The distribution of the nerve growth factor (NGF), the glial fibrillary acidic protein (GFAP) and the ciliary neurotrohic factor (CNTF) was performed in coronal sections of the mesencephalon, rhombencephalon and spinal cord in the developing Mongolian gerbils. Generally, NGF specifically recognizes neurons with the NGF receptor, whereas GFAP does the glia, and CNTF does the motor neurons. The receptor expression was examined separately in gerbils between embryonic days 15 (E15) and postnatal weeks 3 (PNW 3). The NGF-IR was first observed in the spinal cord at E21, which might be related to the maturation. The GFAP reactivity was peaked at the postnatal days 2 (PND2), while the highest CNTF-reaction was expressed at PNW 2. The GFAP stains were observed in the aqueduct and the spinal cord, which appeared to project laterally at E19. The CNTF was observed only after the birth and found in both the neurons and neuroglia of the substantia nigra, mesencephalon, cerebellum and the spinal cord from PND1 to PNW3. These results suggest that NGF, GFAP and CNTF are important for the development of the neurons and the neuroglia in the central nervous system at the late prenatal and postnatal stages.
Animals ; Brain Stem/enzymology/*metabolism ; Ciliary Neurotrophic Factor/*metabolism ; Embryonic and Fetal Development/physiology ; Female ; Gerbillinae/*embryology ; Glial Fibrillary Acidic Protein/*metabolism ; Immunohistochemistry/veterinary ; Mesencephalon/embryology/metabolism ; Nerve Growth Factor/*metabolism ; Pregnancy ; Rhombencephalon/embryology/metabolism ; Spinal Cord/embryology/*metabolism

The regional distributions and relative frequencies of some gastrointestinal endocrine cells in the three portions (cecum, colon and rectum) of the large intestinal tract of C57BL/6 mice were examined with immunohistochemical method using 7 types of specific antisera against chromogranin A (CGA), serotonin, somatostatin, human pancreatic polypeptide (HPP), glucagon, gastrin and cholecyctokinin (CCK)-8. In this study, all 3 types of immunoreactive (IR) cells were identified. Most of these IR cells in the large intestinal portion were generally spherical or spindle in shape (open-typed cell) while cells with a round shape (close-typed cell) were found in the intestinal gland. Their relative frequencies varied according to each portion of the large intestinal tract. CGA-IR cells were found throughout the whole large intestinal tract but were most predominant in the colon. Serotonin-IR cells were detected throughout the whole large intestinal tract and showed highest frequency in the colon. Peculiarly, glucagon-IR cells were restricted to the colon with a low frequency. However, no somatostatin-, HPP-, gastrin- and CCK-8-IR cells were found in the large intestinal tract. In conclusion, some peculiar distributional patterns of large intestinal endocrine cells were identified in C57BL/6 mice.
Animals ; Chromogranin A ; Chromogranins/metabolism ; Enteroendocrine Cells/*metabolism/physiology ; Female ; Gastrins/metabolism ; Glucagon/metabolism ; Immunohistochemistry/veterinary ; Intestine, Large/*cytology/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Pancreatic Polypeptide/metabolism ; Serotonin/metabolism ; Sincalide/metabolism ; Somatostatin/metabolism

To develop a better model of isolated perfused heart, a new apparatus of "coronary artery cannula- fixed-in-aortic tube" was developed for continuous normothermic perfusion and compared to the Casalis apparatus with cold ischemia. Eight mongrel pigs with the body weight of 18 to 24 kg were divided half into two groups.All the continuous perfusion experimental hearts resumed a spontaneous heart beat and stabilized earlier than the control hearts without the need of defibrillator or pacemaker, indicating no reperfusion injury on the heart. All the experimental hearts did not show fibrillation nor stopped beating during the entire experiment, whereas the control hearts fibrillated. Two control hearts stopped beating, and only one of the two survived with the help of pacemaker.The coronary systolic, diastolic, and mean pressures were more stable with low variation in the experimental hearts than the cold ischemic control hearts. The experimental hearts consumed more oxygen than the control hearts, indicating more cardiac output.According to these results, the continuous normothermic perfusion method by the new cannula, even though with a short-period of hypothermic perfusion, provided better myocardial protection than the cold ischemia.
Animals ; Blood Gas Analysis/veterinary ; Blood Pressure ; Cardioplegic Solutions/therapeutic use ; Catheterization/*methods/veterinary ; Heart/*physiology ; Heart Rate ; Perfusion/*methods/veterinary ; Swine/*physiology/surgery

