Lymphocyte-activating gene-3 expression is associated with tumor-infiltrating lymphocyte levels in HER2-positive breast cancers.

ABSTRACT: Lymphocyte-activating gene-3 (LAG-3, CD223) is the third inhibitory receptor targeted for immunotherapy. Several clinical trials investigating the use of interventions targeting LAG-3 are underway. The exact signaling mechanism downstream of LAG-3 is largely unknown, especially in breast cancer. The prognostic significance of tumor-infiltrating lymphocytes (TILs) in breast cancer has been previously determined.Among 167 human epidermal growth factor receptor 2 (HER2)-positive breast cancer patients, 90 and 78 patients were positive and negative for the hormone receptor, respectively. LAG-3 mRNA and protein expression levels in TILs were evaluated by quantitative real-time polymerase chain reaction and immunohistochemistry, respectively, among 12 and 167 HER2-positive breast cancer samples, respectively.High expression of LAG-3 in TILs was significantly correlated with high levels of TILs (P = .003) and an abundance of tertiary lymphoid structures around invasive components (P = .014). In addition, high expression of LAG3 was significantly associated with positivity for programmed death-ligand 1 (PD-L1) in tumor cells, a high immunostaining score of PD-L1 in TILs, and a high total immunostaining score for PD-L1 in tumor cells and TILs (all, P < .001). High expression levels of LAG-3 mRNA were associated with high levels of TILs (P = .091).LAG-3 protein expression was not a prognostic factor in HER2-positive breast cancers, and LAG-3 expression in TILs was significantly associated with the levels of TILs in HER2-positive breast cancer, although it was not a prognostic factor.

Interobserver diagnostic reproducibility in advanced-stage endometrial carcinoma.

BACKGROUND: The accurate pathologic diagnosis and subtyping of high-grade endometrial carcinoma are often problematic, due to its atypical and overlapping histopathological features. METHODS: Three pathologists reviewed 21 surgically resected cases of advancedstage endometrial carcinoma. The primary diagnosis was based only on hematoxylin and eosin stained slides. When a discrepancy arose, a secondary diagnosis was made by additional review of immunohistochemical (IHC) stains. Finally, three pathologists discussed all cases and rendered a consensus diagnosis. RESULTS: The primary diagnoses were identical in 13/21 cases (62%). The secondary diagnosis based on the addition of IHC results was concordant in four of eight discrepant cases. Among four cases with discrepancies occurring in this step, two cases subsequently reached a consensus diagnosis after a thorough discussion between three reviewers. Next-generation sequencing (NGS) study was performed in two cases in which it was difficult to distinguish between serous carcinoma and endometrioid carcinoma. Based on the sequencing results, a final diagnosis of serous carcinoma was rendered. The overall kappa for concordance between the original and consensus diagnosis was 0.566 (moderate agreement). CONCLUSIONS: We investigated stepwise changes in interobserver diagnostic reproducibility in advanced-stage endometrial carcinoma. We demonstrated the utility of IHC and NGS study results in the histopathological diagnosis of advanced-stage endometrial carcinoma.

First draft genome sequence of the rock bream in the family Oplegnathidae.

The rock bream (Oplegnathus fasciatus) is one of the most economically valuable marine fish in East Asia, and due to various environmental factors, there is substantial revenue loss in the production sector. Therefore, knowledge of its genome is required to uncover the genetic factors and the solutions to these problems. In this study, we constructed the first draft genome of O. fasciatus as a reference for the family Oplegnathidae. The genome size is estimated to be 749 Mb, and it was assembled into 766 Mb by combining Illumina and PacBio sequences. A total of 24,053 transcripts (23,338 genes) are predicted, and among those transcripts, 23,362 (97%), are annotated with functional terms. Finally, the completeness of the genome assembly was assessed by CEGMA, which resulted in the complete mapping of 220 (88.7%) core genes in the genome. To the best of our knowledge, this is the first draft genome for the family Oplegnathidae.

