and merits further research.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01320449","term_id":"NCT01320449"}}NCT01320449     

Character of Viscero- and Somatomotor Interactions in Rat Pups at Changes of the Level of Activity of Adrenergic Structures

In the 5–10-day old rat pups, in the course of simultaneous recording of ECG, respiration rate (RR), and spontaneous periodic motor activity (SPMA) there were determined parameters of the studied processes and their correlations at changes of levels of activity of adrenergic structures. As adrenoactive agents, the sympathomimetic amphetamine, - and -adrenolytics phentolamine and propranolol, as well as the dopamine receptor blocker haloperidol were used. It has been established that injection of amphetamine significantly increases intensity of the decasecond rhythm in the activity pattern of excitable structures. However, the action both on the - and -adrenoreceptors and on dopamine receptors produces no significant changes of this rhythm parameters. Based on this, it can be suggested that an increase of the decasecond rhythm is due to the ability of amphetamine to inhibit the pentose phosphate cycle activity. Administration of adrenolytics decreases RR and heart rate (HR). Amphetamine restores them by a simultaneous increase of pathological disturbances of the respiration rhythm, which appear at injections of sympatholytics. Blockade of dopamine receptors leads to replacement of motor activity fires by jerks following in the rhythm close to the previous one. As a rule, at the blockade of -adrenoreceptors the SPMA pattern is deprived of obvious rhythmic components, whereas at the blockade of -adrenoreceptors, they are preserved. Analysis of the obtained data indicates that the respiration frequency modulation that increases at action on adrenoreactive structures correlates with the motor activity changes. However, there are several peculiarities due to action on various types of adrenoreceptors. Of the leading significance in the normal intersystem interaction is preservation of balance between individual adrenergic structures.     

α-Linolenic Acid Dietary Deficiency Alters Age-Related Changes of Dopaminergic and Serotoninergic Neurotransmission in the Rat Frontal Cortex

Abstract: The effects of α-linolenic acid diet deficiency on rat dopaminergic and serotoninergic neurotransmission systems were investigated in the frontal cortex, striatum, and cerebellum of male rats 2, 6, 12, and 24 months of age. The diet deficiency induced a severe decrease in the 22:6n-3 fatty acid levels in all regions and a compensatory increase in n-6 fatty acid levels. A recovery in the levels of 22:6n-3 was observed in deficient rats between 2 and 12 months of age; however, this recovery was lower in frontal cortex than in striatum and cerebellum. In the striatum and the cerebellum, dopaminergic and serotoninergic receptor densities and endogenous dopamine and serotonin levels were affected by aging regardless of the diet. In contrast, a 40–75% lower level of endogenous dopamine in the frontal cortex occurred in deficient rats according to age. The deficiency also induced an 18–46% increase in serotonin 5-HT₂ receptor density in the frontal cortex during aging, without variation in endogenous serotonin level, and a 10% reduction in density of dopaminergic D₂ receptors. Monoamine oxidase-A and -B activities showed specific age-related variations but regardless of the diet. Our results suggest that a chronically α-linolenic-deficient diet specifically affects the monoaminergic systems in the frontal cortex.     

Functional Changes of Rat Brain Microsomal Membrane Surface After Learning Task Depending on Dietary Fatty Acids

Abstract: Biochemical characteristics of brain microsomal membranes were examined before and after the brightness-discrimination learning tasks in rats that were fed either safflower oil (α-linolenate-deficient) or perilla oil (α-linolenate-sufficient) diets. We detected small changes in the chain elongation system for polyunsaturated fatty acids in microsomes, whereas no significant difference was detected in the inositol trisphosphate-induced calcium release and ATP-induced calcium uptake profiles of microsomes between the two dietary groups. The calcium ion-induced aggregation rate of microsomes was determined in both groups. We found that the aggregation rate of microsomes in the safflower oil group was significantly greater than that in the perilla oil group. The difference in susceptibility of microsomal membrane phospholipids to phospholipase A₂ between the groups was obvious, and the amount of released fatty acids by phospholipase A₂ from the perilla oil group microsomes was nearly half of that from the safflower oil group microsomes after the learning task. Susceptibility of sialic acids on the brain microsomal membranes to exogenous sialidase was different only after the learning task in the safflower and perilla oil groups. These results suggest that the biochemical characteristics of membrane surfaces of brain microsomes are affected significantly by the learning task itself in a dietary oil-dependent manner.     

