Molecular Cloning of Plasmodium vivax Calcium-Dependent Protein Kinase 4

A family of calcium-dependent protein kinases (CDPKs) is a unique enzyme which plays crucial roles in intracellular calcium signaling in plants, algae, and protozoa. CDPKs of malaria parasites are known to be key regulators for stage-specific cellular responses to calcium, a widespread secondary messenger that controls the progression of the parasite. In our study, we identified a gene encoding Plasmodium vivax CDPK4 (PvCDPK4) and characterized its molecular property and cellular localization. PvCDPK4 was a typical CDPK which had well-conserved N-terminal kinase domain and C-terminal calmodulin-like structure with 4 EF hand motifs for calcium-binding. The recombinant protein of EF hand domain of PvCDPK4 was expressed in E. coli and a 34 kDa product was obtained. Immunofluorescence assay by confocal laser microscopy revealed that the protein was expressed at the mature schizont of P. vivax. The expression of PvCDPK4-EF in schizont suggests that it may participate in the proliferation or egress process in the life cycle of this parasite.

Therapeutic Considerations of Percutaneous Sacroplasty for the Sacral Insufficiency Fracture

Sacral insufficiency fracture is a debilitating injury not easily found in general radiologic examinations and is rarely diagnosed, since its symptoms are obscure. It is known to frequently occur in patients with osteoporosis, but the treatment has not yet been established and various kinds of treatment methods are being attempted. Sacroplasty is sometimes performed by applying percutaneous vertebroplasty which is known to be a less invasive treatment. Since the course of diagnosis of sacral insufficiency fracture is difficult and clear guidelines for treatments have not yet been established, many spine surgeons fail to diagnose patients or speculate on treatment methods. We report our experience in diagnosing a sacral insufficiency fracture in a 54-year-old healthy female patient using MRI and treating her with sacroplasty. From a therapeutic point of view, we then cover the usefulness, effects and characteristics relating to the complications of sacroplasty, along with literature review.

Characteristics of amniotic fluid derived stem cells with trisomy 21 / 대한산부인과학회지

OBJECTIVE: To assess molecular markers of amniotic fluid derived stem cells (AFSCs) in aspects of increased neurological deficit in Down syndrome. METHODS: Amniotic fluid samples through amniocentesis for prenatal diagnosis from four mid trimester pregnancies; by routine chromosomal analysis, two of them were trisomy 21 (Down syndrome) and others were normal, were selected after informed consent. Cells from two-stage culture protocol were assayed; morphology through phase contrast microscopy, chromosomal analysis, reverse transcriptase-polymerase chain reaction and Western blot analysis. RESULTS: AFSCs were highly proliferative in subcultures and most of them were mononuclear, fibroblast-like, fusiform cells. There were also a few ovoid cells. The chromosomal analysis of amniotic fluid stem cells was identical to that of amniotic fluid cells. Two of four samples were 47,XX,+21, others were 46,XX. Of the proteins related to Down syndrome, the expression of S100beta were increased in AFSCs of Down syndrome, COL6A1 (Collagen IV, alpha 1) was down-regulated in them and insulin like growth factor binding protein-1 was expressed in all AFSCs. Stem cell markers were expressed heterogeneously. Oct4 (POU5F1), nanog, and SOX2 (sex determining region Y) were expressed in both groups. But c-Kit was not expressed in AFSCs of Down syndrome. The neural cell marker, neuron specific enolase was detected in both groups. Other neural cell markers, microtubule associated protein 2, glial fibrillary acidic protein were undetectable in ASFCs of Down syndrome. Bcl-2 gene family proteins related with apoptosis were assayed. The expression of Bcl-XL was increased in Down syndrome more than in normal pregnancy. Bcl-2 and BID were expressed in all AFSCs and Bax was down-regulated in Down syndrome. CONCLUSION: AFSCs are an excellent choice for many future tissue engineering strategies and cell based therapies. Analysis of molecular features of AFSCs from normal and Down syndrome will provide the basis of further experimental study.

