Since pluripotent cells were first derived from the inner cell mass (ICM) of mouse blastocysts, tremendous efforts have been made to establish embryonic stem cell (ESC) lines in several domestic species including the pig; however, authentic porcine ESCs have not yet been established. It has proven difficult to derive pluripotent cells of naïve state that represents full pluripotency, due to the frequent occurrence of spontaneous differentiation into an EpiSC-like state during culture in pigs. We have been able to derive EpiSC-like porcine embryonic stem cell (pESC) lines of a differentiated non-ES cell state from blastocyst stage porcine embryos of various origins, including in vitro fertilized (IVF), in vivo derived, IVF aggregated and parthenogenetic embryos. In addition, we have generated induced pluripotent stem cells (piPSCs) via plasmid transfection of reprogramming factors (Oct4, Sox2, Klf4 and c-Myc) into porcine fibroblast cells. In this study, we analysed characteristics such as marker expression, pluripotency and the X chromosome inactivation (XCI) status of our EpiSC-like pESC lines along with our piPSC line. Our results show that these cell lines demonstrate the expression of genes associated with the Activin/Nodal and FGF2 pathways along with the expression of pluripotent markers Oct4, Sox2, Nanog, SSEA4, TRA 1-60 and TRA 1-81. Furthermore all of these cell lines showed in vitro differentiation potential; female XCI activity and a normal karyotype. Here we provide preliminary results that suggest that, as a nonpermissive species, the porcine species undergoes reprogramming into a primed state during the establishment of pluripotent stem cell lines.

Chronic ulcerations are a physical and financial burden to the health and economic establishment in the United States and Worldwide. Improvements in biotechnology and knowledge in stem cell applications have progressed and basic science results are making their way slowly into the clinical arena. Chronic wounds and diabetic bone healing are the key components in the limb salvage of the common diabetic foot. We have examined the current available literature and present the latest on stem cells applications as a novel clinical technique in the treatment of chronic wound and diabetic bone healing and their impact in the treatment paradigm of patients.

OBJECTIVES The purpose of this study was to assess the microbiological quality of unchlorinated drinking water in Korea, 2010. One hundred and eighty unchlorinated drinking water samples were collected from various sites in Seoul and Gyeonggi province. METHODS To investigate bacterial presence, the pour plate method was used with cultures grown on selective media for total bacteria, total coliforms, and Staphylococcus spp., respectively. RESULTS In the 180 total bacteria investigation, 72 samples from Seoul and 33 samples from Gyeonggi province were of an unacceptable quality (>10(2) CFU/mL). Of all the samples tested, total coliforms were detected in 28 samples (15.6%) and Staphylococcus spp. in 12 samples (6.7%). Most of the coliform isolates exhibited high-level resistance to cefazolin (88.2%), cefonicid (64.7%) and ceftazidime (20.6%). In addition, Staphylococcus spp. isolates exhibited high-level resistance to mupirocin (42%). Species of Pseudomonas, Acinetobacter, Cupriavidus, Hafnia, Rahnella, Serratia, and Yersinia were isolated from the water samples. CONCLUSIONS The results of this study suggest that consumption of unchlorinated drinking water could represent a notable risk to the health of consumers. As such, there is need for continuous monitoring of these water sources and to establish standards.

Zinc Oxide (ZnO) based Thin Film Transistors (TFTs) have been fabricated and analyzed to investigate mechanical characteristics regarding the stress, strain and deformation of electro circuits using the Finite Element Method (FEM). As the best compromise between the stretching and bending abilities, the coating thickness of SU-8 can be as important for bendability as a neutral mechanical plane. The neutral mechanical plane in electro circuits was designed for obtaining flexibility, e.g., bendability, in a previous numerical study. After that, through experimental validation, we observed what degree of SU-8 thickness was attributable for improved mechanical stability. The results suggest that not only numerical but also experimental measurements of the deformation and SU-8 coating thickness in electro circuits are useful for enhancing structural stability.

PURPOSE: To investigate which technique would reduce bending stress at the femoral tunnel aperture and make short tunnel length after ACL reconstruction by comparing the femoral graft bending angle and tunnel length between the single-bundle (SB) transtibial (TT) and double-bundle (DB) transportal (TP) technique using three-dimensional-computed tomography using OsiriX(®) imaging software. METHODS: Forty-nine patients underwent an ACL reconstruction using a SB TT (Group I, 20 patients) and DB TP (Group II, 29 patients) technique. Femoral graft bending angle and femoral tunnel length were measured by CT image using OsiriX(®) imaging software. Groups I and II were compared, and statistical analysis was performed using SPSS software. RESULTS: The mean anteromedial (AM) and posterolateral (PL) femoral graft bending angle of group II (111.5 ± 8.8° and 118.9 ± 9.8°, respectively) was significantly more acute than that of group I (125.3 ± 11.1°)(P < 0.001, P = 0.04). The mean femoral tunnel length of group I was significantly longer than that of group II (P = 0.001). CONCLUSIONS: The femoral graft bending angle and the femoral tunnel length of the TP technique performed in the maximally flexed knee position was more acute and shorter than those of the TT technique after ACL reconstruction. This might increase the bending stress at the femoral tunnel aperture and shorter graft length in the tunnel after an ACL reconstruction using TP technique compared to the TT technique. LEVEL OF EVIDENCE: III.

