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The present study investigated the virulence and expression of innate immunity genes in isolates of infectious hematopoietic necrosis virus (IHNV) in Gangwon province, South Korea, by challenging rainbow trout, Atlantic salmon, and coho salmon. Eight IHNV isolates were used to infect RTG-2 cells for viral replication using plaque assays. Three isolates with the highest replication rates, the RtPc0314g and RtPc0314c isolates of the JRt-Shizuoka type and the RtPc0816g isolate of the JRt-Nagano type, were experimentally infected into the fish. In rainbow trout, both RtPc0314c and RtPc0314g isolates showed 100% cumulative mortality while the RtPc0816g isolate showed 60% cumulative mortality for 14 days. In contrast, all three isolates showed

Background: Giant lip hemangioma is a rare disease that may cause difficulty in preoxygenation and ventilation when using face masks and intubation during general anesthesia induction.Case: A laparoscopic cholecystectomy was planned for a 77-year-old woman. The patient had a giant lower lip hemangioma that was 12 x 5 x 5 cm, which made preoxygenation and ventilation through a face mask impossible and put her at risk of hemangioma rupture. We preoxygenated her through a high-flow nasal cannula (HFNC). Following propofol and succinylcholine administration, we intubated the patient with a video laryngoscope without desaturation, hemangioma rupture, or CO2 retention. Conclusions: HFNC is a useful tool when difficult intubation is expected in patients who have problems using conventional face masks.

Background Postoperative pain control after the minimally invasive repair of pectus excavatum (MIRPE) is essential, but there is a controversy about a better analgesic method between epidural and intravenous (IV) analgesia. This systematic review and meta-analysis aimed to compare the effect of epidural versus IV analgesia following MIRPE. Methods We searched PubMed, MEDLINE, EMBASE, Cochrane Central Register, and ClinicalTrials.gov for randomized control trials (RCTs) dated up to 31st May 2021. The primary outcome was the area under the curve (AUC) of the weighted mean visual analog scale (VAS) after MIRPE. The secondary outcomes were postoperative nausea, operation time, total operating room time, and postoperative length of hospital stay. Results Four RCTs involving 243 patients were finally included in this meta-analysis. The AUC of the weighted mean VAS was 343.62 in the epidural group and 375.24 in IV group. Epidural group showed lower VAS than IV group at 12 hours (mean difference -0.99 [95% CI: -1.52, -0.47], P = 0.001, I2 = 0%), at 24 hours (mean difference -0.65 [95% CI: -1.15, -0.16], P = 0.009, I2 = 0%), and 48 hours (mean difference -0.81 [95% CI: -1.61, -0.01], P = 0.046, I2 = 46%) after the surgery. Conclusion Epidural analgesia after the MIRPE had a better analgesic effect than IV analgesia from 12 hours to 48 hours after surgery, and AUC of VAS was lower in the epidural group. However, IV analgesia may also be a viable option, and physicians should wisely choose analgesic modalities after MIRPE.

Proteins that contain multiple disulfide bonds (SS bonds) expressed in Escherichia coli are usually problematic. This study reports the successful recombinant expression of the antimicrobial peptide β-defensin isolated from olive flounder in E. coli. The native form of β-defensin contained three discrete disulfide bridges: Cys1–Cys5, Cys2–Cys4, Cys3–Cys6. We constructed a periplasmic expression vector using small leading transmembrane protein YoaJ, and eventually, isolated bioactive β-defensin, which was then subjected to mass spectroscopy, circular dichroism spectroscopy, and anti-microbial testing. Results indicated bioactive β-defensin with a properly folded and native structure was formed. To investigate the roles of SS bonds, site-directed mutation method was applied to disrupt one, two, or three disulfide bridges. A dose-dependent effect was observed when more disulfide bridges were broken and a correlation between structure and function was observed, which further illustrated the key roles of SS bonds in maintaining the conserved motif and secondary structure of olive flounder beta-defensin.