Quality of Life in Patients with Osteoporotic Vertebral Compression Fractures.

BACKGROUND: This study aimed to evaluate quality of life (QOL) using the EuroQOL-5 dimensions (EQ-5D) index and to examine factors affecting QOL in patients with an osteoporotic vertebral compression fracture (OVCF). METHODS: This ambispective study used a questionnaire interview. Patients over 50 years old with an OVCF at least 6 months previously were enrolled. Individual results were used to calculate the EQ-5D index. Statistical analysis was performed, and factors related to QOL were examined. RESULTS: Of 196 patients in the study, 84.2% were female, with an average age of 72.7 years. There were 66 (33.7%) patients with multilevel fractures. Conservative management was used in 75.0% of patients, and 56.1% received anti-osteoporosis treatment. The mean EQ-5D index was 0.737±0.221 and was significantly correlated with the Oswestry disability index score (correlation coefficient -0.807, P<0.001). The EQ-5D index was significantly correlated with age (Spearman's rho=-2.0, P=0.005), treatment method (P=0.005), and history of fracture (P=0.044) on univariate analysis and with conservative treatment (P<0.001) and osteoporotic treatment (P=0.017) on multivariate analysis. CONCLUSIONS: OVCF markedly lowers QOL in several dimensions for up to 12 months, even in patients who have healed. Treatment of osteoporosis and conservative treatment methods affect QOL and should be considered in OVCF management.

A Prospective Randomized Study Comparing the Interference Screw and Suture Anchor Techniques for Biceps Tenodesis.

BACKGROUND: Several methods are used to perform biceps tenodesis in patients with superior labrum-biceps complex (SLBC) lesions accompanied by a rotator cuff tear. However, limited clinical data are available regarding the best technique in terms of clinical and anatomic outcomes. PURPOSE: To compare the clinical and anatomic outcomes of the interference screw (IS) and suture anchor (SA) fixation techniques for biceps tenodesis performed along with arthroscopic rotator cuff repair. STUDY DESIGN: Randomized controlled trial; Level of evidence, 2. METHODS: A total of 80 patients who underwent arthroscopic rotator cuff repair with SLBC lesions were prospectively enrolled and randomly divided according to the tenodesis method: the IS and SA groups. Functional outcomes were evaluated with the visual analog scale (VAS) for pain, American Shoulder and Elbow Surgeons (ASES) score, Simple Shoulder Test (SST), Constant score, Korean Shoulder Score (KSS), and long head of the biceps (LHB) score at least 2 years after surgery. The anatomic status of tenodesis was estimated using magnetic resonance imaging or ultrasonography, and the anatomic failure of tenodesis was determined when the biceps tendon was not traced in the intertubercular groove directly from the insertion site of the IS or SA. RESULTS: Thirty-three patients in the IS group and 34 in the SA group were monitored for more than 2 years. All postoperative functional scores improved significantly compared with the preoperative scores (all P < .001) and were not significantly different between the groups, including the LHB score (all P > .05). Nine anatomic failures of tenodesis were observed: 7 in the IS group and 2 in the SA group ( P = .083). In a multivariate analysis using logistic regression, IS fixation ( P = .003) and a higher (ie, more physically demanding) work level ( P = .022) were factors associated with the anatomic failure of tenodesis significantly. In patients with tenodesis failure, the LHB score ( P = .049) and the degree of Popeye deformity by the patient and examiner ( P = .004 and .018, respectively) were statistically different compared with patients with intact tenodeses. CONCLUSION: Care must be taken while performing biceps tenodesis in patients with a higher work level; IS fixation appears to pose a higher risk in terms of the anatomic failure of tenodesis than SA fixation, although functional outcomes were not different.

Transcriptome Analysis Revealed Changes of Multiple Genes Involved in Haliotis discus hannai Innate Immunity during Vibrio parahemolyticus Infection.

