Intermedin ameliorates atherosclerosis in apoE null mice by modifying lipid profiles

Intermedin (IMD) is a recently discovered vasodilator peptide. We studied the role of IMD in the pathogenesis of atherosclerosis by investigating the ability of exogenous IMD to alter lipid profiles and ameliorate the development of atherogenic-diet induced atherosclerosis in ApoE−/− mice. Ten of eight-week-old male C57BL/6J mice were as control. Thirty of eight-week-old male ApoE−/− mice were fed with an atherogenic diet for 18 weeks. After feeding atherogenic diet for 12 weeks, the mice were equally and randomly divided into three groups. Normal saline was given in group A and C57BL/6J mice. Intermedin was given by mini osmotic pumps at the dosage of 100 ng/kg/h and 500 ng/kg/h in group B and group C respectively. After the treatment of IMD for 6 weeks, aortic ultrasonography of group C showed that IMD prevented the progression of atherosclerotic lesions and the increase of wall thickness in the aorta. Oil-red-O staining of the entire aorta and the atherosclerotic aortic root section showed 2 folds decrease atherogenic plaque (p < 0.05). Serum lipid profiles were measured, compared with the group A, in group C TC and LDL-C levels were decreased by 86.32% and 89.68%, respectively (both p < 0.05), meanwhile, HDL-C level was significantly increased by 74.82% (p < 0.05). These data indicate that exogenous administration of IMD could prevent the progression of atherosclerotic plaque. The possible underlying mechanisms may relate to the improvement of lipid profiles.     

Preliminary observations, at the ultrastructural level, on the reactivity of cerebral arteries from New Zealand rabbits to combined atherogenic stimuli (hypercholesterol diet and hypertension)

Both in monkeys (Rhesus and Cynomolgus) and in New Zealand rabbits fed an atherogenic diet, a marked delay in the appearance of atherosclerotic lesions of the cerebral arteries in comparison with other arterial districts has been observed. This appearance has been described in monkeys as relatively earlier if hypertension is added to the atherogenic diet. Preliminary observations on a little group of rabbits on a 3 months hypercholesterolic diet, subjected to Goldblatt aortic coarctation, have shown an increase of blood pressure and a severe gross atherosclerotic involvement of aorta, resembling the one obtainable after 6 months of atherogenic diet. Histologically, the aorta predominantly shows lesions of the fatty streaks type with necrotic areas in the deep; the carotid lesions show some lipid in smooth muscle cells disseminated in a sub-endothelial "edematous" space (rich in protein). The cerebral arteries do not show any lesion. At TEM, the aortic lesions look sometimes as advanced plaques with an initial fibrosis at the surface; the carotid lesions are characterized by a granular deposit in the sub-endothelial space in which some smooth muscle cells (with lipid in the cytoplasm) are present; in the cerebral arteries only the presence of collagen fibers among the smooth muscle cells of the media, never observed in the animals fed the atherogenic diet alone, has sometimes been noted.     

Control of hypercholesterolemia and atherosclerosis using the cholesterol recognition/interaction amino acid sequence of the translocator protein TSPO

The translocator protein (18-kDa) TSPO is an ubiquitous high affinity cholesterol-binding protein reported to be present in the endothelial and smooth muscle cells of the blood vessels; its expression dramatically increased in macrophages found in atherosclerotic plaques. A domain in the carboxy-terminus of TSPO was identified and characterized as the cholesterol recognition/interaction amino acid consensus (CRAC). The ability of the CRAC domain to bind to cholesterol led us to hypothesize that this peptide could be used as an hypocholesterolemic, with potential anti-atherogenic properties, agent. We report herein the therapeutic benefit that resulted for the administration of the VLNYYVWR human CRAC sequence to guinea pigs fed with a high cholesterol diet and ApoE knock-out B6.129P2-Apoetm1Unc/J mice. CRAC treatment (3 and 30 mg/kg once daily for 6 weeks) resulted in reduced circulating cholesterol levels in guinea pigs fed with 2% high cholesterol diet and ApoE knock-out B6.129P2-Apoetm1Unc/J mice. In high cholesterol fed guinea pigs, CRAC treatment administered once daily induced an increase in circulating HDL, decreased total, free and LDL cholesterol, and removed atheroma deposits in the aorta in a dose-dependent manner. The treatment also prevented the high cholesterol diet-induced increase in serum creatine kinase, total and isoforms, markers of neurological, cardiac and muscular damage. No toxicity was observed. Taken together these results support a role of TSPO in lipid homeostasis and atherosclerosis and indicate that CRAC may constitute a novel and safe treatment of hypercholesterolemia and atherosclerosis.     

