The effect of near-infrared fluorescence conjugation on the anti-cancer potential of cetuximab / 한국실험동물학회지

This study investigated the anti-cancer potential of a near-infrared fluorescence (NIRF) molecule conjugated with Cetuximab (Cetuximab-NIRF) in six-week-old female BALB/c athymic (nu+/nu+) nude mice. A431 cells were cultured and injected into the animals to induce solid tumors. Paclitaxel (30 mg/kg body weight (BW)), Cetuximab (1 mg/kg BW), and Cetuximab-NIRF (0.25, 0.5 and 1.0 mg/kg BW) were intraperitoneally injected twice a week into the A431 cell xenografts of the nude mice. Changes in BW, tumor volume and weight, fat and lean mass, and diameter of the peri-tumoral blood vessel were determined after two weeks. Tumor volumes and weights were significantly decreased in the Cetuximab-NIRF (1 mg/kg BW) group compared with the control group (P < 0.001). Lean mass and total body water content were also conspicuously reduced in the Cetuximab-NIRF (1 mg/kg BW) group compared with the vehicle control group. Peri-tumoral blood vessel diameters were very thin in the Cetuximab-NIRF groups compared with those of the paclitaxel group. These results indicate that the conjugation of Cetuximab with NIRF does not affect the anti-cancer potential of Cetuximab and NIRF can be used for molecular imaging in cancer treatments.

Development of osteoporosis animal model using micropigs / 한국실험동물학회지

Osteoporosis is a known major health problem and a serious disease of the bone, there has been a great need to develop more and newer animal models for this disease. Among animal models used for testing drug efficacy, the minipig model has become useful and effective due to its close similarity with humans (validity), particularly with the pharmacokinetics of compounds via subcutaneous administration, the structure and function of the organs, the morphology of bone and the overall metabolic nature. Based on these advantages, we sought to develop a new animal model of osteoporosis using micropig, which differs from other miniature pigs in the genetic background. Female micropigs were used for the induction of a moderate osteoporosis model by bilateral ovariectomy (OVX) and compared with shamoperated animals. For osteoporosis evaluation, clinical biomarkers such as blood osteocalcin (OSC) and parathyroid hormone (PTH) levels were measured, as well as bone mineral density (BMD) using micro-computed tomography (micro-CT). Compared to sham, OVX animals have decreased blood OSC level, while the blood PTH level increased in blood sera. In addition, we observed the significantly decreased BMDs of tibia region in OVX animals. Based on these results, we report that the micropig model developed in this study can be used to develop a new and effective medical method for diagnosis and treatment of osteoporosis.

Differential regulation of Purkinje cell dendritic spines in rolling mouse Nagoya (tg(rol)/tg(rol)), P/Q type calcium channel (alpha1(A)/Ca(v)2.1) mutant / 대한해부학회지

Voltage dependent calcium channels (VDCC) participate in regulation of neuronal Ca2+. The Rolling mouse Nagoya (Cacna1a(tg-rol) ) is a spontaneous P/Q type VDCC mutant, which has been suggested as an animal model for some human neurological diseases such as autosomal dominant cerebellar ataxia (SCA6), familial hemiplegic migraine and episodic ataxia type-2. Morphology of Purkinje cell (PC) dendritic spine is suggested to be regulated by signal molecules such as Ca2+ and by interactions with afferent inputs. The amplitude of excitatory postsynaptic current was decreased in parallel fiber (PF) to PC synapses, whereas apparently increased in climbing fiber (CF) to PC synapses in rolling mice Nagoya. We have studied synaptic morphology changes in cerebella of this mutant strain. We previously found altered synapses between PF varicosity and PC dendritic spines. To study dendritic spine plasticity of PC in the condition of insufficient P/Q type VDCC function, we used high voltage electron microscopy (HVEM). We measured the density and length of PC dendritic spines at tertiary braches. We observed statistically a significant decrease in spine density as well as shorter spine length in rolling mice compared to wild type mice at tertiary dendritic braches. In proximal PC dendrites, however, there were more numerous dendritic spines in rolling mice Nagoya. The differential regulation of rolling PC spines at tertiary and proximal dendrites in rolling mice Nagoya suggests that two major excitatory afferent systems may be regulated reciprocally in the cerebellum of rolling mouse Nagoya.

Establishment of a Research and Assessment System Using High Quality Non-Human Primates / 한국실험동물학회지

At present, eight non-human primate research facilities exist in Korea to examine the validity and safety of new bio-products, and to generate model animal systems using primates of low health status (low quality primates). However specific-pathogen free (SPF) primates (high quality primates) are the preferred choice for emerging disease studies and for numerous other research areas, including cell/gene therapy, stem cell research, regenerative studies, and brain science. Although international primate centers in developed countries have utilized high quality primate resources for many years, there has been little or no collaboration with less developed countries on primate research. Due to this, the establishment of a high quality primate research capacity is a core priority for the advancement of the biomedical research field in less developed countries. In this study, we investigated the demand for, and opportunities to support the development of this research capability.

