The Potential MicroRNA Diagnostic Biomarkers in Oral Squamous Cell Carcinoma of the Tongue.

Oral squamous cell carcinoma (OSCC) of the tongue is a common type of head and neck malignancy with a poor prognosis, underscoring the urgency for early detection. MicroRNAs (miRNAs) have remarkable stability and are easily measurable. Thus, miRNAs may be a promising biomarker candidate among biomarkers in cancer diagnosis. Biomarkers have the potential to facilitate personalized medicine approaches by guiding treatment decisions and optimizing therapy regimens for individual patients. Utilizing data from The Cancer Genome Atlas, we identified 13 differentially expressed upregulated miRNAs in OSCC of the tongue. Differentially expressed miRNAs were analyzed by enrichment analysis to reveal underlying biological processes, pathways, or functions. Furthermore, we identified miRNAs associated with the progression of OSCC of the tongue, utilizing receiver operating characteristic analysis to evaluate their potential as diagnostic biomarkers. A total of 13 upregulated miRNAs were identified as differentially expressed in OSCC of the tongue. Five of these miRNAs had high diagnostic power. In particular, miR-196b has the potential to serve as one of the most effective diagnostic biomarkers. Then, functional enrichment analysis for the target gene of miR-196b was performed, and a protein-protein interaction network was constructed. This study assessed an effective approach for identifying miRNAs as early diagnostic markers for OSCC of the tongue.

Reliability analysis of visual ranking of coronary artery calcification on low-dose CT of the thorax for lung cancer screening: comparison with ECG-gated calcium scoring CT.

Coronary artery calcification (CAC) is frequently detected on low-dose CT (LDCT) of the thorax. Concurrent assessment of CAC and lung cancer screening using LDCT is beneficial in terms of cost and radiation dose reduction. The aim of our study was to evaluate the reliability of visual ranking of positive CAC on LDCT compared to Agatston score (AS) on electrocardiogram (ECG)-gated calcium scoring CT. We studied 576 patients who were consecutively registered for health screening and undergoing both LDCT and ECG-gated calcium scoring CT. We excluded subjects with an AS of zero. The final study cohort included 117 patients with CAC (97 men; mean age, 53.4 ± 8.5). AS was used as the gold standard (mean score 166.0; range 0.4-3,719.3). Two board-certified radiologists and two radiology residents participated in an observer performance study. Visual ranking of CAC was performed according to four categories (1-10, 11-100, 101-400, and 401 or higher) for coronary artery disease risk stratification. Weighted kappa statistics were used to measure the degree of reliability on visual ranking of CAC on LDCT. The degree of reliability on visual ranking of CAC on LDCT compared to ECG-gated calcium scoring CT was excellent for board-certified radiologists and good for radiology residents. A high degree of association was observed with 71.6% of visual rankings in the same category as the Agatston category and 98.9% varying by no more than one category. Visual ranking of positive CAC on LDCT is reliable for predicting AS rank categorization.

Aortic stent-graft for a giant bronchial artery aneurysm with ultrashort neck.

Bronchial artery aneurysms are rarely encountered, but they can cause life-threatening hemorrhages when they rupture. The authors report a case of a giant bronchial artery aneurysm with ultrashort neck in a 73-year-old woman who presented with massive hemoptysis. The aneurysm was successfully treated by a combination of transcatheter arterial embolization and aortic stent-graft placement, and the patient made an uneventful recovery. Follow-up computed tomography (CT) obtained 24 months after aortic stent-graft placement confirmed continued exclusion of the aneurysm, with no evidence of an endoleak.

Molecular characterization of Korean rabies virus isolates.

The nucleoprotein (N) and glycoprotein (G) of 11 Korean rabies virus (RABV) isolates collected from animals diagnosed with rabies between 2008 and 2009 were subjected to molecular and phylogenetic analyses. Six isolates originated from domestic animals (cattle and dogs) and five were obtained from wild free-ranging raccoon dogs. The similarities in the nucleotide sequences of the N gene among all Korean isolates ranged from 98.1 to 99.8%, while those of the G gene ranged from 97.9 to 99.3%. Based on the nucleotide analysis of the N and G genes, the Korean RABV isolates were confirmed as genotype I of Lyssavirus and classified into four distinct subgroups with high similarity. Phylogenetic analysis showed that the Korean isolates were most closely related to the non-Korean NeiMeng1025B and 857r strains, which were isolated from rabid raccoon dogs in Eastern China and Russia, respectively. These findings suggest that the Korean RABV isolates originated from a rabid raccoon dog in Northeastern Asia. Genetic analysis of the Korean RABV isolates revealed no substitutions at several antigenic sites, indicating that the isolates circulating in Korea may be pathogenic in several hosts.

