Anticancer Effects of Mitoquinone via Cell Cycle Arrest and Apoptosis in Canine Mammary Gland Tumor Cells.

Treating female canine mammary gland tumors is crucial owing to their propensity for rapid progression and metastasis, significantly impacting the overall health and well-being of dogs. Mitoquinone (MitoQ), an antioxidant, has shown promise in inhibiting the migration, invasion, and clonogenicity of human breast cancer cells. Thus, we investigated MitoQ's potential anticancer properties against canine mammary gland tumor cells, CMT-U27 and CF41.Mg. MitoQ markedly suppressed the proliferation and migration of both CMT-U27 and CF41.Mg cells and induced apoptotic cell death in a dose-dependent manner. Furthermore, treatment with MitoQ led to increased levels of pro-apoptotic proteins, including cleaved-caspase3, BAX, and phospho-p53. Cell cycle analysis revealed that MitoQ hindered cell progression in the G1 and S phases in CMT-U27 and CF41.Mg cells. These findings were supported using western blot analysis, demonstrating elevated levels of cleaved caspase-3, a hallmark of apoptosis, and decreased expression of cyclin-dependent kinase (CDK) 2 and cyclin D4, pivotal regulators of the cell cycle. In conclusion, MitoQ exhibits in vitro antitumor effects by inducing apoptosis and arresting the cell cycle in canine mammary gland tumors, suggesting its potential as a preventive or therapeutic agent against canine mammary cancer.

The Anti-Inflammatory Effects of Broccoli (Brassica oleracea L. var. italica) Sprout Extract in RAW 264.7 Macrophages and a Lipopolysaccharide-Induced Liver Injury Model.

Brassica oleracea var. italica (broccoli), a member of the cabbage family, is abundant with many nutrients, including vitamins, potassium, fiber, minerals, and phytochemicals. Consequently, it has been used as a functional food additive to reduce oxidative stress and inflammatory responses. In the current study, the effects of sulforaphane-rich broccoli sprout extract (BSE) on the inflammatory response were investigated in vitro and in vivo. Comparative high-performance liquid chromatography analysis of sulforaphane content from different extracts revealed that 70% ethanolic BSE contained more sulforaphane than the other extracts. qPCR and enzyme immunoassay analyses revealed that BSE markedly reduced the expression of proinflammatory cytokines and mediators, including cyclooxygenase 2, interleukin (IL)-1ß, IL-6, IL-1, inducible nitric oxide synthase (iNOS), and tumor necrosis factor-α (TNF-α), in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Pretreatment with BSE improved the survival rate and suppressed alanine aminotransferase and aspartate aminotransferase expression in LPS-induced endotoxemic mice, while proinflammatory cytokines such as IL-1ß, TNF-α, IL-6, cyclooxygenase-2, and iNOS decreased dramatically in the LPS-induced liver injury model via BSE treatment. Additionally, F4/80 immunostaining showed that BSE suppressed hepatic macrophage infiltration in the liver after lipopolysaccharide injection. In conclusion, BSE may be a potential nutraceutical for preventing and regulating excessive immune responses in inflammatory disease.

Cisplatin Induces Apoptosis in Mouse Neonatal Testes Organ Culture.

Chemotherapy is used for childhood cancer but may lead to infertility in patients. Spermatogonia stem cells are present in the testes of prepubertal boys, although they do not produce sperm at this age. Herein, we evaluated the toxicity of cisplatin, a known medicine for cancer treatment, in neonatal mouse testes using in vitro organ culture. Mouse testicular fragments (MTFs) derived from 5.5-d postpartum mouse testes were exposed to 1-10 µg/mL cisplatin. The results showed that cisplatin significantly downregulated the expression of germ cell marker genes, including differentiated and undifferentiated, in a dose-dependent manner. In particular, a high dose of cisplatin (10 µg/mL) led to germ cell depletion. In addition, the expression levels of the Sertoli cell marker gene, the number of SOX9+ Sertoli cells, and the levels of SOX9 protein were markedly decreased in cisplatin-treated MTFs compared to controls. The mRNA expression of steroidogenic enzyme-related genes significantly increased in cisplatin-treated MTFs, except for estrogen receptor 1 (Esr1). Consistently, 3ß-hydroxysteroid dehydrogenase protein was also observed in the interstitial regions of cisplatin-treated MTFs. Altogether, our findings showed a significant impairment in germ cell development, Sertoli cell survival, and steroidogenesis in the MTFs of cisplatin-treated mice.

