Molecular, biochemical, and toxicological characterization of two acetylcholinesterases from the Western flower thrips, Frankliniella occidentalis.

We investigated the molecular and biochemical properties of two acetylcholinesterases (FoAChE1 and FoAChE2) from the Western flower thrips, Frankliniella occidentalis. Polyacrylamide gel electrophoresis and western blotting confirmed the membrane-anchored nature of both FoAChE1 and FoAChE2, which was further supported by hydrophobicity and glycophosphatidylinositol anchor predictions. High expression levels of both enzymes were observed in the head, indicating their predominant distribution in neuronal tissues. FoAChE1 exhibited significantly higher expression levels in all examined tissues compared to FoAChE2, suggesting its major role as a synaptic enzyme. Nonetheless, both recombinant enzymes displayed robust catalytic activity toward acetylthiocholine iodide, and FoAChE1 demonstrated nearly identical catalytic efficiency compared to FoAChE2. FoAChE1 exhibited slightly lower sensitivities to the cholinesterase inhibitors tested, including organophosphates (OPs) and carbamates (CBs), compared to FoAChE2. Field populations of F. occidentalis exhibited polymorphism of alanine vs. serine at position 197 of FoAChE1 within the conserved oxyanion hole. Contrary to common belief, however, functional analysis using recombinant enzymes revealed that neither A197 nor S197 residue was associated with FoAChE1 insensitivity to OPs and CBs. FoAChE2 did not exhibit any polymorphic amino acid substitutions at the positions known to be associated with resistance. Due to the absence of apparent resistance-associated mutations in field populations of F. occidentalis, the judicious use of some OPs or CBs can be suggested for controlling the highly resistant populations to other insecticides. Overall, our findings highlight the significance of both FoAChE1 and FoAChE2 as targets for toxicity assessment, while the specific contribution of each enzyme to toxicity remains unclear.

Transcriptomic identification and characterization of genes commonly responding to sublethal concentrations of six different insecticides in the common fruit fly, Drosophila melanogaster.

Pretreatment with sublethal concentrations (LC10) of six insecticides (chlorantraniliprole, cypermethrin, dinotefuran, indoxacarb, ivermectin, and spinosad) significantly elevated tolerance of the common fruit fly Drosophila melanogaster to lethal concentration of the respective insecticide. Commonly responding genes to sublethal treatments of the six insecticides were identified by transcriptome analysis based on a fold change >1.5 or < -1.5, and p < 0.05 as selection criteria. Following treatment with all the six insecticides, 26 transcripts were commonly over-transcribed, whereas 30 transcripts were commonly under-transcribed. Reliability of the transcriptome data was confirmed by quantitative PCR. A majority of the over-transcribed genes included those related to olfactory behavior, such as odorant-binding proteins, as well as immune-related genes, including attacin, diptericin, and immune-induced molecule 18. In contrast, genes belonging to the mitochondrial respiratory chain, such as mitochondrial NADH-ubiquinone oxidoreductase chain 1/3/4/5 and mitochondrial cytochrome b/c, were commonly under-transcribed. Furthermore, genes related to eggshell formation and motion were also under-transcribed, which may indicate a possible energy trade-off for xenobiotic stress. In summary, most of the differentially expressed genes were not directly related to well-known detoxification genes, suggesting that the roles of commonly expressed tolerance-related genes are not likely related to direct metabolic detoxification, but rather are associated with restoration of homeostasis.

Transcriptomic identification and characterization of genes responding to sublethal doses of three different insecticides in the western flower thrips, Frankliniella occidentalis.

Pretreatment with sublethal concentrations (LC10) of three insecticides (chlorfenapyr, dinotefuran, and spinosad) enhanced tolerance to a lethal dose of the respective insecticide in the Western flower thrips, Frankliniella occidentalis. To identify genes responding to sublethal treatment with insecticides, transcriptome analysis was conducted for thrips treated with LC10 of the three insecticides. When based on a fold change >1.5 or < -1.5 as a selection criterion, 199 transcripts were commonly up-regulated, whereas 31 transcripts were commonly down-regulated following all three insecticide treatments. The differential expression levels of representative genes were validated by quantitative PCR. Most over-transcribed transcripts could be categorized as basic biological processes, such as proteolysis and lipid metabolism. Detoxification genes, such as one glutathione S transferase S1, three UDP-glucuronosyltransferases, four CYP450s, and one ABC transporter G family member 20, were commonly overexpressed in all three insecticide-treated groups. Knockdown of the five representative commonly overexpressed genes via ingestion RNA interference increased mortalities to all the three test insecticides, supporting their common role in tolerance induction. In contrast, three C2H2-type zinc finger-containing proteins were significantly down-regulated in all insecticide-treated thrip groups. Since the tested insecticides have distinct structures and modes of action, the roles of commonly expressed genes in tolerance were discussed.