Bacteria can produce recombinant proteins quickly and cost effectively. However, their physiological properties limit their use for the production of proteins in their native form, especially polypeptides that are subjected to major post-translational modifications. Proteins that rely on disulfide bridges for their stability are difficult to produce in Escherichia coli. The bacterium offers the least costly, simplest, and fastest method for protein production. However, it is difficult to produce proteins with a very large size. Saccharomyces cerevisiae and Pichia pastoris are the most commonly used yeast species for protein production. At a low expense, yeasts can offer high protein yields, generate proteins with a molecular weight greater than 50 kDa, extract signal sequences, and glycosylate proteins. Both eukaryotic and prokaryotic species maintain reducing conditions in the cytoplasm. Hence, the formation of disulfide bonds is inhibited. These bonds are formed in eukaryotic cells during the export cycle, under the oxidizing conditions of the endoplasmic reticulum. Bacteria do not have an advanced subcellular space, but in the oxidizing periplasm, they exhibit both export systems and enzymatic activities directed at the formation and quality of disulfide bonds. Here, we discuss current techniques used to target eukaryotic and prokaryotic species for the generation of correctly folded proteins with disulfide bonds.

Our aim was to study the relationship between the sex-determining genes, sox9a and cyp19a, and water temperature in Gadus chalcogrammus. We assessed the sex ratio based on the expression levels of sox9a and cyp19a at different water temperatures (5, 8, 11, and 14 &deg, C) and at different stages of walleye pollock development (embryos, larvae, and juveniles). Next, we used immature walleye pollock to assess sox9a expression in males and cyp19a and vitellogenin (VTG) expression in females at different water temperatures. Males expressed sox9a in the gonadal tissues, while females expressed cyp19a in the gonadal tissues and VTG in the blood plasma. In the first experiment, cyp19a expression was higher at 5 &deg, C and 8 &deg, C, and sox9a expression was higher at 11 and 14 &deg, C. In the second experiment, sox9a expression remained relatively stable, but cyp19a expression decreased with increasing temperature, decreasing significantly after 14 &deg, C. Similar patterns were also observed for VTG expression. These results indicate that lower water temperatures increase cyp19a expression, which increases the female ratio. Higher water temperatures increase sox9a expression, which increases the male ratio. Therefore, this study highlights the potential of the sex-determining genes and the influence of water temperature.

Hepcidin, an antimicrobial peptide produced by the liver, also controls the iron balance and regeneration in vertebrates. Two types of hepcidin (Hamp1 and Hamp2) have been found in the bodies of black rockfish (Sebastes schlegelii). The full-length cDNA of hepcidin was cloned to enable a study of the antibacterial roles of these two hepcidins (Hamp) in black rockfish. The antimicrobial function of recombinant hepcidins was tested both in vitro and in vivo by the synthesis in Escherichia coli of recombinant hepcidin (approximately 11 kDa) from black rockfish. The recombinant hepcidins inhibited the growth of two bacterial species, Streptococcus iniae FP5228 and Pseudomonas aeruginosa, at various concentrations, in vitro after 6 h post-incubation, respectively. During infection, the production of ferroportin was reduced, suggesting the preservation of iron to prevent microbial proliferation. In vivo administration of Hamp1, but not Hamp2, synthetic peptides induced a substantial reduction in the expression of ferroportin, suggesting that in black rockfish with two forms of hepcidin, ferroportin production is regulated by the iron-regulator Hamp1, and not by the dedicated antimicrobial Hamp2. The findings of this study suggest the various antimicrobial roles of these two types of hepcidin.

Autologous blood cell salvage reduces the need for postoperative allogeneic blood transfusion and alleviates immunologic reactions, so the technique is commonly used in cardiac surgery. The continuous autotransfusion system is a type of blood cell salvage device. Although the processing program of continuous autotransfusion system includes filtering of several materials from suctioned blood, such as clots, leukocytes, cytokines, and complement, we identified some unexpected blood clots in the reinfusion bag. Pathologic examination revealed that the clots were composed of fibrin, red blood cell aggregates, and histiocytes. We report two cases of these abnormal findings during the use of CATSmart in cardiac surgery.

Heat-moisture exchanger (HME) is an inexpensive and effective device used to prevent respiratory complications that can be caused by endotracheal tube insertion during general anesthesia. But, HME can increase airway resistance and be occluded by the patient’s secretions. Whether a HME could be occluded by clear fluids such as condensate in the airway circuit is not certain yet. In vitro, a case of HME occlusion by normal saline was reported. We report a case of HME obstruction by distilled water came from the heated wire circuit which was unintentionally connected to the HME.