The purpose of this study was to estimate predictive markers of intrinsic radiosensitivity in individuals who were exposed to occupational or environmental radiation. Throughout this process, the actual biohazard risks and base-line chromosome damage were evaluated in human population. Further studies were carried out to provide evidence for the existence of individual variations in age-dependent responses through micronuclei (MN) assay.Spontaneous frequencies not only vary greatly between individuals, but also working or living areas. It was shown that the increased level of spontaneous cell with MN was observed with increasing age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in an inverse proportion. Ionizing radiation may be targeted mutagenic effects at the usual exposures of background levels that populations were exposed. Age and gender are the most important demographic variables in determining the MN index with frequencies in females, which were greater than those in males. The main life-style factors influencing the MN index in subjects were correlated significantly and positively with smoke. The results showed that an indicator of the genetic damaged rate in MN index in human populations significantly correlated with age, sex and life-style factors. So far, it is evident that with regard to the application of MN assay all future studies have to take into account the influence of age, gender, and life-style.In Conclusion, using micronuclei assay technique a large population can be easily monitored. This study illustrated that the MN assay may provide a high potential to ensure appropriate quality control and standard documentation protocol that can be used to monitor a large population exposed to radiation epidemiologically.
Adolescent ; Adult ; Age Factors ; Aged ; Background Radiation/*adverse effects ; Chromosomes, Human/radiation effects ; Environmental Exposure/*adverse effects ; Female ; Humans ; Korea ; Life Style ; Lymphocytes/*radiation effects/ultrastructure ; Male ; Micronuclei, Chromosome-Defective/*radiation effects ; *Micronucleus Tests ; Middle Aged ; Occupational Exposure/*adverse effects ; Radiation Tolerance/*physiology ; Sex Factors

Interleukin-2 plays an important role in T lymphocyte proliferation and immune response regulations. In this study, porcine IL-2 cDNA was cloned from peripheral blood mononuclear cells, and recombinant porcine IL-2 (rpIL-2) was expressed in Escherichia coli. The size of rpIL-2 without signal peptides was about 15 kDa when determined by SDS-PAGE and Western blotting analysis. Anti-rpIL-2 antibody was produced from mice immunized with the purified rpIL-2, and its specificity was examined by Western blotting and ELISA. In the Western blotting assay, anti-rpIL-2 and anti-recombinant human IL-2 (rhIL-2) antibodies specifically recognized rpIL-2 and rhIL-2, respectively. However, anti-rpIL-2 antibody did not recognize rhIL-2, and anti-rhIL-2 antibody also did not react with rpIL-2 in the same assay. In ELISA, anti-rpIL-2 antibody strongly interacted with both rpIL-2 and rhIL-2, and anti-rhIL-2 antibody also efficiently recognized both proteins. Taken together, the specificity of anti-rpIL-2 antibody for rpIL-2 was demonstrated by Western blotting and ELISA. It was also shown that ELISA is more efficient than Western blotting in determining the species cross-reactivity of anti-rpIL-2 antibody.
Animals ; Antibodies/diagnostic use/*metabolism ; Antibody Specificity ; Blotting, Western/veterinary ; Cloning, Molecular ; Cross Reactions ; Electrophoresis, Polyacrylamide Gel/veterinary ; Enzyme-Linked Immunosorbent Assay/*methods/veterinary ; Escherichia coli/genetics/metabolism ; Humans ; Interleukin-2/biosynthesis/genetics/*immunology ; Mice ; Recombinant Proteins/biosynthesis/genetics/immunology ; Species Specificity ; Swine

