Live Imaging of Intracranial Lymphatics in the Zebrafish.

RATIONALE: The recent discovery of meningeal lymphatics in mammals is reshaping our understanding of fluid homeostasis and cellular waste management in the brain, but visualization and experimental analysis of these vessels is challenging in mammals. Although the optical clarity and experimental advantages of zebrafish have made this an essential model organism for studying lymphatic development, the existence of meningeal lymphatics has not yet been reported in this species. OBJECTIVE: Examine the intracranial space of larval, juvenile, and adult zebrafish to determine whether and where intracranial lymphatic vessels are present. METHODS AND RESULTS: Using high-resolution optical imaging of the meninges in living animals, we show that zebrafish possess a meningeal lymphatic network comparable to that found in mammals. We confirm that this network is separate from the blood vascular network and that it drains interstitial fluid from the brain. We document the developmental origins and growth of these vessels into a distinct network separated from the external lymphatics. Finally, we show that these vessels contain immune cells and perform live imaging of immune cell trafficking and transmigration in meningeal lymphatics. CONCLUSIONS: This discovery establishes the zebrafish as a important new model for experimental analysis of meningeal lymphatic development and opens up new avenues for probing meningeal lymphatic function in health and disease.

Optimization of a solidified micelle formulation for enhanced oral bioavailability of atorvastatin calcium using statistical experimental design.

To enhance the oral bioavailability of atorvastatin calcium (ATV), a novel solidified micelle (S-micelle) was developed. Two surfactants, Gelucire 48/16 (G48) and Tween 20 (T20), were employed for micelle formation, and two solid carriers (SC), Florite PS-10 (FLO) and Vivapur 105 (VP105), were selected as solid carriers. The S-micelle was optimized using a Box-Behnken design with three independent variables, including G48:T20 (X1, 1.8:1), SC:G48 + T20 (X2, 0.65:1), and FLO:VP105 (X3, 1.4:0.6), resulting in a droplet size (Y1) of 198.4 nm, dissolution efficiency at 15 min in the pH 1.2 medium (Y2) of 47.6%, Carr's index (Y3) of 16.9, and total quantity (Y4) of 562.5 mg. The optimized S-micelle resulted in good correlation showing percentage prediction values less than 10%. The optimized S-micelle formed a nanosized dispersion in the aqueous phase, with a higher dissolution rate than raw ATV and crushed Lipitor®. The optimized S-micelle improved the relative bioavailability of oral ATV (25 mg equivalent/kg) in rats by approximately 509 and 271% compared to raw ATV and crushed Lipitor®, respectively. In conclusion, the optimized S-micelle possesses great potential for the development of solidified formulations for improved oral absorption of poorly water-soluble drugs.

Prognostic Value of Plasma Presepsin and Pneumonia Severity Index in Patients with Community-Acquired Pneumonia in the Emergency Department.

Background and Objectives: Presepsin (PSS) is an independent predictor for estimating disease severity and prognosis in septic patients. Few studies have reported the associations between plasma PSS and the severity and prognosis in patients with community-acquired pneumonia (CAP). We investigated whether a high plasma PSS level was associated with 30-day mortality in CAP patients. Materials and Methods: This retrospective single-center study was conducted in an emergency department. The PSS level was measured in 211 adult CAP patients admitted to the hospital and followed for up to 30 days. We recorded the pneumonia severity index (PSI) and the CURB-65 score. The primary outcome was death from any cause within 30 days. Results: The plasma PSS levels were significantly elevated in the high-risk group (PSI > 130) compared with the low- (PSI < 91) or moderate-risk groups (PSI 91−130). Forty-four patients (20.9%) died within 30 days of admission. Non-survivors had significantly higher plasma PSS levels than survivors among CAP patients: 1083 (697−1736) pg/mL vs. 385 (245−554) pg/mL (p < 0.001). The area under the curve (AUC) to predict 30-day mortality was highest for PSS (0.867), followed by procalcitonin (0.728) and lactate (0.616). The cutoff level of plasma PSS for 30-day mortality was >754 pg/mL. The combination of PSI and plasma PSS level improved the predictive ability for 30-day mortality (AUC = 0.892). Cox regression analysis showed that higher PSS levels (>754 pg/mL) and higher PSI (>126) were associated with 30-day mortality in CAP patients (hazard ratios of 19.472 and 6.375, respectively). Conclusion: Elevated plasma PSS is associated with severity and 30-day mortality in hospitalized CAP patients. Combining plasma PSS level and PSI could significantly improve the predictive ability of PSS for 30-day mortality.