Metabolism of insecticide diazinon by Cunninghamella elegans ATCC36112.

The fungal metabolism of diazinon was investigated and the microbial model (Cunninghamella elegans ATCC36112) could effectively degrade the organophosphorus pesticide (diazinon) mediated by cytochrome P450, which was mainly involved in oxidation and hydrolysis of phase I metabolism. Approximately 89% of diazinon was removed within 7 days and was not observed after 13 days with concomitant accumulation of eight metabolites. Structures of the metabolites were fully or tentatively identified with GC-MS and 1H, 13C NMR. The major metabolites of diazinon were diethyl (2-isopropyl-6-methylpyrimidin-4-yl) phosphate (diazoxon) and 2-isopropyl-6-methyl-4-pyrimidinol (pyrimidinol), and formation of minor metabolites was primarily the result of hydroxylation. To determine the responsible enzymes in diazinon metabolism, piperonyl butoxide and methimazole were treated, and the kinetic responses of diazinon and its metabolites by Cunninghamella elegans were measured. Results indirectly demonstrated that cytochrome P450 and flavin monooxygenase were involved in the metabolism of diazinon, but methimazole inhibited the metabolism less effectively. Based on the metabolic profiling, a possible metabolic pathway involved in phase I metabolism of diazinon was proposed, which would contribute to providing insight into understanding the toxicological effects of diazinon and the potential application of fungi on organophosphorus pesticides.

Successful intubation using video laryngoscope in a child with CHARGE syndrome: A case report

CHARGE syndrome is a rare genetic disorder with CHD7 gene mutation. CHARGE is an acronym for coloboma (C), heart disease (H), atresia of choanae (A), retardation of growth (R), genitourinary malformation (G), and ear abnormalities (E). Patients with CHARGE syndrome need to undergo many surgeries due to their various congenital anomalies. Since airway abnormalities frequently accompany CHARGE syndrome, general anesthesia remains a challenge. Here we report a case of difficult intubation in a 35-month-old boy with CHARGE syndrome during general anesthesia and the experience of successful intubation using D-blade of C-MAC® video laryngoscope.

The effect of gender and age on postoperative pain in laparoscopic cholecystectomy: a prospective observational study

BACKGROUND: Gender and age are known factors that affect postoperative pain. The purpose of this study was to investigate the effect of gender and age on postoperative pain and analgesic consumptions after laparoscopic cholecystectomy. METHODS: We studied 240 adult patients (120 female patients and 120 male patients) who underwent elective 3-port laparoscopic cholecystectomy under general anesthesia. The numerical rating scale (NRS) scores were assessed before surgery, when the patient arrived in the recovery room, and at 8, 16, and 24 h after surgery. Analgesic consumptions in the recovery room and at 24 h postoperatively were evaluated. We compared the NRS scores and quantity of analgesics between the gender groups and the age groups (20–45, 46–64, and ≥ 65 years old). RESULTS: Compared to male patients, female patients had higher NRS scores (6 [5, 7] vs. 5 [4, 6]; P = 0.001) and required a higher dose of fentanyl (0.94 ± 0.47 µg/kg vs. 0.79 ± 0.41 µg/kg; P = 0.011) in the recovery room. The younger female patients (20–45 and 46–64 years old) required a higher dose of fentanyl than those older than 65 years of age in the recovery room. CONCLUSIONS: Female patients exhibit higher NRS scores and greater consumptions of analgesics than male patients immediately postoperatively. Younger female patients require more analgesics than elderly patients in the recovery room.