Preliminary report of Canal Transobturator Tape for Stress Urinary Incontinence in Women Over 65 Years

PURPOSE: To access the feasibility of Canal transobturator tape (Canal TOT) for stress urinary incontinence (SUI) in women over 65 year old. MATERIALS AND METHODS: From August 2006 to December 2008, we reviewed the medical records of 261 patients underwent Canal TOT in Division of Urogynecology, Department of Obstetrics and Gynecology, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine in Seoul. This study is a retrospective analysis of the clinical characteristics (age, gravida, parity, body mass index), previous operation history, comorbidity, surgical procedure and operation outcomes. We performed stress test, one hour pad test, urodynamic study, postvoid residual assessment to all patients for diagnosis of SUI. All patients answered self assessment questionnaires (IIQ-7, UDI-6) at 6 months and 12 months after operation. RESULTS: 55 women over 65 years were underwent Canal TOT. Mean follow up was 11+/-4.5 months. Mean age of patients was 70.2+/-3.9 years, gravida 5.8+/-2.3 times, parity 3.9+/-1.5 times and body mass index (BMI) 25.6+/-3.1Kg/m2. 8 patients had got hysterectomy (14.5%). 16 patients (29%) had sling operation (Canal TOT alone), and 39 patients (71%) had Canal TOT combined with vaginal surgery for pelvic organ prolapse. The cure rate was 96.4% in 6 months follow up. Leakage after operation were reported by 2 patients (3.6%) and 3 patients (5.5%) transiently suffered from postoperative voiding difficulty. Of whom had incontinence complexed with overactive bladder symptoms (frequency, nocturia, and urgency) 12 patients complained of persistent symptoms after Canal TOT (12/33, 36.4%). The scores from self assessment questionnaires (IIQ-7, UDI-6) at6months after operation were improved significantly. CONCLUSION: Canal TOT is feasible and safe method for SUI in old age. The procedure also shows favorable results when combined with other operations for pelvic organ prolapse.

Ex vivo generation of functional dendritic cells from mobilized CD34+ hematopoietic stem cells

The ability to generate dendritic cells (DCs) in sizeable numbers has enormous implications for the development of clinically-effective antigen presentation procedures for cancer immunotherapy. We evaluated the generation of immunostimulatory DCs from peripheral blood CD34+ cells collected from healthy donors. CD34+ cells purified from leukapheresis product were seeded at 1 x 10(4) cells/mL in complete medium supplemented with GM-CSF, TNF alpha, IL-4, c-kit ligand, and flt3 ligand (FL). By day 14 of culture in the presence of GM-CSF + TNF alpha, the total cell number increased by 23.4 +/- 5.4-fold compared to the starting number of CD34+ cells. When the c-kit and FL were added to GM-CSF and TNF alpha, the cell number increased by 109.8 +/- 11.2-fold without affecting the immunophenotype of recovered cells. Flow cytometric analysis indicated that cells with the markers of mature dendritic cells, i.e., CD1a +CD14 -HLA-DR+, and CD80+CD86+HLA-DR+, constituted 49.0% +/- 7.5%, and 38.9% +/- 6.5%, respectively. This pattern of expression of surface antigen was unchanged whether the c-kit ligand and/or FL was added. The irradiated CD1a+HLA-DR+ cells recovered from in vitro cultures elicit a vigorous proliferation of allogeneic peripheral blood T-cells, irrespective of cytokine combinations. These findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment.