Development of the larval lymphatic system in zebrafish.

The lymphatic vascular system is a hierarchically organized complex network essential for tissue fluid homeostasis, immune trafficking and absorption of dietary fats in the human body. Despite its importance, the assembly of the lymphatic network is still not fully understood. The zebrafish is a powerful model organism that enables study of lymphatic vessel development using high-resolution imaging and sophisticated genetic and experimental manipulation. Although several studies have described early lymphatic development in the fish, lymphatic development at later stages has not been completely elucidated. In this study, we generated a new Tg(mrc1a:egfp)y251 transgenic zebrafish that uses a mannose receptor, C type 1 (mrc1a) promoter to drive strong EGFP expression in lymphatic vessels at all stages of development and in adult zebrafish. We used this line to describe the assembly of the major vessels of the trunk lymphatic vascular network, including the later-developing collateral cardinal, spinal, superficial lateral and superficial intersegmental lymphatics. Our results show that major trunk lymphatic vessels are conserved in the zebrafish, and provide a thorough and complete description of trunk lymphatic vessel assembly.

Anti-cancer drugs reactivate tumor suppressor miR-375 expression in tongue cancer cells.

Head and neck cancer is one of the deadliest malignant diseases and chemotherapy is a common treatment option. Despite the development of chemotherapies for several decades, how these drugs affect the dynamics of gene regulation is still largely unknown. In our previous study, miR-375 was shown to be underexpressed in oral cancers and thus unable to serve as a tumor suppressor microRNA to regulate certain putative oncogenes. In this study, we found that common anti-cancer drugs reactivated miR-375 in tongue cancer cells. Incubation of tongue cancer cells CAL 27 and SCC-25 in medium containing doxorubicin, 5-fluorouracil, trichostatin A, or etoposide significantly increased the expression of miR-375 and its primary transcript pri-miR-375. The dose- and time-dependent effects of doxorubicin in CAL 27 were demonstrated by miR-375 increases in response to the drug. Significant suppression of pri-miR-375 expression was observed in human tongue cancer specimens and this decrease was more prominent in advanced stage tumors. Bioinformatics from four publicly available mRNA microarray data sets suggested that these candidate miR-375 targets are mainly involved in cancer biology, indicating that these targets are likely to be suppressed via miR-375 due to the treatment with these drugs. Together, our data suggest that the four anti-cancer drugs examined in this study induce the expression of tumor suppressor miR-375 in tongue cancer.

miR-375 activates p21 and suppresses telomerase activity by coordinately regulating HPV E6/E7, E6AP, CIP2A, and 14-3-3ζ.

BACKGROUND: While microRNAs (miRNAs) are extensively studied in post-transcriptional regulation of gene expressions in many biological processes, cellular miRNA-mediated regulation of viral genes remains unclear. In particular, the interplay between human papillomavirus (HPV) genes and miRNAs and how these interactions contribute to HPV-associated cancers remain elusive. METHODS: Transient transfection of miR-375-mimic was used to compensate the loss-of-function of miR-375 in HPV-positive cancer. Regulation of oncogenic molecules and their downstream molecules via miR-375 in HPV-positive cancer was investigated using qRT-PCR, western blot, dual luciferase assay, indirect immunofluorescence analysis. All experiments were conducted at least three times to achieve statistical significance determined by Student t-test. RESULTS: In this study, we demonstrated how miR-375 negatively regulates HPV16 and 18 transcripts. We also found a cellular protein, E6-associated protein (E6AP), directly regulated by miR-375. miR-375-mediated repression of HPV transcripts and E6AP elevated major tumor suppressors p53, p21, and retinoblastoma protein 1 (RB). Cooperative regulation of miR-375 targets along with the increase of tumor suppressors led to ~60% reduction of telomerase reverse transcriptase (TERT) transcription followed by ~35% decrease of telomerase activity. Furthermore, miR-375-mediated regulation of 14-3-3ζ contributes to decrease telomerase activity by altering nuclear translocation of TERT. CONCLUSION: Taken together, miR-375-mediated suppression of multiple oncogenic components in HPV-associated carcinogenesis generates a cumulative biological response to rescue key tumor suppressors and diminish telomerase activity, which results in cell cycle arrest and cell proliferation inhibition.

