We designed a new electrophoresis apparatus. This was consisted of the body into which agarose could be poured directly and the cover from which the platinium electrodes were suspended. After running the nucleic acid in this electrophoresis, we observed the band of nucleic acid on the ultraviolet transilluminator without touching the agarose gel, keeping the cover with the electrodes and discarding the gel in body. We compared the variuos types of the electrophoresis apparatus. In the electrophoresis apparatus which body was consisted of the flat bottom, the migration of dyes was the fastest among studied types. The high migration velocity of dyes was due to high electric current.
Coloring Agents ; Electrodes ; Electrophoresis* ; Running ; Sepharose

PURPOSE: The purpose of this study was to investigate influences of drinking, smoking, and obesity indices on cardio-cerebrovascular disease risk factors in Korean young men. METHODS: The subjects were 234 young men, aged 20 to 39 years. Body mass index (BMI), percent body fat (PBF), body fat mass (kg), and waist hip ratio (WHR) were measured as obesity indices. Systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood sugar (FBS), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured as cardio-cerebrovascular disease risk factors. The data were analyzed by t-test, ANOVA, Pearson's correlation coefficient, and multiple regression analysis using SAS 9.4. RESULTS: Statistically significant factors were BMI for SEP; age, degree of drinking, body fat mass for DBP; WHR for TC; WHR and age for TG; age, degree of smoking and PBF for HDL-C; WHR, age and degree of smoking for LDL-C. CONCLUSION: Factors identified in this study need to be considered in cardio-cerebrovascular disease prevention programs for young men.

Tailgut cyst of retrorectal space is uncommon and its classification and prognosis are unclear. The lesion usually consists of a multiloculated cyst lined by squamous, transitional, and glandular epithelium. Disorganized fascicles of smooth muscle may be seen in the wall. Glomus coccygeum, normally located at the tip of coccyx, is incidentally discovered in the presacral mass including tailgut cyst. We report a case of tailgut cyst in a 35 year old female who had internal hemorrhoid and intermittent anal pain for 3 years. Pelvic computerized tomography revealed a 3x2 cm sized mass in the presacral area. The mass was multicystic and lined by squamous, transitional, and peudostratified ciliated columnar epithelium with inflammatory cells and scattered smooth muscle in the wall. Dermal adnexal structures were not present, which excluded dermoid cyst. A glomus body was identified in the surrounding soft tissue.
Female ; Humans ; Cysts

No abstract available.
Cleft Lip* ; Diagnosis* ; Ultrasonography*

Toxoplasma gondii infections occur throughout the world, and efforts are needed to develop various vaccine candidates expressing recombinant protein antigens. In this study, influenza matrix protein (M1) virus-like particles (VLPs) consisting of T. gondii rhoptry antigen 4 (ROP4 protein) were generated using baculovirus (rBV) expression system. Recombinant ROP4 protein with influenza M1 were cloned and expressed in rBV. SF9 insect cells were coinfected with recombinant rBVs expressing T. gondii ROP4 and influenza M1. As the results, influenza M1 VLPs showed spherical shapes, and T. gondii ROP4 protein exhibited as spikes on VLP surface under transmission electron microscopy (TEM). The M1 VLPs resemble virions in morphology and size. We found that M1 VLPs reacted with antibody from T. gondii-infected mice by western blot and ELISA. This study demonstrated that T. gondii ROP4 protein can be expressed on the surface of influenza M1 VLPs and the M1 VLPs containing T. gondii ROP4 reacted with T. gondii-infected sera, indicating the possibility that M1 VLPs could be used as a coating antigen for diagnostic and/or vaccine candidate against T. gondii infection.
Animals ; Baculoviridae ; Blotting, Western ; Clone Cells ; Cloning, Organism ; Enzyme-Linked Immunosorbent Assay ; Influenza, Human* ; Insects ; Mice ; Microscopy, Electron, Transmission ; Toxoplasma* ; Toxoplasmosis ; Virion

Familial juvenile polyposis is a rare intestinal polyposis characterized by the occurrence of multiple juvenile polyps in the gastrointestinal tract. We report a case of familial juvenile polyposis in a 17-year-old man with a review of the literature. This patient underwent total colectomy due to a 6 years history of rectal bleeding. Grossly, the colon showed 36 variable sized pedunculated polyps, measuring 2.5cm x 2cm from the largest size and 0.2cm x 0.2cm to the smallest size. Histologically, the polyps consisted of cystically dilated glands, lined by normal colonic epithelial cells, scattered in loose, edematous stroma containing inflammatory cell infiltration. There were no areas of tubular adenoma or malignancy in any of the polyp.
Adenoma ; Adolescent ; Colectomy ; Colon ; Epithelial Cells ; Gastrointestinal Tract ; Hemorrhage ; Humans ; Intestinal Polyposis ; Polyps