Lipoic acid effects on established atherosclerosis

AimsAlpha-lipoic acid (LA) is a commonly used dietary supplement that exerts anti-oxidant and anti-inflammatory effects in vivo and in vitro. We investigated the mechanisms by which LA may confer protection in models of established atherosclerosis.Main methodsWatanabe heritable hyperlipidemic (WHHL) rabbits were fed with high cholesterol chow for 6 weeks and then randomized to receive either high cholesterol diet alone or combined with LA (20 mg/kg/day) for 12 weeks. Vascular function was analyzed by myography. The effects of LA on T cell migration to chemokine gradients was assessed by Boyden chamber. NF-κB activation was determined by measuring translocation and electrophoresis migration shift assay (EMSA).Key findingsLA decreased body weight by 15 ± 5% without alterations in lipid parameters. Magnetic Resonance Imaging (MRI) analysis demonstrated that LA reduced atherosclerotic plaques in the abdominal aorta, with morphological analysis revealing reduced lipid and inflammatory cell content. Consistent with its effect on atherosclerosis, LA improved vascular reactivity (decreased constriction to angiotensin II and increased relaxation to acetylcholine and insulin), inhibited NF-κB activation, and decreased oxidative stress and expression of key adhesion molecules in the vasculature. LA reduced T cell content in atherosclerotic plaque in conjunction with decreasing ICAM and CD62L (l-selectin) expression. These effects were confirmed by demonstration of a direct effect of LA in reducing T cell migration in response to CCL5 and SDF-1 and decreasing T cell adhesion to the endothelium by intra-vital microscopy.SignificanceThe present findings offer a mechanistic insight into the therapeutic effects of LA on atherosclerosis.     

Anti-obesity potential of almond (Prunus dulcis) in experimental animals under cafeteria and atherogenic diets

Background & objectivesNatural dietary supplements are progressively getting famous to supplant synthetic substances particularly in chronic morbidities. The aim of this study was to evaluate the anti-obesity potential of almond on the normal, Cafeteria, and Atherogenic diets.Materials and methodsParameters such as change in body weight, body temperature, lipid profile, organ weights, and fat pad weights were assessed. Central Nervous System related studies (Despair Swim test and Elevated Plus maze test) were also performed to comprehend the effect of the diets, and almond on the brain. All of the experimental animals were randomly assigned to one of three diet categoriesregular, cafeteria, or atherogenic, and fed those diets for 40 days. Each diet had the control group, standard drug group and three almond groups (low dose: 50; medium dose: 100 and high dose: 200 mg/kg body weight). Body weight was recorded every alternate day. On 40th day, body temperature was measured. On day 41, lipid parameters, organ weights, fat pad weights and the CNS parameters were evaluated. ANOVA followed by Duncans Multiple Range Test were used for statistical analysis.ResultsTreatment of animals with either a low or high dose of almond as well as a standard herb prevented a rise in body weight significantly (p = 0.01) in all three diet groups. When a regular diet was replaced with a cafeteria and atherogenic diet, the serum levels of triglycerides and LDL increased significantly, while HDL levels decreased significantly. Overall, almond preparation reduced lipid parameters, organ weights, fat-pad weights, and stabilized CNS parameters substantially.Interpretation & conclusionThe almond high dose was the most effective of all the almond preparations. Our study suggests that chronic administration of almond independently reduces the body weight in experimental animals.     

