Viral load and the duration of viral shedding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are important determinants of the transmission of coronavirus disease 2019.In this study, we examined the effects of viral doses on the lung and spleen of K18-hACE2 transgenic mice by temporal histological and transcriptional analyses. Approximately, 1×105 plaque-forming units (PFU) of SARS-CoV-2 induced strong host responses in the lungs from 2 days post inoculation (dpi) which did not recover until the mice died, whereas responses to the virus were obvious at 5 days, recovering to the basal state by 14 dpi at 1×102 PFU. Further, flow cytometry showed that number of CD8+ T cells continuously increased in 1×102 PFU-virusinfected lungs from 2 dpi, but not in 1×105 PFU-virus-infected lungs. In spleens, responses to the virus were prominent from 2 dpi, and number of B cells was significantly decreased at 1×105PFU; however, 1×102 PFU of virus induced very weak responses from 2 dpi which recovered by 10 dpi. Although the defense responses returned to normal and the mice survived, lung histology showed evidence of fibrosis, suggesting sequelae of SARS-CoV-2 infection. Our findings indicate that specific effectors of the immune response in the lung and spleen were either increased or depleted in response to doses of SARS-CoV-2. This study demonstrated that the response of local and systemic immune effectors to a viral infection varies with viral dose, which either exacerbates the severity of the infection or accelerates its elimination.

Klippel-Trenaunay syndrome is a rare congenital mesodermal abnormality characterized by varicose veins, cutaneous hemangiomas, soft tissue and bony hypertrophy of limb. Potential complications such as deep venous thrombosis and pulmonary thromboembolism have not been reported in Korea to date. We demonstrate the case of a 48-year-old woman with Klippel-Trenaunay syndrome with extensive varicose veins on right lower limb, hypertrophy of left big toe and basilar artery tip aneurysm, complicated with acute submassive pulmonary thromboembolism treated successfully with intravenous thrombolytic therapy.
Aneurysm ; Basilar Artery ; Extremities ; Female ; Heart Failure ; Hemangioma ; Humans ; Hypertrophy ; Intracranial Aneurysm ; Klippel-Trenaunay-Weber Syndrome* ; Korea ; Lower Extremity ; Mesoderm ; Middle Aged ; Pulmonary Embolism* ; Thrombolytic Therapy* ; Toes ; Varicose Veins ; Venous Thromboembolism ; Venous Thrombosis

PURPOSE: We designed a water-based bolus device for radiation therapy in Kaposi's sarcoma. This study evaluated the usefulness of this new device and compared it with the currently used rice-based bolus. MATERIALS AND METHODS: We fashioned a polystyrene box and cut a hole in order to insert patient's extremities while the patient was in the supine position. We used a vacuum-vinyl based polymer to reduce water leakage. Next, we eliminated air using a vacuum pump and a vacuum valve to reduce the air gap between the water and extremities in the vacuum-vinyl box. We performed CT scans to evaluate the density difference of the fabricated water-based bolus device when the device in which the rice-based bolus was placed directly, the rice-based bolus with polymer-vinyl packed rice, and the water were all put in. We analyzed the density change with the air gap volume using a planning system. In addition, we measured the homogeneity and dose in the low-extremities phantom, attached to six TLD, and wrapped film exposed in parallel-opposite fields with the LINAC under the same conditions as the set-up of the CT-simulator. RESULTS: The density value of the rice-based bolus with the rice put in directly was 14% lower than that of the water-based bolus. Moreover, the value of the other experiments in the rice-based bolus with the polymer-vinyl packed rice showed an 18% reduction in density. The analysis of the EDR2 film revealed that the water-based bolus shows a more homogeneous dose plan, which was superior by 4.0~4.4% to the rice-base bolus. The mean TLD readings of the rice-based bolus, with the rice put directly into the polystyrene box had a 3.4% higher density value. Moreover, the density value in the case of the rice-based bolus with polymer-vinyl packed rice had a 4.3% higher reading compared to the water-based bolus. CONCLUSION: Our custom-made water-based bolus device increases the accuracy of the set-up by confirming the treatment field. It also improves the accuracy of the therapy owing to the reduction of the air gap using a vacuum pump and a vacuum valve. This set-up represents a promising alternative device for delivering a homogenous dose to the target volume.

