In this study, we examined the effects of a two-step culture system, which involves the use of different culture media for early cleavage and later stage embryos, on the in vitro development of bovine embryos. We also investigated the effect of glucose, phosphate and citrate on the in vitro early developmental period of bovine embryos in a two-step culture system. Moreover, the supplementation of different protein sources (BSA-V, BSA-FAF and FBS) during IVC did not affect the frequency of blastocyst development. Using two-step culture, embryos were cultured in protein-free media for an initial 5 days. This was then followed by the same culture media or an FBS supplemented media. The developmental rates of blastocysts in the FBS containing group were significantly higher than in the replaced with no serum containing group. Embryos cultured in mSOF supplemented with 1.5 mM glucose plus 1.2 mM phosphate were significantly inhibited. The inhibition of developmental competence by glucose plus phosphate was consistent with the existence of 0.5 mM sodium citrate. This study indicates that a two-step culture system, which applies different conditions for early cleavage embryos, i.e., serum-free media, vs. later stage embryos, with serum containing media, may be effective for in vitro production systems. In addition, the developmental competence of bovine embryos was depressed in the presence of glucose plus phosphate as compared to either alone or the absence of both. Therefore, the avoidance of this negative effect should allow more optimal conditions to be developed for in vitro production.
Animals ; Blastocyst/drug effects/metabolism ; Cattle/*embryology ; Citric Acid/pharmacology ; Culture Media/*chemistry ; Culture Techniques/*methods ; Ectogenesis/drug effects ; Embryo/*drug effects/embryology/metabolism ; Embryonic and Fetal Development/drug effects ; *Energy Metabolism ; Female ; Fertilization in Vitro ; Glucose/pharmacology ; Male ; Phosphates/pharmacology ; Proteins/*pharmacology ; Zygote/drug effects/metabolism

OBJECTIVES: Posttraumatic stress disorder(PTSD) has been primarily associated with emotional problems. Recently, however, the impact of PTSD on cognitive processes has interested a growing number of researchers. The current study is aimed at investigating the cognitive aspects of PTSD at both behavioral and neurological levels. METHODS: We recruited individuals with PTSD who survived the Daegu subway explosion in 2003 as well as non-PTSD individuals as a control group. To evaluate the inhibitory processes and the neural mechanisms, we had these individuals perform the negative priming task simultaneously with functional MRI scanning. RESULTS: Behaviorally, the negative priming effect was intact in the control group but was not evident in the PTSD group. In the imaging results, only the PTSD group showed the negative priming effect (i.e., increased activation of the negative priming condition as opposed to the neutral condition) in the dorsolateral prefrontal cortex, anterior cingulate cortex, and inferior temporal gyrus. The PTSD group also showed increased activity for the positive priming condition as opposed to the neutral condition in the claustrum. These results confirm and extend the previous findings that the integrity of the ACC is compromised in the trauma survivors due to disrupted white matter tract. CONCLUSIONS: The current results suggest that deteriorated performance of the PTSD group may be due to the functional problem as well as the structural abnormalities.
Basal Ganglia ; European Continental Ancestry Group ; Explosions ; Gyrus Cinguli ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Magnetics ; Magnets ; Prefrontal Cortex ; Railroads ; Stress Disorders, Post-Traumatic ; Survivors