Damage of Guinea Pig Heart and Arteries by a Trioleate-Enriched Diet and of Cultured Cardiomyocytes by Oleic Acid

Background

Mono-unsaturated fatty acids (MUFAs) like oleic acid have been shown to cause apoptosis of cultured endothelial cells by activating protein phosphatase type 2C α and β (PP2C). The question arises whether damage of endothelial or other cells could be observed in intact animals fed with a trioleate-enriched diet.

Methodology/Principal Findings

Dunkin-Hartley guinea pigs were fed with a trioleate-enriched diet for 5 months. Advanced atherosclerotic changes of the aorta and the coronary arteries could not be seen but the arteries appeared in a pre-atherosclerotic stage of vascular remodelling. However, the weight and size of the hearts were lower than in controls and the number of apoptotic myocytes increased in the hearts of trioleate-fed animals. To confirm the idea that oleic acid may have caused this apoptosis by activation of PP2C, cultured cardiomyocytes from guinea pigs and mice were treated with various lipids. It was demonstrable that oleic acid dose-dependently caused apoptosis of cardiomyocytes from both species, yet, similar to previous experiments with cultured neurons and endothelial cells, stearic acid, elaidic acid and oleic acid methylester did not. The apoptotic effect caused by oleic acid was diminished when PP2C α and β were downregulated by siRNA showing that PP2C was causally involved in apoptosis caused by oleic acid.

Conclusions/Significance

The glycerol trioleate diet given to guinea pigs for 5 months did not cause marked atherosclerosis but clearly damaged the hearts by activating PP2C α and β. The diet used with 24% (wt/wt) glycerol trioleate is not comparable to human diets. The detrimental role of MUFAs for guinea pig heart tissue in vivo is shown for the first time. Whether it is true for humans remains to be shown.     

Relationship between the amino acid release kinetics of feed proteins and nitrogen balance in finishing pigs

In current nutrition requirements of swine, although the protein diets are formulated based on the ileal digestibility of protein and amino acid (AA), there is a difference in nitrogen utilisation among various protein diets, which might be related to the AA release kinetics. To evaluate the relationship between AA release kinetics of feed proteins and nitrogen balance in finishing pigs, pigs were fed diets based on casein (CAS) or corn gluten meal (CGM) at normal or low-protein concentrations, and the AA release patterns were assessed. A 2 × 2 full factorial experimental design was used. 24 pigs (Duroc × Landrace × Yorkshire) with an initial weight of 67.0 ± 1.8 kg were randomly assigned to consume a normal-protein casein-based diet (N.CAS, 10% CP), normal-protein corn gluten meal-based diet (N.CGM, 10% CP), low-protein casein-based diet (L.CAS, 8.5% CP), or low-protein corn gluten meal-based diet (L.CGM, 8.5% CP) for 14 days (n = 6 per group; pigs housed and fed separately). The low-protein diets were associated with a more rapid release of AAs in the early stages of gastric digestion than the normal-protein diets. The N.CAS and L.CAS diets were associated with a peak AA release at approximately 4 h during trypsin digestion, whereas N.CGM and L.CGM were at approximately 16 h. The N.CAS diet was associated with the least dispersed release curves and lowest synchronisation indexes, implying that it was associated with the best AA release synchronism, whereas the L.CGM diet was on the contrary. The nitrogen intake (NI), faecal nitrogen, urine nitrogen (UN), total nitrogen, net protein utilisation and apparent biological value (ABV) of protein of pigs fed the L.CAS or L.CGM diets were lower than those fed the N.CAS or N.CGM diets (P < 0.05). Notably, there was a difference in NI (P < 0.05) and trends with respect to UN and ABV (0.05 < P < 0.1), but no differences in retained nitrogen or apparent nitrogen digestibility between pigs fed the N.CAS or L.CAS diets and those fed the N.CGM or L.CGM diets. Pigs fed the N.CAS or N.CGM diets had higher serum concentrations of UN than pigs fed the L.CAS or L.CGM diets (P < 0.05), but there were no differences in serum total protein, albumin, triglyceride, glucose, alanine transaminase, or aspartate aminotransferase between the groups. In addition, there was an interaction between protein level and protein source on serum globulin (P < 0.05). Therefore, the diet with a better AA release synchronism can improve protein utilisation efficiency in finishing pigs and to reduce environmental pollution.     