Serum Magnesium Level is Negatively Associated with Fasting Serum Glucose Level in Korean Adults

Several studies have observed an inverse association between serum magnesium and fasting glucose levels. However, there have been sparse data on Koreans. The present study was designed to evaluate the association between serum magnesium and fasting serum glucose levels in Korean adults. A total of 949 Korean adults who visited an outpatient at a university hospital were included in this study. We compared mean values of clinical data according to the tertile of serum magnesium level using analysis of covariance. Association between serum magnesium and fasting glucose levels was evaluated using multiple regression analysis. Odds ratio (OR) was calculated to determine the association between hypomagnesemia and abnormal glucose metabolism. Fasting serum glucose levels decreased significantly from the lowest to the highest tertile of serum magnesium level (108.0 ± 1.4, 106.5 ± 2.1, 102.5 ± 1.7 mg/dl, respectively, p = 0.015), whereas blood pressure and lipid profile showed no significant associations. Serum magnesium level was negatively associated with fasting serum glucose level (β = −0.114, p = 0.001). Participants with hypomagnesemia had significantly higher OR for abnormal glucose metabolism compared to those with normomagnesaemia (OR = 2.28, 95%CI 1.29–4.02). There was a negative association between serum magnesium and fasting glucose levels in Korean adults. In addition, hypomagnesemia was associated with abnormal glucose metabolism.     

Accelerated Myelinogenesis by Dietary Lipids in Rat Brain

Abstract: Our previous work showed an early development of behavioral reflexes in rats whose mothers had been fed, during pregnancy and lactation, a lipid fraction extracted from yeast grown on n -alkanes (which contain 50% odd-chain fatty acids) in comparison with controls fed a margarine diet. To clarify whether the observed changes might be linked to an early myelination, we have investigated mRNAs involved in myelin synthesis in the brains of offspring at 5 days of age by northern blot and in situ hybridization. Northern blot analysis showed that proteolipid protein (PLP) and myelin oligodendrocyte glycoprotein (MOG) mRNAs were higher in animals on the lipid diet compared with controls. In situ hybridization with probes specific for PLP, myelin basic protein, and MOG mRNA showed significantly higher numbers of positive cells in test animals compared with controls in all brain regions. This study shows an acceleration of myelinogenesis induced by dietary lipids. These data can give a new insight in the therapeutical approaches involved to promote repair in demyelinating diseases.     

Protein-induced bending of the simian virus 40 origin of replication

A 3.5 S protein, isolated from mammalian nuclei, specifically binds to DNA fragments containing the simian virus 40 (SV40) origin of replication. Two distinct nucleoprotein complexes are formed, a complex with high electrophoretic mobility carrying probably only one protein molecule, and a complex with reduced electrophoretic mobility carrying probably two protein molecules per DNA fragment. Band shift competition as well as methylation interference assays locate the binding site of the protein in the A + T-rich "late" region of the origin between SV40 nucleotides 13 and 35. The late origin binding (LOB) protein and T antigen bind simultaneously to adjacent sites in the origin. Using circularly permuted DNA fragments of identical lengths we show that the LOB protein induces pronounced bending of the origin fragment. The bending center maps at the 5' end of the adenine tract with one bound protein molecule and at the 3' end when two LOB proteins are bound to one origin fragment.     