Clinical Utility of the Tokyo Guidelines 2018 for Acute Cholangitis in the Emergency Department and Comparison with Novel Markers (Neutrophil-to-Lymphocyte and Blood Nitrogen Urea-to-Albumin Ratios).

Introduction: The Tokyo Guidelines 2018 (TG2018) is a scoring system used to recommend the clinical management of AC. However, such a scoring system must incorporate a variety of clinical outcomes of acute cholangitis (AC). In an emergency department (ED)-based setting, where efficiency and practicality are highly desired, clinicians may find the application of various parameters challenging. The neutrophil-to-lymphocyte ratio (NLR) and blood urea nitrogen-to-albumin ratio (BAR) are relatively common biomarkers used to assess disease severity. This study evaluated the potential value of TG2018 scores measured in an ED to predict a variety of clinical outcomes. Furthermore, the study also compared TG2018 scores with NLR and BAR scores to demonstrate their usefulness. Methods: This retrospective observational study was performed in an ED. In total, 502 patients with AC visited the ED between January 2016 and December 2021. The primary endpoint was to evaluate whether the TG2018 scoring system measured in the ED was a predictor of intensive care, long-term hospital stays (≥14 days), percutaneous transhepatic biliary drainage (PTBD) during admission care, and endotracheal intubation (ETI). Results: The analysis included 81 patients requiring intensive care, 111 requiring long-term hospital stays (≥14 days), 49 requiring PTBD during hospitalization, and 14 requiring ETI during hospitalization. For the TG2018 score, the adjusted OR (aOR) using (1) as a reference was 23.169 (95% CI: 9.788-54.844) for (3) compared to (1). The AUC of the TG2018 for the need for intensive care was 0.850 (95% CI: 0.815-0.881) with a cutoff of >2. The AUC for long-term hospital stays did not exceed 0.7 for any of the markers. the AUC for PTBD also did not exceed 0.7 for any of the markers. The AUC for ETI was the highest for BAR at 0.870 (95% CI: 0.837-0.899) with a cutoff value of >5.2. Conclusions: The TG2018 score measured in the ED helps predict various clinical outcomes of AC. Other novel markers such as BAR and NLR are also associated, but their explanatory power is weak.

Keratinization-associated miR-7 and miR-21 regulate tumor suppressor reversion-inducing cysteine-rich protein with kazal motifs (RECK) in oral cancer.

MicroRNAs (miRNAs) are small non-coding RNAs that posttranscriptionally regulate gene expression during many biological processes. Recently, the aberrant expressions of miRNAs have become a major focus in cancer research. The purpose of this study was to identify deregulated miRNAs in oral cancer and further focus on specific miRNAs that were related to patient survival. Here, we report that miRNA expression profiling provided more precise information when oral squamous cell carcinomas were subcategorized on the basis of clinicopathological parameters (tumor primary site, histological subtype, tumor stage, and HPV16 status). An innovative radar chart analysis method was developed to depict subcategories of cancers taking into consideration the expression patterns of multiple miRNAs combined with the clinicopathological parameters. Keratinization of tumors and the high expression of miR-21 were the major factors related to the poor prognosis of patients. Interestingly, a majority of the keratinized tumors expressed high levels of miR-21. Further investigations demonstrated the regulation of the tumor suppressor gene reversion-inducing cysteine-rich protein with kazal motifs (RECK) by two keratinization-associated miRNAs, miR-7 and miR-21. Transfection of miR-7 and miR-21-mimics reduced the expression of RECK through direct miRNA-mediated regulation, and these miRNAs were inversely correlated with RECK in CAL 27 orthotopic xenograft tumors. Furthermore, a similar inverse correlation was demonstrated in CAL 27 cells treated in vitro by different external stimuli such as trypsinization, cell density, and serum concentration. Taken together, our data show that keratinization is associated with poor prognosis of oral cancer patients and keratinization-associated miRNAs mediate deregulation of RECK which may contribute to the aggressiveness of tumors.

