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2.  Hereditary Risk Evaluation for Borderline Ovarian Tumors Based on Immunohistochemistry 
Journal of Menopausal Medicine  2014;20(1):14-20.
Borderline ovarian tumors (BOT) are premalignant lesions. Approximately 10% of all epithelial ovarian cancers are known to be hereditary with hereditary breast and ovarian cancer (HBOC) accounting for approximately 90% of cases; the remaining 10% are attributable to Lynch syndrome, also known as hereditary nonpolyposis colorectal cancer (HNPCC). The aim of our study is to estimate this risk based on screening immunohistochemistry (IHC).
Thirty-four patients diagnosed with BOT were identified. Family history, clinical characteristics, and IHC data (breast cancer 1, early onset [BRCA1], breast cancer 2, early onset [BRCA2], mutS homolog 2 [MSH2], mutL homolog 1 [MLH1]) were collected for all cases from the patients' medical charts. Nuclear staining of the tumor was scored as negative and positive.
Among 32 patients, 14 (44%) had serous type and 18 (56%) had mucinous type. The mean patient age was 44 years (range 19-86).The number of patients with weak IHC staining for MSH2 and BRCA2 was 1 (3%) and 6 (19%) respectively. The median follow up was 21.8 months.
According to the results, we discovered that 3% and 19% of patients with BOT had a risk of hereditary cancer based on IHC analysis respectively. This pilot study may help clinician to counsel effectively for confirmative tests.
PMCID: PMC4217565  PMID: 25371887
Lynch syndrome; Ovarian neoplasmsms
3.  Perimenopausal Ovarian Carcinoma Patient with Subclavian Node Metastasis Proven by Immunohistochemistry 
Journal of Menopausal Medicine  2014;20(1):43-46.
Ovarian cancer is the seventh most common cancer in the world and the fifth most common cause of death from cancer; it is responsible for over half of all deaths related to gynecological cancers. The presence of lymphatic metastasis is an important prognostic factor in ovarian cancer. Nodal metastases to the pelvic and the para-aortic lymph nodes are common, particularly in an advanced of the disease (stages III-IV). The finding of distant nodal metastasis, especially subclavian lymph node metastasis, from ovarian carcinoma is very uncommon. 18F-fluorodeoxyglucose-positron emission tomography (FDG-PET) or FDG-PET/computed tomography (CT) provides an improved imaging for detecting metastatic lymph nodes in patients with ovarian cancer. Immunohistochemically, ovarian carcinoma cells are positive for estrogen receptor, progesterone receptor, cancer antigen 125, Wilms' tumor 1 protein, and p53; they are negative for thyroid transcription factor (TTF-1) and caudal-related homeobox 2 (CDX-2). This report describes a Korean woman diagnosed with ovarian cancer with subclavian lymph node metastasis revealed by FDG PET/CT and verified by an immunohistochemical staining. Differentiating between the primary ovarian lesion and the metastatic lesion will allow the initiation of an appropriate treatment and help predict the prognosis.
PMCID: PMC4217566  PMID: 25371892
Lymphatic metastasis; Ovary cancer
4.  A Case of Perimenopausal Endometrial Cancer in a Woman with MSH2 Germline Mutation 
Journal of Menopausal Medicine  2013;19(3):143-146.
Lynch syndrome is a genetic malignancy syndrome affecting the colon, endometrium, and other organs. It is difficult to find a Lynch syndrome patient without any family history of cancer. We have recently examined an endometrial cancer patient with a MSH2 gene mutation without a family history of cancer. A 55-year old Korean woman was admitted to a local clinic for vaginal bleeding. An endometrial biopsy revealed the presence of adenocarcinoma (endometrioid type, grade 1). After surgical staging, no further adjuvant therapy was required. Analysis of the tissue using immunohistochemistry (IHC) showed the endometrium stained negatively for MSH2. Microsatellite instability (MSI) was analyzed for five markers. The patient was scored as unstable. Further, additional gene sequencing revealed one missense mutation in c.23C > T (p.Thr8Met). This is the first case of Lynch syndrome endometrial cancer in Korea in which the patient does not have any family history of cancer.
PMCID: PMC4217557  PMID: 25371881
Endometrial neoplasms; Lynch syndrome
5.  One case of endometrial cancer occurrence: Over 10 years after colon cancer in Lynch family 
Obstetrics & Gynecology Science  2013;56(6):408-411.