A comparison of clinical performance between i-gel and endotracheal tube in pediatric laparoscopic surgeries

BACKGROUND: The current evidence on the safe use of supraglottic airway for pediatric laparoscopic surgeries is limited. Although i-gel has been successfully used in adult laparoscopic surgeries, to our knowledge, no studies have compared it to the endotracheal tube (ETT) in pediatric laparoscopic surgeries. This study evaluated the effectiveness of i-gel over ETT with regards to the respiratory and hemodynamic parameters during pediatric laparoscopic surgeries. METHODS: A total of 60 pediatric patients undergoing elective laparoscopic surgeries were randomly allocated to either the i-gel or ETT groups. Anesthetics used included ketamine, sevoflurane, and rocuronium. The primary outcome measured was the peak airway pressure (PAP) and the secondary outcomes measured were leak fraction, end-tidal CO₂, respiratory rate, insertion time, heart rate, blood pressure and perioperative complications. RESULTS: The PAP during surgeries was higher in the ETT group than in the i-gel group. There were no statistically significant differences in the leak fraction, end-tidal CO₂, and respiratory rate. The i-gel group had a shorter insertion time compared with the ETT group. The changes in heart rate were comparable in both groups. However, systolic and diastolic pressures were higher in the ETT group following intubation, before and after the creation of pneumoperitoneum. The incidence of perioperative complications was similar in both groups. CONCLUSIONS: The i-gel provided adequate ventilation with lower PAP compared with ETT. In addition, it provided minimal hemodynamic changes compared with ETT. Therefore, the i-gel may provide a suitable alternative to ETT in pediatric laparoscopic surgeries.

Advantages of Using Ultrasound in Regional Anesthesia for a Super-Super Obese Patient / 고신대학교의과대학학술지

In this case report, we describe the use of ultrasound in the administration regional anesthesia for a super-super obese patient. A 23-year-old female patient (height 167.2 cm, weight 191.5 kg, body mass index 68.6 kg/m²) was admitted to the hospital for surgical repair of an anterior talofibular ligament rupture. We used ultrasound to help facilitate the administration of regional anesthesia. In the sagittal view of the lumbar spine, (with the patient in a sitting position) we were able to identify the border between the sacrum and the lumbar vertebral; in the transverse view, we were able to identify the transverse process, posterior dura, vertebral body, and the distance from the skin to the posterior dura. After skin marking, regional anesthesia was successfully performed. Based on this case study, we suggest that ultrasound can be very useful in regional anesthesia for severely obese patients.

An investigation of the molecular and biochemical basis underlying chlorantraniliprole-resistant Drosophila strains and their cross-resistance to other insecticides.

Chlorantraniliprole is an anthranilic diamide insecticide that binds to the insect ryanodine receptor (RyR) and induces an uncontrolled release of Ca2+ , resulting in paralysis and ultimately death of the target insects. Recently, it was reported that chlorantraniliprole-resistant diamondback moths, Plutella xylostella Linnaeus, have mutations in their RyR. In this study, we developed two different chlorantraniliprole-resistant Drosophila melanogaster strain. The resistance ratio (RR) of the low-concentration chlorantraniliprole-treated resistant (Low-Res) strain was 2.3, while that of the high-concentration chlorantraniliprole-treated resistant (High-Res) strain was 21.3. The LC 50 of the untreated control (Con) strain was 23.8~25.9 ppm, which was significantly higher than that reported for the susceptible diamondback moth (0.03~0.51 ppm). The high LC 50 of the Con may be because the helix S2 amino acid sequence of D. melanogaster RyR ( DmRyR) is identical to the I4790M mutation of the chlorantraniliprole-resistant diamondback moths, resulting in a lower binding affinity of DmRyR for chlorantraniliprole. Among the tested detoxification enzymes, the activity of esterase was significantly increased in the two Res strains, but glutathione S-transferases and acetylcholinesterase were significantly decreased in the two Res strains. The cross-resistance of the High-Res strain to other insecticides with different modes of actions (MoAs) revealed that the RRs of the neuronal acetylcholine receptor allosteric and competitive modulators were significantly increased, while those of the Na 2+ channel modulators were significantly reduced. Our studies showed that RRs against the same insecticide vary with the treatment concentration, and that RRs against other insecticides with different MoAs can be altered.

Anti-BACE1 and Antimicrobial Activities of Steroidal Compounds Isolated from Marine Urechis unicinctus.

