Proteome profiling of different rat brain regions reveals the modulatory effect of prolonged maternal separation on proteins involved in cell death-related processes

BACKGROUND: Early-life stress in the form of maternal separation can be associated with alterations in offspring neurodevelopment and brain functioning. Here, we aimed to investigate the potential impact of prolonged maternal separation on proteomic profiling of prefrontal cortex, hippocampus and cerebellum of juvenile and young adult rats. A special attention was devoted to proteins involved in the process of cell death and redox state maintenance. METHODS: Long-Evans pups were separated from their mothers for 3 h daily over the first 3 weeks of life (during days 2-21 of age). Brain tissue samples collected from juvenile (22-day-old) and young adult (90-day-old) rats were used for label-free quantitative (LFQ) proteomic analysis. In parallel, selected oxidative stress markers and apoptosis-related proteins were assessed biochemically and by Western blot, respectively. RESULTS: In total, 5526 proteins were detected in our proteomic analysis of rat brain tissue. Approximately one tenth of them (586 proteins) represented those involved in cell death processes or regulation of oxidative stress balance. Prolonged maternal separation caused changes in less than half of these proteins (271). The observed alterations in protein expression levels were age-, sex- and brain region-dependent. Interestingly, the proteins detected by mass spectrometry that are known to be involved in the maintenance of redox state were not markedly altered. Accordingly, we did not observe any significant differences between selected oxidative stress markers, such as the levels of hydrogen peroxide, reduced glutathione, protein carbonylation and lipid peroxidation in brain samples from rats that underwent maternal separation and from the corresponding controls. On the other hand, a number of changes were found in cell death-associated proteins, mainly in those involved in the apoptotic and autophagic pathways. However, there were no detectable alterations in the levels of cleaved products of caspases or Bcl-2 family members. Taken together, these data indicate that the apoptotic and autophagic cell death pathways were not activated by maternal separation either in adolescent or young adult rats. CONCLUSIONS: Prolonged maternal separation can distinctly modulate expression profiles of proteins associated with cell death pathways in prefrontal cortex, hippocampus and cerebellum of juvenile rats and the consequences of early-life stress may last into adulthood and likely participate in variations in stress reactivity.

Anti-Proliferative Effects of Rutin on OLETF Rat Vascular Smooth Muscle Cells Stimulated by Glucose Variability

PURPOSE: Proliferation of vascular smooth muscle cells (VSMCs) plays a crucial role in atherosclerosis. Rutin is a major representative of the flavonol subclass of flavonoids and has various pharmacological activities. Currently, data are lacking regarding its effects on VSMC proliferation induced by intermittent hyperglycemia. Here, we demonstrate the effects of rutin on VSMC proliferation and migration according to fluctuating glucose levels. MATERIALS AND METHODS: Primary cultures of male Otsuka Long-Evans Tokushima Fatty (OLETF) rat VSMCs were obtained from enzymatically dissociated rat thoracic aortas. VSMCs were incubated for 72 h with alternating normal (5.5 mmol/L) and high (25.0 mmol/L) glucose media every 12 h. Proliferation and migration of VSMCs, the proliferative molecular pathway [including p44/42 mitogen-activated protein kinases (MAPK), mitogen-activated protein kinase kinase 1/2 (MEK1/2), p38 MAPK, phosphoinositide 3-kinase (PI3K), c-Jun N-terminal protein kinase (JNK), nuclear factor kappa B (NF-kappaB), and Akt], the migratory pathway (big MAPK 1, BMK1), reactive oxygen species (ROS), and apoptotic pathway were analyzed. RESULTS: We found enhanced proliferation and migration of VSMCs when cells were incubated in intermittent high glucose conditions, compared to normal glucose. These effects were lowered upon rutin treatment. Intermittent treatment with high glucose for 72 h increased the expression of phospho-p44/42 MAPK (extracellular signal regulated kinase 1/2, ERK1/2), phospho-MEK1/2, phospho-PI3K, phospho-NF-kappaB, phospho-BMK1, and ROS, compared to treatment with normal glucose. These effects were suppressed by rutin. Phospho-p38 MAPK, phospho-Akt, JNK, and apoptotic pathways [B-cell lymphoma (Bcl)-xL, Bcl-2, phospho-Bad, and caspase-3] were not affected by fluctuations in glucose levels. CONCLUSION: Fluctuating glucose levels increased proliferation and migration of OLETF rat VSMCs via MAPK (ERK1/2), BMK1, PI3K, and NF-kappaB pathways. These effects were inhibited by the antioxidant rutin.