The Usefulness of Fetal Fibronectin in Pregnant Women / 대한임상병리학회지

BACKGROUND: The purpose of our study is to evaluate the usefulness of cervicovaginal fetal fibronectin assay for the prediction of rupture of membrane and preterm labor. METHODS: A group of 39 pregnant women was involved in this prospective study. Out of 139 pregnant women, 96 were clinically diagnosed as ruptured membranes (group A). The remaining 43 of 139 pregnant women were clinically diagnosed as preterm labor(group B). The assay was performed by using the ROMCheckTM kit (Adeza Biomedical Corp., Sunnyvale, CA). RESULTS: In group 4, fetal fibronectin (fFN) positive rate is 55% (53 patients) and negative rate is 45% (43 patients). In group B, fFN positive rate is 56% (24 patients) and negative rate is 44% (19 patients). Both group of fFN positive patients show a significantly shorter interval from sampling to delivery than fFN negative patients. Also in group A, the percentage of fFN positive patients who delivered at less than 48 hours after sampling is greater than those with fFN negative patients and in group B, the preterm delivery rate is 79% with positive fFN and 37% with negative fFN. As a predictor for preterm delivery, the presence of fFN has the sensitivity 79%, the specificity 84%, the positive Predictive value 76% and the negative predictive value 86%. CONCLUSIONS: The result suggests that a positive fFN in pregnant women who have uterine contractions and ruptured membrane indicates a significant risk for preterm delivery and labor onset, and a negative fFN is a reassuring sign.

Prevalence Rate of Irregular Antibodies in Transfusion Candidates and Pregnant Women in Korea / 대한임상병리학회지

BACKGROUND: The authors studied to estimate the frequency of irregular antibodies and their significance in blood transfusion and antenatal care in Korea. METHODS: Irregular antibodies were tested by immediate saline spin, 37degrees C albumin and antiglobulin test for 2,008 transfusion candidates and 1,047 pregnant women at Severance hospital using commercial screening and identification cells (Dade, U.S.A.). RESULTS: The irregular antibodies were detected in 38 (1.24%) of total 3,055 subjects (transfusion candidates: 0.9%, pregnant women: 1.91%) . In transfusion candidates, the detected antibodies were Lewis antibodies, cold antibodies (anti-M, anti-P 1), Rh antibodies and unspeified warm antibodies, and their distributions were 56%, 22%, 17%, and 5%, respectively. In pregnant women, the detected antibodies were Lewis antibodies, Rh antibodies, anti-Jra, and unspeified warm antibodies, and their distributions were 45%, 45%, 5% and 5%, respectively. At immediate saline phase, 58% of irregular antibodies were detected. At 37degrees C albumin phase, 90% of irregular antibodies were detected and only 10% of irregular antibodies were detected at antiglobulin phase. CONCLUSIONS: Although the prevalence rates of clinically important irregular antibodies were low, 1/1000 of irregular antibodies could not be detected. Therefore, irregular antibody screening should be performed in all pretransfusion test. And, if antibody detection tests are negative, immediate saline crossmaching methods are acceptable in Korea.

Prevalence Rate of Irregular Antibodies in Transfusion Candidates and Pregnant Women in Korea / 대한임상병리학회지

BACKGROUND: The authors studied to estimate the frequency of irregular antibodies and their significance in blood transfusion and antenatal care in Korea. METHODS: Irregular antibodies were tested by immediate saline spin, 37degrees C albumin and antiglobulin test for 2,008 transfusion candidates and 1,047 pregnant women at Severance hospital using commercial screening and identification cells (Dade, U.S.A.). RESULTS: The irregular antibodies were detected in 38 (1.24%) of total 3,055 subjects (transfusion candidates: 0.9%, pregnant women: 1.91%) . In transfusion candidates, the detected antibodies were Lewis antibodies, cold antibodies (anti-M, anti-P 1), Rh antibodies and unspeified warm antibodies, and their distributions were 56%, 22%, 17%, and 5%, respectively. In pregnant women, the detected antibodies were Lewis antibodies, Rh antibodies, anti-Jra, and unspeified warm antibodies, and their distributions were 45%, 45%, 5% and 5%, respectively. At immediate saline phase, 58% of irregular antibodies were detected. At 37degrees C albumin phase, 90% of irregular antibodies were detected and only 10% of irregular antibodies were detected at antiglobulin phase. CONCLUSIONS: Although the prevalence rates of clinically important irregular antibodies were low, 1/1000 of irregular antibodies could not be detected. Therefore, irregular antibody screening should be performed in all pretransfusion test. And, if antibody detection tests are negative, immediate saline crossmaching methods are acceptable in Korea.