Chronic-arginine supplementation improves endothelial cell vasoactive functions in hypercholesterolemic and atherosclerotic monkeys

Chronic exposure to l-arginine results in regression of atherosclerotic lesions and reversal of endothelial dysfunction. We investigated whether chronic l-arginine supplementation induces regression of atherosclerotic lesions and reversal of endothelial dysfunction in atherogenic rhesus monkeys and the mechanism which leads to these effects. About 12 male rhesus monkeys were fed 1% cholesterol and 18 g butter for 6 months to create an experimental model of hypercholesterolaemia and atherosclerosis (Group I) and 12 monkeys were fed standard stock diet for 6 months (Group II). After, 6 months these two groups were further divided into 2 sub-groups which in addition to their respective diets were fed 2.5% l-arginine in drinking water for additional 6 months (Group III and Group IV). Systemic nitric oxide (NO) formation was assessed as plasma nitrite and cGMP formation every 3 months. Oxygen free radical (OFR) generation and malondialdehyde production as an index of lipid peroxidation were determined. Changes in isometric tension were compared in isolated ring segments of thoracic aorta from normal and hypercholesterolemic animals.Cholesterol feeding progressively reduced plasma nitrite and cGMP generation (p<0.05). Dietary l-arginine partly restored the levels of plasma nitrite and cGMP (p<0.05) but did not change plasma cholesterol levels. l-arginine significantly reduced aortic intimal thickening, blocked the production of carotid and coronary intimal plaques and completely preserved endothelium-dependent vasodilator function. Further, l-arginine significantly inhibited generation of the reactive oxygen species (ROS) generation and lipid peroxidation.Chronic oral supplementation with l-arginine blocks the progression of plaques via restoration of nitric oxide synthase substrate availability and reduction of vascular oxidative stress. (Mol Cell Biochem 269: 1–11, 2005)     

Targeted Disruption of LDLR Causes Hypercholesterolemia and Atherosclerosis in Yucatan Miniature Pigs

Recent progress in engineering the genomes of large animals has spurred increased interest in developing better animal models for diseases where current options are inadequate. Here, we report the creation of Yucatan miniature pigs with targeted disruptions of the low-density lipoprotein receptor (LDLR) gene in an effort to provide an improved large animal model of familial hypercholesterolemia and atherosclerosis. Yucatan miniature pigs are well established as translational research models because of similarities to humans in physiology, anatomy, genetics, and size. Using recombinant adeno-associated virus-mediated gene targeting and somatic cell nuclear transfer, male and female LDLR+/− pigs were generated. Subsequent breeding of heterozygotes produced LDLR−/− pigs. When fed a standard swine diet (low fat, no cholesterol), LDLR+/− pigs exhibited a moderate, but consistent increase in total and LDL cholesterol, while LDLR−/− pigs had considerably elevated levels. This severe hypercholesterolemia in homozygote animals resulted in atherosclerotic lesions in the coronary arteries and abdominal aorta that resemble human atherosclerosis. These phenotypes were more severe and developed over a shorter time when fed a diet containing natural sources of fat and cholesterol. LDLR-targeted Yucatan miniature pigs offer several advantages over existing large animal models including size, consistency, availability, and versatility. This new model of cardiovascular disease could be an important resource for developing and testing novel detection and treatment strategies for coronary and aortic atherosclerosis and its complications.     