The human ß-site amyloid cleaving enzyme (BACE1) has been considered as an effective drug target for treatment of Alzheimer's disease (AD). In this study, Urechis unicinctus (U. unicinctus), which is a Far East specialty food known as innkeeper worm, ethanol extract was studied by bioassay-directed fractionation and isolation to examine its potential ß-site amyloid cleaving enzyme inhibitory and antimicrobial activity. The following compounds were characterized: hecogenin, cholest-4-en-3-one, cholesta-4,6-dien-3-ol, and hurgadacin. These compounds were identified by their mass spectrometry, ¹H, and 13C NMR spectral data, comparing those data with NIST/EPA/NIH Mass spectral database (NIST11) and published values. Hecogenin and cholest-4-en-3-one showed significant inhibitory activity against BACE1 with EC50 values of 116.3 and 390.6 µM, respectively. Cholesta-4,6-dien-3-ol and hurgadacin showed broad spectrum antimicrobial activity, particularly strongly against Escherichia coli (E. coli), Salmonella enterica (S. enterica), Pasteurella multocida (P. multocida), and Physalospora piricola (P. piricola), with minimal inhibitory concentration (MIC) ranging from 0.46 to 0.94 mg/mL. This is the first report regarding those four known compounds that were isolated from U. unicinctus and their anti-BACE1 and antimicrobial activity, highlighting the fact that known natural compounds may be a critical source of new medicine leads. These findings provide scientific evidence for potential application of those bioactive compounds for the development of AD drugs and antimicrobial agents.

Transcriptome-based identification and characterization of genes commonly responding to five different insecticides in the diamondback moth, Plutella xylostella.

When the 3rd instar larvae of the diamondback moth (DBM), Plutella xylostella, were pretreated with sublethal doses (LC10) and then subsequently exposed to lethal doses (LC50) of chlorantraniliprole, cypermethrin, dinotefuran, indoxacarb and spinosad via leaf dipping, their tolerance to insecticides was significantly enhanced. To identify genes that commonly respond to the treatment of different insecticides and are responsible for the tolerance enhancement, transcriptomic profiles of larvae treated with sublethal doses of the five insecticides were compared with that of untreated control. A total of 117,181 transcripts with a mean length of 662bp were generated by de novo assembly, of which 35,329 transcripts were annotated. Among them, 125, 143, 182, 215 and 149 transcripts were determined to be up-regulated whereas 67, 45, 60, 60 and 38 genes were down-regulated following treatments with chlorantraniliprole, cypermethrin, dinotefuran, indoxacarb and spinosad, respectively. Gene ontology (GO) analysis of differentially expressed genes (DEGs) revealed little differences in their GO profiles between treatments with different insecticides except for spinosad. Finally, the DEGs commonly responding to all insecticides were selected for further characterization, and some of their over-transcription levels were confirmed by quantitative PCR. The most notable examples of commonly responding over-transcribed genes were two cytochrome P450 genes (Cyp301a1 and Cyp9e2) and nine cuticular protein genes. In contrast, several genes composing the mitochondrial energy generation system were significantly down-regulated in all treated larvae. Considering the distinct structure and mode of action of the five insecticides tested, the differentially expressed genes identified in this study appear to be involved in general chemical defense at the initial stage of intoxication. Their possible roles in the tolerance/resistance development were discussed.

A double-knotted pulmonary artery catheter with large loop in the right internal jugular vein: A case report / 고신대학교의과대학학술지

Knotting of a pulmonary artery catheter (PAC) is a rare, but well-known complication of pulmonary artery (PA) catheterization. We report a case of a double-knotted PAC with a large loop in a patient with hepatocellular carcinoma (HCC) undergoing liver transplantation, which has been rarely reported in the literature. A PAC was advanced under pressure wave form guidance. PAC insertion was repeatedly attempted and the PAC was inserted 80 cm deep even though PAC should be normally inserted 45 to 55 cm deep. However, since no wave change was observed, we began deflating and pulling the balloon. At the 30-cm mark, the PAC could no longer be pulled. Fluoroscopy confirmed knotting of the PAC after surgery (The loop-formed PAC was shown in right internal jugular vein); thus, it was removed. For safe PA catheterization, deep insertion or repeated attempts should be avoided when the catheter cannot be easily inserted into the pulmonary artery. If possible, the insertion of PACs can be performed more safely by monitoring the movement of the catheter under fluoroscopy or transesophageal echocardiography.