Protective Effects of Curcumin on Renal Oxidative Stress and Lipid Metabolism in a Rat Model of Type 2 Diabetic Nephropathy

PURPOSE: Diabetic nephropathy is a serious complication of type 2 diabetes mellitus, and delaying the development of diabetic nephropathy in patients with diabetes mellitus is very important. In this study, we investigated inflammation, oxidative stress, and lipid metabolism to assess whether curcumin ameliorates diabetic nephropathy. MATERIALS AND METHODS: Animals were divided into three groups: Long-Evans-Tokushima-Otsuka rats for normal controls, Otsuka-Long-Evans-Tokushima Fatty (OLETF) rats for the diabetic group, and curcumin-treated (100 mg/kg/day) OLETF rats. We measured body and epididymal fat weights, and examined plasma glucose, adiponectin, and lipid profiles at 45 weeks. To confirm renal damage, we measured albumin-creatinine ratio, superoxide dismutase (SOD), and malondialdehyde (MDA) in urine samples. Glomerular basement membrane thickness and slit pore density were evaluated in the renal cortex tissue of rats. Furthermore, we conducted adenosine monophosphate-activated protein kinase (AMPK) signaling and oxidative stress-related nuclear factor (erythroid-derived 2)-like 2 (Nrf2) signaling to investigate mechanisms of lipotoxicity in kidneys. RESULTS: Curcumin ameliorated albuminuria, pathophysiologic changes on the glomerulus, urinary MDA, and urinary SOD related with elevated Nrf2 signaling, as well as serum lipid-related index and ectopic lipid accumulation through activation of AMPK signaling. CONCLUSION: Collectively, these findings indicate that curcumin exerts renoprotective effects by inhibiting renal lipid accumulation and oxidative stress through AMPK and Nrf2 signaling pathway.

Emblica officinalis improves glycemic status and oxidative stress in STZ induced type 2 diabetic model rats

OBJECTIVE@#To evaluate the antidiabetic and antioxidant potential of Emblica officinalis (E. officinalis) fruit on normal and type 2 diabetic rats.@*METHODS@#Type 2 diabetes was induced into the male Long-Evans rats. The rats were divided into nine groups including control groups receiving water, type 2 diabetic controls, type 2 diabetic rats treated with glibenclamide (T2GT) and type 2 diabetic rats treated with aqueous extract of fruit pulp of E. officinalis. They were fed orally for 8 weeks with a single feeding. Blood was collected by cutting the tail tip on 0 and 28 days and by decapitation on 56 day. Packed red blood cells and serum were used for evaluating different biochemical parameters.@*RESULTS@#Four weeks administration of aqueous extract of E. officinalis improved oral glucose tolerance in type 2 rats and after 8 weeks it caused significant (P<0.007) reduction in fasting serum glucose level compared to 0 day. Triglycerides decreased by 14% but there was no significant change in serum ALT, creatinine, cholesterol and insulin level in any group. Furthermore, reduced erythrocyte malondialdehyde level showed no significant change (P<0.07) but reduced glutathione content was found to be increased significantly (P<0.05).@*CONCLUSIONS@#The aqueous extract of E. officinalis has a promising antidiabetic and antioxidant properties and may be considered for further clinical studies in drug development.

COMP-Angiopoietin-1 Promotes Cavernous Angiogenesis in a Type 2 Diabetic Rat Model