Increased production of prostacyclin (PGI2) by vascular intimal smooth muscle cells from atherosclerotic rabbit aorta

PGI₂ synthesis by aortic strips obtained from thoracic aorta of rabbits fed a high cholesterol diet was examined and compared with that of control rabbits fed a normal diet. In this report, the amounts of PGI₂ produced were shown as 6-keto-PGF_1α per μg of aortic tissue DNA instead of per mg wet weight. We also investigated PGI₂ synthesis by cultured smooth muscle cells (SMC) obtained from atherosclerotic intima.Basal PGI₂ production by aortic strips from atherosclerotic rabbit aorta was significantly augmented compared with that of controls. Arachidonic acid (AA)-induced PGI₂ production by atherosclerotic aorta was also significantly higher than that of controls. PGI₂ producing capacities of intimal and medial layers, separated from atherosclerotic aorta, were examined and the intimal layer was found to elicit a significantly greater PGI₂ production than the medial layer.Furthermore, cultured intimal SMC obtained from atherosclerotic rabbit aorta produced a greater amount of PGI₂ than medial SMC from normal rabbit aorta at various cultured conditions. These results suggest that the possibility of enhanced PGI₂ production by atherosclerotic aorta may well be considered as a defence mechanism of the vessel wall against damaging stimuli.     

Dietary preference for methionine sources in weaned pigs

The objective of the current study was to investigate the preference of weaned pigs given the choice of diets supplemented with dl-methionine (DLM) or liquid dl-methionine hydroxy analog-free acid (MHA-FA). A basal diet (BD) was formulated to contain 2.5 g methionine (Met) per kilogram of diet. The experimental diets included: (1) BD, (2) BD + 1 g DLM/kg, (3) BD + 2 g DLM/kg, (4) BD + 1.13 g MHA-FA/kg, (5) BD + 1.52 g MHA-FA/kg, (6) BD + 2.25 g MHA-FA/kg and (7) BD + 3.05 g MHA-FA/kg. Sixty weaned mixed-sex pigs were allotted to 5 treatment groups with 12 pig replicates per treatment in a randomized complete block design. During a 35-day experimental period, pigs in treatment group 1 received the BD whereas pigs in the other 4 treatment groups were allowed to choose between a pair of diets with either added 1 g DLM/kg in combination with either 1.13 or 1.52 g MHA-FA/kg or 2 g DLM/kg in combination with 2.25 or 3.05 g MHA-FA/kg, respectively. Pigs were housed in individual pens and had free access to feed and water. Daily feed intake (FI) was used as an indicator of diet preference. Cumulatively, pigs showed a preference (% of total FI) for the diet added with 1 g/kg DLM at 74% (P<0.05) in group 2, and 65% in group 3. Irrespective of the level of MHA-FA supplementation (2.25 or 3.05 g/kg), a preference for the diet supplemented with 2 g DLM/kg was 84% (P<0.05) in groups 4 and 5. During the entire period, pigs consistently (P<0.05) consumed more and preferred the diets supplemented with DLM more than the diets supplemented with MHA-FA in groups 2, 4 and 5. The preference for diets supplemented with DLM was more pronounced at higher Met supplementation levels. Feeding pigs a pair of diets supplemented with DLM or MHA-FA improved (P<0.05) the final body weight, daily weight gain, and FCR. The performance of pigs among the Met-supplemented groups was not different. In conclusion, when given a choice pigs preferred the diets supplemented with DLM more than the diets supplemented with liquid MHA-FA.     

Sand accumulation in the digestive tract of rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus): The role of the appendix

We determined location and amount of accumulated sand in the gastrointestinal tract (GIT) of rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus) fed diets containing external (silicate) abrasives. Computed tomographic abdominal images of rabbits (n = 44) and guinea pigs (n = 16) that each received varying numbers (4–7) of different diets for 14 days each (total n = 311 computed tomographs), and radiographs of dissected GIT and presence of silica in GIT content (n = 46 animals) were evaluated. In rabbits, the majority of accumulated sand was located in the caecal appendix, an elongated, intestinal structure in the left side of the abdomen. The ‘wash-back’ colonic separation mechanism in rabbits may be partly responsible for a retrograde transport of sand back to the caecum, where dense, small particles accumulate in the appendix. The appendix likely acted as a reservoir of these particles, leading to significant effects not only of the momentary but also of the previous diet on recorded sand volumes in the rabbits. Guinea pigs have no caecal appendix and a colonic separation mechanism not based on a ‘wash-back’. Less sand accumulation was found in their GIT without a specific location pattern, and there were less previous diet effects in this species. None of the rabbits or guinea pigs developed clinical signs of obstruction during the study, and the recorded sand volumes represented 1.0 ± 1.2% of the 14-d sand intake in rabbits and 0.2 ± 0.2% in guinea pigs. Accumulation of sand in volumes up to 10 cm³ in the GIT of rabbits does not seem to cause clinical health impairment. Large inter-individual differences in rabbits indicate inter-individual variation in proneness to sand accumulation. The reason for the presence of a sand-trapping caecal appendix in animals that are, due to their burrowing lifestyle and feeding close to the ground, predestined for accidental sand ingestion, remains to be unveiled.     