We have recently experienced an endometrial cancer 12 years after the diagnosis of colon cancer with Lynch syndrome. A 49-year-old Korean woman had a family history of colon cancer. Her mother had colon cancer at 56-year-old, and her brother had colon cancer at 48 years old. The patient received surgery for endometrial cancer at the same hospital 12 years after being treated for colon cancer. Immunohistochemistry showed that her endometrial tissue stained negative for MSH2. A microsatellite instability test was performed and showed the presence of instability high microsatellite instability. An hMLH2 gene mutation was detected at codon 629 codon of exon 12, in which a glutamine was replaced with an arginine (1886A>G [p.Gln629Arg]). To our knowledge, this is the first case of metachronous cancer in a Lynch syndrome family in Korea with a gap of more than ten years between cancer diagnoses.
PMCID: PMC3859014  PMID: 24396821
Colon cancer; Endometrial cancer; Lynch syndrome
6.  Identification of RUNX3 as a Component of the MST/Hpo Signaling Pathway 
Journal of cellular physiology  2012;227(2):839-849.
Recent genetic screens of fly mutants and molecular analysis have revealed that the Hippo (Hpo) pathway controls both cell proliferation and cell death. Deregulation of its human counterpart (the MST pathway) has been implicated in human cancers. However, how this pathway is linked with the known tumor suppressor network remains to be established. RUNX3 functions as a tumor suppressor of gastric cancer, lung cancer, bladder cancer, and colon cancer. Here, we show that RUNX3 is a principal and evolutionarily conserved component of the MST pathway. SAV1/WW45 facilitates the close association between MST2 and RUNX3. MST2, in turn, stimulates the SAV1–RUNX3 interaction. In addition, we show that siRNA-mediated RUNX3 knockdown abolishes MST/Hpo-mediated cell death. By establishing that RUNX3 is an endpoint effector of the MST pathway and that RUNX3 is capable of inducing cell death in cooperation with MST and SAV1, we define an evolutionarily conserved novel regulatory mechanism loop for tumor suppression in human cancers.
PMCID: PMC3632350  PMID: 21678419
7.  Is a Metallic Microcoil Really a Permanent Embolic Agent for the Management of Distal Guidewire-Induced Coronary Artery Perforation? 
Korean Circulation Journal  2011;41(8):474-478.
Coronary artery perforation (CAP) after percutaneous coronary intervention is a rare, but serious complication. It can cause cardiac tamponade, acute myocardial infarction or death. The treatments of CAP involve prolonged balloon inflation, emergent surgery, coil embolization, and implantation of covered stent. We have successfully performed the emergent microcoil embolization in a patient with uncontrolled Ellis grade 3 guidewire-induced CAP resulting in delayed cardiac tamponade. Contrasting our usual expectation, the 1-year follow-up angiography showed a patent flow at the embolized site.
PMCID: PMC3173669  PMID: 21949533
Embolization, therapeutic; Cardiac tamponade; Angioplasty, balloon, coronary
8.  Knock-in of Enhanced Green Fluorescent Protein or/and Human Fibroblast Growth Factor 2 Gene into β-Casein Gene Locus in the Porcine Fibroblasts to Produce Therapeutic Protein 
Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine β-casein gene locus using a knock-in vector for the β-casein gene locus. We developed the knock-in vector on the porcine β-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5′ arm and 3′ arm from the porcine β-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using β-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous β-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.
PMCID: PMC4213711  PMID: 25358326
Knock-in; Homologous Recombination; Therapeutic Proteins; Porcine β-casein Gene
9.  Impact of Framingham Risk Score, Flow-Mediated Dilation, Pulse Wave Velocity, and Biomarkers for Cardiovascular Events in Stable Angina 
Journal of Korean Medical Science  2014;29(10):1391-1397.