Cartilage oligomeric matrix protein-angiopoietin-1 (COMP-Ang1) is an angiogenic factor for vascular angiogenesis. The aim was to investigate the effect of an intracavernosal injection of COMP-Ang1 on cavernosal angiogenesis in a diabetic rat model. Male Otsuka Long-Evans Tokushima Fatty (OLETF) rats made up the experimental group (1 yr old) and Long-Evans Tokushima Otsuka (LETO) rats made up the control group. The experimental group was divided into vehicle only, 10 microg COMP-Ang1, and 20 microg COMP-Ang1. COMP-Ang1 was injected into the corpus cavernosum of the penis. After 4 weeks, the penile tissues of the rats were obtained for immunohistochemistry and Western blot analysis. The immunoreactivity of PECAM-1 and VEGF was increased in the COMP-Ang1 group compared with the vehicle only group. Moreover, the expression of PECAM-1 and VEGF was notably augmented in the 20 microg Comp Ang-1 group. In the immunoblotting study, the expression of PECAM-1 and VEGF protein was significantly less in the OLEFT rats than in the control LETO rats. However, this expression was restored to control level after intracavernosal injection of COMP-Ang1. These results show that an intracavernosal injection of COMP-Ang1 enhances cavernous angiogenesis by structurally reinforcing the cavernosal endothelium.

Effect of acupuncture on pattern-visual evoked potential in rats with monocular visual deprivation / 中国针灸

<p><b>OBJECTIVE</b>To explore electrophysiology mechanism of acupuncture for treatment and prevention of visual deprivation effect.</p><p><b>METHODS</b>Eighteen healthy 15-day Evans rats were randomly divided into a normal group, a model group and an acupuncture group, 6 rats in each one. Deprivation amblyopia model was established by monocular eyelid suture in the model group and acupuncture group. Acupuncture was applied at "Jingming" (BL 1), "Chengqi" (ST 1), "Qiuhou" (EX-HN 7) and "Cuanzhu" (BL 2) in the acupuncture group. The bilateral acupoints were selected alternately, one side for a day, and totally 14 days were required. The effect of acupuncture on visual evoked potential in different spatial frequencies was observed.</p><p><b>METHODS</b>Under three different kinds of spatial frequencies of 2 X 2, 4 X 4 and 8 X 8, compared with normal group, there was obvious visual deprivation effect in the model group where P1 peak latency was delayed (P<0.01) while N1 -P1 amplitude value was decreased (P<0.01). Compared with model group, P1 peak latency was obviously ahead of time (P<0.01) while N1-P1 amplitude value was increased (P<0.01) in the acupuncture group, there was no statistical significance compared with normal group (P>0.05). Under spatial frequency of 4 X 4, N1-P1 amplitude value was maximum in the normal group and acupuncture group. With this spatial frequency the rat's eye had best resolving ability, indicating it could be the best spatial frequency for rat visual system.</p><p><b>CONCLUSION</b>The visual system has obvious electrophysiology plasticity in sensitive period. Acupuncture treatment could adjust visual deprivation-induced suppression and slow of visual response in order to antagonism deprivation effect.</p>

Taurine ameliorates hyperglycemia and dyslipidemia by reducing insulin resistance and leptin level in Otsuka Long-Evans Tokushima fatty (OLETF) rats with long-term diabetes

This study aimed to determine whether taurine supplementation improves metabolic disturbances and diabetic complications in an animal model for type 2 diabetes. We investigated whether taurine has therapeutic effects on glucose metabolism, lipid metabolism, and diabetic complications in Otsuka Long-Evans Tokushima fatty (OLETF) rats with long-term duration of diabetes. Fourteen 50-week-old OLETF rats with chronic diabetes were fed a diet supplemented with taurine (2%) or a non-supplemented control diet for 12 weeks. Taurine reduced blood glucose levels over 12 weeks, and improved OGTT outcomes at 6 weeks after taurine supplementation, in OLETF rats. Taurine significantly reduced insulin resistance but did not improve beta-cell function or islet mass. After 12 weeks, taurine significantly decreased serum levels of lipids such as triglyceride, cholesterol, high density lipoprotein cholesterol, and low density lipoprotein cholesterol. Taurine significantly reduced serum leptin, but not adiponectin levels. However, taurine had no therapeutic effect on damaged tissues. Taurine ameliorated hyperglycemia and dyslipidemia, at least in part, by improving insulin sensitivity and leptin modulation in OLETF rats with long-term diabetes. Additional study is needed to investigate whether taurine has the same beneficial effects in human diabetic patients.