Divergent GW182 functional domains in the regulation of translational silencing.

MicroRNA (miRNA)-mediated gene regulation has become a major focus in many biological processes. GW182 and its long isoform TNGW1 are marker proteins of GW/P bodies and bind to Argonaute proteins of the RNA induced silencing complex. The goal of this study is to further define and distinguish the repression domain(s) in human GW182/TNGW1. Two non-overlapping regions, Δ12 (amino acids 896-1219) containing the Ago hook and Δ5 (amino acids 1670-1962) containing the RRM, both induced comparable silencing in a tethering assay. Mapping data showed that the RRM and its flanking sequences in Δ5, but not the Ago hook in Δ12, were important for silencing. Repression mediated by Δ5 or Δ12 was not differentially affected when known endogenous repressors RCK/p54, GW182/TNGW1, TNRC6B were depleted. Transfected Δ5, but not Δ12, enhanced Ago2-mediated repression in a tethering assay. Transfected Δ12, but not Δ5, released endogenous miRNA reporter silencing without affecting siRNA function. Alanine substitution showed that GW/WG motifs in Δ12 (Δ12a, amino acids 896-1045) were important for silencing activity. Although Δ12 appeared to bind PABPC1 more efficiently than Δ5, neither Δ5 nor Δ12 significantly enhanced reporter mRNA degradation. These different functional characteristics of Δ5 and Δ12 suggest that their roles are distinct, and possibly dynamic, in human GW182-mediated silencing.

Fabrication of Assembled FeS2 Nanosheet and Application for High-Performance Supercapacitor Electrodes.

To overcome the low-energy-density limitation of supercapacitors, we aimed to achieve a material with a high specific capacitance by manipulating the nanostructure of FeS2, which comprises the most abundant and affordable elements. In this study, nanosheet-assembled FeS2 (NSA-FeS2) was fabricated using a novel method. Sub-micron droplets of sulfur particles stabilized with polyvinylpyrrolidone were formed in silicone oil medium, and Fe(CO)5 was absorbed and reacted on the surface to form core-shell particles, ES/[Fe], with a sulfur core and an iron-containing outer shell. The high temperature treatment of ES/[Fe] produced NSA-FeS2, in which pyrite FeS2 nanosheets grew and were partially interconnected. In a three-electrode system, the as-prepared NSA-FeS2 and NSA-FeS2/polyaniline (PANI) composites exhibited specific capacitances of 763 and 976 Fg-1, respectively, at a current density of 0.5 Ag-1, with corresponding capacitance retentions of 93 and 96% after 3000 charge-discharge cycles. The capacitance retention of the NSA-FeS2/PANI composites was 49% when the current density was increased from 0.5 to 5 Ag-1. Notably, the obtained specific capacitances exhibited the highest values in pure FeS2 and FeS2-based composites, indicating the significant potential for the utilization of iron sulfide in pseudocapacitive electrode materials.

Functionalized Lipid Nanocarriers for Simultaneous Delivery of Docetaxel and Tariquidar to Chemoresistant Cancer Cells.