The Symptom Frequency Characteristics of the Hamilton Depression Rating Scale and Possible Symptom Clusters of Depressive Disorders in Korea: The CRESCEND Study.

OBJECTIVE: This study analyzed the symptom frequencies of 17-item Hamilton Depression Rating Scale (HDRS-17) to understand the characteristics of each item and to propose the possible symptoms clusters. METHODS: From psychiatric clinics of 18 Hospitals in Korea, 1,183 patients, diagnosed with major depressive disorder (psychotic or non-psychotic), dysthymia or depressive disorder not otherwise specified. according to DSM-IV criteria, participated in this study from January 2006 to August 2008. The frequencies of each item of HDRS-17 were analyzed according to sex and severity. In addition, we compared this study with a previous study performed in England by Hamilton and with two studies performed in Korea by Kim et al. RESULTS: The frequencies of HDRS-17 items varied widely in this study, ranging from 95.8% in work and activities to 37.4% in loss of weight. But, depressed mood, psychic anxiety and work and activities items exhibited constant and higher frequency or rank regardless of study, the severity of depression or sex. Insomnia early, somatic gastrointestinal, genital symptoms and insight showed relatively constant but lower frequency or rank in disregard of studies or the clinical variables. Other symptoms had variable frequencies or ranks according to the variable clinical situations (culture, time, sex, severity of depression). CONCLUSION: WE PROPOSE THREE CLUSTERS OF SYMPTOMS IN DEPRESSIVE DISORDERS: core symptoms cluster, an associated symptoms, and a situation-specific symptoms. We can use these possible symptom clusters of depression in simplifying diagnosis of depression, increasing diagnostic specificity in special situation and indexing disease severity.

Simple immunohistochemical staining method using large sized gold colloid conjugated secondary antibody.

To test the feasibility of semiquantitative immunohistochemical staining (IHC) with large sized gold conjugated secondary antibody (gold-2 degrees Ab), we used beads covered with a known amount of primary antibody and a secondary antibody conjugated with gold colloid particles (20 and 40 nm in diameter), and we compared the results to those obtained by enzyme IHC. Beads coated with 6 graded amounts of mouse IgG molecules showed 6 levels of color intensity. The graded color intensities could readily be distinguished. The color developed as soon as we added gold-2 degrees Ab, and the intensities were stable for 1 wk. Enzyme IHC using identical beads showed dregs of pigment after incubation in DAB for 5 min. The large sized gold-2 degrees Ab showed strong signals on cell surfaces; application of the large sized gold-2 degrees Ab to paraffin-embedded tissue sections was also feasible. The color was bright red and was easier to differentiate from hemosiderin pigment than the color developed by enzyme IHC. In conclusion, gold IHC with large sized gold-2 degrees Ab is superior to enzyme IHC for quantification of antigens via IHC. Gold IHC is especially recommended for tissues with many macrophages, such as bone marrow and spleen.

Protein disulfide isomerase-like proteins play opposing roles during retrotranslocation.

Misfolded proteins in the endoplasmic reticulum (ER) are retained in the organelle or retrotranslocated to the cytosol for proteasomal degradation. ER chaperones that guide these opposing processes are largely unknown. We developed a semipermeabilized cell system to study the retrotranslocation of cholera toxin (CT), a toxic agent that crosses the ER membrane to reach the cytosol during intoxication. We found that protein disulfide isomerase (PDI) facilitates CT retrotranslocation, whereas ERp72, a PDI-like protein, mediates its ER retention. In vitro analysis revealed that PDI and ERp72 alter CT's conformation in a manner consistent with their roles in retrotranslocation and ER retention. Moreover, we found that PDI's and ERp72's opposing functions operate on endogenous ER misfolded proteins. Thus, our data identify PDI family proteins that play opposing roles in ER quality control and establish an assay to further delineate the mechanism of CT retrotranslocation.

Mixed-disulfide folding intermediates between thyroglobulin and endoplasmic reticulum resident oxidoreductases ERp57 and protein disulfide isomerase.