Peroxisome proliferator-activated receptor delta agonist attenuates hepatic steatosis by anti-inflammatory mechanism

Although peroxisome proliferator receptor (PPAR)-alpha and PPAR-gamma agonist have been developed as chemical tools to uncover biological roles for the PPARs such as lipid and carbohydrate metabolism, PPAR-delta has not been fully investigated. In this study, we examined the effects of the PPAR-delta agonist GW0742 on fatty liver changes and inflammatory markers. We investigated the effects of PPAR-delta agonist GW0742 on fatty liver changes in OLETF rats. Intrahepatic triglyceride contents and expression of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and monocyte chemo-attractant protein-1 (MCP-1) and also, PPAR-gamma coactivator (PGC)-1alpha gene were evaluated in liver tissues of OLETF rats and HepG2 cells after GW0742 treatment. The level of TNF-alpha and MCP-1 was also examined in supernatant of Raw264. 7 cell culture. To address the effects of GW0742 on insulin signaling, we performed in vitro study with AML12 mouse hepatocytes. Rats treated with GW0742 (10 mg/kg/day) from 26 to 36 weeks showed improvement in fatty infiltration of the liver. In liver tissues, mRNA expressions of TNF-alpha, MCP-1, and PGC-1alpha were significantly decreased in diabetic rats treated with GW0742 compared to diabetic control rats. We also observed that GW0742 had inhibitory effects on palmitic acid-induced fatty accumulation and inflammatory markers in HepG2 and Raw264.7 cells. The expression level of Akt and IRS-1 was significantly increased by treatment with GW0742. The PPAR-delta agonist may attenuate hepatic fat accumulation through anti-inflammatory mechanism, reducing hepatic PGC-1alpha gene expression, and improvement of insulin signaling.

L-NAC protect hair cells in the rat cochlea from injury of exposure to styrene / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the effects of N-acetyl-L-cysteine (L-NAC) protect hair cells in the rat cochlea from injury of exposure to styrene.@*METHOD@#Seventeen adult Long Evans rats were used in present study. The animals were randomly assigned into test group (n=9) and control group (n=8). The animals were exposed to styrene by gavage at 400 mg/kg (2 g styrene was mixed with 1 ml olive oil). Test group animals received styrene exposure plus L-NAC 325 mg/kg (L-NAC was dissolved in physiological saline solution) by intraperitoneal injection. Treatment was performed once a day, 5 days per week for 3 weeks. Control group animals received the same volume of saline injection on an identical time schedule used for the test group. The auditory brainstem response (ABR) thresholds of both ears elicited with clicks were measured before and at the end of the 3-week styrene or styrene plus L-NAC treatment. After hearing was re-assessed, animals were sacrificed and cochleae were quickly removed from the skull. Following fixation, whole specimens comprising the basilar membrane with Corti's organ were separated from the modiolus. The organs of Corti were stained with propidium iodide (PI) and the TUNEL assay to visualize the morphologic viability of hair cell nuclei, FITC-labeled phalloidin, a F-actin intercalating fluorescent probe used to visualize the morphologic viability of cuticular plate and the stereocilia in the hair cells. Each organ of Corti was thoroughly examined using fluorescence microscopy. The numbers of damaged OHCs (apoptotic, necrotic and missing OHCs) were documented.@*RESULT@#There was a statistically significant decrease in ABR threshold shift (P0.05).@*CONCLUSION@#The results indicate that the treatment with L-NAC may effectively protect against the styrene-induced hair cells damage and preferably reduce the number of apoptotic OHCs.

Hypoadiponectinemia leads to insulin resistance in OLETF rats: a preliminary study / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association between plasma adiponectin and insulin resistance in OLETF rats.</p><p><b>METHODS</b>Twenty male Otsuka Long-Evans Tokushima Fatty (OLETF) rats and 10 male Long-evans Tokushima Otsuka (LETO) rats underwent oral glucose tolerance test (OGTT) at 13 and 40 weeks of age. At 8, 32 and 40 weeks of age, the rats were sacrificed to measure the blood glucose, plasma insulin and adiponectin levels, and serum levels of TG, CHOL and FFA.</p><p><b>RESULTS</b>The plasma adiponectin level was significantly decreased in 8-week-old OLETF rats compared with that of LETO rats (P<0.05). The plasma insulin level, TG, CHOL, and FFA were significantly higher in OLETF rats than in LETO rats at 32 and 40 weeks of age.</p><p><b>CONCLUSION</b>A decreased plasma level of adiponectin preludes insulin resistance and is inversely correlated to insulin sensitivity. Hypoadiponectinemia may be an important reason leading to insulin resistance.</p>