BACKGROUND: As the result of renal transplantation improving, also increasing the number of graft failure which will be a candidate for second renal transplantation. The purpose of this study is to evaluate the factors that influence the survival of retransplanted kidney. METHODS: Among 775 renal transplantations that have been performed in Dongsan Medical Center until August 2007, 225 cases were failed their graft function and 59 of them were retransplanted during their follow up period. Graft survival of retransplanted kidney was compared with primary renal transplantation and factors that affecting the survival of kidney retransplantation were evaluated. RESULTS: Main causes of graft failure of first kidney transplantation were chronic rejection, followed by recurrence of original disease of recipient and acute vascular rejection. Mean survival time was 72.6 months (15 days~161 months). One and 5 years graft survivals were 94.6%, 90.7%, and patient survivals were 100.0%, 97.8%, respectively. Among the factors which showed significance in univariate analysis, short interval between failure of first transplantation and retransplantation, and graft failure due to chronic rejection were statistically significant unfavorable factors for survival of retransplanted kidney. CONCLUSIONS: Kidney retransplantation showed similar graft and patient survival compare to the first one. However, retransplantation should be performed after enough time after graft failure and should be cautious in a patient who lost their graft due to chronic rejection.
Follow-Up Studies ; Graft Survival ; Humans ; Kidney ; Kidney Transplantation ; Recurrence ; Rejection (Psychology) ; Survival Rate ; Transplants

BACKGROUND AND OBJECTIVES: Tympanic membrane perforation is an important clinical problem found in various populations of the world. In large number of cases, acute traumatic perforations heal spontaneously, and in the healing process, stem cells appear to play an important role. However, no studies have been reported regarding somatic stem cells in the tympanic membrane. Herein, we tried to show that guinea pig's tympanic membrane contains cells that display the characteristic features of stem cells. MATERIALS AND METHOD: The tympanic membrane was obtained from the guinea pig. The cells were cultured in a medium with epidermal growth factor (EGF) and fibroblast growth factor (FGF). Proliferating cells were checked with stem cell markers, bromodeoxyuridine (BrdU) and nestin. Differentiated cells from stem cells are checked with betaIII tubulin and S-100. RESULTS: We observed that some of the cultured cells from the tympanic membrane were stained with both stem cell markers, BrdU and nestin. And we observed that these cells differentiated into neuron and gilal cells, which expressed betaIII tubulin and S-100, respectively. CONCLUSION: These results suggest that the tympanic membrane of guinea pigs may have neural stem cells. Further study is needed for finding the origin of stem cells.
Adult ; Adult Stem Cells ; Animals ; Bromodeoxyuridine ; Cells, Cultured ; Epidermal Growth Factor ; Fibroblast Growth Factors ; Guinea ; Guinea Pigs ; Humans ; Intermediate Filament Proteins ; Nerve Tissue Proteins ; Neural Stem Cells ; Neurons ; Stem Cells ; Tubulin ; Tympanic Membrane ; Tympanic Membrane Perforation

BACKGROUND AND OBJECTIVES: Neural cell adhesion molecule (NCAM) and polysialic acid (PSA) function basically in cell adhesion and migration. In neural development, they are closely associated with axon pathfinding, synaptogenesis, neural cell migration, differentiation and myelination. The purpose of this study is to assess expression of NCAM and PSA expression in spiral ganglion neurons and Schwann cells and to postulate their functions. MATERIALS AND METHOD: Guinea pig spiral ganglion cells were harvested and cultured in vitro. The cells were grown and differentiated in culture medium together with brain derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3) and glial cell derived neurotrophic factor (GDNF). After 1 week of culturing, the cells were fixed and immunocytochemical staining with beta-III tubulin, S-100, polysialic acid (PSA) and neural cell adhesion molecule (NCAM) were performed. We then checked axon growth rate with Axon Analyzer System(R). RESULTS: In the spiral ganglion culture, cultured neurons showed positive staining for beta-III tubulin, NCAM, and different expressions of PSA. S-100 positive glial cells (Schwann cells) showed different expressions of NCAM and no expression of PSA. Some NCAM positive neurons and Schwann cells were in contact each other. The growth rate of neuron was about 10-30 micrometer/h using Axon Analyzer System(R). CONCLUSION: We postulated that NCAM may play an important role in neural cell adhesion, myelination, fasciculation and ganglion formation. But PSA did not express the adhesive function of NCAM ; its absence may have been due to developmental reason. The differential expression of NCAM in the Schwann cells may indicate its different immunocytochemical characteristics and functions as shown in the CNS glial cells, astrocytes and oligodendrocytes.
Adhesives ; Animals ; Astrocytes ; Axons ; Brain-Derived Neurotrophic Factor ; Cell Adhesion ; Cell Movement ; Fasciculation ; Ganglion Cysts ; Guinea Pigs ; Myelin Sheath ; Neural Cell Adhesion Molecules* ; Neuroglia ; Neurons* ; Neurotrophin 3 ; Oligodendroglia ; Schwann Cells ; Spiral Ganglion* ; Tubulin

