Mechanical regulation of glycolysis via cytoskeleton architecture.

The mechanics of the cellular microenvironment continuously modulates cell functions such as growth, survival, apoptosis, differentiation and morphogenesis via cytoskeletal remodelling and actomyosin contractility1-3. Although all of these processes consume energy4,5, it is unknown whether and how cells adapt their metabolic activity to variable mechanical cues. Here we report that the transfer of human bronchial epithelial cells from stiff to soft substrates causes a downregulation of glycolysis via proteasomal degradation of the rate-limiting metabolic enzyme phosphofructokinase (PFK). PFK degradation is triggered by the disassembly of stress fibres, which releases the PFK-targeting E3 ubiquitin ligase tripartite motif (TRIM)-containing protein 21 (TRIM21). Transformed non-small-cell lung cancer cells, which maintain high glycolytic rates regardless of changing environmental mechanics, retain PFK expression by downregulating TRIM21, and by sequestering residual TRIM21 on a stress-fibre subset that is insensitive to substrate stiffness. Our data reveal a mechanism by which glycolysis responds to architectural features of the actomyosin cytoskeleton, thus coupling cell metabolism to the mechanical properties of the surrounding tissue. These processes enable normal cells to tune energy production in variable microenvironments, whereas the resistance of the cytoskeleton in response to mechanical cues enables the persistence of high glycolytic rates in cancer cells despite constant alterations of the tumour tissue.

Development and Clinical Evaluation of a Rapid Diagnostic Test for Yellow Fever Non-Structural Protein 1.

A rapid diagnostic test (RDT) kit was developed to detect non-structural protein 1 (NS1) of yellow fever virus (YFV) using monoclonal antibody. NS1 protein was purified from the cultured YFV and used to immunize mice. Monoclonal antibody to NS1 was selected and conjugated with colloidal gold to produce the YFV NS1 RDT kit. The YFV RDTs were evaluated for sensitivity and specificity using positive and negative samples of monkeys from Brazil and negative human blood samples from Korea. Among monoclonal antibodies, clones 3A11 and 3B7 proved most sensitive, and used for YFV RDT kit. Diagnostic accuracy of YFV RDT was fairly high; Sensitivity was 0.0% and specificity was 100% against Dengue viruses type 2 and 3, Zika, Chikungunya and Mayaro viruses. This YFV RDT kit could be employed as a test of choice for point-of-care diagnosis and large scale surveys of YFV infection under clinical or field conditions in endemic areas and on the globe.

An integrative somatic mutation analysis to identify pathways linked with survival outcomes across 19 cancer types.

MOTIVATION: Identification of altered pathways that are clinically relevant across human cancers is a key challenge in cancer genomics. Precise identification and understanding of these altered pathways may provide novel insights into patient stratification, therapeutic strategies and the development of new drugs. However, a challenge remains in accurately identifying pathways altered by somatic mutations across human cancers, due to the diverse mutation spectrum. We developed an innovative approach to integrate somatic mutation data with gene networks and pathways, in order to identify pathways altered by somatic mutations across cancers. RESULTS: We applied our approach to The Cancer Genome Atlas (TCGA) dataset of somatic mutations in 4790 cancer patients with 19 different types of tumors. Our analysis identified cancer-type-specific altered pathways enriched with known cancer-relevant genes and targets of currently available drugs. To investigate the clinical significance of these altered pathways, we performed consensus clustering for patient stratification using member genes in the altered pathways coupled with gene expression datasets from 4870 patients from TCGA, and multiple independent cohorts confirmed that the altered pathways could be used to stratify patients into subgroups with significantly different clinical outcomes. Of particular significance, certain patient subpopulations with poor prognosis were identified because they had specific altered pathways for which there are available targeted therapies. These findings could be used to tailor and intensify therapy in these patients, for whom current therapy is suboptimal. AVAILABILITY AND IMPLEMENTATION: The code is available at: http://www.taehyunlab.org CONTACT: jhcheong@yuhs.ac or taehyun.hwang@utsouthwestern.edu or taehyun.cs@gmail.com SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Intercellular transfer of apoptotic signals via electrofusion.