This study was performed to examine the influence of isoflurane anesthesia on the pulsatility index (PI) and the peak systolic velocity (PSV) of the blood flow in the basilar artery of dogs by duplex Doppler ultrasonography. Twelve healthy dogs were used to measure the PI and the PSV under the conscious state and isoflurane anesthesia. The pulsatility index (PI) and the peak systolic velocity (PSV) in the basilar artery were measured five times with random intervals. The blood pressure was measured. The PI and PSV values in dogs under isoflurane anesthesia were 1.37 +/- 0.32 and 72 +/- 19 cm/sec, whereas those in the conscious dogs were 1.37 +/- 0.13 and 81 +/- 16 cm/sec, respectively. The indirect mean arterial systolic and diastolic pressures under isoflurane anesthesia were 107 and 51 mmHg, whereas those in the conscious dogs were 133 and 74 mmHg. Though the isoflurane is generally known to induce hypotension, there were no significant differences in the PI and PSV between the isoflurane-anesthetized and the conscious dogs. In conclusion, the isoflurane anesthesia did not influence the PI and PSV in the basilar artery of dogs.
Anesthetics, Inhalation/*pharmacology ; Animals ; Basilar Artery/drug effects/*ultrasonography ; Blood Flow Velocity/drug effects/veterinary ; Blood Pressure/drug effects ; Cerebrovascular Circulation/drug effects ; Dogs/*physiology ; Female ; Isoflurane/*pharmacology ; Male ; Pulsatile Flow/drug effects ; Ultrasonography, Doppler, Duplex/veterinary

The effects of electroacupuncture (EA) on the minimum alveolar concentration (MAC) and on the cardiovascular system were evaluated with dogs under isoflurane anesthesia. Eight healthy male beagles were randomly assigned to six study groups (five heads/group) with washout intervals of 7 ~ 31 days between experiments for recovery and anesthetic clearance. MAC of isoflurane and cardiovascular parameters were determined after EA at nonacupoint and and at acupoints LI-4, SP-6, ST-36 and TH-8. Electroacupuncture for 30 minutes at LI-4, SP-6, ST-36 and TH-8 acupoints lowered the MAC of isoflurane by 17.5 +/- 3.1%, 21.3 +/- 8.0%, 20.5 +/- 8.2% and 15.6 +/- 3.1%, respectively (p < 0.05). However, electrical stimulation of nonacupoint did not induce a significant change in MAC of isoflurane. In the cardiovascular system, the ST-36 group did not induce any significant change in cardiovascular parameters. In the TH-8 group, the mean and diastolic arterial pressure and the systemic vascular resistance were decreased. In the LI-4 group, cardiac output and cardiac index decreased after EA. These results indicate that EA at LI-4, SP-6 and ST-36 have advantages in isoflurane anesthesia in terms of reducing the dose of anesthetics and minimizing cardiovascular side effects.
Anesthesia, Inhalation/adverse effects/*veterinary ; Anesthetics, Inhalation/*pharmacokinetics/pharmacology ; Animals ; Blood Pressure/drug effects ; Cardiac Output ; Dogs/*metabolism/physiology ; Electroacupuncture/*veterinary ; Heart Rate/drug effects ; Isoflurane/*pharmacokinetics/pharmacology ; Male ; Pulmonary Alveoli/*metabolism ; Pulmonary Wedge Pressure/drug effects ; Random Allocation ; Vascular Resistance/drug effects