Association between vitamin D level and bronchopulmonary dysplasia: A systematic review and meta-analysis.

Neonatal vitamin D deficiency is common and is associated with development of pulmonary disease in children and adults. While the role of vitamin D in normal lung development is well established, the association between vitamin D deficiency and bronchopulmonary dysplasia (BPD) remains unclear. The present meta-analysis was conducted to evaluate the relationship between vitamin D and BPD. We identified relevant studies (n = 8) using the PubMed, EMBASE, Cochrane Library, and KoreaMed databases and applied the Newcastle-Ottawa Scale to assess the methodological components of each study, and used I2 statistic to evaluate heterogeneity. Comprehensive Meta-Analysis software version 3.3 was used for the statistical analysis. A total of 909 infants were included, of whom 251 (27.6%) were diagnosed with BPD. We found that both vitamin D deficiency at birth (four studies; OR 2.405; 95% CI 1.269 to 4.560; p = 0.007) and low levels of vitamin D at birth (four studies; standardized mean difference -1.463; 95% CI -2.900 to -0.027; p = 0.046) were associated with BPD. The compiled data suggest that antenatal vitamin D deficiency and low vitamin D levels are associated with neonatal BPD.

Transdifferentiation of α-1,3-galactosyltransferase knockout pig bone marrow derived mesenchymal stem cells into pancreatic ß-like cells by microenvironment modulation.

OBJECTIVE: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. METHODS: The BM-MSCs have been differentiated into pancreatic ß-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. RESULTS: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic ß-like cells in the N2B27-based media than in the ADMEM-based media. CONCLUSION: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic ß-like cells.

Oncology nurses' knowledge and awareness on metabolic syndrome in cancer survivors and the perceived barriers to the provision of related care: A mixed-method study.

OBJECTIVE: To identify oncology nurses' level of knowledge and awareness of metabolic syndrome (MetS) in cancer survivors and the perceived barriers to the provision of MetS-related care. METHODS: In this mixed-method study, 196 participants responded to a structured modified questionnaire that included items pertaining to MetS-related knowledge and awareness. Concurrently, 24 semi-structured interviews were conducted. A qualitative survey and quantitative interview were conducted between October 2018 and December 2018. RESULTS: While oncology nurses had a high level of knowledge of MetS in terms of its individual components, they failed to accurately differentiate MetS cases from non-MetS ones. Further, they showed a high level of awareness of MetS-related care for cancer survivors but did not apply their knowledge in clinical settings. In the qualitative survey, the nurses cited various factors pertaining to their perceived barriers to the provision of MetS-related care, including the fact that cancer survivors are distinguished by the specificity of the subject and inpatient environmental constraints. CONCLUSIONS: Oncology nurses had a high level of knowledge of MetS but failed to accurately identify MetS cases. Thus, their level of knowledge should be improved, and strategies are needed to overcome the perceived barriers to the provision of MetS-related care.

Novel Germline Mutations of BRCA1 and BRCA2 in Korean Familial Breast Cancer Patients.