Anti-Atherogenic Effect of Hydrogen Sulfide by Over-Expression of Cystathionine Gamma-Lyase (CSE) Gene

Hydrogen sulfide (H₂S) is an important gaseous signaling molecule that functions in physiological and pathological conditions, such as atherosclerosis. H₂S dilates vessels and therefore has been suggested as an anti-atherogenic molecule. Since cystathionine gamma-lyase (CSE) enzyme is responsible for producing H₂S in the cardiovascular system, we hypothesized that up-regulation of CSE expression in vivo with preservation of H₂S bioactivity can slow down plaque formation and, can serve as a therapeutic strategy against atherosclerosis. In this study, C57BL/6 wild type mice (WT), ApoE knockout mice (KO) and transgenic ApoE knockout mice overexpressing CSE (Tg/KO) at four weeks of age were weaned. They were then fed with either normal or atherogenic diet for 12 weeks. At week 16, serial plasma lipid levels, body weight, and blood pressure were measured prior to euthanization of the mice and the size of atherosclerotic plaques at their aortic roots was measured. Tg/KO mice showed an increase in endogenous H₂S production in aortic tissue, reduced atherosclerotic plaque sizes and attenuation in plasma lipid profiles. We also showed an up-regulation in plasma glutathionine peroxidase that could indicate reduced oxidative stress. Furthermore, there was an increase in expression of p-p53 and down regulation of inflammatory nuclear factor-kappa B (NF-κB) in aorta. To conclude, alteration of endogenous H₂S by CSE gene activation was associated with reduced atherosclerosis in ApoE-deficient mice. Up-regulation of CSE/H₂S pathway attenuates atherosclerosis and this would be a potential target for therapeutic intervention against its formation.     

Overexpression of CXCL16 promotes a vulnerable plaque phenotype in Apolipoprotein E-Knockout Mice

BackgroundCXCL16 has been shown to be involved in atherosclerotic lesion development, but its role in preexisting lesions is still unclear. This study aims to assess the effect of CXCL16 on the stability of preexisting lesions.MethodsWe firstly measured plasma CXCL16 level in Apolipoprotein E–Knockout (ApoE KO) mice with either high-cholesterol diet (HCD) or normal diet (ND) by enzyme-linked immunosorbent assay (ELISA). Then, silastic collars were placed around the carotid arteries in HCD-ApoE KO mice to accelerate atherosclerotic lesions. Five weeks later, CXCL16 was overexpressed by intravenous injection of lentivirus carrying CXCL16 transgene. Two weeks after infection, lesions were stained with hematoxylin and eosin (HE) and with oil red O. Biomarkers in the lesions, such as MMPs, CCL2, VCAM-1 and TNF-α were measured by real-time polymerase chain reaction (RT-PCR), which indicate the instability of plaques.ResultsThe level of CXCL16 in plasma was higher in HCD-ApoE KO mice as compared to ND-ApoE KO mice. Circulating CXCL16 overexpression does not affect the size of preexisting plaques, but it leads to vulnerable plaque morphology and increases the expression of markers of plaque destabilization.ConclusionSystemic CXCL16 becomes much higher in atherosclerosis, and it could be a potential atherogenic biomarker. Overexpression of CXCL16 promotes the evolution of preexisting lesions to vulnerable plaques in ApoE KO mice.     