Using site-directed mutagenesis technique, I have replaced serine 285 and serine 292 with the alanine, and assessed the binding of agonist and signaling such as the inhibition of adenylyl cyclase activity.I have found that serine 292 has an important role in the signal transduction of cannabinoid agonists, HU-210 and CP55940, but not in that of aminoalkylindoles derivatives WIN55,212-2. All mutants express well in protein level determined by western blot using monoclonal antibody HA 11 as compared with the wild type receptor.Interestingly, binding affinity of S285A and S292A mutants with classical cannabinoid agonist HU-243 was somewhat decreased. In signaling assay, the inhibition of adenylyl cyclase by HU-210, CP55940 and WIN55, 212-2 is the same order in both wild type receptor and S285A mutant receptor. However, S292A have been shown that the inhibition curves of adenylyl cyclase activity moved to the right by HU-210 and CP55940, but those of adenylyl cyclase activity did not by aminoalkylindole WIN55,212-2, which is indicating that this residue is closely related to the binding site with HU-210 and CP55940. In addition, serine 292 might take more important role in CB2 receptor and G-protein signaling than serine 285.
Adenylate Cyclase/*metabolism ; Animals ; Binding, Competitive ; Blotting, Western ; COS Cells ; Cannabinoids/metabolism ; Cercopithecus aethiops ; Cyclohexanols/metabolism ; Excitatory Amino Acid Antagonists/metabolism ; Mutagenesis, Site-Directed ; Protein Conformation ; Protein Structure, Tertiary ; Receptors, Cannabinoid ; Receptors, Drug/genetics/metabolism/*physiology ; Serine/metabolism/*physiology ; Signal Transduction/physiology ; Tetrahydrocannabinol/*analogs&derivatives/metabolism ; Transfection

It is now well established that several G protein- coupled receptors can signal without agonist stimulation (constitutive receptors). Inverse agonists have been shown to inhibit the activity of such constitutive G protein-coupled receptor signaling. Agonist activation of the Gi/o-coupled peripheral cannabinoid receptor CB2 normally inhibits adenylyl cyclase type V and stimulates adenylyl cyclase type II. Using transfected COS cells, we show here that application of SR144528, an inverse agonist of CB2, leads to a reverse action (stimulation of adenylyl cyclase V and inhibition of adenylyl cyclase II). This inverse agonism of SR144528 is dependent on the temperature, as well as on the concentration of the cDNA of CB2 transfected. Pertussis toxin blocked the regulation of adenylyl cyclase activity by SR 144528.
Adenylate Cyclase/antagonists&inhibitors/genetics/metabolism ; Animals ; Binding, Competitive ; Bornanes/metabolism/*pharmacology ; COS Cells ; Cannabinoids/metabolism ; Cercopithecus aethiops ; Isoenzymes/antagonists&inhibitors/genetics/metabolism ; Pyrazoles/metabolism/*pharmacology ; Rats ; *Receptor, Cannabinoid, CB2 ; Receptors, Cannabinoid ; Receptors, Drug/agonists/*antagonists&inhibitors/genetics/metabolism ; Signal Transduction/drug effects/physiology ; Transfection

Secretion of neurotransmitters is initiated by voltagegated calcium influx through presynaptic, voltage- gated N-type calcium channels. However, little is known about their cellular distribution in the mouse cerebellum. In the cerebellum, alpha1B immunoreactivity is found mainly on the cell bodies of all Purkinje cells. In addition, the immunoreactivity was detected on a subset of Purkinje cell dendrites, clustered to form a parasagittal array of bands. In the anterior lobe vermis, immunoreactive Purkinje cell dendrites form narrow stripes separated by broad bands of unstained dendrites. Moving caudally through the vermis, these stripes become thicker as a larger fraction of the Purkinje cell dendrites become immunoreactive. This localization study of the alpha1B pore-forming subunits in mouse cerebellum may guide future investigations of the role of calcium channels in neurological pathways.
Animals ; Calcium Channels, N-Type/*metabolism ; Cerebellum/cytology/*metabolism ; Dendrites/metabolism ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Purkinje Cells/metabolism