Heat shock proteins (HSPs) are induced in response to various physiological or environmental stressors. However, the transcriptional activation of HSPs is regulated by a family of heat shock factors (HSFs). Fish models provide an ideal system for examining the biochemical and molecular mechanisms of adaptation to various temperatures and water environments. In this study, we examined the pattern differentials of heat shock factor 1 (HSF1) and expression of heat shock protein 70 (HSP70) in response to thermal stress in goldfish and mouse hepatocyte cultures by immune-blot analysis. Goldfish HSF1 (gfHSF1) changed from a monomer to a trimer at 33°C and showed slightly at 37°C, whereas mouse HSF1 (mHSF1) did so at 42°C. This experiment showed similar results to a previous study, indicating that gfHSF1 and mHSF1 play different temperature in the stress response. We also examined the activation conditions of the purified recombinant proteins in human HSF1 (hmHSF1) and gfHSF1 using CD spectroscopy and immune-blot analysis. The purified recombinant HSF1s were treated from 25°C to 42°C. Structural changes were observed in hmHSF1 and gfHSF1 according to the heat-treatment conditions. These results revealed that both mammal HSF1 (human and mouse HSF1) and fish HSF1 exhibited temperature-dependent changes; however, their optimal activation temperatures differed.

Peptides containing multiple disulfide bonds are usually problematic when expressed in Escherichia coli. We conducted the expression of β-defensin with three disulfide bonds, hepcidin with four disulfide bonds, and DkTx with six disulfide bonds using a small leader protein mosaic expression in the periplasm of E. coli, and purified them by affinity chromatography and characterized them by mass spectroscopy. The result showed that the expression level was high. A large amount of the pure recombinant peptide was also recovered after purification with a Ni2+ affinity column. The mass results by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) indicated that the recombinant peptides had a folding structure, with the native status of all of the cysteines participating in disulfide bond formation. Moreover, after removing the tags, the result was identical to its natural form as well. We thus provide a method for producing large amounts of soluble peptides containing multiple disulfide bonds in E. coli.

Objective A randomized, double-blind, prospective study to evaluate the effect of anticholinergic drugs on thermoregulation in paediatric patients undergoing ambulatory anaesthesia with ketamine. Methods Patients were randomized to receive either 0.005 mg/kg glycopyrrolate or the equivalent volume of normal saline (placebo) at 30 min before ketamine anaesthesia. Body temperature was measured tympanically at baseline and at 0, 30, 60 and 90 min postoperatively. The quantity of saliva prodiced during surgery and incidence of fever were recorded. Results Body temperature was significantly higher in the glycopyrrolate group ( n = 42) than the placebo group ( n = 42) at 30, 60 and 90 min after surgery, and higher than baseline at 0, 30, 60 and 90 min after surgery. In the placebo group, body temperature was significantly higher than baseline at 0 and 30 min after surgery. Saliva secretion was significantly lower in the glycopyrrolate group than the placebo group. Conclusion Routine premedication with adjunctive anticholinergics should not be considered in paediatric patients receiving ketamine sedation due to the increased risk of fever. Trial registration number, Clinicaltrials.gov: NCT02430272

The expression of eukaryotic molecular chaperones (heat shock proteins, HSPs) is triggered in response to a wide range of environmental stresses, including: heat shock, hydrogen peroxide, heavy metal, low-pH, or virus infection. Biochemical and genetic studies have clearly shown the fundamental roles of heat shock factor 1 (HSF1) in stress-inducible HSP gene expression, resistance to stress-induced cell death, carcinogenesis, and other biological phenomena. Previous studies show that acidic pH changes within the physiological range directly activate the HSF1 function in vitro. However, the detailed mechanism is unclear. Though computational pKa-predications of the amino acid side-chain, acidic-pH induced protonation of a histidine residue was found to be most-likely involved in this process. The histidine 83 (His83) residue, which could be protonated by mild decrease in pH, causes mild acidic-induced HSF1 activation (including in-vitro trimerization, DNA binding, in-vivo nuclear accumulation, and HSPs expression). His83, which is located in the loop region of the HSF1 DNA binding domain, was suggested to enhance the intermolecular force with Arginine 79, which helps HSF1 form a DNA-binding competent. Therefore, low-pH-induced activation of HSF1 by the protonation of histidine can help us better to understand the HSF1 mechanism and develop more therapeutic applications (particularly in cancer therapy). J. Cell. Biochem. 116: 977-984, 2015. © 2015 Wiley Periodicals, Inc.

