Differential effects of treadmill exercise on calretinin immunoreactivity in type 2 diabetic rats in early and chronic diabetic stages

In this study, we investigated the effects of treadmill exercise on calretinin (CR), a marker of early postmitotic neurons, immunoreactivity in the dentate gyrus (DG) of Zucker diabetic fatty (ZDF) rats, before or after diabetes onset, and Zucker lean control (ZLC) rats. For this study, 6-week-old ZLC and prediabetic ZDF rats, and 22-week-old ZLC and ZDF rats were exercised on the treadmill. Sedentary ZLC and ZDF rats of the same age were used as exercise experiment controls. The exercised prediabetic ZDF rats did not show diabetes onset, while the sedentary prediabetic ZDF rats showed significantly increased blood glucose levels. The exercised diabetic ZDF rats exhibited a decrease in their blood glucose levels compared to the sedentary diabetic ZDF rats, but the levels were still above 20 mmol/l. ZLC rats of both ages were in the normoglycemic range. CR immunoreactivity was detected throughout the DG, including the subgranular zone and the polymorphic layer. Diabetic rats exhibited a significant decrease in the number of CR-immunoreactive cells and fibers in the DG. Exercise in the prediabetic ZDF rats significantly increased the number of CR-immunoreactive cells and fibers in the subgranular zone of the DG. In the ZLC and ZDF rats of chronic diabetic phase, exercise increased CR-immunoreactive neurons in the hilar region. These results suggest that diabetes significantly reduces the number of postmitotic CR-immunoreactive neurons and the intensity of immunoreactivity and that exercise increases these CR-related parameters in a diabetic stage-dependent manner.     

Mitochondrial dysfunction influences apoptosis and autophagy in porcine parthenotes developing in vitro

Mitochondria are important regulators of both apoptosis and autophagy. One of the triggers for mitochondrial-mediated apoptosis is the production of reactive oxygen species (ROS), which include hydrogen peroxide, superoxide, hydroxyl radical, nitric oxide and peroxynitrite. Recently, several studies have indicated that ROS may also be involved in the induction of autophagy. In the present study, we used H(2)O(2) to induce mitochondrial stress, examined apoptotic- and autophagic-related gene expression and observed LC3 protein (autophagosome presence marker) expression in porcine parthenotes developing in vitro. In porcine four-cell parthenotes cultured for 5 days in NCSU37 medium containing 0.4% BSA, the developmental rate and mitochondrial distribution did not differ from that of the group supplemented with 100 μM H(2)O(2) but was significantly decreased in the group supplemented with 500 μM H(2)O(2) (P<0.05). Transmission electron microscopy (TEM) indicated that whereas normal shaped mitochondria were observed in blastocysts from the control group, abnormal mitochondria (mitophagy) and autophagic vacuoles were observed in blastocysts from the group that received 500 μM H(2)O(2). Furthermore, addition of H(2)O(2) (100 μM and 500 μM) decreased cell numbers (P<0.05) and increased both apoptosis (P<0.05) and LC3 protein expression in the blastocysts. Real-time RT-PCR showed that H(2)O(2) significantly decreased mRNA expression of anti-apoptotic gene Bcl-xL but increased pro-apoptotic genes, Caspase 3 (Casp3) and Bak, and autophagy-related genes, microtubule-associated protein 1 light chain 3 (Map1lc3b) and lysosomal-associated membrane protein 2 (Lamp2). However, the addition of H(2)O(2) had no effect on mRNA expression levels in nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1, in blastocysts. These results suggest that H(2)O(2) leads to mitochondrial dysfunction that results in apoptosis and autophagy, which is possibly related to porcine early embryo development.     

Reduced immunoreactivity of tyrosine hydroxylase in the hypothalamic paraventricular nucleus of the seizure sensitive gerbil

We compared the immunoreactivity and numbers of tyrosine hydroxylase (TH) immunoreactive neurons and neuropil in the paraventricular nucleus (PVN) of the hypothalamus between the seizure sensitive (SS) and seizure resistant (SR) gerbils. The distributional pattern of TH immunoreactivity was similar in both groups: TH immunoreactivity was seen mainly in magnocellular neurons of the PVN. However, total TH immunoreactivity in the neurons and neuropil in the SS gerbils was significantly lower than that in the SR gerbils. In addition, the number of TH immunoreactive neurons in the SS gerbils was also much lower than those in the SR gerbils. These results indicate that SS gerbils have a low TH immunoreactivity in the hypothalamic PVN compared with that in SR gerbils.     

Changes in glial fibrillary acidic protein immunoreactivity in the dentate gyrus and hippocampus proper of adult and aged dogs

Astrocytes perform neuron-supportive tasks, repair and scarring process in the central nervous system. In this study, we observed glial fibrillary acidic protein (GFAP), a marker for astrocytes, immunoreactivity in the dentate gyrus and hippocampus proper (CA1-3 region) of adult (2-3 years of age) and aged (10-12 years of age) dogs. In the adult group, GFAP immunoreactive astrocytes were distributed in all layers of the dentate gyrus and CA1-3 region, except in the stratum pyramidale of the CA1-3 region. In the aged group, GFAP immunoreactivity decreased markedly in the molecular layer of the dentate gyrus. However, GFAP immunoreactivity in the CA1-3 region increased in all layers, and the cytoplasm of GFAP immunoreactive astrocytes was hypertrophied. GFAP protein levels in the aged dentate gyrus decreased; however, GFAP levels in the CA1-3 region increased. These results suggest that the morphology of astrocytes and GFAP protein levels in the hippocampal dentate gyrus and CA1 region are changed, respectively, with age.     