Breast cancer is the second most common cancer in Korean women. Germline mutations in the BRCA1 and BRCA2 genes cause hereditary breast cancer and are detected in 15-20% of hereditary breast cancer. We investigated the BRCA1 and BRCA2 mutations in 114 familial breast cancer patients using next-generation sequencing. We confirmed 20 different mutations of BRCA1 and BRCA2 in 25 subjects (21.9%). Two such mutations in eight patients were novel (not reported in any variant database or previous study). Six mutations have been reported as disease-causing mutations in public databases. Seven mutations were found only in a single nucleotide polymorphism database and one mutation has been reported in Korea. The BRCA1/2 mutation frequency was similar to that of other studies on familial breast cancer patients in the Korean population. Further studies should examine more cases and mutations of whole exons.

Cytokine levels in febrile seizure patients: A systematic review and meta-analysis.

PURPOSE: Febrile seizures (FSs) are the most common form of childhood seizures. During infection, both pro-inflammatory and anti-inflammatory cytokines are produced. Complex interactions among immune-inflammatory process, cytokine activation, and genetic factors are involved in the pathogenesis of FSs. The association between cytokines and FSs during childhood is inconclusive due to inconsistent results reported in different studies. We performed a systematic review and meta-analysis to determine an association between cytokines and FS in children. METHODS: We searched PubMed, EMBASE, and Cochrane databases for studies published up to January 2017 using the following key words: ["cytokine" OR "interleukin" OR "tumor necrosis factor alpha" OR "interferon-gamma" OR "single nucleotide polymorphism"] AND ["febrile seizure" OR "febrile convulsion"] AND ["pediatric" OR "infant" OR "child"]. Standardized mead difference (SMD) and 95% confidence intervals (CI) were calculated using standard meta-analysis techniques. RESULTS: A total of 6 studies enrolling 243 children with FS and 234 controls were included in the meta-analysis. A total of 4 different inflammatory mediators were. The results indicated that CSF IL-1ß level and serum IL-6 level were significantly associated with FS (CSF IL-1ß: SMD, 1.064; 95% CI, 0.217-1.611; P < 0.01, serum IL-6 SMD, 2.654; 95% CI, 2.332-2.975; P < 0.01). CONCLUSION: The results of this meta-analysis suggest that CSF IL-1ß level and serum IL-6 level are associated with an increased risk of FSs in children. Based on these results, it is expected that a therapeutic agent for specific cytokines could be developed in the future to prevent FS.

Interleukin-4 and tumor necrosis factor-alpha levels in children with febrile seizures.

PURPOSE: The pathophysiology of febrile seizures remains unclear. Cytokines have been suggested to play a role in the pathogenesis of febrile seizures. We compared TNF-α and IL-4 levels in patients with febrile seizure (FS) with those in controls and identified the relationship between cytokines and various other factors. METHOD: Fifty FS patients who visited Konkuk University Hospital from December 2015 to December 2016 were included. Thirty-nine patients who had fever without seizures were enrolled as the control group. Serum samples from febrile patients with a history of febrile seizures without present seizures (FPH) (N = 12) and from the afebrile seizure (AF) group (N = 13) were also analyzed. In the FS group, we compared cytokine levels among patients stratified by sex, family history, seizure recurrence, duration of seizure and serum lactate levels. RESULTS: The median serum TNF-α level in the FS group (19.54 pg/mL) was significantly higher than that in the control group (15.86 pg/mL). Higher median serum IL-4 levels were detected in the FS group (3.38 pg/mL) than in the control group (3.30 pg/mL). In the FS group, the serum IL-4 and TNF-α levels correlated with seizure recurrence and serum lactate levels, but they did not correlate with family history, duration of seizures or sex. CONCLUSIONS: Our study supports the hypothesis that TNF-α production is involved in the pathogenesis of febrile seizures. IL-4 is believed to be involved in the pathogenesis of febrile seizures. The number of seizures and lactate levels were correlated with IL-4 and TNF-α levels.

Germ cell-specific apoptosis by extracellular clusterin in cryptorchid dog testes.

