A simple and accurate SNP scoring strategy based on typeIIS restriction endonuclease cleavage and matrix-assisted laser desorption/ionization mass spectrometry

<p>Abstract</p> <p>Background</p> <p>We describe the development of a novel matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)-based single nucleotide polymorphism (SNP) scoring strategy, termed Restriction Fragment Mass Polymorphism (RFMP) that is suitable for genotyping variations in a simple, accurate, and high-throughput manner. The assay is based on polymerase chain reaction (PCR) amplification and mass measurement of oligonucleotides containing a polymorphic base, to which a typeIIS restriction endonuclease recognition was introduced by PCR amplification. Enzymatic cleavage of the products leads to excision of oligonucleotide fragments representing base variation of the polymorphic site whose masses were determined by MALDI-TOF MS.</p> <p>Results</p> <p>The assay represents an improvement over previous methods because it relies on the direct mass determination of PCR products rather than on an indirect analysis, where a base-extended or fluorescent report tag is interpreted. The RFMP strategy is simple and straightforward, requiring one restriction digestion reaction following target amplification in a single vessel. With this technology, genotypes are generated with a high call rate (99.6%) and high accuracy (99.8%) as determined by independent sequencing.</p> <p>Conclusion</p> <p>The simplicity, accuracy and amenability to high-throughput screening analysis should make the RFMP assay suitable for large-scale genotype association study as well as clinical genotyping in laboratories.</p

Plasma Cell Granuloma Associated with Pulmonary Actinomycosis: A Case Report

Plasma cell granuloma (PCG) of the lung is a rare disease that usually presents as a pulmonary nodule or mass on incidental radiographic examination without symptoms. Although the etiology of PCG is still controversial, many findings have lent support to the lesion being a reactive inflammatory process rather than a neoplastic one. We describe a 53-yr-old male who presented with a hemoptysis and have a lung mass at the left upper lobe on chest radiograph. The lung mass was primarily diagnosed as PCG by percutaneous needle aspiration and biopsy, and the patient was treated with oral steroid because he and relatives refused the operation. However, the size of the lung mass did not change and open thoracotomy and lobectomy were done therefore. He was confirmed as having pulmonary actinomycosis with PCG after surgery. To our knowledge, this is the first report of PCG associated with actinomycosis in Korea

Bleeding complications associated with the molecular adsorbent recirculating system: a retrospective study

Background The molecular adsorbent recirculating system (MARS) is a hepatic replacement system that supports excretory liver function in patients with liver failure. However, since MARS has been employed in our hospital, bleeding complications have occurred in many patients during or after MARS. The objective of this study was to determine how MARS affects coagulopathy and identify specific factors associated with bleeding complications. Methods We retrospectively analyzed data from 17 patients undergoing a total of 41 MARS sessions. Complete blood count, coagulation profiles, and blood chemistry values were compared before and after MARS. To identify pre-MARS factors associated with increased bleeding after MARS, we divided patients into bleeder and non-bleeder groups and compared their pre-MARS laboratory values. Results MARS significantly reduced bilirubin and creatinine levels. MARS also increased prothrombin time and reduced platelet and fibrinogen, thus negatively impacting coagulation. Pre-MARS hemoglobin was significantly lower in the bleeder group than in the non-bleeder group (P=0.015). When comparing the upper and lower 33% of MARS sessions based on the hemoglobin reduction rate, hemoglobin reduction was significantly greater in MARS sessions involving patients with low pre-MARS international normalized ratio of prothrombin time (PT-INR) and factor V (P=0.038 and P=0.023, respectively). Conclusions MARS could appears to alter coagulation-related factors such as factor V and increase the risk of bleeding complications particularly in patient with low hemoglobin. However, individual differences among patients were large, and various factors, such as low hemoglobin, PT-INR, and factor V levels, appear to be involved