The simultaneous drug delivery efficiency of a co-loaded single-carrier system of docetaxel (DTX)- and tariquidar (TRQ)-loaded nanostructured lipid carrier (NLC) functionalized with PEG and RIPL peptide (PRN) (D^T-PRN) was compared with that of a physically mixed dual-carrier system of DTX-loaded PRN (D-PRN) and TRQ-loaded PRN (T-PRN) to overcome DTX mono-administration-induced multidrug resistance. NLC samples were prepared using the solvent emulsification evaporation technique and showed homogeneous spherical morphology, with nano-sized dispersion (<220 nm) and zeta potential values of -15 to -7 mV. DTX and/or TRQ was successfully encapsulated in NLC samples (>95% encapsulation efficiency and 73-78 µg/mg drug loading). In vitro cytotoxicity was concentration-dependent; D^T-PRN exhibited the highest MDR reversal efficiency, with the lowest combination index value, and increased the cytotoxicity and apoptosis in MCF7/ADR cells by inducing cell-cycle arrest in the G2/M phase. A competitive cellular uptake assay using fluorescent probes showed that, compared to the dual nanocarrier system, the single nanocarrier system exhibited better intracellular delivery efficiency of multiple probes to target cells. In the MCF7/ADR-xenografted mouse models, simultaneous DTX and TRQ delivery using D^T-PRN significantly suppressed tumor growth as compared to other treatments. A single co-loaded system for PRN-based co-delivery of DTX/TRQ (1:1, w/w) constitutes a promising therapeutic strategy for drug-resistant breast cancer cells.

Soft-capsule formulation of a re-esterified triglyceride omega-3 employing self-emulsifying technology and bioavailability evaluation in healthy volunteers.

Despite the superior clinical efficacy of the re-esterified triglyceride (rTG) form compared to the ethylester form, few studies have been conducted on improving the bioavailability of the rTG form of omega-3 oil. The aim of study was to evaluate the effect of self emulsifying formulation on the improvement of bioavailability of rTG form of omega-3 oil. To develop a re-esterified triglyceride (rTG) soft capsule, an rTG-loaded self-emulsifying delivery system (SEDS) was designed using coconut oil, polysorbate 80, and lecithin. Candidate formulations were designed from a phase-diagram study and optimal SEDS formulations containing 85% of high omega-3 (ω-3) oils were screened from the evaluation of droplet size distribution, measurement of oil floating area and emulsion turbidity. The selected, optimized rTG SEDS formulation was filled into a soft capsule (NOVASEDS) and applied to a sequence-randomized, double-blind, single-dose, and two-way crossover clinical study (n = 44), and the the bioavailability of NOVASEDS was compared with that of a 'raw' rTG capsule (rTG OMEGA3) as control. The droplet size (D50) formed from the candidate formulations was approximately 30-45 µm, and the optimal formulation showed a unimodal particle distribution with the smallest oil floating area and small changes in turbidity after 24 h. Cmax and AUC from 0 to 24 h for NOVASEDS, calculated from docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and as the sum of DHA and EPA, were significantly higher (P < 0.05) than corresponding values for rTG OMEGA3. In conclusion, NOVASEDS formulated by SEDS technology enabled the manufacture of a high rTG payload soft capsule with improved bioavailability in human subjects.

Plasma presepsin level predicts acute kidney injury in patients with sepsis in the emergency department.

Acute kidney injury (AKI) is a common complication in patients with sepsis. We evaluated the potential prognostic value of plasma presepsin to predict AKI in patients with sepsis in the emergency department. A total of 193 patients diagnosed with sepsis based on the Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3) were included in this observational study. AKI was defined according to the Kidney Disease Improving Global Outcomes clinical practice guideline. Plasma presepsin levels were measured on admission to the emergency department. We compared plasma presepsin levels between patients who did and those who did not develop AKI. AKI occurred in 100 (51.8%) patients. The median plasma presepsin level was significantly higher in patients with AKI than in those without AKI (1061 pg/mL vs 495 pg/mL, P <.001). Plasma presepsin levels were significantly increased in patients with AKI stage 3 compared with those with AKI stages 1 and 2 (P =.001). The area under the curve of presepsin for predicting AKI was 0.793 (95% confidence interval: 0.729-0.848). The optimal presepsin cutoff value for predicting AKI was >572 pg/mL, with a sensitivity of 77.0% and specificity of 81.7%. Plasma presepsin level is a valuable biomarker for the prediction of AKI in patients with sepsis in the emergency department.

Synergistic Effects of Fe2O3 Nanotube/Polyaniline Composites for an Electrochemical Supercapacitor with Enhanced Capacitance.