Immunolocalization of anion exchanger 1 (Band 3) in the renal collecting duct of the common marmoset

The purpose of this study was to determine the expression and distribution of band 3 in the collecting duct and connecting tubules of the kidney of the marmoset monkey (Callithrix jacchus), and to establish whether band 3 is expressed in type A intercalated cells. The intracellular localization of band 3 in the different populations of intercalated cells was determined by double-labeling immunohistochemistry. Immunohistochemical microscopy demonstrated that band 3 is located in the basolateral plasma membranes of all type A intercalated cells in the connecting tubule (CNT), cortical collecting duct (CCD), and outer medullary collecting duct (OMCD) of the marmoset. However, type B intercalated cells and non-A/ non-B intercalated cells did not show band 3 labeling. Electron microscopy of the CNT, CCD and OMCD confirmed the light microscopic observation of the basolateral plasma membrane staining for band 3 in a subpopulation of interacted cells. Basolateral staining was seen on the plasma membrane and small coated vesicles in the perinuclear structure, some of which were located in the Golgi region. In addition, there was no labeling of band 3 in the mitochondria of the CNT, CCD and in OMCD cells. The intensity of the immunostaining of the basolateral membrane was less in the CNT than in the CCD and OMCD. In contrast, band 3 immunoreactivity was greater in the intracellular vesicles of the CNT. From these results, we suggest that the basolateral Cl-/HCO3- exchanger in the monkey kidney is in a more active state in the collecting duct than in the CNT.

Glutamate and GABA concentrations in the cerebellum of novel ataxic mutant Pogo mice

The Pogo mouse is an autosomal recessive ataxic mutant that arose spontaneously in the inbred KJR/MsKist strain derived originally from Korean wild mice. The ataxic phenotype is characterized by difficulty in maintaining posture and side to side stability, faulty coordination between limbs and trunk, and the consequent inability to walk straight. In the present study, the cerebellar concentrations of glutamate and GABA were analyzed, since glutamate is a most prevalent excitatory neurotransmitter whereas gammar-aminobutyric acid (GABA) is one of the most abundant inhibitory neurotransmitters, which may be the main neurotransmitters related with the ataxia and epilepsy. The concentration of glutamate of cerebellum decreased significantly in ataxic mutant Pogo mouse compared to those of control mouse. However, GABA concentration was not decrease. These results suggested that the decrease in glutamate concentration may contribute to ataxia in mutant Pogo mouse.

Pirfenidone Suppressed the Development of Glomerulosclerosis in the FGS/Kist mouse

Pirfenidone(PFD) is a newly developed anti-fibrotic agent. We evaluated the effect of PFD for the prevention of renal fibrosis using a spontaneous progressive glomerulosclerosis animal model, FGS/Kist mice. Male and female FGS/Kist mice were fed a diet containing 0.5% PFD or the same control diet (CD) without PFD, for 1, 2, or 3-month periods. Body weight was monitored for the general effect of PFD on the mice. Proteinuria and glomerular filtration rate (GFR) were evaluated for renal function. The sclerosis index was examined for the morphological changes. There were no significant changes in body weight between the PFD and control groups in both sexes. Proteinuria levels were low in all the PFD groups compared to the corresponding CD groups. The sclerosis scores were also reduced in both sexes of the 3-month PFD groups (p<0.05), and glomerular filtration rates were increased in both sexes of the 3-month PFD groups compared to the CD groups. The treatment of PFD for 1 or 2-month periods did not have statistic significances but the treatment for 3 months had statistic significances in sclerosis and GFR compared to CD groups. These results suggested that long-term administration of PFD sup-pressed the progression of glomerulosclerosis and improved renal function of the renal function of the FGS/Kist mice.

Morphological Studies of the Neuropeptide Y Immunoreactiive Neurons in the Cerebral Cortex and the Corpus Striatum of Striped Field Mouse (apodemus agrarius coreae) / 대한해부학회지

Neuropeptide Y(NPY) was first isolated from porcine brain. This discovery has led some workers to study the distribution of this peptide in the central nervous system of various mammals. In this study examined the distribution pattern of NPY-immunoreactive (NPY-IR) neurons in the Striped Field Mouse (Apodemus agrarius coreae) cerebral cortex and striatum, using immunohistochemical method. The results obtained in this study were summarized as followings. 1. NPY-IR neurons distributed in all layer of cerebral cortex. The number of neurons were higher in layer V and VI than in layer I and IV. 2. The shape of neurons was predominantly round or oval in layer I and II, and triangular in layer V and VI. And the processes were parallel to pia mater in layer I and II and were vertical in layer III. 3. The highest number of NPY-IR neurons were found in the perirhinal cortex but a few distinct population were found in the retrosplenial cortex. 4. In stiatum NPY-IR neurons were observed only in caudate-putamen nucleus. 5. The Immunoreactive neurons in caudate-putamen varied in their shape, but most of them were triangular or multiform neurons had omnidirectional processes.