We determined whether cells that are induced to undergo anoikis by matrix detachment can initiate apoptosis in healthy cells following electroporation-induced fusion. Separate populations of MDCK cells undergoing anoikis and stained with FITC-annexin or viable MDCK cells that were labeled with spectrally discrete fluorescent beads were electroporated. Cells were analyzed by flow cytometry for enumeration of viable cells with beads, apoptotic cells or fused cells. Electroporation promoted a 49-fold increase of the percentage of viable cells that had fused with apoptotic cells. Apoptotic cell-viable cell fusions were 8-fold more likely to not attach to cell culture plastic and 2.3-fold less likely to proliferate after 24hr incubation than viable cell fusion controls. These data demonstrate that apoptotic signals can be transferred between cells by electrofusion, possibly suggesting a novel investigative approach for optimizing targeted cell deletion in cancer treatment.

Reputation Incongruence and the Preference of Stakeholder: Case of MBA Rankings.

In this paper, we examine the effect of an organization's multi-dimensional reputation on the external stakeholders' preference for an organization in the notions of reputation incongruence. We propose that an organization's incongruent reputation, or large variations among the reputations of each dimension, can be an unfavorable signal to its stakeholders based on theoretical ideas that claim reputation incongruence induces the ambiguity and risk of an organization perceived by stakeholders. We also investigate the moderating effect of reputation incongruence by positing that this incongruence may nullify the influences of reputation dimensions on the preferences of stakeholders. These propositions about reputation incongruence are empirically examined in the context of MBA programs of the global business schools which have three dimensions of reputation-career development, globalization, and research performance.

Product Creativity as an Identity Issue: Through the Eyes of New Product Development Team Members.

In this study, we introduce a concept of product identification that denotes the overlap between identities of a new product and its developer. As creativity is the most important identity dimension in the new product, we draw on two dimensions of creativity: novelty and meaningfulness. According to the argument that novelty represents exploration, whereas meaningfulness represents exploitation, we hypothesize that product novelty is associated with an explorative behavior of new product team members, while product meaningfulness is associated with exploitative behavior. More importantly, product identification is proposed as the mechanism that explains the amplification effect of product identity on team members. Based on survey data collected from 200 Japanese new product development (NPD) team members, we conduct a statistical analysis to test the hypotheses. The findings demonstrate the alignment between the identity of a new product and the behaviors of the NPD members, which is amplified by product identification but not by organizational identification.

Clinical and Technical Guideline for Endoscopic Ultrasound (EUS)-Guided Tissue Acquisition of Pancreatic Solid Tumor: Korean Society of Gastrointestinal Endoscopy (KSGE).

Endoscopic ultrasound (EUS)-guided tissue acquisition of pancreatic solid tumor requires a strict recommendation for its proper use in clinical practice because of its technical difficulty and invasiveness. The Korean Society of Gastrointestinal Endoscopy (KSGE) appointed a task force to draft clinical practice guidelines for EUS-guided tissue acquisition of pancreatic solid tumor. The strength of recommendation and the level of evidence for each statement were graded according to the Minds Handbook for Clinical Practice Guideline Development 2014. The committee, comprising a development panel of 16 endosonographers and an expert on guideline development methodology, developed 12 evidence-based recommendations in eight categories intended to help physicians make evidence-based clinical judgments with regard to the diagnosis of pancreatic solid tumor. This clinical practice guideline discusses EUS-guided sampling in pancreatic solid tumor and makes recommendations on circumstances that warrant its use, technical issues related to maximizing the diagnostic yield (e.g., needle type, needle diameter, adequate number of needle passes, sample obtaining techniques, and methods of specimen processing), adverse events of EUS-guided tissue acquisition, and learning-related issues. This guideline was reviewed by external experts and suggests best practices recommended based on the evidence available at the time of preparation. This guideline may not be applicable for all clinical situations and should be interpreted in light of specific situations and the availability of resources. It will be revised as necessary to cover progress and changes in technology and evidence from clinical practice.

[Clinical and Technical Guideline for Endoscopic Ultrasound-guided Tissue Acquisition of Pancreatic Solid Tumor: Korean Society of Gastrointestinal Endoscopy].