A Sporadic Outbreak of Human Brucellosis in Korea

Eleven cases of human brucellosis occurred among livestock workers and a veterinarian who lived and worked in a rural area around Jeongeup City, Jeollabuk-Do, Korea from February 2003 to August 2003. Eight of the patients had taken care of Korean native cattle that were infected with bovine brucellosis and had already been slaughtered. Two of the patients had taken care of dairy cattle, and one case was a veterinarian who acquired the disease through an accidental contact with infected cattle while assisting in calf delivery. Eleven cases were identified by serologic work ups and four cases were identified via positive blood cultures. This study shows that the Republic of Korea is no longer free of human brucellosis, Brucella abortus biotype 1. We reviewed the patients' characteristics and serologic data during the one-year follow up period, and we also discuss on the efficacy and side effects of the rifampin and doxycyline regimen used for the treatment of human brucellosis

Seroprevalence of Mycoplasma pneumoniae and Chlamydia pneumoniae in Stable Asthma and Chronic Obstructive Pulmonary Disease

Mycoplasma pneumoniae and Chlamydia pneumoniae have been suggested to take part in the acute exacerbation of bronchial asthma and chronic obstructive pulmonary disease (COPD). Several studies have questioned whether they may play pathogenic roles in connection with bronchial asthma and COPD. This study was designed to evaluate the seroprevalences of M. pneumoniae and C. pneumoniae in stable asthma and COPD patients, and to compare with control patients. The medical records of one hundred forty patients who underwent M. pneumoniae and C. pneumoniae serology were retrospectively reviewed. Seroprevalences of M. pneumoniae and C. pneumoniae in the asthma group (11.1% and 8.3%, respectively) were higher than in the control group (4.4% and 2.2%, respectively) without statistical significance. The seroprevalence of M. pneumoniae in the COPD group (16.9%) was significantly higher than in the control group, and the seroprevalence of C. pneumoniae in the COPD group (3.4%) was higher than in the control group without statistical significance. This study raises important questions about the relation of M. pneumoniae and C. pneumoniae infection with stable asthma or COPD

Molecular Analysis of X-linked Chronic Granulomatous Disease in Five Unrelated Korean Patients

Chronic granulomatous disease (CGD) is a fatal genetic disorder in which phagocytes fail to produce antimicrobial superoxide because of NADPH oxidase deficiency. Molecular defects in CYBB gene causing X-linked CGD are responsible for about 70% of all cases. This study was done to confirm genetic defects of CYBB gene in five Korean patients who were highly suggestive of having CGD by clinical history. We performed initial screening for five unrelated Korean patients using single strand conformation polymorphism (SSCP) and then selective sequencing for the regions involving the abnormal bands. Activated NBT tests revealed that all patients were X-linked. SSCP analysis for CYBB gene showed abnormal bands in all patients. The molecular defects of five patients were as follows: c.1663insT, c.1111-1G>T, c.39_40insG, c.927delC and c.434T>C mutation. This result will help the families with prenatal diagnosis or genetic counseling

Cranial-first approach for laparoscopic extended right hemicolectomy

Complete mesocolic excision and central vascular ligation with D3 lymphadenectomy are important surgical principles for improving oncological outcomes in colon cancer. The cranial-first approach is a colonic mobilization–first approach for radical right hemicolectomy, which has several advantages, including early feasibility assessment, safe dissection from surrounding organs, preestablished inferior margin of lymph node dissection, and revelation of the tangible anatomy of the tributaries of the gastrocolic trunk. This video demonstrates the cranial-first approach for laparoscopic extended right hemicolectomy in a 66-year-old man with locally advanced cecal cancer

Clear Cell "Sugar" Tumor of the Lung: A Well-Enhanced Mass with an Early Washout Pattern on Dynamic Contrast-Enhanced Computed Tomography

Clear cell tumor of the lung is a rare and very unusual benign pulmonary tumor. As clear cell tumor of the lung contains abundant cytoplasmic glycogen, this tumor is called "sugar tumor". We report a case of sugar tumor in a 64-yr-old man presenting as a round pulmonary nodule. On dynamic computed tomography (CT) scans, the solitary pulmonary nodule showed early wash-in enhancement with an early washout pattern like a lung malignancy. The patient underwent wedge resection for the tumor. Pathologic examination, including immunohistochemical studies, revealed that the nodule was a benign clear cell tumor, so-called "sugar tumor". Because only a small number of cases have been reported previously, clinical aspects, radiological characteristics on dynamic contrast-enhanced CT, and differential diagnosis of the tumor are not well established. Herein we present a clear cell tumor of the lung and discuss the clinical, radiological, and pathological features of the tumor