Systemic Treatment with Erythropoietin Protects the Neurovascular Unit in a Rat Model of Retinal Neurodegeneration

Rats expressing a transgenic polycystic kidney disease (PKD) gene develop photoreceptor degeneration and subsequent vasoregression, as well as activation of retinal microglia and macroglia. To target the whole neuroglialvascular unit, neuro- and vasoprotective Erythropoietin (EPO) was intraperitoneally injected into four –week old male heterozygous PKD rats three times a week at a dose of 256 IU/kg body weight. For comparison EPO-like peptide, lacking unwanted side effects of EPO treatment, was given five times a week at a dose of 10 µg/kg body weight. Matched EPO treated Sprague Dawley and water-injected PKD rats were held as controls. After four weeks of treatment the animals were sacrificed and analysis of the neurovascular morphology, glial cell activity and pAkt localization was performed. The number of endothelial cells and pericytes did not change after treatment with EPO or EPO-like peptide. There was a nonsignificant reduction of migrating pericytes by 23% and 49%, respectively. Formation of acellular capillaries was significantly reduced by 49% (p<0.001) or 40% (p<0.05). EPO-treatment protected against thinning of the central retina by 10% (p<0.05), a composite of an increase of the outer nuclear layer by 12% (p<0.01) and in the outer segments of photoreceptors by 26% (p<0.001). Quantification of cell nuclei revealed no difference. Microglial activity, shown by gene expression of CD74, decreased by 67% (p<0.01) after EPO and 36% (n.s.) after EPO-like peptide treatment. In conclusion, EPO safeguards the neuroglialvascular unit in a model of retinal neurodegeneration and secondary vasoregression. This finding strengthens EPO in its protective capability for the whole neuroglialvascular unit.     

Multi-Modal Assessment of Long-Term Erythropoietin Treatment after Neonatal Hypoxic-Ischemic Injury in Rat Brain

Erythropoietin (EPO) has been recognized as a neuroprotective agent. In animal models of neonatal brain injury, exogenous EPO has been shown to reduce lesion size, improve structure and function. Experimental studies have focused on short course treatment after injury. Timing, dose and length of treatment in preterm brain damage remain to be defined. We have evaluated the effects of high dose and long-term EPO treatment in hypoxic-ischemic (HI) injury in 3 days old (P3) rat pups using histopathology, magnetic resonance imaging (MRI) and spectroscopy (MRS) as well as functional assessment with somatosensory-evoked potentials (SEP). After HI, rat pups were assessed by MRI for initial damage and were randomized to receive EPO or vehicle. At the end of treatment period (P25) the size of resulting cortical damage and white matter (WM) microstructure integrity were assessed by MRI and cortical metabolism by MRS. Whisker elicited SEP were recorded to evaluate somatosensory function. Brains were collected for neuropathological assessment. The EPO treated animals did not show significant decrease of the HI induced cortical loss at P25. WM microstructure measured by diffusion tensor imaging was improved and SEP response in the injured cortex was recovered in the EPO treated animals compared to vehicle treated animals. In addition, the metabolic profile was less altered in the EPO group. Long-term treatment with high dose EPO after HI injury in the very immature rat brain induced recovery of WM microstructure and connectivity as well as somatosensory cortical function despite no effects on volume of cortical damage. This indicates that long-term high-dose EPO induces recovery of structural and functional connectivity despite persisting gross anatomical cortical alteration resulting from HI.     

Serum Selenium Level in Healthy Koreans

Of trace elements in the serum of living organisms, selenium (Se) is an essential mineral and plays the role of an antioxidant as selenoproteins protecting the organism against oxidative damage induced by hydrogen peroxide, other lipid hydroperoxides, and their derivatives. The aim of this study was to determine the mean serum Se levels in healthy Korean volunteers (50 males and 50 females) by using an inductively coupled plasma-mass spectrometry method. The samples were collected at the Health Promotion Centre of Kangnam St. Mary’s Hospital, College of Medicine, the Catholic University of Korea, Kangnam District, Seoul in accordance with procedures approved by the Institutional Review Board of the Catholic University of Korea. The mean serum Se level in healthy subjects was 112.05 ± 30.42 μg/l. For gender, it was 120.81 ± 27.37 μg/l for females and 103.29 ± 31.05 μg/l for males. From the study result, there was a significant difference between the mean Se concentrations of gender groups (p = 0.0035). Also, the study indicated no effect of age on Se levels (p > 0.05) in the healthy individuals.     

Effect of Endotoxin on Serum Zinc Concentrations in the Rat

Serum zinc concentrations decreased significantly in a dose-dependent response after endotoxin administration in the rat. The reproducibility and sensitivity of the biological response offer a potential bioassay of endotoxin.