Ion channel inhibitors are widely used for pharmacological discrimination between the different channel types as well as for determination of their functional role. In the present study, we tested the hypothesis that 4-aminopyridine (4-AP) could affect the large conductance Ca2+ -activated K+ channel (BKCa) currents using perforated-patch or cell-attached configuration of patch-clamp technique in the rabbit pulmonary arterial smooth muscle. Application of 4-AP reversibly inhibited the spontaneous transient outward currents (STOCs). The reversal potential and the sensitivity to charybdotoxin indicated that the STOCs were due to the activation of BKCa. The BKCa currents were recorded in single channel resolution under the cell-attached mode of patch-clamp technique for minimal perturbation of intracellular environment. Application of 4-AP also inhibited the single BKCa currents reversibly and dose-dependently. The membrane potential of rabbit pulmonary arterial smooth muscle cells showed spontaneous transient hyperpolarizations (STHPs), presumably due to the STOC activities, which was also inhibited by 4-AP. These results suggest that 4-AP can inhibit BKCa currents in the intact rabbit vascular smooth muscle. The use of 4-AP as a selective voltage-dependent K+ (KV) channel blocker in vascular smooth muscle, therefore, must be reevaluated.
4-Aminopyridine* ; Charybdotoxin ; Discrimination (Psychology) ; Ion Channels ; Membrane Potentials ; Muscle, Smooth* ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle* ; Patch-Clamp Techniques ; Pulmonary Artery

BACKGROUND: This study was designed to determine the relationship of propranolol pharmacokinetic parameters with portosystemic shunt in CCl4-induced cirrhotic rats. METHODS: Cirrhotic rats(n=6) were induced by intramuscular injection of CCl4 in olive oil(two time per weeks) for 12 weeks. Controls (n=6) were injected intramuscularly with the same dose of olive oil for 12 weeks. We evaluated the amount of portosystemic shunt by thallium-201 per rectal scintigraphy. After intravenous bolus injection of propranolol (2mg/kg) to rats, the serum propranolol concentrations were analyzed by a HPLC-fluorimetric detector system. Pharmacokinetic parameters such as C0, AUC, t(1/2(beta)), and CLp were determined in each group. Then, a small amount of heptic tissue was obtained and subjected to determination of the hepatic collagen content by quantitating 4-hydroxyproline and were inspected by microscope after hematoxylin and eosin stain. RESULTS: In liver biopsy, liver fibrosis progressed in CCl4-induced cirrhotic rats. The serum concentrations of propranolol were significantly (p < 0.01) elevated in CCl4-induced cirrhotic rats. Mean amount of 4-hydroxyproline, mean H/L ratio, and mean AUC in CCl4-induced cirrhotic rats was significantly (p < 0.01) higher than that in control rats. There was a relationship between AUC, H/L ratio, and amount of 4-hydroxyproline. CONCLUSION: H/L ratio may help in the selection of drug dosage (especially blood flow dependent drug) in pre-clinical studies for chronic liver disease during the drug development process.
Animals ; Carbon Tetrachloride Poisoning/*complications ; Chromatography, High Pressure Liquid ; English Abstract ; Liver Cirrhosis, Experimental/*metabolism/physiopathology ; Portal System/physiopathology ; Propranolol/*pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Thallium Radioisotopes/diagnostic use