Although the age-adjusted Framingham risk score (AFRS), flow-mediated dilation (FMD), brachial-ankle pulse wave velocity (baPWV), high-sensitivity C-reactive protein (hsCRP), fibrinogen, homocysteine, and free fatty acid (FFA) can predict future cardiovascular events (CVEs), a comparison of these risk assessments for patients with stable angina has not been reported. We enrolled 203 patients with stable angina who had been scheduled for coronary angiography (CAG). After CAG, 134 patients showed significant coronary artery disease. During 4.2 yr follow-up, 36 patients (18%) showed CVEs, including myocardial infarction, de-novo coronary artery revascularization, in-stent restenosis, stroke, and cardiovascular death. ROC analysis showed that AFRS, FMD, baPWV, and hsCRP could predict CVEs (with AUC values of 0.752, 0.707, 0.659, and 0.702, respectively, all P<0.001 except baPWV P=0.003). A Cox proportional hazard analysis showed that AFRS and FMD were independent predictors of CVEs (HR, 2.945; 95% CI, 1.572-5.522; P=0.001 and HR, 0.914; 95% CI, 0.826-0.989; P=0.008, respectively). However, there was no difference in predictive power between combining AFRS plus FMD and AFRS alone (AUC 0.752 vs. 0.763; z=1.358, P=0.175). In patients with stable angina, AFRS and FMD are independent predictors of CVEs. However, there is no additive value of FMD on the AFRS in predicting CVEs.
Graphical Abstract
PMCID: PMC4214940  PMID: 25368493
Framingham Risk Score; Flow-Mediated Dilation; Cardiovascular Event
10.  Crystallization and preliminary X-ray crystallographic analysis of the putative NADP(H)-dependent oxidoreductase YncB from Vibrio vulnificus  
In order to investigate its structure and function, the medium-chain dehydrogenase/reductase superfamily YncB-like protein from V. vulnificus was expressed, purified and crystallized. X-ray diffraction analyses of YncB and the YncB–NADP(H) complex are reported at resolutions of 1.85 and 1.81 Å, respectively.
The yncB gene product from Vibrio vulnificus, which belongs to the medium-chain dehydrogenase/reductase (MDR) superfamily, was crystallized using the microbatch crystallization method at 295 K. Diffraction data sets were collected using synchrotron radiation. Crystals of selenomethionine-substituted YncB protein belonged to space group P212121, with unit-cell parameters a = 90.52, b = 91.56, c = 104.79 Å. Assuming the presence of two molecules in the asymmetric unit, the solvent content was estimated to be about 57%. Crystals of the YncB–NADP(H) complex belonged to space group P41212 or P43212, with unit-cell parameters a = b = 90.14, c = 105.61 Å. Assuming the presence of one molecule in the asymmetric unit, the solvent content was estimated to be about 56.42%.
PMCID: PMC3433207  PMID: 22949204
MDR superfamily; Vibrio vulnificus; YncB
11.  Holmes' Tremor Associated with Bilateral Hypertrophic Olivary Degeneration Following Brain Stem Hemorrhage: A Case Report 
Holmes' tremor is a condition characterized by a mixture of postural, rest, and action tremors due to midbrain lesions in the vicinity of the red nucleus. Hypertrophic olivary degeneration (HOD) is a rare type of neuronal degeneration involving the dento-rubro-olivary pathway and may present clinically as Holmes tremor. We report on a 59-year-old female patient who developed Holmes tremor in association with bilateral HOD, following brain stem hemorrhage.
PMCID: PMC4205259  PMID: 25340035
Tremor; Red nucleus; Olivary Nucleus; Midbrain
12.  In-House Zinc SAD Phasing at Cu Kα Edge 
Molecules and Cells  2013;36(1):74-81.
De novo zinc single-wavelength anomalous dispersion (Zn-SAD) phasing has been demonstrated with the 1.9 Å resolution data of glucose isomerase and 2.6 Å resolution data of Staphylococcus aureus Fur (SaFur) collected using in-house Cu Kα X-ray source. The successful in-house Zn-SAD phasing of glucose isomerase, based on the anomalous signals of both zinc ions introduced to crystals by soaking and native sulfur atoms, drove us to determine the structure of SaFur, a zinc-containing transcription factor, by Zn-SAD phasing using in-house X-ray source. The abundance of zinc-containing proteins in nature, the easy zinc derivatization of the protein surface, no need of synchrotron access, and the successful experimental phasing with the modest 2.6 Å resolution SAD data indicate that in-house Zn-SAD phasing can be widely applicable to structure determination.
PMCID: PMC3887929  PMID: 23686432
anomalous scattering; experimental phasing; protein crystallography; SAD phasing; zinc
13.  The potential of mesenchymal stem cells derived from amniotic membrane and amniotic fluid for neuronal regenerative therapy 
BMB Reports  2014;47(3):135-140.