Endoscopic ultrasound (EUS)-guided tissue acquisition of pancreatic solid tumor requires a strict recommendation for its proper use in clinical practice because of its technical difficulty and invasiveness. The Korean Society of Gastrointestinal Endoscopy appointed a Task Force to draft clinical practice guidelines for EUS-guided tissue acquisition of pancreatic solid tumor. The strength of recommendation and the level of evidence for each statement were graded according to the Minds Handbook for Clinical Practice Guideline Development 2014. The committee, comprising a development panel of 16 endosonographers and an expert on guideline development methodology, developed 12 evidence-based recommendations in eight categories intended to help physicians make evidence- based clinical judgments with regard to the diagnosis of pancreatic solid tumor. This clinical practice guideline discusses EUS-guided sampling in pancreatic solid tumor and makes recommendations on circumstances that warrant its use, technical issues related to maximizing the diagnostic yield (e.g., needle type, needle diameter, adequate number of needle passes, sample obtaining techniques, and methods of specimen processing), adverse events of EUS-guided tissue acquisition, and learning-related issues. This guideline was reviewed by external experts and suggests best practices recommended based on the evidence available at the time of preparation. This guideline may not be applicable for all clinical situations and should be interpreted in light of specific situations and the availability of resources. It will be revised as necessary to cover progress and changes in technology and evidence from clinical practice.

Clinical and Technical Guideline for Endoscopic Ultrasound (EUS)-Guided Tissue Acquisition of Pancreatic Solid Tumor: Korean Society of Gastrointestinal Endoscopy (KSGE).

Endoscopic ultrasound (EUS)-guided tissue acquisition of pancreatic solid tumor requires a strict recommendation for its proper use in clinical practice because of its technical difficulty and invasiveness. The Korean Society of Gastrointestinal Endoscopy (KSGE) appointed a Task Force to draft clinical practice guidelines for EUS-guided tissue acquisition of pancreatic solid tumor. The strength of recommendation and the level of evidence for each statement were graded according to the Minds Handbook for Clinical Practice Guideline Development 2014. The committee, comprising a development panel of 16 endosonographers and an expert on guideline development methodology, developed 12 evidence-based recommendations in 8 categories intended to help physicians make evidence-based clinical judgments with regard to the diagnosis of pancreatic solid tumor. This clinical practice guideline discusses EUS-guided sampling in pancreatic solid tumor and makes recommendations on circumstances that warrant its use, technical issues related to maximizing the diagnostic yield (e.g., needle type, needle diameter, adequate number of needle passes, sample obtaining techniques, and methods of specimen processing), adverse events of EUS-guided tissue acquisition, and learning-related issues. This guideline was reviewed by external experts and suggests best practices recommended based on the evidence available at the time of preparation. This guideline may not be applicable for all clinical situations and should be interpreted in light of specific situations and the availability of resources. It will be revised as necessary to cover progress and changes in technology and evidence from clinical practice.

Regulation of gene expression by altered promoter methylation using a CRISPR/Cas9-mediated epigenetic editing system.

Despite the increased interest in epigenetic research, its progress has been hampered by a lack of satisfactory tools to control epigenetic factors in specific genomic regions. Until now, many attempts to manipulate DNA methylation have been made using drugs but these drugs are not target-specific and have global effects on the whole genome. However, due to new genome editing technologies, potential epigenetic factors can now possibly be regulated in a site-specific manner. Here, we demonstrate the utility of CRISPR/Cas9 to modulate methylation at specific CpG sites and to elicit gene expression. We targeted the murine Oct4 gene which is transcriptionally locked due to hypermethylation at the promoter region in NIH3T3 cells. To induce site-specific demethylation at the Oct4 promoter region and its gene expression, we used the CRISPR/Cas9 knock-in and CRISPR/dCas9-Tet1 systems. Using these two approaches, we induced site-specific demethylation at the Oct4 promoter and confirmed the up-regulation of Oct4 expression. Furthermore, we confirmed that the synergistic effect of DNA demethylation and other epigenetic regulations increased the expression of Oct4 significantly. Based on our research, we suggest that our proven epigenetic editing methods can selectively modulate epigenetic factors such as DNA methylation and have promise for various applications in epigenetics.