The study of dopaminergic immunoreactive cell change in mesencephalon and pons of mongolian gerbil by water deprived day / 대한해부학회지

Mongolian gerbil (Meriones unguiculatus) has been as an model animal for studing the neurologic disease because of the long-term survival in the condition of water-deprived desert condition. In order to accomplish the this research, first of all another divided the laboratory animals 10groups. In this study of the long term water deprived condition investigated catecholamine synthetic enzymes, tyrosine hydroxylase(TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N- methyltransferase(PNMT) in the brain by using immunohistochemical stain. The results obtained in this study were summarized as following. 1. It were observed TH-IR cells in substantia nigra pars compacta, ventral tegmental area and substantia nigra pars reticular of Midbrian. Most of them were presented in pars compacta and ventral tegmental area, but a few in pars reticular. TH-IR cell decreased until the 5th water-deprived day, increased from the 10th water-deprived day to the 15th water-deprived day and redecreased in the 20th water-deprived day 2. In locus ceruleus and rubrospinal tract were observed TH-IR cells and a few DBH-IR cell. Therefore there was composed of dopaminergic neuron and noradrenergic neuron. 3. The quantity of dopamin in serum were decreased until the 4th water-deprived day, increased from the 5th water-deprived day, redecreased on the 15th water-deprived day and reincreased from the 20th water-deprived day.

Prevention of Virus - induced Diabetes by Single Immunization with Recombinant BCG in SJL/J Male Mice / 대한면역학회지

D variant of encephalomyocarditis (EMC-D) virus causes diabetes in susceptible mice by direct infection and cytolysis of pancreatic beta cells. cDNA covering the major outer capsid protein (VP1) of EMC-D virus was cloned into Mycobacterium bovis bacillus Calmette-Guerin (BCG). None of the SJL/J male mice, immunized with live recombinant BCG-VP1, became diabetic when challenged with highly diabetogenic EMC-D virus. But the control mice inoculated with normal BCG or rBCG transformed with vector alone developed diabetes in the same challenge. VP1-specific antibodies including neutralizing antibodies were markedly increased as time went on and reached to the maximum titer at week 10 after a single immunization. The plateau of the titer lasted longer than following 4 weeks. Guinea pigs immunized with the live rBCG-VP1 showed strong delayed type hypersensitivity (DTH) to the VP1of EMC-D virus. It means that the live rBCG-VP1 elicit efficient humoral and cell-mediated imrnune responses against EMC-D virus, resulting in prevention of virus-induced diabetes in susceptible mice.

Changes In The Distribution of Oxytocin and Vasopressin-Immunoreactive Neurons In the Hypothalamic Area of Normal and Hypophysectomized Rats / 체질인류학회지

The localization and number of oxytocin- and vasopressin-immunoreactive neurons (OXY-IR & VP-IR) and their fibers in the hypothalamic areas (supraoptic nucleus, paraventricular nucleus, lateral hypothalamic area and median eminence) of the hypophysectomized rat were compared with normal rats at 6 months of survival after surgery at the light microscopic level. The number of VP-IR neurons was markedly decreased in the supraoptic nucleus (SON) and paraventricular nucleus (PVN) in the hypophysectomized rats as compared to normal rats. Moreover, The number of VP-IR fibers was decresed in the SON, PVN, lateral hypothalamic area (LHA) and median eminence in the hypophysectomized rats. The number of OXY-IR neurons and thier fibers were also decreased in the SON and PVN in the hypophysectomized rats. The present results demonstrate that hypophysectomy induces a significant decrease in the number of OXY- and VPIR neurons and fibers within hypothalamic areas (SON, PVN, and LHA at 6 months of post-hypophysectomy) are decreased.

Immunohistochemical Study on the Vasopressinergic and Oxytocinergic Neurons in the Hypothalamus of water-deprived mongolian gerbil (Meriones unguiculatus) / 대한해부학회지