Antidiabetic and hypolipidemic effects of different fractions of coccinia cordifolia L. on normal and streptozotocin-induced diabetic rats

The present study was carried out to observe the antidiabetic and hypolipidemic effects of petroleum-ether, ethyl acetate and chloroform fractions isolated from ethanolic extract of the leaves of Coccinia cordifolia Linn. [150 mg/kg body weight] on normal and streptozotocin [STZ]-induced diabetic rats for one day experiment. Single doses [150 mg/kg, i.p.] of C. cordifolia extracts were given to normal and diabetic rats. The fasting blood glucose [FBG], serum triglyceride [TG] and serum total cholesterol [TC] levels were investigated in normal and STZ-diabetic rats on 0, 1, 2, 3, 6, 10, 16, and 24[th] hours. In normoglycemic rats the pet-ether and ethyl acetate fractions of C. cordifolia reduced blood glucose level significantly [39.66% and 40.68% at 16[th] and 24[th] hour respectively]. In the STZ-diabetic rats pet-ether and ethyl acetate fractions also reduced blood glucose level significantly [50.39% and 50% at 10[th] and 24[th] hour respectively]. Ethyl acetate fraction is most effective which reduced total cholesterol level by 31.04% and 36.69% in normal and STZ-diabetic rats respectively. Ethyl acetate fraction reduced triglyceride level by 43.82% and 42.01% in normal and STZ-diabetic rats respectively. Our results indicate that pet-ether and ethyl acetate fractions of C. cordifolia have potentiality against diabetes

Peroxisome Proliferator-Activated Receptor-Gamma Expression in the Lung Tissue of Obese Rats

PURPOSE: Obesity is a risk factor for asthma and type II diabetes. Peroxisome proliferator-activated receptor (PPAR)-gamma has been suggested to regulate inflammatory responses in diabetes and asthma. We investigated whether PPAR-alpha, PPAR-gamma, adiponectin receptors (AdipoR1, AdipoR2), leptin, and tumor necrosis factor (TNF)-alpha are expressed in rat lung tissues and whether the expression differs between obese Otsuka Long-Evans Tokushima Fatty (OLETF) and lean Long Evans Tokushima Otsuka (LETO) rats. MATERIALS AND METHODS: Obese and lean rats were given with a high fat diet or a 30% restricted diet for 32 weeks, and their blood glucose levels and weights were monitored. After 32 weeks, mRNA levels of PPAR-alpha, PPAR-gamma, AdipoR1, AdipoR2, leptin, and TNF-alpha in lung tissues were measured using real time PCR. RESULTS: PPAR-alpha, PPAR-gamma, AdipoR1, AdipoR2, leptin, and TNF-alpha were expressed in both obese and lean rat lung tissues. Increased serum glucose levels on intraperitoneal glucose tolerance testing and a higher weight gain at 32 weeks were observed in OLETF control rats compared to OLETF diet restricted rats. PPAR-gamma expression was markedly elevated in obese control and diet restricted rats compared to lean rats, although PPAR-gamma expression in obese rats was not affected by diet restriction. Leptin was highly expressed in OLETF rats compared to LETO rats. TNF-alpha expression was enhanced in OLETF control rats compared LETO diet restricted rats, and decreased by diet restriction. PPAR-alpha, AdipoR1, and AdipoR2 expression were not significantly different between obese and lean rats. CONCLUSION: PPAR-gamma was highly expressed in the lung tissues of obese rats and may be a novel treatment target for regulating lung inflammation associated with obesity.