Mammalian testes are maintained at a relatively lesser temperature than the abdominal region so that normal spermatogenesis can occur. Germ cell apoptosis has resulted in heat-damaged testes that occurs as a result of cryptorchidism, but the mechanism is not yet fully understood. To elucidate the cause of germ-cell death by cryptorchidism, cryptorchidism was surgically induced in dog testes and histological and molecular analyses were performed. Histological data indicated that the seminiferous tubules of cryptorchid testes and epididymis contained fewer germ cells. Total RNA sequencing was performed to screen for overexpressed genes in cryptorchid dog testes. Clusterin RNA was in greater abundance (approximately 12.8-fold) in cryptorchid testes than in normal testes. In addition, cleaved caspase-3 and -8 were detected in greater abundance in cryptorchid dog testes. Real time RT-PCR and western blotting analysis indicated there was a greater abundance of clusterin in cryptorchid dog testes. Furthermore, clusterin was detected in extracellular regions of cryptorchid dog testes during the 4 weeks after surgery. Thus, germ-cell specific apoptosis and expression of clusterin genes occur with a resulting presence of this protein in extracellular regions of cryptorchid dog testes. This result will facilitate further study of spermatogenesis and the specific mechanisms by which cryptorchidism results in male infertility.

Stage-specific expression of DDX4 and c-kit at different developmental stages of the porcine testis.

Spermatogenesis begins with spermatogonial stem cells (SSCs), which are located in the basement membrane of the adult testes. Previous studies have described specific biomarkers for undifferentiated porcine spermatogonia or SSCs; however, these markers are not sufficient to understand spermatogenesis at different developmental stages. The objective of this study was characterize the expression of DEAD-Box polypeptide 4 (DDX4, also known as VASA) and tyrosine-protein kinase kit (c-kit), as potential markers of male germ cells in the porcine testis. In porcine testis tissue at prepubertal stages (5, 30, and 60 days), DDX4 and c-kit protein expression was detected in the most undifferentiated spermatogonia, which also express protein gene product 9.5 (PGP9.5). However, in porcine testis tissues from pubertal and postpubertal stages (90, 120, and 150 days), DDX4 and c-kit were not detected in PGP9.5-positive undifferentiated spermatogonia. The DDX4 expression pattern was similar to that of c-kit in the porcine testis. In adult porcine testes, DDX4-expressing cells were located on the lumenal side, compared to synaptonemal complex protein 3-positive primary spermatocytes, but DDX-4 was not co-expressed with acrosin, a known acrosome marker. In addition, DDX4 was detected in PGP9.5-expressing porcine SSCs in culture. Based on our results, we suggest that DDX4 and c-kit are putative markers of undifferentiated spermatogonia in the prepubertal porcine testis. While in the postpubertal porcine testis, they are markers of differentiated spermatocytes. These findings may facilitate future studies of porcine spermatogenesis.

Dose-dependent effects of busulfan on dog testes in preparation for spermatogonial stem cell transplantation.

Successful male germ cell transplantation requires depletion of the host germ cells to allow efficient colonization of the donor spermatogonial stem cells. Although a sterilizing drug, busulfan, is commonly used for the preparation of recipient models before transplantation, the optimal dose of this drug has not yet been defined in dogs. In this study, 1-year-old mongrel dogs were intravenously injected with three different concentrations of busulfan (10, 15, or 17.5 mg/kg). Four weeks after busulfan treatment, no fully matured spermatozoa were detected in any of the busulfan-treated groups. However, small numbers of PGP9.5-positive spermatogonia were detected in all treatment groups, although no synaptonemal complex protein-3-positive spermatocytes were detected. Of note, acrosin-positive spermatids were not detected in the dogs treated with 15 or 17.5 mg/kg busulfan, but were detected in the other group. Eight weeks after busulfan treatment, the dogs treated with 10 mg/kg busulfan fully recovered, but those in the other groups did not. PGP9.5-positive spermatogonia were detected in the 10 mg/kg group, and at a similar level as in the control group, but these cells were rarely detected in the 15 and 17.5 mg/kg groups. These results suggest that a dose of 15-17.5 mg/kg is optimal for ablative treatment with busulfan to prepare the recipient dogs for male germ cell transplantation. At least eight weeks should be allowed for recovery. The results of this study might facilitate the production of recipient dogs for male germ cell transplantation and can also contribute to studies on chemotherapy.