Toxoplasma gondii infection induces parasite infiltration and apoptosis in the spleen. However, dose-dependent parasite infiltration, apoptosis, body weight alternations and survival in mice remain largely unknown. In this study, mice were intraperitoneally infected with 10, 30 or 100 tachyzoites of T. gondii, respectively. Parasite infiltration and apoptosis in the spleen were analyzed on days 3, 7, and 9 post-infection by immunohistochemistry and flow cytometry. Significantly higher levels of T. gondii infiltration and apoptosis in the spleen were found in 30 and 100 tachyzoites infected mice compared to 10 tachyzoites infected mice on days 7 and 9 post-infection. Although 30 and 100 tachyzoites infected mice showed significant body weight loss compared to 10 tachyzoites infected mice, all of the 100, 30, and 10 tachyzoites infected mice died by days 12, 15, and 17, each respectively. Interestingly, T. gondii infiltration in 10 tachyzoites infected mice were limited to capsule area of the spleen on day 9 post-infection. Several areas of parasite infiltrations were found in the 30 tachyzoites infected mice, where noticeable levels of splenic capsule de-adhesion occurred. These results indicated that parasite infiltration and apoptosis in the spleen, as well as body weight loss (survival) are closely correlated with infection dosage. The level of T. gondii infiltration and apoptosis in the spleen and splenic de-adhesion were dependent on the parasite dose.
Animals ; Apoptosis ; Body Weight ; Flow Cytometry ; Immunohistochemistry ; Mice ; Parasites ; Spleen ; Toxoplasma ; Toxoplasmosis

Encystation mediating cyst specific cysteine proteinase (CSCP) of Acanthamoeba castellanii is expressed remarkably during encystation. However, the molecular mechanism involved in the regulation of CSCP gene expression remains unclear. In this study, we focused on epigenetic regulation of gene expression during encystation of Acanthamoeba. To evaluate methylation as a potential mechanism involved in the regulation of CSCP expression, we first investigated the correlation between promoter methylation status of CSCP gene and its expression. A 2,878 bp of promoter sequence of CSCP gene was amplified by PCR. Three CpG islands (island 1–3) were detected in this sequence using bioinformatics tools. Methylation of CpG island in trophozoites and cysts was measured by bisulfite sequence PCR. CSCP promoter methylation of CpG island 1 (1,633 bp) was found in 8.2% of trophozoites and 7.3% of cysts. Methylation of CpG island 2 (625 bp) was observed in 4.2% of trophozoites and 5.8% of cysts. Methylation of CpG island 3 (367 bp) in trophozoites and cysts was both 3.6%. These results suggest that DNA methylation system is present in CSCP gene expression of Acanthamoeba. In addition, the expression of encystation mediating CSCP is correlated with promoter CpG island 1 hypomethylation.
Acanthamoeba castellanii* ; Acanthamoeba* ; Computational Biology ; CpG Islands ; Cysteine Proteases ; DNA Methylation* ; DNA* ; Epigenomics ; Gene Expression Regulation ; Gene Expression* ; Methylation ; Negotiating ; Polymerase Chain Reaction ; Trophozoites

Protein arginine methyltransferase (PRMT) is an important epigenetic regulator in eukaryotic cells. During encystation, an essential process for Acanthamoeba survival, the expression of a lot of genes involved in the encystation process has to be regulated in order to be induced or inhibited. However, the regulation mechanism of these genes is yet unknown. In this study, the full-length 1,059 bp cDNA sequence of Acanthamoeba castellanii PRMT1 (AcPRMT1) was cloned for the first time. The AcPRMT1 protein comprised of 352 amino acids with a SAM-dependent methyltransferase PRMT-type domain. The expression level of AcPRMT1 was highly increased during encystation of A. castellanii. The EGFP-AcPRMT1 fusion protein was distributed over the cytoplasm, but it was mainly localized in the nucleus of Acanthamoeba. Knock down of AcPRMT1 by synthetic siRNA with a complementary sequence failed to form mature cysts. These findings suggested that AcPRMT1 plays a critical role in the regulation of encystation of A. castellanii. The target gene of AcPRMT1 regulation and the detailed mechanisms need to be investigated by further studies.
Acanthamoeba castellanii* ; Acanthamoeba* ; Amino Acids ; Clone Cells ; Cytoplasm ; DNA, Complementary ; Epigenomics ; Eukaryotic Cells ; Protein-Arginine N-Methyltransferases* ; RNA, Small Interfering

Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.