Oral glucose tolerance test [OGTT] in normal control and glucose induced hyperglycemic rats with Coccinia Cordifolia L. and Catharanthus roseus L

The aim of this study is to investigate the hypoglycemic effects of petroleum ether, chloroform and ethyl acetate fractions isolated from ethanolic extracts of Coccinia cordifolia and Catharanthus roseus on normal control and orally glucose-induced hyperglycemic rats. Single doses [150 mg/kg] of different fractions of C. cordifolia and C. roseus extracts were intraperitonelly administered. The serum blood glucose level was obtained by pricking the tail vein using glucometer at time 0, 30, 60, 90, 150 and 270 minutes.In the orally glucose induced hyperglycemic rats, chloroform-coccinia [CHCl[3]-CC] fraction showed maximum reduction of blood glucose level by 21.94% on 60 minute of the experiment. On the other hand maximum reduction [p<0.05] of 17.92% was observed for petroleum ether-catharanthus [PET-CR] on 30 minute of the experiment. Metformin HCl was used as standard drug.Our results indicate that the CHCl[3]-CC fraction is relatively more potent than other fractions of C. cordifolia. Similarly the PET-CR is found to be better than other fractions of catharanthus. Phytochemical screening test results showed that chloroform fraction of C. cordifolia contain saponins and flavonoids compounds, which are known to be hypoglycemic. On the other hand petroleum ether fraction of C. roseus contains tannins, flavonoids and alkaloid compounds produced varying degree of blood sugar reduction. On the pharmacological point of view C. cordifolia and C. roseus appears to be a valuable plant, which can be useful, at least as an adjunct, in the therapy of diabetes

Ultrasound-mediated microbubble delivery of pigment epithelium-derived factor gene into retina inhibits choroidal neovascularization / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Many studies have suggested that the imbalance of angiogenic factor and anti-angiogenic factor expression contributes significantly to the development of choroidal neovascularization (CNV), and ultrasound microbubble combination system can increase the gene transfection efficiency successfully. This study was designed to investigate whether ultrasound-mediated microbubble destruction could effectively deliver therapeutic plasmid into the retina of rat, and whether gene transfer of pigment epithelium-derived factor (PEDF) could inhibit CNV.</p><p><b>METHODS</b>Human retinal pigment epithelial cells were isolated and treated either with ultrasound or plasmid alone, or with a combination of plasmid, ultrasound and microbubbles to approach feasibility of microbubble-enhanced ultrasound enhance PEDF gene expression; For in vivo animal studies, CNV was induced by argon lasgon laser in rats. These rats were randomly assigned to five groups and were treated by infusing microbubbles attached with the naked plasmid DNA of PEDF into the vitreous of rats followed by immediate ultrasound exposure (intravitreal injection); infusing liposomes with the naked plasmid DNA of PEDF into the vitreous (lipofectamine + PEDF); infusing microbubbles attached with PEDF into the orbit of rats with ultrasound irradiation immediately (retrobular injection); infusing microbubbles attached with PEDF into the femoral vein of rats with exposed to ultrasound immediately (vein injection). The CNV rats without any treatment served as control. Rats were sacrificed and eyes were enucleated at 7, 14, and 28 days after treatment. Gene and protein expression of PEDF was detected by quantitative real-time RT-PCR, Western blotting and immunofluorescence staining, respectively. The effect of PEDF gene transfer on CNV was examined by fluorescein fundus angiography.</p><p><b>RESULTS</b>In vitro cell experiments showed that microbubbles with ultrasound irradiation could significantly enhance PEDF delivery as compared with microbubbles or ultrasound alone. In the rat CNV model, transfection efficiency mediated by ultrasound/microbubbles was significantly higher than that by lipofectamine-mediated gene transfer at 28 days after treatment. The study also showed that with the administration of ultrasound-mediated microbubbles destruction, the CNV of rats was inhibited effectively.</p><p><b>CONCLUSIONS</b>Ultrasound-microbubble technique could increase PEDF gene transfer into rats' retina and chorioid, in association with a significant inhibition of the development of CNV, suggesting that this noninvasive gene transfer method may provide a useful tool for clinical gene therapy.</p>

Effect of Diamine Bridges Using L-lysine in Glutaraldehyde Treated Porcine Pericardium / 대한흉부외과학회지