Relationship between iron deficiency anemia and febrile seizures in children: A systematic review and meta-analysis.

PURPOSE: The association between iron deficiency anemia (IDA) and febrile seizures (FS) during childhood is inconclusive due to inconsistent results reported in different studies. We performed a systematic review and meta-analysis to determine an association between IDA and FS in children. METHODS: We searched PubMed, EMBASE, and Cochrane Library databases for studies published up to August 2015 using the following key words: ["iron deficiency" OR "iron status"] AND ["febrile seizure" OR "febrile convulsion"] AND ["pediatric" OR "infant" OR "child"]. Pooled odds ratios (OR) and 95% confidence intervals (CI) were calculated using standard meta-analysis techniques. Subgroup analysis also was performed. RESULTS: A total of 17 studies enrolling 2416 children with FS and 2387 controls were included in the meta-analysis. The results indicated that IDA was significantly associated with FS (OR, 1.98; 95% CI, 1.26-3.13; P=0.003). Subgroup analyses evaluated the diagnostic indices for IDA including serum iron, plasma ferritin, and mean corpuscular volume (MCV). The results indicated that IDA diagnosed on the basis of plasma ferritin (OR, 3.78; 95% CI, 1.80-7.94; P<0.001) or MCV (OR, 2.08; 95% CI, 1.36-3.17; P=0.001) was modestly associated with FS, whereas IDA diagnosed on the basis of two serum iron studies was not associated with FS (OR, 0.57; 95% CI, 0.24-1.37; P=0.210). CONCLUSION: The results of this meta-analysis suggest that IDA is associated with an increased risk of FS in children.

Relationship between plasma pentraxin 3 level and risk of chronic kidney disease in the Korean elderly: the Dong-gu study.

Purpose This study investigated the cross-sectional association between chronic kidney disease (CKD) and plasma pentraxin 3 (PTX3) levels in a Korean population, in a community-based cohort study. METHODS: A total of 1816 (891 men, 925 women) subjects were randomly selected from the cohort of participants for the final analyses. Plasma PTX3 concentration was determined using enzyme-linked immunosorbent assays. The participants were divided into four quartiles according to the PTX3 levels. Multivariate logistic regression was performed to evaluate the association between plasma PTX3 level and CKD. Covariates inserted into the multivariate model included smoking status, systolic blood pressure, body mass index, waist circumference, high-density lipoprotein, low-density lipoprotein uric acid, white blood cell count, and carotid intima-media thickness. RESULTS: Compared to the lowest PTX3 group (Q1), a significantly higher risk of CKD was found in the highest group (Q4), with an odds ratio of 1.58 and 95% confidence interval of 1.18-2.11 (P for trend <0.001). CONCLUSIONS: This study showed that higher plasma PTX3 levels are significantly associated with CKD risk. The biological mechanism remains unclear; therefore, further molecular investigation of association between CKD and PTX3 is needed.

Acute Kidney Injury Following Cardiopulmonary Bypass in Children　- Risk Factors and Outcomes.