α-Fe2O3, which is an attractive material for supercapacitor electrodes, has been studied to address the issue of low capacitance through structural development and complexation to maximize the use of surface pseudocapacitance. In this study, the limited performance of α-Fe2O3 was greatly improved by optimizing the nanotube structure of α-Fe2O3 and its combination with polyaniline (PANI). α-Fe2O3 nanotubes (α-NT) were fabricated in a form in which the thickness and inner diameter of the tube were controlled by Fe(CO)5 vapor deposition using anodized aluminum oxide as a template. PANI was combined with the prepared α-NT in two forms: PANI@α-NT-a enclosed inside and outside with PANI and PANI@α-NT-b containing PANI only on the inside. In contrast to α-NT, which showed a very low specific capacitance, these two composites showed significantly improved capacitances of 185 Fg-1 for PANI@α-NT-a and 62 Fg-1 for PANI@α-NT-b. In the electrochemical impedance spectroscopy analysis, it was observed that the resistance of charge transfer was minimized in PANI@α-NT-a, and the pseudocapacitance on the entire surface of the α-Fe2O3 nanotubes was utilized with high efficiency through binding and conductivity improvements by PANI. PANI@α-NT-a exhibited a capacitance retention of 36% even when the current density was increased 10-fold, and showed excellent stability of 90.1% over 3000 charge-discharge cycles. This approach of incorporating conducting polymers through well-controlled nanostructures suggests a solution to overcome the limitations of α-Fe2O3 electrode materials and improve performance.

Association of Plasma Glucose to Potassium Ratio and Mortality After Aneurysmal Subarachnoid Hemorrhage.

Objectives: Hyperglycemia and hypokalemia are common problems in patients with aneurysmal subarachnoid hemorrhage (aSAH). The aim of this study was to determine whether the plasma glucose to potassium ratio (GPR) predicts mortality due to aSAH. Methods: We prospectively recruited aSAH patients and healthy controls between March 2007 and May 2017. Clinical outcomes included mortality and poor outcome (modified Rankin scale score of 3-6) after 3 months. Multivariable analysis was used to determine the association between plasma GPR and 3-month mortality in aSAH patients. Results: A total of 553 patients were recruited, and the mortality rate was 11%. The GPR was significantly elevated in aSAH patients compared with controls, in patients with a poor outcome than with a good outcome and in non-survivals than in survivals. Multivariable analysis showed that the plasma GPR was an independent factor associated with 3-month mortality. The area under the curve of the GPR was 0.747 in predicting 3-month mortality. Conclusion: The plasma GPR on admission has potential as a predictor of 3-month mortality in patients with aSAH.

Comparison of ultrasonography and computed tomography for measuring optic nerve sheath diameter for the detection of elevated intracranial pressure.

OBJECTIVE: The main aim of this study was to compare optic nerve sheath diameter (ONSD) measured using ultrasonography (USG) and computed tomography (CT) almost simultaneously in the same patients with suspected elevated intracranial pressure. The other aim of this study was to evaluate the diagnostic ability for detecting elevated intracranial pressure using ONSD measured by USG (USG-ONSD) and by CT (CT-ONSD). PATIENTS AND METHODS: This prospective, observational study was undertaken from June to October 2020 in the emergency department (ED) of a tertiary medical center in Seoul. ONSD was measured by USG and CT at 3 mm behind the posterior aspect of the globe. RESULT: A total of 199 patients were enrolled. The median USG-ONSD and CT-ONSD were significantly higher in patients with elevated intracranial pressure than in patients with normal intracranial pressure. The interclass correlation coefficient between USG-ONSD and CT-ONSD was 0.785 (95% CI 0.715-0.837). A Bland-Altman plot showed significant agreement between USG and CT measurements. The optimal cutoff for detecting elevated intracranial pressure was >5.3 mm (sensitivity of 75.4% and specificity of 90.8%) for USG and >5.0 mm (sensitivity of 68.4% and specificity of 85.2%) for CT. CONCLUSION: The ONSD measured using USG and CT were increased in patients with elevated intracranial pressure. Measurement of ONSD by USG and CT showed very high agreement.

Increased immortalization-upregulated protein 2 (IMUP-2) by hypoxia induces apoptosis of the trophoblast and pre-eclampsia.