BACKGROUND: Various studies and experimental trials have been done to develop bioprosthetic devices to treat complex congenital heart disease due to the limited usage of homograft tissue. The purpose of the present study was to evaluate the effect of diamine bridges with using L-lysine, as compared with using ethanol. MATERIAL AND METHOD: Porcine pericardium was fixed at 0.625% GA (commercial fixation). An interim step of ethanol (80%; 1 day at room temperature) or L-lysine (0.1 M; 2 days at 37degrees C) was followed by completion of the GA fixation (2 days at 4degrees C and 7 days at room temperature). The tensile strength and thickness of the porcine percardium were measured, respectively. The treated pericardiums were implanted subcutaneously into three-week old Long-Evans rats for 8 weeks. The calcium content of the implants was assessed by atomic absorption spectroscopy and the histology. RESULT: Ethanol pretreatment (13.6+/-10.0 ug/mg, p=0.008), L-lysine pretreatment (15.3+/-1.0 ug/mg, p=0.002), and both treatment (16.1+/-11.1 ug/mg, p=0.012) significantly inhibited calcification, as compared with the controls (51.2+/-8.5 ug/mg). L-lysine pretreatment (0.18+/-0.02 mm, 1.20+/-0.30 kg f/5 mm) significantly increased the thickness and tensile strength, as compared with ethanol pretreatment (0.13+/-0.03 mm, 0.85+/-0.36 1.0 kg f/5 mm) (p<0.01, p=0.035). CONCLUSION: The diamine bridges using L-lysine seemed to decrease the calcification of porcine pericardium fixed with glutaraldehyde, and this was comparable with Ethanol. Additionally, it seemed to enhance the thickness and tensile strength.

Topography and axon arbor architecture in the visual callosal pathway: effects of deafferentation and blockade of TV-methyl-D-aspartate receptors

Visual callosal fibers link cortical loci in opposite hemispheres that represent the same visual field but whose locations are not mirror-symmetric with respect to the brain midline. Presence of the eyes from postnatal day 4 (P4) to P6 is required for this map to be specified. We tested the hypothesis that specification of the callosal map requires the activation of A'-methyl-D-aspartate receptors (NMDARs). Our results show that blockade of NMDARs with MK-801 during this critical period did not induce obvious abnormalities in callosal connectivity patterns, suggesting that retinal influences do not operate through NMDAR-mediated processes to specify normal callosal topography. In contrast, we found that interfering with NMDAR function either through MK801-induced blockade of NMDARs starting at P6 or neonatal enucleation significantly increases the length of axon branches and total length of arbors, without major effects on the number of branch tips. Our results further suggest that NMDARs act by altering the initial elaboration of arbors rather than by inhibiting a later-occurring remodeling process. Since the callosal map is present by P6, just as axonal branches of simple architecture grow into gray matter, we suggest that regulation of arbor development by NMDAR-mediated processes is important for maintaining the precision of this map.

A neutral cue facilitates detection of a visual target by modulating attention

Twelve rats were trained to perform a two-choice visual detection task in which a right or left light was presented and the animals were required to press the lever located under the illuminated light for a food reward. In seventy percent of the trials the target light was preceded by presentation of a neutral cue (a central light). Relevance of the neutral cue for detection of the target was analyzed by comparing behavioral indices of attention in its presence and absence. Accuracy was significantly higher in presence than in absence of the neutral cue, while mean response latencies were lower in presence than in absence of the neutral cue. These results indicate that the animals allocated attentional resources on the target detection during a high expectancy period after the onset of the neutral cue. This could facilitate target detection and improve the performance in the presence of the neutral cue.

Effects of Liuweidihuang pills on plasma adiponectin level in OLETF rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effects of Liuweidihuang (LWDH) pills on plasma adiponectin level in Otsuka Long-Evans Tokushima Fatty (OLETF) rats.</p><p><b>METHODS</b>Forty male OLETF rats were randomized into LWDH and control OLETF groups (n=20), and 10 male Long-Evans Tokushima Otsuka (LETO) rats served as the normal control group (LETO group). The rats in LWDH group were given LWDH pills (daily dose of 2.4 mg/kg) intragastrically since the age of 8 weeks, and the two control groups received water only. Regular blood glucose test was performed using oral glucose tolerance test, and the body weight and food intake of the rats were recorded on a weekly basis. At 8, 32 and 40 weeks of age, respectively, the rats were sacrificed for measurement of plasma adiponectin and plasma insulin.</p><p><b>RESULTS</b>The food intake of the OLETF rats in both groups were significantly greater than the LETO rats (P<0.01). The rats in LWDH group developed diabetes since 30 weeks of age with an incidence of 28.6% at 40 weeks, which was significantly lower than that in the control OLETF rats (P< 0.01).</p><p><b>CONCLUSION</b>Plasma adiponectin level is positively correlated to insulin sensitivity in OLETF rats, in which LWDH pills can increase the plasma level of adiponectin and improve the status of insulin resistance.</p>