The mesenchymal stem cells (MSCs), which are derived from the mesoderm, are considered as a readily available source for tissue engineering. They have multipotent differentiation capacity and can be differentiated into various cell types. Many studies have demonstrated that the MSCs identified from amniotic membrane (AM-MSCs) and amniotic fluid (AF-MSCs) are shows advantages for many reasons, including the possibility of noninvasive isolation, multipotency, self-renewal, low immunogenicity, anti-inflammatory and nontumorigenicity properties, and minimal ethical problem. The AF-MSCs and AM-MSCs may be appropriate sources of mesenchymal stem cells for regenerative medicine, as an alternative to embryonic stem cells (ESCs). Recently, regenerative treatments such as tissue engineering and cell transplantation have shown potential in clinical applications for degenerative diseases. Therefore, amnion and MSCs derived from amnion can be applied to cell therapy in neuro-degeneration diseases. In this review, we will describe the potential of AM-MSCs and AF-MSCs, with particular focus on cures for neuronal degenerative diseases. [BMB Reports 2014; 47(3): 135-140]
PMCID: PMC4163884  PMID: 24499672
Amniotic fluid; Amniotic membrane; Mesenchymal stem cell; Neuronal disease; Regeneration therapy
14.  Value of Ultrasonography in the Diagnosis of Carpal Tunnel Syndrome: Correlation with Electrophysiological Abnormalities and Clinical Severity 
To investigate a diagnostic value of ultrasonography in carpal tunnel syndrome (CTS) patients and to evaluate a correlation of sonographic measurements with the degree of electrodiagnostic abnormalities and clinical severity.
Two-hundred-forty-six symptomatic hands in 135 patients and 30 asymptomatic hands in 19 healthy individuals as control group were included. In ultrasonographic study, we measured the cross-sectional area (CSA) and flattening ratio (FR) of the median nerve at the pisiform as well as palmar bowing (PB) of the flexor retinaculum. Sensitivity and specificity of ultrasonographic measurements were evaluated and ultrasonographic data from the symptomatic and control hands were compared to the grade of electrodiagnostic and clinical severity.
The mean CSA was 13.7±4.2 mm2 in symptomatic hands and 7.9±1.3 mm2 in asymptomatic hands. The mean FR was 4.2±1.0 in symptomatic hands and 3.4±0.4 in asymptomatic hands. The mean PB was 3.5±0.5 mm in symptomatic hands and 2.6±0.3 mm in asymptomatic hands. Statistical analysis showed differences of the mean CSA, FR and PB between groups were significant. A cut-off value of 10 mm2 for the mean CSA was found to be the upper limit for normal value. Both the mean CSA and PB are correlated with the grade of electrophysiological abnormalities and clinical severity, respectively.
Ultrasographic measurement of the CSA and PB is helpful to diagnose CTS as a non-invasive and an alternative modality for the evaluation of CTS. In addition, ultrasonography also provides a reliable correlation with the grade of electrodiagnostic abnormalities and clinical severity.
PMCID: PMC3958577  PMID: 24653800
Carpal tunnel syndrome; Ultrasonography; Diagnosis; Severity
15.  Develop to Term Rat Oocytes Injected with Heat-Dried Sperm Heads 
PLoS ONE  2013;8(11):e78260.
This study investigated the development of rat oocytes in vitro and in vivo following intracytoplasmic injection of heads from spermatozoa heat-dried at 50°C for 8 h and stored at 4°C in different gas phases. Sperm membrane and chromosome are damaged by the process of heat-drying. Oocyte activation and cleavage of oocytes were worse in oocytes injected with spermatozoa heat-dried and stored for 1 week than unheated, fresh spermatozoa, but in heat-dried spermatozoa, there were no differences in these abilities of oocytes between the samples stored in nitrogen gas and in air. The oocytes injected with heat-dried spermatozoa stored for 1 week could develop to the morula and blastocyst stages without difference between the samples stored in nitrogen gas and in air after artificial stimulation. Cleavage of oocytes and development of cleaved embryos were higher when heat-dried spermatozoa were stored for 3 and 6 months in nitrogen gas than in air. However, the ability of injected oocytes to develop to the morula and blastocyst stages was not inhibited even when heat-dried spermatozoa stored in both atmosphere conditions for as long as 6 months were used. When 2-cell embryos derived from oocytes injected with heads from spermatozoa heat-dried and stored for 1 week and 1 month were transferred, each 1 of 4 recipients was conceived, and the conceived recipients delivered 1 live young each. These results demonstrate that rat oocytes can be fertilized with heat-dried spermatozoa and that the fertilized oocytes can develop to term.