PURPOSE: The purpose of this study was to examine a developed surgical technique by performing a mid-term evaluation of clinical and stability results and complications. METHODS: Thirty patients who underwent transtibial posterior cruciate ligament (PCL) reconstruction using a bioabsorbable cross-pin tibial back side fixation method were enrolled in this prospective study. Lysholm and International Knee Documentation Committee (IKDC) knee scales were used to evaluate clinical outcomes. Stability was evaluated using a Telos device with a 150 N force at 90 degrees of knee flexion. Follow-up magnetic resonance imaging (MRI) was also performed in 20 (66.7%) patients, and complications were evaluated. Those with complication by MRI were assigned to an abnormal MRI group. RESULTS: The follow-up period was 47 (range, 25-62) months. On comparing preoperative and final follow-up clinical results, Lysholm and IKDC knee scale scores were found to have improved significantly (P < 0.001). The mean side-to-side difference in posterior translation measured using a Telos device was 13.4 ± 3.1 mm (range 10-20 mm) preoperatively and 3.2 ± 1.5 mm (range 1-7 mm) at last follow-up, which represented a significant improvement in stability (P < 0.001). Five patients showed cyst formation in the tibial tunnel and two patients showed a significant signal increase at the anterior portion of the tibial tunnel, which was believed to indicate a pro-cystic status. The normal and abnormal MRI groups had similar Lysholm and IKDC knee scale scores and stress radiographs (P > 0.05). CONCLUSIONS: Single-bundle transtibial PCL reconstruction using a bioabsorbable cross-pin tibial back side fixation was found to produce satisfactory clinical and stability results. However, despite these satisfactory results, a potential complication of tibial cyst formation was observed. LEVEL OF EVIDENCE: Case series, Level IV.

Round ligament varicosities during pregnancy are rare, and can easily be mistaken for an inguinal hernia. On physical examination, round ligament varicosities and groin hernia are difficult to distinguish. The diagnosis of round ligament varicosities can be established on gray-scale and color Doppler sonography. We experienced a case of round ligament varicosities in which a 29-year-old woman presented symptoms at 36 weeks gestation. The patient was diagnosed using Doppler sonography, managed with conservative therapy, and had an uncomplicated vaginal delivery at 40 weeks. The symptoms were resolved completely by two weeks postpartum. We report a case of round ligament varicosities that was diagnosed at 36 weeks of gestation with a review of the literature.

Pseudomonas aeruginosa exists in various environments, and can cause mild or serious infections resulting in a wide range of symptoms. In this study, we collected bacterial isolates from hospitalized patients and unchlorinated drinking water, in Korea, 2010. The water-borne and clinical isolates were compared using colony morphology, antimicrobial susceptibility testing, and random amplification of polymorphic DNA analysis. We first compared morphological features of the water-borne and clinical isolates. The clearest difference in colony morphology was colony shape; five water-borne isolate colonies (83%) had a smooth, circular morphology, while nine (75%) clinical isolate colonies had a rough, irregular morphology. Minimum inhibitory concentrations analyses were performed to determine antimicrobial resistant patterns; using ceftazidime, gentamicin, tigecycline, chloramphenicol, meropenem, and tobramycin according to Clinical and Laboratory Standard Institute (CLSI, 2009) methodology. All waterborne isolates were not resistant to gentamicin, tobramycin, and meropenem. The clinical isolates were resistant to every antibiotic except chloramphenicol. Genotyping was performed using the repetitive extragenic palindromic polymerase-chain-reaction. The DNA fingerprinting patterns did not reveal genetic similarity between the water-borne and clinical P. aeruginosa isolates. On the contrary, they showed that genetically distinct populations have been established in each of these environments. We have revealed significant morphological, clinical and genetic differences between water-borne and clinical isolates of the same bacterial species.

Imiquimod is a synthetic Toll-like receptor 7 (TLR7) agonist approved for the topical treatment of actinic keratoses, superficial basal cell carcinoma, and genital warts. Imiquimod leads to an 80-100% cure rate of lentigo maligna; however, studies of invasive melanoma are lacking. We conducted a pilot study to characterize the local, regional, and systemic immune responses induced by imiquimod in patients with high-risk melanoma. After treatment of the primary melanoma biopsy site with placebo or imiquimod cream, we measured immune responses in the treated skin, sentinel lymph nodes (SLNs), and peripheral blood. Treatment of primary melanomas with 5% imiquimod cream was associated with an increase in both CD4+ and CD8+ T cells in the skin, and CD4+ T cells in the SLN. Most of the CD8+ T cells in the skin were CD25 negative. We could not detect any increases in CD8+ T cells specifically recognizing HLA-A(*)0201-restricted melanoma epitopes in the peripheral blood. The findings from this small pilot study demonstrate that topical imiquimod treatment results in enhanced local and regional T-cell numbers in both the skin and SLN. Further research into TLR7 immunomodulating pathways as a basis for effective immunotherapy against melanoma in conjunction with surgery is warranted.

The purpose of this study was to fabricate porous poly(lactide-co-glycolide)(PLGA) microparticles for efficient pulmonary deposition and increased therapeutic duration of the antioxidant anthocyanin (ATH). These microparticles were prepared by a water-in-oil-in-water (W(1)/O/W(2)) multi-emulsion method with vaporizing ammonium bicarbonate (AB) as a porogen and starch as a viscous additive. High porosity achieved by the decomposition reaction of AB to the base of ammonia, carbon dioxide, and water vapor at 50°C enabled efficient deposition of ATH throughout the entire lung in BALB/c mice. In addition, the porous microparticles incorporating starch showed sustained ATH release characteristics (up to 5 days) and protracted antioxidant activity (up to 5 days) for 2,2-diphenyl-1-pikryl-hydrazyl (DPPH) radicals, which was comparable to that of the porous microparticles without starch which completely released ATH in 2h. Furthermore, these porous microparticles incorporating starch led to longer ATH residence (up to 20 days) in in vivo lung epithelium. We believe that this system has great pharmaceutical potential as a long-acting antioxidant for continuously relieving oxidative stress in pulmonary diseases like chronic obstructive pulmonary disease (COPD).