Efecto del factor de crecimiento endotelio vascular (VEGF) sobre perforaciones timpánicas en ratas Long-Evans / Effect of vascular endothelial growth factor (VEGF) on tympanic perforations in Long-Evans rats

Introducción: La anglogenesis es el desarrollo de nuevos vasos sanguíneos desde una red vascular existente, contempla una secuencia de eventos complejos y es fundamental en el proceso reparativo. Existen múltiples factores estimulantes de la angiogénesls, entre ellos se encuentran factores de crecimiento como el VEGF (factor de crecimiento endotelio vascular). Debido a su rol reparativo se han utilizado factores proanglogénicos para reparar perforaciones timpánicas. Objetivo: Estudiar el efecto del VEGF sobre perforaciones timpánicas de ratas Long-Evans. Material y método: Se usan 15 ratas adultas, se realizan perforaciones timpánicas bilaterales, se instilan al azar las perforaciones con solución fisiológica y VEGF, se realiza visualización microscópica de los tímpanos a los días 9,15 y 21 posperforación. Las ratas son sacrificadas el día 21 y se realiza estudio histológico del grosor timpánico. Resultados: No se aprecia un efecto inductivo del VEGF sobre el cierre de las perforaciones timpánicas, se produce un aumento en el grosor timpánico de las ratas tratadas con VEGF.

Evaluación del efecto protector de 2 inmunógenos de Trchinella spiralis en ratas Long Evans con modificación nutricional e infectado con Trichinella spiralis / Evaluation of the protective effect of 2 inmunogenos of Trchinella spiralis in rats Long Evans with nutritional modification and infected with Trichinella spiralis

La Trichinellosis es una zoonosis endémica, cosmopolita, sus huéspedes son, ratas, cerdo y otros mamíferos entre ellos el hombre, la presencia de Trichinellosis se debe a la ingestión de carne de cerdo insuficientemente cocinada, afecta a países con bajos recursos económicos. Se han caracterizado Inmunógenos, siendo el inmunodominante el de 45 kDa, efectivo contra T. spiralis, desafortunadamente no ha cristalizado en una vacuna. Se han descrito efectos de desnutrición (DN) sobre órganos linfáticos. Los mecanismos de defensa del huésped son alterados con DN proteico-calórico (DPC). Evaluar el efecto protector de 2 inmunógenos de T. spiralis en ratas Long Evans con modificación Nutricional e infectado con T. spiralis. 80 ratas Long Evans de 30 días de edad, divididas en 2 grupos: 40 Nut con 24 de proteína y 40 DN con 12 de proteína de los cuales se subdividieron en 8 sub grupos con tratamiento: a) 10 animales control b)10 animales infectados con T. spiralis, c) 10 animales inmunizados con antígeno soluble total (AST), d) 10 animales inmunizados con inmunógeno de 45 kDa de T. spiralis (esquema de inmunización una aplicación cada semana por 4 ocasiones), retados a la 1era semana de la culminación de inmunización, sacrificadas a la 6 sexta semana post-infección. Parámetros a evaluar: Determinación de la carga parasitaria mediante la Digestión Artificial (D/A), Determinación de las características morfológicas de la célula nodriza por la técnica de compresión de tejidos, detección de la respuesta inmune por WB. Con la técnica de D/A, se obtuvieron en los diferentes tratamientos, de las ratas Nut, DN, Inmunizadas e Infectadas. Mostró diferencia significativa en la cantidad de Ll que se presentaron en 30gr de músculos estriado, los grupos controles de ratas Nut y DN sin infección, ausencia de Ll, y en los grupos de ratas Nut infectadas, se recolectaron 200 µl de Ll, para el grupo de DN infectado se recolecto 400 µl de Ll, en el grupo de Nut inmunizadas con AST.