PMCID: PMC3817241  PMID: 24223784
16.  Neuronal cell differentiation of mesenchymal stem cells originating from canine amniotic fluid 
Human Cell  2013;27(2):51-58.
The amniotic fluid contains mesenchymal stem cells (MSCs) and can be readily available for tissue engineering. Regenerative treatments such as tissue engineering, cell therapy, and transplantation show potential in clinical trials of degenerative diseases. Disease presentation and clinical responses in the Canis familiaris not only are physiologically similar to human compared with other traditional mammalian models but is also a suitable model for human diseases. The aim of this study was to investigate whether canine amniotic-fluid-derived mesenchymal stem cells (cAF-MSCs) can differentiate into neural precursor cells in vitro when exposed to neural induction reagent. During neural differentiation, cAF-MSCs progressively acquire neuron-like morphology. Messenger RNA (mRNA) expression levels of neural-specific genes, such as NEFL, NSE, and TUBB3 (βIII-tubulin) dramatically increased in the differentiated cAF-MSCs after induction. In addition, protein expression levels of nestin, βIII-tubulin, and tyrosine hydroxylase remarkably increased in differentiated cAF-MSCs. This study demonstrates that cAF-MSCs have great potential for neural precursor differentiation in vitro. Therefore, amniotic fluid may be a suitable alternative source of stem cells and can be applied to cell therapy in neurodegenerative diseases.
PMCID: PMC3964299  PMID: 24166061
Amniotic fluids; Mesenchymal stem cells; Degenerative diseases; Canine; Neural precursor; Neurodegenerative diseases
17.  Poor Prognosis in Elderly Patients Who Refused Surgery Because of Economic Burden and Medical Problem After Hip Fracture 
Journal of Korean Medical Science  2013;28(9):1378-1381.
Although many studies have assessed mortality and morbidity of conservative treatment after hip fracture in elderly patients, the mortality of conservative treatment done because of economic burden is unclear. Among 451 patients diagnosed with displaced hip fracture during 3 yr, 28 patients (Group I) were enrolled as conservative treatment. Fifty-six patients matched in age, gender, ASA score, and diagnosis (Group II) who had undergone surgical treatment were used as the control group. The causal factors of non-operative treatment and mortality rate and functional recovery were evaluated according to the causal factors of patients with surgical procedure. Ten patients (36%) in Group I involved medical problems and 18 (64%) by economic burdens. The cumulative mortality rate over 3, 6, 12, and 24 months was 54%, 61%, 64%, and 82% in Group I and 9%, 11%, 14%, and 21% in Group II, respectively. At the latest follow-up, all five patients in Group I displayed a nonfunctional ambulatory state, whereas only seven of 44 patients in Group II were in a nonfunctional ambulatory state. Non-surgical treatment following hip fracture that is done because of the economic burden is associated with substantially high mortality and serious functional loss.
PMCID: PMC3763115  PMID: 24015046
Hip Fractures; Conservative Treatment; Economic Burden; Mortality; Functional Loss
18.  Osteonecrosis of the Knee after Arthroscopic Partial Meniscectomy 
Knee Surgery & Related Research  2013;25(3):150-154.
Osteonecrosis of the femoral condyle is known as an uncommon complication after arthroscopic meniscectomy. The lesion of osteonecrosis can be irreversible, thus early detection of the disease is crucial for treatment. A 50-year-old male patient without known risk factors of osteonecrosis developed increasing knee pain after arthroscopic partial meniscectomy. Magnetic resonance imaging showed rapid progression of osteonecrosis of the medial femoral condyle. Unicompartmental knee arthroplasty was performed after 9 months of conservative therapy. The patient is now free from pain during daily activities. It might be important to remind that if the patient's pain after arthroscopic partial meniscectomy is severe than expected, clinical doctors should pay attention to the possibility of ongoing osteonecrosis of the femoral condyle.
PMCID: PMC3767902  PMID: 24032105
Knee; Arthroscopy; Meniscectomy; Osteonecrosis
19.  Dissection of the Dimerization Modes in the DJ-1 Superfamily 
Molecules and Cells  2012;33(2):163-171.