Increased survival of neonatal pigs by supplementing medium-chain triglycerides in late-gestating sow diets

Two experiments were conducted to study the efficacy and causes of medium-chain triglycerides (MCT) in sow diet in improving the survival of neonatal pigs. In Experiment 1, beginning on d84 of gestation and continuing through d28 of lactation, 51 sows were fed corn–soybean meal diet mixed with either soybean oil (SO; n=17), coconut oil (CO; n=18), or MCT (n=16) in a proportion of 9 : 1 by weight. The highest improvement in survival of pigs by sows fed MCT (p<0.01) or CO (p<0.05) was observed during the first three days after birth in pigs weighing <1100 g at birth, compared with sows fed SO. Their three-day survival was 98.6, 80.0 and 47.6%, respectively, for MCT, CO and SO groups. In Experiment 2, beginning on d84 of gestation and continuing through farrowing, 24 sows, 8 sows per treatment, were fed diets as in Experiment 1. Liver glycogen content of pigs 4 h after born from sows fed MCT (p<0.10) and CO (p<0.01), and muscle glycogen of pigs from sows fed MCT (p<0.01) and CO (p<0.10) were increased, compared to those of pigs from sows fed SO. Plasma albumin was increased by MCT and CO (p<0.01), relative to SO. The results suggest that MCT or CO in sow diets may enhance the body glycogen stores and maturity of pigs at birth and, hence, their survival, particularly in pigs with low birth weight during the first three days of life.     

Dietary Manipulation and Social Isolation Alter Disease Progression in a Murine Model of Coronary Heart Disease

Background

Mice with a deficiency in the HDL receptor SR-BI and low expression of a modified apolipoprotein E gene (SR-BI KO/ApoeR61^h/h) called ‘HypoE’ when fed an atherogenic, ‘Paigen’ diet develop occlusive, atherosclerotic coronary arterial disease (CHD), myocardial infarctions (MI), and heart dysfunction and die prematurely (50% mortality ∼40 days after initiation of this diet). Because few murine models share with HypoE mice these cardinal, human-like, features of CHD, HypoE mice represent a novel, small animal, diet-inducible and genetically tractable model for CHD. To better describe the properties of this model, we have explored the effects of varying the composition and timing of administration of atherogenic diets, as well as social isolation vs. group housing, on these animals.

Methodology/Principal Findings

HypoE mice were maintained on a standard lab chow diet (control) until two months of age. Subsequently they received one of three atherogenic diets (Paigen, Paigen without cholate, Western) or control diet for varying times and were housed in groups or singly, and we determined the plasma cholesterol levels, extent of cardiomegaly and/or survival. The rate of disease progression could be reduced by lowering the severity of the atherogenic diet and accelerated by social isolation. Disease could be induced by Paigen diets either containing or free of cholate. We also established conditions under which CHD could be initiated by an atherogenic diet and then subsequently, by replacing this diet with standard lab chow, hypercholesterolemia could be reduced and progression to early death prevented.

Conclusions/Significance

HypoE mice provide a powerful, surgery-free, diet-‘titratable’ small animal model that can be used to study the onset of recovery from occlusive, atherosclerotic CHD and heart failure due to MI. HypoE mice can be used for the analysis of the effects of environment (diet, social isolation) on a variety of features of cardiovascular disease.     

Effects of Physical Properties of Feed on Microbial Ecology and Survival of Salmonella enterica Serovar Typhimurium in the Pig Gastrointestinal Tract