Coronary artery spasm as the probable cause of cardiac arrest immediately after the induction of spinal anesthesia: A case report

A 72-year-old man underwent spinal anesthesia for artificial urinary sphincter placement for urinary incontinence. After the block level was confirmed below T6, 1 g of cefotetan, which had not shown any reaction on skin test, was administered as a prophylactic antibiotic. The patient began complaining of chest discomfort and dyspnea shortly after injection. ST elevation appeared on the electrocardiogram and the patient's pulse could not be palpated. Accordingly, cardiopulmonary resuscitation was performed for 5 minutes; the patient recovered spontaneous circulation. The patient was diagnosed as experienced coronary artery spasm by coronary angiography with spasm test. Because coronary artery spasm can also develop in patients with no history of coronary artery disease and under spinal anesthesia, careful observation, suspicion of coronary artery spasm and prompt response to hemodynamic and electrocardiogram changes are necessary.

Metabolism of an Insecticide Fenitrothion by Cunninghamella elegans ATCC36112.

In this study, the detailed metabolic pathways of fenitrothion (FNT), an organophosphorus insecticide by Cunninghamella elegans, were investigated. Approximately 81% of FNT was degraded within 5 days after treatment with concomitant accumulation of four metabolites (M1-M4). The four metabolites were separated by high-performance liquid chromatography, and their structures were identified by mass spectroscopy and/or nuclear magnetic resonance. M3 is confirmed to be an initial precursor of others and identified as fenitrothion-oxon. On the basis of their metabolic profiling, the possible metabolic pathways involved in phase I and II metabolism of FNT by C. elegans was proposed. We also found that C. elegans was able to efficiently and rapidly degrade other organophosphorus pesticides (OPs). Thus, these results will provide insight into understanding of the fungal degradation of FNT and the potential application for bioremediation of OPs. Furthermore, the ability of C. elegans to mimic mammalian metabolism would help us elucidate the metabolic fates of organic compounds occurring in mammalian liver cells and evaluate their toxicity and potential adverse effects.

Isolation and Characterization of 2,4-D Butyl Ester Degrading Acinetobacter sp. ZX02 from a Chinese Ginger Cultivated Soil.

Strain ZX02 was isolated from Chinese ginger cultivated soil contaminated with various pesticides, which could utilize 2,4-dichlorophenoxyacetic acid butyl ester (2,4-D butyl ester) as the sole carbon source. On the basis of the sequence analysis of 16S rRNA gene as well as the morphological, biochemical, and physiological characteristics of strain ZX02, the organism belonged to Gram-negative bacterium and was identified as Acinetobacter sp. ZX02. The strain ZX02 showed a remarkable performance in 2,4-D butyl ester degradation (100% removal in <96 h) in pure culture. Strain ZX02 was sensitive to tetracycline and resistant to amoxicillin and chloramphenicol in an antibiotic sensitivity test. The curing study indicates that the gene for degradation of 2,4-D butyl ester was encoded on a single plasmid of 23 kb. The gene encoding resistance to polymixin B sulfate was also located on this plasmid. On the basis of its greater biodegradation activity, this bacterium is a potential candidate as a bioremediation agent in soils contaminated with 2,4-D butyl ester.

Risk factors of emergence agitation after general anesthesia in adult patients

BACKGROUND: Emergence agitation (EA) is one of the most common complications after general anesthesia. The goal of this retrospective study was to determine the risk factors of EA in adult patients who underwent general anesthesia. METHODS: We retrospectively investigated the medical records of 5,358 adult patients who stayed in the postanesthesia care unit (PACU) of our hospital after general anesthesia during the 1-year period from January 2014 to December 2014. Psychological and behavioral status in the PACU was determined by the Aono four-point scale. Grade of 3 or 4 were considered as manifestations of EA. Multiple variables assessed EA risk factors. RESULTS: Two-hundred-forty-five patients (4.6%) developed EA. In multivariate analysis, male gender (OR = 1.626, P = 0.001), older age (OR = 1.010, P = 0.035), abdominal surgery (OR = 1.633, P = 0.002), spine surgery (OR = 1.777, P = 0.015), longer duration of anesthesia (OR = 1.002, P < 0.001), postoperative nausea and vomiting (OR = 20.164, P < 0.001) and postoperative pain (OR = 3.614, P < 0.001) were risk factors of EA. CONCLUSIONS: Male gender and older patients were risk factors of EA after general anesthesia in adult patients. Careful attention is needed for patients who receive abdominal or spine surgery, and who receive prolonged anesthesia. Adequate postoperative analgesia and antiemetic therapy should be provided to reduce the incidence of EA.