We present the first identification of transient folding intermediates of endogenous thyroglobulin (Tg; a large homodimeric secretory glycoprotein of thyrocytes), which include mixed disulfides with endogenous oxidoreductases servicing Tg folding needs. Formation of disulfide-linked Tg adducts with endoplasmic reticulum (ER) oxidoreductases begins cotranslationally. Inhibition of ER glucosidase activity blocked formation of a subgroup of Tg adducts containing ERp57 while causing increased Tg adduct formation with protein disulfide isomerase (PDI), delayed adduct resolution, perturbed oxidative folding of Tg monomers, impaired Tg dimerization, increased Tg association with BiP/GRP78 and GRP94, activation of the unfolded protein response, increased ER-associated degradation of a subpopulation of Tg, partial Tg escape from ER quality control with increased secretion of free monomers, and decreased overall Tg secretion. These data point towards mixed disulfides with the ERp57 oxidoreductase in conjunction with calreticulin/calnexin chaperones acting as normal early Tg folding intermediates that can be "substituted" by PDI adducts only at the expense of lower folding efficiency with resultant ER stress.

A prospective study of a new immunochemical fecal occult blood test in Korean patients referred for colonoscopy.

OBJECTIVES: The exquisite performance characteristics of an immunochemical fecal occult blood test (IFOBT) are well understood. We evaluated the diagnostic validity of a new IFOBT for colorectal neoplasia in patients undergoing colonoscopy and compared its results with two other commercially available IFOBTs. DESIGN AND METHODS: Eighty-five consecutive patients referred for colonoscopy were studied. We performed three different IFOBTs, namely, HM-Jack, Instant-View, and a newly developed OcculTech on each fecal specimen. RESULTS: OcculTech was easy to perform and had a sensitivity, specificity, and positive predictive value of 58.3%, 76.3%, and 27.9%, respectively, for the detection of colorectal cancers and >or=1 cm adenomas. OcculTech showed higher sensitivity than the automatic analyzer, HM-Jack. No improvement was obtained by combining tests. CONCLUSIONS: The OcculTech test had performance characteristics comparable to the two other IFOBTs. This study confirms the usefulness of the OcculTech test for colorectal neoplasia screening.

Deficiency of ATP2C1, a Golgi ion pump, induces secretory pathway defects in endoplasmic reticulum (ER)-associated degradation and sensitivity to ER stress.

Relatively few clues have been uncovered to elucidate the cell biological role(s) of mammalian ATP2C1 encoding an inwardly directed secretory pathway Ca2+/Mn2+ pump that is ubiquitously expressed. Deficiency of ATP2C1 results in a human disease (Hailey-Hailey), which primarily affects keratinocytes. ATP2C1-encoded protein is detected in the Golgi complex in a calcium-dependent manner. A small interfering RNA causes knockdown of ATP2C1 expression, resulting in defects in both post-translational processing of wild-type thyroglobulin (a secretory glycoprotein) as well as endoplasmic reticulum-associated protein degradation of mutant thyroglobulin, whereas degradation of a nonglycosylated misfolded secretory protein substrate appears unaffected. Knockdown of ATP2C1 is not associated with elevated steady state levels of ER chaperone proteins, nor does it block cellular activation of either the PERK, ATF6, or Ire1/XBP1 portions of the ER stress response. However, deficiency of ATP2C1 renders cells hypersensitive to ER stress. These data point to the important contributions of the Golgi-localized ATP2C1 protein in homeostatic maintenance throughout the secretory pathway.

The acetylcholinesterase homology region is essential for normal conformational maturation and secretion of thyroglobulin.

Secretion of thyroglobulin (Tg, a large homodimeric glycoprotein) is essential to deliver Tg to its site of iodination for thyroxine biosynthesis. An L2263P missense mutation in Tg has been proposed as the molecular defect causing congenital goitrous hypothyroidism in cog/cog mice due to perturbed Tg homodimerization, resulting in its retention within the endoplasmic reticulum. The mutation falls within a carboxyl-terminal region of Tg with high structural similarity to the entirety of acetylcholinesterase (AChE), a secretory protein that also forms homodimers. We provide new evidence that authentic AChE and the cholinesterase-like domain of Tg share a common tertiary structure. Moreover, we find that a Tg truncation, deleted of the cholinesterase-like region (but not a comparably sized deletion of internal Tg regions), blocks Tg export. Appending to this truncation a cDNA encoding authentic AChE results in translation of a chimeric protein in which AChE is present in a native, enzymatically active (albeit latent) conformation, and this fully rescues Tg secretion. Introduction of the cog mutation inhibits AChE enzyme activity, and established denaturing mutations of AChE block secretion of the Tg. Additional studies show that the native structure of the AChE region functions as a "dimerization domain," facilitating intracellular transport of Tg to the site of thyroid hormonogenesis.