Abalone (Haliotis discus hannai) is one of the most valuable marine aquatic species in Korea, Japan and China. Tremendous exposure to bacterial infection is common in aquaculture environment, especially by Vibrio sp. infections. It's therefore necessary and urgent to understand the mechanism of H. discus hannai host defense against Vibrio parahemolyticus infection. However studies on its immune system are hindered by the lack of genomic resources. In the present study, we sequenced the transcriptome of control and bacterial challenged H. discus hannai tissues. Totally, 138 MB of reference transcriptome were obtained from de novo assembly of 34 GB clean bases from ten different libraries and annotated with the biological terms (GO and KEGG). A total of 10,575 transcripts exhibiting the differentially expression at least one pair of comparison and the functional annotations highlight genes related to immune response, cell adhesion, immune regulators, redox molecules and mitochondrial coding genes. Mostly, these groups of genes were dominated in hemocytes compared to other tissues. This work is a prerequisite for the identification of those physiological traits controlling H. discus hannai ability to survive against Vibrio infection.

Discovery of Gene Sources for Economic Traits in Hanwoo by Whole-genome Resequencing.

Hanwoo, a Korean native cattle (Bos taurus coreana), has great economic value due to high meat quality. Also, the breed has genetic variations that are associated with production traits such as health, disease resistance, reproduction, growth as well as carcass quality. In this study, next generation sequencing technologies and the availability of an appropriate reference genome were applied to discover a large amount of single nucleotide polymorphisms (SNPs) in ten Hanwoo bulls. Analysis of whole-genome resequencing generated a total of 26.5 Gb data, of which 594,716,859 and 592,990,750 reads covered 98.73% and 93.79% of the bovine reference genomes of UMD 3.1 and Btau 4.6.1, respectively. In total, 2,473,884 and 2,402,997 putative SNPs were discovered, of which 1,095,922 (44.3%) and 982,674 (40.9%) novel SNPs were discovered against UMD3.1 and Btau 4.6.1, respectively. Among the SNPs, the 46,301 (UMD 3.1) and 28,613 SNPs (Btau 4.6.1) that were identified as Hanwoo-specific SNPs were included in the functional genes that may be involved in the mechanisms of milk production, tenderness, juiciness, marbling of Hanwoo beef and yellow hair. Most of the Hanwoo-specific SNPs were identified in the promoter region, suggesting that the SNPs influence differential expression of the regulated genes relative to the relevant traits. In particular, the non-synonymous (ns) SNPs found in CORIN, which is a negative regulator of Agouti, might be a causal variant to determine yellow hair of Hanwoo. Our results will provide abundant genetic sources of variation to characterize Hanwoo genetics and for subsequent breeding.

Pancreatic pseudocystocolonic fistula treated without surgical or endoscopic intervention.

We report here a case of pancreatic pseudocystocolic fistula that was treated without surgical or endoscopic intervention. A 76-year-old woman, presenting with a fever and epigastric pain, was referred to our institution. Three months prior to this admission, the patient had been admitted to the hospital for acute pancreatitis. Abdominal computerized tomography (CT) revealed a 9 cm pseudocyst containing air, and a fistular opening was observed via colonoscopy. After colonoscopy, the abdominal pain was slightly improved, the fever subsided and laboratory results showed decreased C-reactive protein levels. The observed improvement was likely due to the cleansing of the bowel, which induced spontaneous drainage from the pseudocyst into the colon. Antibiotic therapy was administered and daily bowel cleansing was performed using a polyethylene glycol solution. After three weeks, a follow-up CT revealed that the size of the pseudocyst had decreased significantly from 9 to 5.3 cm. In addition, laboratory tests were improved. The patient was able to resume a normal diet and was discharged in good overall health from the hospital, without aggravation of the symptoms. A colonoscopy performed 3 mo later and a follow-up CT performed 6 mo later confirmed that both the fistula and pseudocyst had completely disappeared.

Comparison of functional gene annotation of Toxascaris leonina and Toxocara canis using CLC genomics workbench.