PURPOSE: FP(5-FU, Cisplatin) combination is one of the most active regimen for the advanced gastric cancer with a response rate of 50~60%. In spite of this high response rate, there is little evidence that FP regimen results in survival benefit for patients with advanced gastric cancer. This study was performed to evaluate the efficacy and toxicity of this regimen with the addition of levamisole, an immunomodulatory agent, known as enhancing the antitumor effects of 5-FU in other cancer. MATERIALS AND METHODS: Previously untreated patients with metastatic or recurrent gastric cancer were treated with 5-FU(1000 mg/M2 civ, D1~5), cisplatin(60 mg/M2 iv, Dl) every 3 weeks, and levamisole(150 mg/day, Dl~3) every 2 weeks. The major endpoints were response rate, response duration, and toxicities. RESULTS: Between June 1992 and Aug. 1996, thirty three patients were included in this study. Patients received 2~18 cycles of chemotherapy(median 5). Among the evaluable 31 patients, 18 patients(58%, 95% C.I. 40.4~75.7) showed objective responses including one(3.2%) clinical complete response. The median response duration was 7.7 months(95% C.I. 3.6~11.8). During total of 189 cycles of chemotherapy, 79 episodes(41.7%) of leucopenia were observed. There was no death from concurrent infection. CONCLUSION: FPL combination therapy is at least as effective as conventional FP chemotherapy, but resulted in somewhat more myelosuppression.
Cisplatin* ; Drug Therapy ; Drug Therapy, Combination* ; Fluorouracil* ; Humans ; Levamisole* ; Stomach Neoplasms

This study was conducted to determine the effect of treatment with intravenous human immunoglobulin G (hIVIgG) on outcome in dogs with idiopathic immune-mediated hemolytic anemia (IMHA), and to identify prognostic variables that determine outcome in affected dogs. Thirty-seven dogs that met the inclusion criteria were enrolled in a retrospective study. The dogs were categorized into two groups based on their having received hIVIgG. There was no significant difference in survival between the hIVIgG group and the non-hIVIgG group. Mortality during hospitalization and at 1 month, 1 year, or 2 years after discharge was not significantly different between the hIVIgG and the non-hIVIgG groups. Hemoglobinuria was significantly less prevalent in dogs that lived more than 1 year than in those who lived less than 1 year, and was less prevalent in dogs that lived more than 2 years than in those who lived less than 2 years. However, there was no difference in the presence of hemoglobinuria between dogs that lived less than 1 month and those that lived more than 1 month. Overall, there was no evidence of a beneficial effect of hIVIgG in dogs with idiopathic IMHA.
Anemia, Hemolytic* ; Anemia, Hemolytic, Autoimmune ; Animals ; Dogs* ; Hemoglobinuria ; Hospitalization ; Humans* ; Immunoglobulin G* ; Immunoglobulins* ; Mortality ; Retrospective Studies*

The level of P4 at the time of embryo transfer (ET) is important. P4 concentrations and numbers of corpora lutea for 126 recipients were evaluated. Nuclear transfer embryos were transferred into 126 surrogates. 11 maintained their pregnancy until full-term delivery, 17 miscarried, and implantation failed in 98 animals. P4 levels in the full-term group were significantly different from those of the pigs that aborted or in which implantation failed (p < 0.05). However, the numbers of corpora lutea were not significantly different. These findings indicate that the concentration of progesterone can be an important factor for successful ET in pigs.
Animals ; Corpus Luteum/*physiology ; Embryo Transfer/*veterinary ; Embryo, Mammalian/*physiology ; Female ; Nuclear Transfer Techniques ; Pregnancy ; *Pregnancy Rate ; Progesterone/*blood ; Retrospective Studies ; Sus scrofa/*physiology

Quercetin is a plant-derived flavonoid found in fruits or vegetables that has antioxidant properties and acts as a free radical scavenger. We investigated the effects of quercetin on porcine oocyte nuclear maturation and embryonic development after parthenogenetic activation. We then evaluated the antioxidant activities of quercetin by measuring reactive oxygen species (ROS) levels in matured oocytes. Immature oocytes were untreated or treated with 1, 10, and 50 microg/mL quercetin during in vitro maturation (IVM). Quercetin treatment did not improve oocyte nuclear maturation, but significantly higher blastocyst rates (p < 0.05) of parthenogenetically activated oocytes were achieved when the IVM medium was supplemented with an adequate concentration of quercetin (1 microg/mL). However, cleavage rates and blastocyst cell numbers were not affected. Oocytes treated with 1 or 10 microg/mL quercetin had significantly lower (p < 0.05) levels of ROS than the control and group treated with the highest concentration of quercetin (50 microg/mL). Moreover, this highest concentration was detrimental to oocyte nuclear maturation and blastocyst formation. Based on our findings, we concluded that exogenous quercetin reduces ROS levels during oocyte maturation and is beneficial for subsequent embryo development.
Animals ; Antioxidants/administration & dosage/*pharmacology ; Dose-Response Relationship, Drug ; In Vitro Oocyte Maturation Techniques/*veterinary ; Oocytes/cytology/*drug effects/physiology ; Quercetin/administration & dosage/*pharmacology ; Reactive Oxygen Species/*metabolism ; *Swine