Development of real-time RT-PCR for evaluation of JEV clearance during purification of HPV type 16 L1 virus-like particles.

Insect cell culture has greatly increased in part due to the widespread use of insect virus-based vectors for efficient expression of foreign proteins. Insect cells such as Sf9 cells are susceptible to arboviruses which may pose a safety concern by adventitious introduction during the production process. The objective of this study was to establish techniques for viral clearance validation of insect cell-derived biotechnological products using Japanese encephalitis virus (JEV) as a model, since JEV is a member of arthropod-borne flaviviruses that are known to be infectious in insect cells. Here we report the development of a quantitative assay for JEV RNA using real-time reverse transcription-polymerase chain reaction (RT-PCR). The assay was performed using LightCycler and RNA amplification kit SYBR Green I. The JEV specific primer was selected from the 3' untranslated region, and the expected band size was 323 base pairs (bp). The sensitivity of the assay was calculated to be approximately 15 TCID(50)per reaction. Highly reproducible standard curves were obtained from experiments performed on three different days. JEV clearance was determined during the purification process of rHPV-16 L1 VLPs by CsCl equilibrium density centrifugation. The comparative results obtained by real-time RT-PCR assay for JEV and infectivity titrations suggested that the real-time RT-PCR assay could have an additive effect on the interpretation and evaluation of virus clearance, especially during the virus removal process.

Effect of immobilization stress on brain polyamine levels in spontaneously hypertensive and Wistar-Kyoto rats.

The present study aimed to compare the basal brain polyamine levels and stress-induced brain polyamine level changes in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. For immobilization stress, both strains underwent acute (3 h per day immobilization for 2 days), chronic (3 h per day immobilization for 15 consecutive days), or no immobilization stress (control group). Basal putrescine (PU) levels in frontal cortex and hippocampus of SHR (11.03 +/- 0.81 and 11.36 +/- 0.33 nmol/g tissue, respectively) were significantly higher than WKY rats (6.90 +/- 1.44 and 7.82 +/- 0.71 nmol/g tissue, respectively). However, there were no strain differences in basal spermidine and spermine levels between the two. After acute stress, the PU levels in frontal cortex and hippocampus (15.99 +/- 0.45 and 14.10 +/- 0.95 nmol/g tissue, respectively) were significantly increased in SHR as compared to the non-stressed SHR (11.03 +/- 0.81 and 11.36 +/- 0.33 nmol/g tissue, respectively). In WKY rats, the PU level was significantly increased by acute stress in frontal cortex (11.68 +/- 1.12 nmol/g tissue) as compared to the non-stressed WKY (6.90 +/- 1.44 nmol/g tissue). After chronic stress, the PU levels in frontal cortex and hippocampus of SHR (12.44 +/- 0.54 and 11.34 +/- 0.66 nmol/g tissue, respectively) significantly decreased as compared to acute-stressed groups (15.99 +/- 0.45 and 14.01 +/- 0.95 nmol/g tissue, respectively). In WKY rats, after chronic stress, the PU level was significantly decreased in frontal cortex (5.73 +/- 0.36 nmol/g tissue) as compared to acute-stressed groups (11.68 +/- 1.12 nmol/g tissue). The PU levels in frontal cortex and hippocampus of acute-stressed (15.99 +/- 0.45 nmol/g tissue and 14.10 +/- 0.95 nmol/g tissue, respectively) and chronic-stressed (12.44 +/- 0.54 and 11.34 +/- 0.66 nmol/g tissue, respectively) SHR were significantly higher than acute-stressed (11.68 +/- 1.12 and 9.76 +/- 0.45 nmol/g tissue, respectively) and chronic-stressed (5.73 +/- 0.36 and 8.44 +/- 0.71 nmol/g tissue, respectively) WKY rats. The present study provides the higher basal PU level and stress-induced PU response in SHR as compared to WKY rats may be related to enhanced response of hypothalamic-pituitary-adrenocortical axis and sympathetic influence that may significantly contribute to the development of hypertension in SHR.