BACKGROUND: Acute kidney injury (AKI) is the most common and most serious complication following heart surgery. We aimed to determine the prevalence of, and risk factors for, AKI following pediatric cardiac surgery.Methods and Results:We retrospectively analyzed 135 patients aged ≤18 years who underwent cardiac surgery for congenital heart defects; by RACHS-1 category, 58 patients (43%) had an operative risk score ≥3. AKI was defined and classified using the pediatric pRIFLE criteria (Pediatric Risk, Injury, Failure, Loss, and End-stage Kidney Disease); 19 patients (14.1%) developed AKI: 17 had AKI with a severity classified as risk (R) and 2 had AKI classified as injury (I). Body weight, height, body surface area, and preoperative mechanical ventilation were all independently associated with AKI development (P=0.038, 0.040, 0.033 and 0.008, respectively). Preoperative ventilation strongly correlated with AKI severity. Higher pRIFLE classification positively correlated with increased incidence of peritoneal dialysis, increased postoperative mechanical ventilation duration, and longer hospital stay (P=0.009, 0.039 and 0.042, respectively). CONCLUSIONS: In this study, we found a low prevalence of postoperative AKI in pediatric patients undergoing severe cardiac surgery. AKI was associated with worse early postoperative outcomes. Early prediction and appropriate treatment of AKI during the postoperative period are emphasized.

Association between the TCF7L2 polymorphism and colorectal cancer does not differ by diabetes and obesity statuses.

This study evaluated the association between polymorphism in a newly identified locus, rs11196172, located in transcription factors 7-like 2 (TCF7L2) and colorectal cancer (CRC) risk according to diabetes and obesity statuses. A study enrolled 6138 CRC patients and 4367 community controls. The adjusted odds ratios (aORs) with age, sex, smoking, and body mass index of the A allele, compared with the G allele, was 1.08 (95% CI 1.01-1.16). The significantly higher risk of CRC with the A allele remained after adjusting for diabetic status (aOR 1.07, 95% CI 1.01-1.15). When stratified by diabetic or obesity status, significant associations between TCF7L2 polymorphism and CRC risk were limited to non-diabetic or normal-weight subjects. No significant interactions between the A/G allele and diabetes status or the A/G allele and overweight status were found. The results indicated that the TCF7L2 rs11196172 polymorphism increases the risk of CRC independently, with no evidence of an interaction with diabetes or obesity.

Biochanin A Ameliorates Arsenic-Induced Hepato- and Hematotoxicity in Rats.

Biochanin A (BCA) is a natural organic compound of the phytoestrogenic isoflavone class that has antioxidant and metal chelator properties in the presence of transition metal ions, however, its efficacy in animal models is still obscure. Therefore, the objective of this study was to investigate the protective effects of BCA against arsenic-induced hepatic injury and hematotoxicity in rats. The results suggest that arsenic intoxicated rats showed significantly higher levels of plasma hepatic markers than normal control rats. Furthermore, an increase in lipid peroxidation with depletion of reduced glutathione (GSH) and activities of superoxide dismutase (SOD) and catalase (CAT) occurred in the livers of rats exposed to arsenic. Administration of BCA (20 mg/kg·bw/day) and selenium (3 mg/kg·bw/day) resulted in a significant reversal of hepatic and oxidative stress markers in arsenic-intoxicated rats. A low dose of BCA (10 mg/kg·bw/day) did not show any preventive effect, while a high dose of BCA (40 mg/kg·bw/day) partially prevented all hepatotoxicity events. These biochemical perturbations were supported by histopathological observations of the liver. Our results suggest that administration of BCA (20 mg/kg·bw/day) attenuated the arsenic hepatotoxicity, a property that could contribute to the therapeutic approaches for chronic liver diseases.

Establishment of a surgically induced cryptorchidism canine recipient model for spermatogonial stem cell transplantation.