Cell forces meet cell metabolism.

Epithelial cells form energetically costly cell-cell adhesions in response to mechanical forces. How cells obtain their energy during this event is unclear. Activity of a key regulator of cell metabolism, the AMP-activated protein kinase (AMPK), is now shown to be mechanoresponsive, and thus can bridge adhesion mechanotransduction and energy homeostasis.

Lectin from Sambucus sieboldiana abrogates the anoikis resistance of colon cancer cells conferred by N-acetylglucosaminyltransferase V during hematogenous metastasis.

Anoikis is a form of anchorage-dependent apoptosis, and cancer cells adopt anokis-resistance molecular machinery to conduct metastasis. Here, we report that N-acetylglucosaminyltransferase V gene expression confers anoikis resistance during cancer progression. Overexpression of N-acetylglucosaminyltransferase V protected detached cancer cells from apoptotic death, and suppression or knockout of the gene sensitized cancer cells to the apoptotic death. The gene expression also stimulated anchorage-dependent as well as anchorage-independent colony formation of cancer cells following anoikis stress treatments. Importantly, treatment with the lectin from Sambucus sieboldiana significantly sensitized anoikis-induced cancer cell deaths in vitro as well as in vivo. We propose that the lectin alone or an engineered form could offer a new therapeutic treatment option for cancer patients with advanced tumors.

A 2-month-old boy with hemolytic anemia and reticulocytopenia following intravenous immunoglobulin therapy for Kawasaki disease: a case report and literature review.

Herein, we report a rare case of hemolytic anemia with reticulocytopenia following intravenous immunoglobulin therapy in a young infant treated for Kawasaki disease. A 2-month-old boy presented with fever lasting 3 days, conjunctival injection, strawberry tongue, erythematous edema of the hands, and macular rash, symptoms and signs suggestive of incomplete Kawasaki disease. His fever resolved 8 days after treatment with aspirin and high dose infusion of intravenous immunoglobulin. The hemoglobin and hematocrit decreased from 9.7 g/dL and 27.1% to 7.4 g/dL and 21.3%, respectively. The patient had normocytic hypochromic anemia with anisocytosis, poikilocytosis, immature neutrophils, and nucleated red blood cells. The direct antiglobulin test result was positive, and the reticulocyte count was 1.39%. The patient had an uneventful recovery. However, reticulocytopenia persisted 1 month after discharge.

Ultrasound-guided Aspiration of the Iatrogenic Pneumothorax Caused by Paravertebral Block -A Case Report-.

Thoracic paravertebral block is performed for the treatment of patients with chronic pain, such as complex regional pain syndrome (CRPS) and post-herpetic neuralgia. Thoracic paravertebral block can result in iatrogenic pneumothorax. Because pneumothorax can develop into medical emergencies and needle aspiration or chest tube placement may be needed, early diagnosis is very important. Recently, thoracic ultrasonography has begun to be used to diagnose pneumothorax. In addition, ultrasound-guided aspiration can be an accurate and safe technique for treatment of pneumothorax, as the needle position can be followed in real time. We report a case of iatrogenic pneumothorax following thoracic paravertebral block for the treatment of chronic pain due to CRPS, treated successfully by ultrasound-guided aspiration.

Effects of pneumoperitoneal pressure and position changes on respiratory mechanics during laparoscopic colectomy.