According to recent statistics published by the National Industrial Security Center, former and current employees are responsible for 80.4% of companies' technology leakages, and employees of cooperative firms are responsible for another 9.6%. This means that 90% of technology leakages are intentionally or mistakenly caused by insiders. In a recent incident, a credit card company leaked private information, and the person responsible was an employee of a cooperative firm. These types of incidents have an adverse effect not only on a company's assets but also on its reputation. Therefore, most institutions implement various security solutions to prevent information from being leaked. However, security solutions are difficult to analyze and distinguish from one another because their logs are independently operated and managed. A large number of logs are created from various security solutions. This thesis investigates how to prevent internal data leakage by setting up individual scenarios for each security solution, analyzing each scenario's logs, and applying a monitoring system to each scenario.

Recombinant expression in Escherichia coli allows the simple, economical, and effective production of bioactive peptides. On the other hand, the production of native peptides, particularly those rich in disulfide bonds, is a major problem. Previous studies have reported that the use of carrier proteins for fusion expression can result in good peptide yields, but few are folded correctly. In this study, two transmembrane small proteins in E. coli, YoaJ and YkgR, which both orientate with their N-termini in cytoplasm and their C-termini in periplasm, were used for fusion expression. The recombinant production of two peptides, asteropsin A (ASPA) and β-defensin (BD), was induced in the periplasm of E. coli using a selected carrier protein. Both peptides were expressed at high levels, at yields of approximately 5-10 mg/L of culture. Mass spectrometry showed that the resulting peptide had the same molecular weight as their natural forms. After purification, single peaks were observed by reversed phase high-performance liquid chromatography (RP-HPLC), demonstrating the absence of isoforms. Furthermore, cytoplasmically expressed fusion proteins with a carrier at their C-termini did not contain disulfide bonds. This study provides new carrier proteins for fusion expression of disulfide bond-rich peptides in E. coli.

We experienced a case of venous vessel wall entrapment between the introducer needle and the guide wire during an attempt to perform right internal jugular vein (IJV) catheterization. The guide wire was introduced with no resistance but could not be withdrawn. We performed ultrasonography and C-arm fluoroscopy to confirm the entrapment location. We assumed the introducer needle penetrated the posterior vessel wall during the puncture and that only the guide wire entered the vein; an attempt to retract the wire pinched the vein wall between the needle tip and the guide wire. Careful examination with various diagnostic tools to determine the exact cause of entrapment is crucial for reducing catastrophic complications and achieving better outcomes during catheterization procedures.

The estrogen receptor (ER) and aryl hydrocarbon receptor (AhR) are ligand-activated transcription factors involved in estrogen or xenobiotic exposure, whereas the 90-kDa heat shock protein (HSP90), which is a ubiquitously expressed molecular chaperone, is involved in the signal transduction process. Although the interactions between these pathways have been under investigation, the mechanisms are unclear and the potential role of HSP90 in these interactions has not been reported. The results of goldfish primary hepatocytes showed that exposure to PCB77 and 17β-estradiol (E2) alone induced significant protein expression of cytochrome P450 1A (CYP1A) and vitellogenin (VTG), respectively. On the other hand, the combined exposure to PCB77 and E2 led to the reduction of CYP1A and VTG compared to the single treatments. Although the AhRs and ERs were naturally induced during the co-treatment, the total amount of HSP90 binding to the receptors was not changed. Furthermore, while the HSP90 chaperon activity was blocked by the specific inhibitor (geldanamycin), reciprocal inhibition between AhR and ER pathways was not observed. These findings indicate a potential role of HSP90 where competition between AhR and ER for binding to HSP90 can occur and cause reciprocal inhibition.

This study examined the influence of water temperature on the induction of vitellogenin (VTG) in walleye pollocks (Gadus chalcogrammus) in aquaculture. The full-length VTG-A (or VTG-I) gene of walleye pollock was cloned using RACE (Rapid amplification of cDNA ends)-PCR method from liver tissue and recombinant VTG protein (wpVTG) was expressed in E. coli. The wpVTG protein was used to construct polyclonal antibodies to specifically detect VTG in blood; the antibodies were verified using western blotting. The VTG in blood plasma was determined by ELISA. The relationship between GSI (gonadosomatic index), VTG and body length were all positively correlated, and the results indicated that VTG can be used as an indicator of sex and/or maturation of female G. chalcogrammus in aquaculture. When VTG was measured in by the aqua-cultured female pollocks raised in water temperature of 5 °C, 8 °C, 11 °C, and 14 °C, VTG was detected only at 5 °C and 8 °C. In the histological analysis of VTG-inducing individuals, vitellogenesis was found in ovaries. In those same samples, E2 (17beta-estradiol) was detected in the blood by LC-MS. On the other hand, there was no significant difference in level of VTG induction when E2 of same concentration was treated to hepatocyte culture in different water temperatures. These results seemed to indicate that VTG is not directly affected by water temperature. However, we considered that the water temperature indirectly affect VTG induction by affecting the E2 concentration level.