Mongolian gerbil has been as an model animal for studing the neurological diseases such as stroke and epilepsy because of the congenital incompleteries in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other animal species. In order to accomplish this research, first of all another divided the laboratory animals 5 groups of which each group include the 5 animals. In this study of the long term water deprived condition author investigatied the vasopressinergic and oxytocinergic magnocellular neurons of the hypothalamus by using a quantitative immunohistochemistry, measured the plasma osmolalities at the time of sacrifice of indivisual animals, and the body weights every day during water-deprived. The results obtained in this study were summarized as followings: 1. The body weights and decreasing rates of the body weight in water-deprived animal groups were continuosly decreased. 2. The plasma osmolalities were increased from the 5th water-deprived day, after then the gradually increase reached nearly its equilibrium state at the 10th water-deprived day. 3. Vasopressin and oxytocin immunoreactive cells were mainly observed in PVN, SON and a few in the lateral magnocellular area of hypothalamus. 4. The number of VP immunoreactive cells in paraventricular and supraoptic nucleus were abruptly decreas-ed until the 5th day in the supraoptic nucleus in number and until the 10th day in the paraventricular nucleus of water-deprived. 5. The OT secreting cells were severely decreased on the 5th water deprived day in paraventricular and supraoptic nucleus, after than these cells were very slowly decreased until to the 38th water deprived day.

The Effects of Angiotensin Converting Enzyme Inhibitor on Progressive Glomerular Sclerosis

Almost all advanced glomerular diseases have glomerular sclerotic changes to varying degrees whatever causes their primary glomerular disease are. Pathogenesis of these sclerosis has been thought of as the hyperfiltration in the primary glomerulosclerosis due to development of glomerular hypertension in each insulted glomeruli. This background gave the theoretical bases for antihypertensive therapies for supporting chronic renal insufficient patients. Angiotensin converting enzyme (ACE) inhibitor, one of the antihypertensive drugs, has received attention recently for its effectiveness. The aims of this study determined the effects and mechanism of the ACE inhibitor, enalapril, on the glomerulosclerosis in FGS/NgaKist mice, which was an animal model of chronic renal failure by generating spontaneously heavy proteinuria and progressive glomerulosclerosis. Five-week-old FGS/NgaKist mice (n=38) were assigned to four groups. Group 1a (n=6) and group 2a (n=8) fed with a vehicle, were sacrificed at the end of 10 weeks and 15 weeks, respectively. Group 1b (n=12) and 2b (n=12) received enalapril (100 mg/L) in drinking water for 5 weeks and 10 weeks from 6th week of age respectively, and were sacrified on the same day as the control groups. Doses of enanapril were maintained to 2 mg/kg/day by measuring the amount of water consumption. In enalapril groups 1b and 2b, systemic blood pressure (74.7 14.0 mm Hg, 74.3 15.9 mmHg) were significantly lower than control group 2a (116.1 4.6 mmHg, P<0.001). Similarly, degree of proteinuria lowered in enalapril group 2b versus control group 2a (0% and 50.0%, P<0.001). Glomerulosclerosis percentage significantly decreased (P<0.001) (group 1b and 2b; 1.9 6.5, 5.6 7.0 vs control 1a and 2a; 32.8 15.5, 31.4 13.8). Glomerulosclerosis score also decreased (P<0.001) (group 1b and 2b; 0.02 0.08 vs control 1a and 2a; 0.48 0.12, 0.30 0.14). The immunofluorescent staining of enalapril groups showed negative for mesangial deposition of IgG, IgA, IgM, and C3 which were positive in control groups. Immunohistochemical staining with TGF-beta1 was negative in enalapril groups and sclerotic glomeruli both enalapril groups and control groups. These results support that the ACE inhibitor has a renoprotective effect on glomerulosclerosis not only by decreasing the blood pressure but also by suppressing the immune deposits on glomeruli.

Expression of Transforming Growth Factor-beta and Morphologic Changes of Glomerulosclerosis in FGS/NgaKist Mouse

Focal segmental glomerulosclerosis (FSGS) is presented as not only one of the primary glomerular diseases but also as a secondary phenomenon for chronic irreversible renal diseases. The main pathological feature of FSGS is the accumulation of extracellular matrix in the glomeruli, for which overexpression of transforming growth factor-beta (TGF-beta) may be responsible for the accumulation of pathological matrix. A new animal model (FGS/NgaKist mouse) of renal failure by spontaneously generating glomerulosclerosis was developed. To elucidate the role of TGF-beta for FSGS, authors observed glomeruli of FGS/NgaKist mouse periodically. FGS/NgaKist mouse strain showed progression of proteinuria and focal glomerular sclerosis with the aging. The glomeruli showed anti IgG, IgA, IgM and complement complex deposits and extracellular matrix accumulation in the mesangium. TGF-beta mRNA and beta2antibody expressions were increased with the advance of glomerular sclerosis. The results suggest the following; FSGS of FGS/NgaKist strain is immune mediated disease and this stimuli on mesangial or endothelial cells may activate TGF-beta gene in their nuclei. This activation, in turn, can cause sclerosis by increasing TGF-beta mRNA transcription followed by secretion of TGF-beta and its action as cytokine for making collagen fibrils.

Antitumor Effect of Methotrexate in SCID Mice with Human Leukemia CCRF-CEM Cell Line / 대한암학회지

No abstract available