BACKGROUND: This study was designed to assess the effects of pneumoperitoneal pressure (PP) and positional changes on the respiratory mechanics during laparoscopy assisted colectomy. METHODS: Peak inspiratory pressure, plateau pressure, lung compliance, and airway resistance were recorded in PP of 10 mmHg and 15 mmHg, with the position change in 5 steps: head-down at 20°, head-down at 10°, neutral position, head-up at 10° and head-up at 20°. RESULTS: When the patient was placed head-down, the position change accentuated the effects of pneumoperitoneum on respiratory mechanics. However, when the patient was placed in a head-up position during pneumoperitoneum the results showed no pattern. In the 20° head-up position with the PP being 10 mmHg, the compliance increased from 30.6 to 32.6 ml/cmH(2)O compared with neutral position (P = 0.002). However with the PP being 15 mmHg, the compliance had not changed compared with neutral position (P = 0.989). In 20° head-down position with the PP of 10 mmHg, the compliance was measured as 24.2 ml/cmH(2)O. This was higher than that for patients in the 10° head-down position with a PP of 15 mmHg, which was recorded as 21.2 ml/cmH(2)O. Also in the airway resistance, the patient in the 20° head-down position with the PP of 10 mmHg showed 15.8 cmH(2)O/L/sec, while the patient in the 10° head-down position with the PP of 15 mmHg showed 16.2 cmH(2)O/L/sec of airway resistance. These results were not statistically significant but still suggested that the head-down position accentuated the effects of pneumoperitoneum on respiratory mechanics. CONCLUSIONS: Our results suggest that respiratory mechanics are affected by the patient position and the level of PP - the latter having greater effect.

Total intravenous anesthesia with propofol and remifentanil in a patient with MELAS syndrome -A case report-.

A 23-year-old woman with MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) underwent a laparoscopy-assisted appendectomy. MELAS syndrome is a multisystemic disease caused by mitochondrial dysfunction. General anesthesia has several potential hazards to patients with MELAS syndrome, such as malignant hyperthermia, hypothermia, and metabolic acidosis. In this case, anesthesia was performed with propofol, remifentanil TCI, and atracurium without any surgical or anesthetic complications. We discuss the anesthetic effects of MELAS syndrome.

[Studies on the Metagonimus fluke in the Daecheong reservoir and the upper stream of Geum river, Korea]

The prevalences of the fluke belonging to genus Metagonimus have been reported along the upper stream of inhabitants by several workers since 1980, however the taxonomical problems of the fluke was not yet settled. The larval flukes; cercaria and metacercaria as well as their intermediate hosts, and adult were studied in order to identify the Metagonimus in the areas. The results obtained are summarized as follows: The snails, Semisulcospira globus were collected from the three different localities along the upper stream of the River. The cercariae were found from 125(7.2%) out of 1,730 snails by natural emerging method, and were identified into 5 species including Metagonimus sp. (3.7%), Pseudexorchis major(1.4%), Cercaria nipponensis (0.9), Cercaria incerta (0.6%) and Cercaria yoshidae(0.6%). Cercariae of Metagonimus species had four to five oral spines on its anterior of the first line. The cercariae of Metagonimus were experimentally exposed to goldfish. Infection rate was 22.9% out of 105 goldfish, and the encysted metacercariae were found in fins(86.7%) and on scales (13.7%) of the fishes, but not in their muscle, head or visceral organs. Seven species of fish were caught in the Daecheong reservoir and the upper stream. Infestations with metacercaria of Metagonimus were found 100% in Opsariichtys bidens and the parasitized numbers of the metacercariae were observed from 250 to 2,400 per fish. In the upper stream, Zacco temmincki, Z. platypus and Pseudogobio esocinus were infected 100% with the metacercaria, on the other hand, the fishes caught in the reservoir showed the lower infestation rates, and a few metacercariae found in the fishes Carassius carassius and Cyprinus carpio in the reservoir and the stream. The majority of metacercariae was detected only on the scales of fishes. In order to know the infectivity and the distribution patterns in the intestine of hosts, rats and dogs were infected with the metacercariae obtained from O. bidens and Z. platypus. In addition the metacercariae obtained from Z. temmincki, P. esocinus and goldfish were given to the rats. The recovery rates of the worms in the small intestine of dogs were higher (63.3-65.8%) than those of the rats (3.5-31.6%). The flukes were found mostly in the middle and the lower part of small tntestine of the rats and the dogs, but no worm was collected in the upper part of the intestine of rats. The size of adult flukes varied by the hosts. In the adult flukes, oral sucker was smaller than ventral sucker, and the right and left testes were located diagonally, the uterine tubules circled around the upper left testis. The average egg size was 29.1 x 17.7 micro-meter. According to the above results, the flukes belonging to genus Metagonimus distributed along the Geum River was concluded to be identical with Miyata type of M. yokogawai as that Saito had proposed.