Quercetin is a plant-derived flavonoid found in fruits or vegetables that has antioxidant properties and acts as a free radical scavenger. We investigated the effects of quercetin on porcine oocyte nuclear maturation and embryonic development after parthenogenetic activation. We then evaluated the antioxidant activities of quercetin by measuring reactive oxygen species (ROS) levels in matured oocytes. Immature oocytes were untreated or treated with 1, 10, and 50 microg/mL quercetin during in vitro maturation (IVM). Quercetin treatment did not improve oocyte nuclear maturation, but significantly higher blastocyst rates (p < 0.05) of parthenogenetically activated oocytes were achieved when the IVM medium was supplemented with an adequate concentration of quercetin (1 microg/mL). However, cleavage rates and blastocyst cell numbers were not affected. Oocytes treated with 1 or 10 microg/mL quercetin had significantly lower (p < 0.05) levels of ROS than the control and group treated with the highest concentration of quercetin (50 microg/mL). Moreover, this highest concentration was detrimental to oocyte nuclear maturation and blastocyst formation. Based on our findings, we concluded that exogenous quercetin reduces ROS levels during oocyte maturation and is beneficial for subsequent embryo development.
Animals ; Antioxidants/administration & dosage/*pharmacology ; Dose-Response Relationship, Drug ; In Vitro Oocyte Maturation Techniques/*veterinary ; Oocytes/cytology/*drug effects/physiology ; Quercetin/administration & dosage/*pharmacology ; Reactive Oxygen Species/*metabolism ; *Swine

PURPOSE: The purpose of this study is to determine the relationship between the repetitive flexion or extension posture on the cervical spine on labor and degenerative change of the cervical spine, and the factors affecting degenerative change of the cervical spine. MATERIALS AND METHODS: To determine the factors affecting degenerative change of the cervical spine, age, sex, height, weight, body mass index, smoking, diabetes mellitus (DM), time engaging in labor, and cervical spine posture (flexion or extension) required repetitively on labor were investigated in the subjects. In addition, to evaluate the level of degenerative change of the cervical spine on 83 people in the flexion group (flexion strain) and 83 people in the extension group (extension strain), cervical degenerative index (CDI) in the simple cervical spine lateral radiograph was used to score (0-60 points) the degenerative severity. RESULTS: A total of 166 subjects (flexion group: 83 people, extension group: 83 people) participated in this study, and for the CDI, the cervical spine flexion group scored 7.8±6.2 points, and the cervical spine extension group scored 12.2±6.0 points to show that the cervical spine extension group had significant degenerative change in the cervical spine. In the multiple linear regression test performed to verify the risk factors affecting the degenerative change of the cervical spine, age (p=0.004), contraction of DM (p=0.029), and extension posture of cervical spine (p<0.001) influenced the degenerative change of the cervical spine. CONCLUSION: Repetitive extension posture on the cervical spine on labor and contraction of diabetes affected degenerative change of the cervical spine, therefore, training in medical care and posture on labor are required to prevent the progression of degenerative change in the cervical spine.
Agriculture* ; Body Weight ; Cervical Vertebrae ; Diabetes Mellitus ; Female ; Linear Models ; Neck* ; Posture ; Risk Factors* ; Smoke ; Smoking ; Spine ; Spondylosis*