Transplantation of spermatogonial stem cells (SSCs) in experimental animal models has been used to study germ line stem cell biology and to produce transgenic animals. The species-specific recipient model preparation is important for the characterization of SSCs and the production of offspring. Here, we investigated the effects of surgically induced cryptorchidism in dog as a new recipient model for spermatogonial stem cell transplantation. Artificially unilateral or bilateral cryptorchidism was induced in ten mature male dogs by surgically returning the testis and epididymis to the abdominal cavity. The testes and epididymides were collected every week after the induction of artificial cryptorchidism (surgery) for one month. To determine the effect of surgical cryptorchidism, the seminiferous tubule diameter was measured and immunohistochemistry using PGP9.5 and GATA4 antibodies was analyzed. The diameters of the seminiferous tubules of abdominal testes were significantly reduced compared to those of the scrotal testes. Immunohistochemistry results showed that PGP9.5 positive undifferentiated spermatogonia were significantly reduced after surgical cryptorchidism induction, but there were no significant changes in GATA-4 positive sertoli cells. To evaluate the testis function recovery rate, orchiopexy was performed on two dogs after 30 days of bilateral cryptorchidism. In the orchiopexy group, SCP3 positive spermatocytes were detected, and spermatogenesis was recovered 8 weeks after orchiopexy. In this study, we provided optimum experimental conditions and time for surgical preparation of a recipient canine model for SSC transplantation. Additionally, our data will contribute to recipient preparation by using surgically induced cryptorchidism in non-rodent species.

Association between ABO genotype and risk of hepatocellular carcinoma in Koreans.

BACKGROUND: Associations between ABO blood groups and risk of several malignancies have been reported, although there are limited data regarding hepatocellular carcinoma (HCC). The aim of this study was to investigate any possible association between the ABO genotype, especially blood group A, and HCC risk in Koreans. MATERIALS AND METHODS: We conducted a case-control study of 1,538 patients with newly diagnosed HCC at Chonnam National University Hwasun Hospital and 1,305 randomly selected members of the general population. The ABO genotype was determined by multicolor real-time polymerase chain reaction (PCR) using displacing probes. Adjusted odds ratios (aORs) and their 95% confidence intervals (CIs) were calculated using logistic regression models with adjustment for gender, age, smoking, alcohol drinking, and hepatitis B and C status. RESULTS: The risk of HCC in genotype AA was significantly higher than in OO (aOR=1.773, 95% CI=1.161-2.705). The risk in blood group A was also higher than in blood group O (aOR=1.448, 95% CI=1.005 1.897). No significant difference was found for the AA, BO, BB, and AB genotypes, or blood group B and AB. CONCLUSIONS: Blood group A and genotype AA showed the highest risks of HCC in a Korean population. No significant difference was found for the AO, BO, BB, and AB genotypes, or blood group B and AB.

Identification and in vitro derivation of spermatogonia in beagle testis.

BACKGROUND: In vitro culture of spermatogonial stem cells (SSCs) is important for exploration of SSCs self-renewal, differentiation, and manipulation. There are several reports on rodent SSC cultures; however, data on SSC cultures in domestic animals are limited. To provide basic scientific information on canine SSC cultures, we report canine testes development, and the development of spermatogonia-derived colonies (SDCs) for in vitro cultures. METHODOLOGY/PRINCIPAL FINDINGS: Testes from 2-, 3-, and 12-month-old beagles were used for histology, immunohistochemistry, in vitro culture, immunocytochemistry, and PCR. Protein gene product 9.5 (PGP9.5)-positive spermatogonia, both single and paired, were found to be abundant in the testes of 2-month-old beagles. stempro-34 and Dulbecco's modified Eagle medium with 5% fetal bovine serum provided as useful substrates for culture of SDCs, and fibroblast growth factor (FGF) played a key role in colony formation. Colonies were positive for alkaline phosphatase and anti-PGP9.5 staining. The early spermatogonia and stem cell markers such as octamer binding protein 4 (Oct4), Nanog homeobox (Nanog), promyelocytic leukemia zinc finger (PLZF), PGP9.5, and GDNF family receptor alpha-1 (GFRα-1) were expressed in the colonies at higher levels than in the testis tissue. CONCLUSIONS: Testes of the 2-month-old beagles had abundant single and paired spermatogonia, which can be used for derivation of SDCs, and FGF was important for colony formation.