A two-by-two factorial experiment with pigs was conducted to study the effect of feed grinding (fine and coarse) and feed processing (pelleted and nonpelleted) on physicochemical properties, microbial populations, and survival of Salmonella enterica serovar Typhimurium DT12 in the gastrointestinal tracts of pigs. Results demonstrated a strong effect of diet on parameters measured in the stomachs of the pigs, whereas the effect was less in the other parts of the gastrointestinal tract. Pigs fed the coarse nonpelleted (C-NP) diet showed more solid gastric content with higher dry matter content than pigs fed the fine nonpelleted (F-NP), coarse pelleted (C-P), or fine pelleted (F-P) diet. Pigs fed the C-NP diet also showed significantly increased number of anaerobic bacteria (P < 0.05), increased concentrations of organic acids, and reduced pH in the stomach. In addition, pigs fed the C-NP diet showed increased in vitro death rate of S. enterica serovar Typhimurium DT12 in content from the stomach (P < 0.001). Pigs fed the C-NP diet had a significantly higher concentration of undissociated lactic acid in gastric content than pigs fed the other diets (P < 0.001). A strong correlation between the concentration of undissociated lactic acid and the death rate of S. enterica serovar Typhimurium DT12 was found. In the distal small intestine, cecum, and midcolon, significantly lower numbers of coliform bacteria were observed in pigs fed the coarse diets than in pigs fed the fine diets (P < 0.01). Pigs fed the C-NP diet showed the lowest number of coliform bacteria in these segments of the gastrointestinal tract. Pigs fed the coarse diets showed increased concentration of butyric acid in the cecum (P < 0.05) and colon (P < 0.10) compared with pigs fed the fine diets. It was concluded that feeding a coarsely ground meal feed to pigs changes the physicochemical and microbial properties of content in the stomach, which decreases the survival of Salmonella during passage through the stomach. In this way the stomach acts as a barrier preventing harmful bacteria from entering and proliferating in the lower part of the gastrointestinal tract.     

High fat diets and pathology in the guinea pig. Atherosclerosis or liver damage?

Animal models have been widely used to investigate the relationship between diet and atherosclerosis and also to study disease etiology and possible interventions. Guinea pigs have been suggested to be a more “realistic” model for atherosclerosis due to their many similarities to humans. However, few published studies actually reported observations of characteristic atherosclerotic lesions and even fewer of advanced lesions. Studies, by our group, of guinea pigs fed on a high-fat diet revealed similar observations, with indications primarily of fatty streaks but little evidence of atherosclerotic plaques. This review discusses the feasibility of the guinea pig as a model for dietary-induced atherosclerosis. As it stands, current evidence raises doubt as to whether guinea pigs could serve as a realistic model for atherosclerosis. However, our own data and the literature suggest that they could be useful models for studying lipoprotein metabolism, non-alcoholic fatty liver disease, and dietary interventions which may help regulate these conditions.     

Differential relaxant responses of pulmonary arteries and veins in lung explants of guinea pigs

Shi, Weibin, David H. Eidelman, and René P. Michel.Differential relaxant responses of pulmonary arteries and veins inlung explants of guinea pigs. J. Appl.Physiol. 83(5): 1476-1481, 1997.The endotheliumregulates vascular tone through release of relaxing or contractingfactors, with nitric oxide (NO) being a major endothelium-derivedrelaxing factor. In the present study, we used a lung explant techniqueto determine the differential abilities and mechanisms of pulmonaryarteries and veins of normal guinea pigs to relax after precontraction.Excised lungs of 15 guinea pigs were filled through the airways with1% agarose, cut into 1-mm-thick slices, and cultured overnight.Luminal areas of vascular cross sections were measured with animage-analysis system. Vessels were precontracted with U-46619, andresponses to histamine, acetylcholine (ACh), sodium nitroprusside, andpapaverine were examined. We also determined the effects ofN-nitro-L-arginineand of indomethacin on ACh-induced responses. We found that histaminerelaxed arteries more than veins and that ACh relaxed only arteries.N-nitro-L-arginine pretreatmentabolished ACh-induced relaxation of arteries and caused ACh-inducedcontraction of veins, whereas indomethacin markedly augmentedACh-induced relaxation of arteries (maximal relaxation: 48.5 ± 4.7 vs. 19.2 ± 5.1% without it) and induced a dose-dependentrelaxation of veins (maximal relaxation: 17.0 ± 4.1%). Sodiumnitroprusside induced a significantly greater relaxation of arteriesthan veins, whereas papaverine relaxed them equally. We conclude thatin guinea pigs endothelial NO-mediated relaxation is greater inpulmonary arteries than in veins and that ACh-induced NO-mediatedrelaxation is reduced by the simultaneous production ofcyclooxygenase-derived vasoconstrictors.