Paralytic shellfish poison as an attractant for toxic snails

The attracting effect of paralytic shell poison (PSP) on eight toxic snail species (Polinices didyma, Natica lineata, N. vitellus, Zeuxis sufflatus, Niotha clathrata, Oliva miniacea, O. mustelina, O. hirasei) and two non-toxic species (Pomacea canaliculata, Satsuma bairdi) was investigated. Eight toxic species were reported to contain tetrodotoxin, and three out of these contained minor PSP. The minimum lethal dose of PSP in most toxic gastropods was estimated to be more than 150 MU PSP/20 g body weight, but in non-toxic gastropods was less than 15 MU PSP/20 g body weight. After the attracting test, all toxic gastropods showed significant positive linear relationship (Y=3.716X+0.363, r=0.8427) between comparative attracting variation and the toxicity reported. The relationship between PSP resistance ability and the toxicity also had a positive correlation (Y=0.91X+210.090, r=0.6803). However, the non-toxic species showed a negative response. This indicates that PSP is an attractant for toxic snails.     

Validation of multi-frequency inversion method by using dummy scatterers of zooplankton

ABSTRACT:   High- and multi-frequency acoustic measurement systems and the multi-frequency inversion (MFI) method have been used to measure spatial distributions and abundances of zooplankton by size. In this study, the calibration method for high- and multi-frequency systems was developed and the validation of MFI method was carried out by scatterer measurement. The standard sphere calibration method that has not been applied to such high- and multi-frequencies was applied to calibrate our high- and multi-frequency system, TAPS-6 (Tracor Acoustic Profiling System, BAE Systems). An optimum size of standard sphere of tungsten carbide of 1 mm radius was derived to have a small target strength variation for the six frequencies of TAPS-6, and the practicability and precision of the standard sphere calibration method was confirmed for those frequencies. A school or cluster of dummy scatterers of zooplankton with small tungsten carbide spheres were designed to validate the MFI method, and volume back-scattering strength values were measured by the multi-frequency system. By comparing the result of the inversion with their real composition, the features of the MFI method could be validated and examined.     

Comparison of arylalkylamine N-acetyltransferase and melatonin receptor type 1B immunoreactivity between young adult and aged canine spinal cord

Melatonin affects diverse physiological functions through its receptor and plays an important role in the central nervous system. In the present study, we compared immunoreactivity patterns of arylalkylamine N-acetyltransferase (AANAT), an enzyme essential for melatonin synthesis, and melatonin receptor type 1B (MT2) in the spinal cord of young adult (2~3 years) and aged (10~12 years) beagle dogs using immunohistochemistry and Western blotting. AANAT-specific immunoreactivity was observed in the nuclei of spinal neurons, and was significantly increased in aged dog spinal neurons compared to young adult spinal neurons. MT2-specific immunoreactivity was found in the cytoplasm of spinal neurons, and was predominantly increased in the margin of the neuron cytoplasm in aged spinal cord compared to that in the young adult dogs. These increased levels of AANAT and MT2 immunoreactivity in aged spinal cord might be a feature of normal aging and associated with a feedback mechanism that compensates for decreased production of melatonin during aging.     

Effect of muscle type and vacuum chiller ageing period on the chemical compositions,meat quality,sensory attributes and volatile compounds of Korean native cattle beef

The present study demonstrates the effects of different muscle types and chiller ageing periods on the chemical composition, meat quality parameters, sensory characteristics and volatile compounds of Karean native cattle beed. Longissimus dorsi (LD) and Semitendinosus (ST) muscles aged for 7 days and 28 days were used. Moisture, cooking loss, total collagen and Warner‐Bratzler shear force (WBSF) values for the ST were higher than the LD muscle regardless of ageing period (P < 0.05). The LD muscle had higher intramuscular fat (IMF) (P < 0.05). Ageing for 28 days decreased WBSF values whereas it increased thiobarbituric acid of both muscles. Moreover, tenderness, juiciness and flavor scores were significantly higher for the LD muscle at both ageing periods. Increased ageing time improved tenderness of both muscles, and increased juiciness of the LD muscle, whereas there was decreased flavor score of ST muscle (P < 0.05). The majority of the volatile compounds formed from the oxidation of lipids showed differences between the two muscles. Ageing for 28 days increased in the amounts of many volatile compounds; however, the amounts of some important volatile compounds were decreased. These results clearly demonstrate that muscle type and ageing have a potential effect on meat quality, sensory characteristics and volatile profile.     

Cell proliferation and neuroblast differentiation in the dentate gyrus of high-fat diet-fed mice are increased after rosiglitazone treatment

In this study, we determined how rosiglitazone (RSG) differentially affected hippocampal neurogenesis in mice fed a low-fat diet (LFD) or high-fat diet (HFD; 60% fat). LFD and HFD were given to the mice for 8 weeks. Four weeks after initiating the LFD and HFD feeding, vehicle or RSG was administered orally once a day to both groups of mice. We measured cell proliferation and neuroblast differentiation in the subgranular zone of the dentate gyrus using Ki67 and doublecortin (DCX), respectively, as markers. In addition, we monitored the effects of RSG on the levels of DCX and brain-derived neurotrophic factor (BDNF) in hippocampal homogenates. At 8 weeks after the LFD feeding, the numbers of Ki67- and DCX-positive cells as well as hippocampal levels of DCX and BDNF were significantly decreased in the RSG-treated group compared to the vehicle-treated animals. In contrast, the numbers of Ki67- and DCX-positive cells along with hippocampal levels of DCX and BDNF in the HFD fed mice were significantly increased in the RSG-treated mice compared to the vehicle-treated group. Our data demonstrate that RSG can modulate the levels of BDNF, which could play a pivotal role in cell proliferation and neuroblast differentiation in the hippocampal dentate gyrus.