Recombination activating gene-2 (RAG-2) plays a crucial role in the development of lymphocytes by mediating recombination of T cell receptors and immunoglobulins, and loss of RAG-2 causes severe combined immunodeficiency (SCID) in humans. RAG-2 knockout mice created using homologous recombination in ES cells have served as a valuable immunodeficient platform, but concerns have persisted on the specificity of RAG-2-related phenotypes in these animals due to the limitations associated with the genome engineering method used. To precisely investigate the function of RAG-2, we recently established a new RAG-2 knockout FVB mouse line (RAG-2(−/−)) manifesting lymphopenia by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease. In this study, we further characterized their phenotypes focusing on histopathological analysis of lymphoid organs. RAG-2(−/−) mice showed no abnormality in development compared to their WT littermates for 26 weeks. At necropsy, gross examination revealed significantly smaller spleens and thymuses in RAG-2(−/−) mice, while histopathological investigation revealed hypoplastic white pulps with intact red pulps in the spleen, severe atrophy of the thymic cortex and disappearance of follicles in lymph nodes. However, no perceivable change was observed in the bone marrow. Moreover, our analyses showed a specific reduction of lymphocytes with a complete loss of mature T cells and B cells in the lymphoid organs, while natural killer cells and splenic megakaryocytes were increased in RAG-2(−/−) mice. These findings indicate that our RAG-2(−/−) mice show systemic lymphopenia with the relevant histopathological changes in the lymphoid organs, suggesting them as an improved Rag-2-related immunodeficient model.
Animals ; Atrophy ; B-Lymphocytes ; Bone Marrow ; Genome ; Homologous Recombination ; Humans ; Immunoglobulins ; Killer Cells, Natural ; Lymph Nodes ; Lymphocytes ; Lymphopenia* ; Megakaryocytes ; Methods ; Mice* ; Mice, Knockout ; Negotiating ; Phenotype ; Receptors, Antigen, T-Cell ; Recombination, Genetic ; Sensitivity and Specificity ; Severe Combined Immunodeficiency ; Spleen ; T-Lymphocytes ; Thymus Gland

STUDY DESIGN: Retrospective study. OBJECTIVES: The purpose of this study was to evaluate risk factors for subsidence after posterior lumbar interbody fusion (PLIF). SUMMARY OF LITERATURE REVIEW: Body mass index (BMI), bone mineral density (BMD), cage characteristics (titanium or poly-ether-ether-ketone (PEEK)) and degree of disc distraction are risk factors for cage subsidence after PLIF. MATERIALS AND METHODS: From January 2010 to January 2015, a total of 69 patients (93 segments) who were diagnosed with degenerative lumbar spine disease at the current authors' institution and underwent follow-up for at least 1 year were included in this retrospective study. Data on all factors related to cage subsidence were taken into consideration. The degree of association for each of the factors was determined through the calculation of odds ratios (ORs) with a 95% confidence interval. Logistic regression analyses were performed. The P-value that represented statistical significance was set below 0.05. RESULTS: There were no significant associations between fused segment level and cage subsidence (p=0.588), nor were there any significant associations between the kind of cage (titanium or PEEK) and cage subsidence (p=0.371). Using logistic regression, the factors with a P-value below the 0.20 level in univariate analyses were included in the multivariate analyses. In multivariate analyses, diabetes mellitus (DM) (p=0.029, OR, 4.524), osteoporosis (p=0.026, OR, 6.046), and degree of disc distraction (p=0.010, OR, 1.446) had significant associations with cage subsidence. In addition, there were significant associations between cage subsidence and instrument failure (p=0.008, OR, 8.235). CONCLUSIONS: DM and osteoporosis, which may affect bony structures, have significant associations with cage subsidence after PLIF. Cage insertion with excessive disc distraction during surgery may also affect cage subsidence after PLIF.
Body Mass Index ; Bone Density ; Diabetes Mellitus ; Follow-Up Studies ; Humans ; Logistic Models ; Multivariate Analysis ; Odds Ratio ; Osteoporosis ; Retrospective Studies ; Risk Factors* ; Spine