L-type Ca2+ channels play an important role in regulating cytosolic Ca2+ and thereby regulating hormone secretions in neuroendocrine cells. Since hormone secretions are also regulated by various kinds of protein kinases, we investigated the role of some kinase activators and inhibitors in the regulation of the L-type Ca2+ channel currents in rat pituitary GH3 cells using the patch-clamp technique. Phorbol 12,13-dibutyrate (PDBu), a protein kinase C (PKC) activator, and vanadate, a protein tyrosine phosphatase (PTP) inhibitor, increased the Ba2+ current through the L-type Ca2+ channels. In contrast, bisindolylmaleimide I (BIM I), a PKC inhibitor, and genistein, a protein tyrosine kinase (PTK) inhibitor, suppressed the Ba2+ currents. Forskolin, an adenylate cyclase activator, and isobutyl methylxanthine (IBMX), a non-specific phosphodiesterase inhibitor, reduced Ba2+ currents. The above results show that the L-type Ca2+ channels are activated by PKC and PTK, and inhibited by elevation of cyclic nucleotides such as cAMP. From these results, it is suggested that the regulation of hormone secretion by various kinase activity in GH3 cells may be attributable, at least in part, to their effect on L-type Ca2+ channels.
Adenylyl Cyclases ; Animals ; Cell Line* ; Colforsin ; Cytosol ; Genistein ; Neuroendocrine Cells ; Nucleotides, Cyclic ; Patch-Clamp Techniques ; Phorbol 12,13-Dibutyrate ; Phosphotransferases ; Protein Kinase C ; Protein Kinases* ; Protein Tyrosine Phosphatases ; Protein-Tyrosine Kinases ; Rats* ; Vanadates

The aim of this study was to investigate the efficacy of simvastatin to improved lipid profiles in hypercholesterolemic Korean patients. METHODS: From 25 hospitals in Korea, 478 hypercholesterolemic patients were enrolled from November 1996 to April 1998. The inclusion criteria was hypercholesterolemia over 240 mg/dl after diet therapy for 1 month or hypercholesterolemia over 220 mg/dl in patients with definite evidence of ischemic heart disease. Simvastatin 10mg was started and doubled up to 40mg if total cholesterol level remained higher than 200 mg/dl at monthly check. Of 478 subjects, 344 patients in whom study protocol was not violated were analyzed. RESULTS: Male to female ratio was 27:73 and 47% of the subjects were in 6th decade. Hypertension, coronary artery disease, and diabetes mellitus were present in 30, 10, and 4% of the subjects. Baseline lipid profile (mean of total cholesterol-LDL-HDL-triglyceride mg/dl) was 274-185-52-188. The dose of simvastatin for 3 months was 10/10/10mg in 61% of subjects, 10/20/20mg in 21%, 10/10/20mg in 7%, and 10/20/40mg in 12%. The change of total cholesterol level(before-4wk-8wk-12wk-withdrawal 4wk) was 274-209- 205-198-250, and the maximal reduction rate was 27%. The change of LDL-cholesterol was 185-123-116-110-159, with maximal reduction rate 39%. The change of HDL-cholesterol was 52-54-56-55-54, with maximal increase rate 9%. The change of tryglyceride was 188-161- 164-162-189, with maximal reduction rate 15%. The value before/after treatment of ApoA1, ApoB, and Lp(a) was 129/129, 138/83, and 9.3/10.7, respectively. The level of LDL-cholesterol at the end of treatment was below 100mg/dl in 36% of subjects, 100-130 in 45%, 130-160 in 16%, and over 160mg/dl in 4%. The reduction rate of LDL-cholesterol was different between subjects whose LDL decreased below 100 and those whose LDL did not decrease below 130mg/dl, which suggests the existence of the individual difference of responsiveness to simvastatin. There were only 3 subjects (0.9%) who showed increase of liver enzyme over 3 times as the upper normal limit. Conclusion: Simvastatin is effective in improving lipid profiles in hypercholesterolemic Korean patients without serious side effects.
Apolipoproteins B ; Cholesterol ; Coronary Artery Disease ; Diabetes Mellitus ; Diet Therapy ; Female ; Humans ; Hypercholesterolemia ; Hypertension ; Individuality ; Korea ; Liver ; Male ; Myocardial Ischemia ; Simvastatin*