The DJ-1 superfamily (DJ-1/ThiJ/PfpI superfamily) is distributed across all three kingdoms of life. These proteins are involved in a highly diverse range of cellular functions, including chaperone and protease activity. DJ-1 proteins usually form dimers or hexamers in vivo and show at least four different binding orientations via distinct interface patches. Abnormal oligomerization of human DJ-1 is related to neurodegenerative disorders including Parkinson’s disease, suggesting important functional roles of quaternary structures. However, the quaternary structures of the DJ-1 superfamily have not been extensively studied. Here, we focus on the diverse oligomerization modes among the DJ-1 superfamily proteins and investigate the functional roles of quaternary structures both computationally and experimentally. The oligomerization modes are classified into 4 types (DJ-1, YhbO, Hsp, and YDR types) depending on the distinct interface patches (I–IV) upon dimerization. A unique, rotated interface via patch I is reported, which may potentially be related to higher order oligomerization. In general, the groups based on sequence similarity are consistent with the quaternary structural classes, but their biochemical functions cannot be directly inferred using sequence information alone. The observed phyletic pattern suggests the dynamic nature of quaternary structures in the course of evolution. The amino acid residues at the interfaces tend to show lower mutation rates than those of non-interfacial surfaces.
PMCID: PMC3887719  PMID: 22228183
DJ-1 superfamily; DJ-1/ThiJ/PfpI superfamily; quaternary structure
20.  Crystallization and preliminary X-ray crystallographic analysis of the NmrA-like DDB_G0286605 protein from the social amoeba Dictyostelium discoideum  
In order to investigate its structure and function, the NmrA-like domain-containing DDB_G0286605 protein from D. discoideum was expressed, purified and crystallized. X-ray diffraction analysis is reported to a resolution of 1.64 Å.
The DDB_G0286605 gene product from Dictyostelium discoideum, an NmrA-like protein that belongs to the short-chain dehydrogenase/reductase family, has been crystallized by the hanging-drop vapour-diffusion method at 295 K. A 1.64 Å resolution data set was collected using synchrotron radiation. The DDB_G0286605 protein crystals belonged to space group P21, with unit-cell parameters a = 67.598, b = 54.935, c = 84.219 Å, β = 109.620°. Assuming the presence of two molecules in the asymmetric unit, the solvent content was estimated to be about 43.25% with 99% probability. Molecular-replacement trials were attempted with three NmrA-like proteins, NmrA, HSCARG and QOR2, as search models, but failed. This may be a consequence of the low sequence identity between the DDB_G0286605 protein and the search models (DDB_G0286605 has a primary-sequence identity of 28, 32 and 19% to NmrA, HCARG and QOR2, respectively).
PMCID: PMC3079982  PMID: 21206034
SDR superfamily; NmrA-like proteins; Dictyostelium discoideum
21.  Crystallization and preliminary X-ray crystallographic analysis of the short-chain dehydrogenase/reductase-type DDB_G0291732 protein from Dictyostelium discoideum  
In order to investigate its structure and function, the NmrA-like short-chain dehydrogenase/reductase-type DDB_G0291732 protein from D. discoideum was expressed, purified and crystallized. X-ray diffraction analysis is reported to a resolution of 1.65 Å.
The DDB_G0291732 gene product from Dictyostelium discoideum, which is a NmrA-like protein that belongs to the short-chain dehydrogenase/reductase superfamily but shows deviations in conserved sequence regions, has been crystallized by the hanging-drop vapour-diffusion method at 295 K. A 1.65 Å resolution data set was collected using synchrotron radiation. The crystals of DDB_G0291732 protein belonged to space group P21, with unit-cell parameters a = 38.5, b = 63.7, c = 56.0 Å, β = 91.7°. Assuming the presence of one molecule in the asymmetric unit, the solvent content was estimated to be about 38.1%.
PMCID: PMC3079983  PMID: 21206035
SDR superfamily; Rossmann fold; Dictyostelium discoideum
22.  Is It a Simple Stress Fracture or Bisphosphonate-related Atypical Fracture? 
Journal of Bone Metabolism  2012;19(2):129-132.