The original version of this article unfortunately contained a typo in the author name and in Acknowledgement section. The author name should be Youg Ok Kim instead it was published incorrectly as Yong Ok Kim.

The activation of heat shock factor 1 (HSF1) can be induced by the changes in environmental pH, but the mechanism of HSF1 activation by acidification is not completely understood. This paper reports that a low pH (pH~6.0) can trigger human HSF1 activation. Considering the involvement of the imidazole group of histidine residues under acid pH stress, an in vitro EMSA experiment, Trp-fluorescence spectroscopy, and protein structural analysis showed that the residue, His83, is the essential for pH-dependent human HSF1-activation. To determine the roles of His83 in the HSF1-mediated stress response affecting the cellular acid resistance, mouse embryo fibroblasts with normal wild-type or mutant mouse HSF1 expression were preconditioned by heating or pH stress. The results suggest that His83 is essential for HSF1 activation or the HSF1-mediated transcription of heat shock proteins, in protecting cells from acid pH stress.

We report a case of significant reduction in bispectral index (BIS) associated with suspected amniotic fluid embolism (AFE) that occurred prior to change in haemodynamic variables. The patient was a 29-year-old nulliparous, who was admitted for Caesarean section under general anaesthesia in the 33rd week of pregnancy. After the baby was born, the BIS value suddenly decreased to 0, with suppression ratio of 100. One minute later, saturation decreased abruptly to 85%, end-tidal carbon dioxide (EtCO2) decreased to 5 mmHg, peak inspiratory pressure increased to 35 cm H2O, and non-invasive blood pressure (BP) failed to obtain a reading. After administration of vasoactive drugs, the systolic BP was maintained at 100 mmHg or higher, the BIS value rose to 10-20, and the EtCO2 increased to 24-33 mmHg. In this case, the BIS monitoring may provide an earlier warning of impending cardiovascular collapse in the case of AFE.

Objectives Impedance cardiography (ICG) is a noninvasive technique that provides reasonably accurate measurements of cardiac output, but the usefulness of ICG in patients undergoing open abdominal surgery has not been validated. Methods Cardiac output was measured while patients underwent open gastrectomy using an ICG monitor ( niccomo™; ICG-CO); the results were compared with those measured using a FloTrac™/Vigileo™ monitor (Flo-CO), which measures cardiac output by analysing the arterial waveform. Data collection commenced at the beginning of anaesthetic induction and continued until the patient was awake. Data were compared using the Bland–Altman analysis, and the clinical significance of the difference between the two methods was evaluated by calculating the percentage error (%). Results Eleven patients were monitored during surgery. The bias of the Flo-CO and ICG-CO values was −0.45 l/min. The upper and lower limits of agreement were 0.96 l/min and −1.85 l/min, respectively. The percentage error was 28.5%. Electrocautery induced interference that transiently impaired the performance of the ICG monitor. Conclusions ICG provided useful information in evaluating the cardiac output of patients during abdominal surgery.

The identification of the sex and sexual maturity of geoducks (Panopea japonica) is difficult because the outer shape of the males and females is indistinguishable. Moreover, depending on their size or age, the annual maturation cycle can vary. Vitellogenin (VTG) is a precursor protein of egg yolk. It has been studied as a biomarker for sex identification in geoducks. This article reports on the cDNA cloning of the P. japonica VTG gene for the VTG fragment. It was found to exhibit homology to known VTGs isolated from other marine molluscs, with more than 50% homology with oyster VTG. An enzyme-linked immunosorbent assay was developed to quantitatively detect the VTG level in geoducks. The VTG polyclonal antibody could recognize the extracted VTG in females, but the level of VTG in males was undetectable, providing a basis for an immunological assay for the identification of the sex and female maturational status of geoducks. Thus, this study established a simple and quantitative technique for the rapid identification of sex and/or maturation of females by using VTG in the hemolymph as a biomarker for the sampled geoducks without dissection or seriously injuring them.