The ascarids, Toxocara canis and Toxascaris leonina, are probably the most common gastrointestinal helminths encountered in dogs. In order to understand biological differences of 2 ascarids, we analyzed gene expression profiles of female adults of T. canis and T. leonina using CLC Genomics Workbench, and the results were compared with those of free-living nematode Caenorhabditis elegans. A total of 2,880 and 7,949 ESTs were collected from T. leonina and T. canis, respectively. The length of ESTs ranged from 106 to 4,637 bp with an average insert size of 820 bp. Overall, our results showed that most functional gene annotations of 2 ascarids were quite similar to each other in 3 major categories, i.e., cellular component, biological process, and molecular function. Although some different transcript expression categories were found, the distance was short and it was not enough to explain their different lifestyles. However, we found distinguished transcript differences between ascarid parasites and free-living nematodes. Understanding evolutionary genetic changes might be helpful for studies of the lifestyle and evolution of parasites.

Inhibition of P-glycoprotein-induced multidrug resistance by a clerodane-type diterpenoid from Sindora sumatrana.

The aim of the present study was to investigate the effects of di- and sesquiterpenoids isolated from the pods of Sindora sumatrana Miq. (Leguminosae) on P-glycoprotein (P-gp) function in an adriamycin-resistant human breast cancer cell line, MCF-7/ADR. Over-expression of P-gp is known to be one of the mechanisms involved in multidrug resistance (MDR), which is a major obstacle in clinical cancer treatment. Among six di- and sesquiterpenoids extracted from S. sumatrana, (+)-7beta-acetoxy-15,16-epoxycleroda-3,13(16),14-trien-18-oic acid (1) showed a strong P-gp inhibitory effect, as great as that of verapamil, a representative P-gp inhibitor. Compound 1 enhanced daunomycin accumulation more than fourfold and significantly decreased daunomycin efflux compared with control, resulting in a decrease in the IC(50) value for daunomycin. These results suggest that compound 1 inhibits the functioning of P-gp and, therefore, can be developed as an MDR-reversing agent.

Potent modulation of P-glycoprotein activity by naturally occurring phenylbutenoids from Zingiber cassumunar.

Five phenylbutenoid derivatives from the rhizomes of Zingiber cassumunar Roxb. (Zingiberaceae) were evaluated for their P-glycoprotein (P-gp) inhibitory effects in a P-gp over-expressing multidrug resistant (MDR) human breast cancer cell line, MCF-7/ADR. As a result, a phenylbutenoid dimer, (+/-)-trans-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene (1), exhibited highly potent P-gp inhibitory activity, decreasing the IC(50) value of daunomycin (DNM) to 4.31 +/- 0.40 microm in the cells (DNM IC(50) = 37.1 +/- 0.59 microm). The positive control, verapamil decreased the IC(50) value of DNM to 6.94 +/- 0.40 microm. Three phenylbutenoid monomers, 2-4 from this plant, also resulted in a significant decrease in the IC(50) values of DNM compared with the control. In particular, compound 1 markedly enhanced [(3)H]-DNM accumulation and significantly reduced [(3)H]-DNM efflux compared with the control, and this effect was more potent than that of verapamil, a well-known P-gp inhibitor. These results suggest that compound 1 of Z. cassumunar can be developed as a potent chemo-sensitizing agent that reverses P-gp-mediated MDR in human cancer chemotherapy.

Theory for the capillary electrophoretic separation of DNA in polymer solutions.

We present a mathematical model based on the models of Hubert et al. [Macromolecules 29 (1996) 1006] and Sunada and Blanch [Electrophoresis, 19 (1998) 3128] to describe the electrophoretic mobility of DNA by a transient entanglement coupling mechanism. The proposed model takes into account the interactions between molecules in the capillary and the cross-section of collision between DNA and polymer molecules. The results show that the calculated values agree remarkably well with our electrophoretic mobility data.