A number of reports regarding atypical fractures of the femur have raised questions concerning the possible correlation between long-term bisphosphonate treatment and the occurrence of insufficiency fractures in the proximal femur. However, clinically, it is often confused whether is it a fatigue fracture because of implant induced stress concentration or a bisphosphonate-related atypical fracture, especially in a patient with a subtrochanteric fracture who receive bisphosphonate therapy after open reduction and internal fixation, such as dynamic hip screw (DHS) fixation for previous ipsilateral femoral neck or intertrochanteric fracture. The authors experienced a case of a progressive femoral insufficiency fracture in a woman who had been on Fosamax (Alendronic acid with Vitamin D; Merck & Co. Inc, NJ, USA) therapy for four years after ipsilateral femoral neck fracture treated with a two hole DHS system. Despite a high suspicion of an insufficiency femoral subtrochanteric fracture by bone scan, the occult fracture progressed to a displaced femoral subtrochanteric fracture one year after. The fracture site was fixed with a 6 hole DHS plate, and six months after reoperation the patient had no symptoms and the fracture site had united without any complication.
PMCID: PMC3780922  PMID: 24524043
Bisphosphonate; Early diagnosis; Femoral fractures; Fractures stress; Osteoporosis
23.  Left Atrial Mechanical Function and Stiffness in Patients with Paroxysmal Atrial Fibrillation 
Abnormalities in the left atrial (LA) structure and function may develop in patients with paroxysmal atrial fibrillation (AF). We sought to determine the contribution of LA mechanical function, including LA stiffness, to AF by comparing patients with paroxysmal AF with normal control subjects, and to evaluate whether LA mechanical function and stiffness are related with the structural changes of LA.
Sixty-four paroxysmal AF patients (57 ± 13 years, 59% male) were studied, using a speckle tracking echocardiography, and were compared with 34 age-, gender-, and left ventricular (LV) mass-matched controls (53 ± 14 years, 61% male). LA volume indices, expansion index for reservoir function, active emptying fraction for contractile function, mitral annular velocities, and global longitudinal LA strain were measured. The ratio of E/e' to LA strain was used as an index of LA stiffness.
Patients with paroxysmal AF had similar LV volume indices, ejection fraction, and diastolic function, when compared with that of the normal controls. However, paroxysmal AF patients showed increased LA volume indices and decreased LA reservoir function, but similar contractile function. LA stiffness was increased in patients with paroxysmal AF than in the control subjects (0.40 ± 0.25 vs. 0.29 ± 0.10, p = 0.002), and was related with LA volume indices and reservoir function.
Patients with paroxysmal AF have decreased LA reservoir function and increased stiffness, in comparison with that of the control subjects. LA stiffness was significantly related with LA volume indices and reservoir function. LA stiffness can be used for the assessment of LA function in patients with paroxysmal AF.
PMCID: PMC3498311  PMID: 23185657
Atrial fibrillation; Left atrial function; Left atrial stiffness
24.  Experimental phasing using zinc anomalous scattering 
The surface of proteins can be charged with zinc ions and the anomalous signals from these zinc ions can be used for structure determination of proteins.
Zinc is a suitable metal for anomalous dispersion phasing methods in protein crystallography. Structure determination using zinc anomalous scattering has been almost exclusively limited to proteins with intrinsically bound zinc(s). Here, it is reported that multiple zinc ions can easily be charged onto the surface of proteins with no intrinsic zinc-binding site by using zinc-containing solutions. Zn derivatization of protein surfaces appears to be a largely unnoticed but promising method of protein structure determination.
PMCID: PMC3489106  PMID: 22948927
zinc anomalous scattering; phasing; Zn derivatization
25.  SRY-positive 78, XY ovotesticular disorder of sex development in a wolf cloned by nuclear transfer 
Journal of Veterinary Science  2012;13(2):211-213.
Recently, we reported the three wolves cloning with normal karyotype from somatic cells of endangered male gray wolves (Canis lupus), but one wolf had female external genitalia. In this study, we conducted further clinical, histological, and genetic analyses. This cloned wolf had a normal uterus but developed ovotestis. Through molecular analysis of the SRY gene, a mutation in the coding sequence of SRY gene could be excluded as a cause of intersexuality. This is the first report of a cloned wolf with a 78, XY ovotesticular disorder affecting sexual development characterized by bilateral ovotestes.
PMCID: PMC3386349  PMID: 22705746
hermaphrodite; intersexuality; wolf

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