In regulation of the developmental process, the balance between cellular proliferation and cell death is critical. Placental development tightly controls this mechanism, and increased apoptosis of placental trophoblasts can cause a variety of gynecological diseases. Members of the immortalization-upregulated protein (IMUP) family are nuclear proteins implicated in SV40-mediated immortalization and cellular proliferation; however, the mechanisms by which their expression is regulated in placental development are still unknown. We compared IMUP-2 expression in normal and pre-eclamptic placental tissues and evaluated the function of IMUP-2 in HTR-8/SVneo trophoblast cells under hypoxic conditions. IMUP-2 was expressed in syncytiotrophoblasts and syncytial knots of the placental villi. IMUP-2 expression was significantly higher in preterm pre-eclampsia patients than in patients who went to term (P < 0.001); however, we observed no differences in IMUP-2 expression between normal term patients with and without pre-eclampsia. Hypoxic conditions increased apoptosis of HTR8/SVneo trophoblast cells and induced IMUP-2 expression. Also, apoptosis of HTR-8/SVneo trophoblast cells was increased after IMUP-2 gene transfection. These results suggest that IMUP-2 expression is specifically elevated in preterm pre-eclampsia and under hypoxic conditions, and that IMUP-2 induces apoptosis of the trophoblast. Therefore, IMUP-2 might have functional involvement in placental development and gynecological diseases such as pre-eclampsia.

Identification and characterization of WSG, a fusion gene associated with the proliferation of the WI-38 VA13 cells.

It is well known that human fibroblasts can be immortalized using simian virus 40 (SV40) T antigen. However, the mechanisms of the SV40-immortalization processes remain unclear. In the present study, the authors identified and characterized a fusion gene, WSG (WI-38 VA13 Specific Gene), which has an integrated sequence of SV40 and chromosome 16p13. WSG is only detectable in WI-38 VA13 cells and not in other human cell lines or tissues. Transient transfection of the constructed pEGFP-WSG certified the WSG localization at the nuclear of HeLa cells. The growth assays and the knockdown experiment indicate that WSG is involved in the WI-38 VA13 cell proliferation. These results support potential capacities of WSG to be a candidate gene involved in proliferation of the WI-38 VA13 cells.

Expression profiles of SV40-immortalization-associated genes upregulated in various human cancers.

Immortalization is an early and essential step of human carcinogenesis which is associated with alterations in gene expression and regulation. Suppression subtractive hybridization (SSH) was successfully performed to identify immortalization-associated genes upregulated in SV40-immortalized lung fibroblasts. We identified 116 known genes which were related to diverse functions, with 32.8% relevant for cell cycle or proliferation indicating the potential involvement of these genes in immortalization. We chose eight known genes located on the overrepresented chromosomes of non-small-cell lung cancers (NSCLCs). ASPM, RFC4, C3orf26, BXDC2, C15orf44, AURKA, C20orf77, and RBMX were upregulated in immortalized cells, cancer cells, and non-small-cell lung cancer (NSCLC) tissues. We additionally cloned two novel genes (CHA-V-97 and CHA-V-165) which showed similar upregulated expression patterns in cells and tissues examined. Identification and further characterization of these genes may provide insights of novel players for immortalization and human carcinogenesis.

Tracheal Rupture Related to Endotracheal Intubation during Cardiopulmonary Resuscitation.

Postintubation tracheal rupture is rare, but serious. Emergency intubation is often conducted during cardiopulmonary resuscitation (CPR), and the risk of postintubation tracheal rupture can be increased during CPR. We describe here a case of postintubation tracheal rupture in a 65-year-old female who was transferred from another hospital after CPR. Postintubation tracheal rupture in this case is thought to have been related to malposition of the endotracheal tube (ETT), elevation of the intratrachea pressure due to chest compression, and an overinflated cuff. However, the most important factor is considered to be the overinflated cuff, which is often caused by manual palpation. Therefore, emergency physicians should consider using a manometer to check the cuff pressure of the ETT, even during CPR. When spontaneous circulation is restored, the pressure of the cuff must be measured with a manometer.