Exploration of Mycobiota in Cypripedium japonicum, an Endangered Species.

Orchids live with mycorrhizal fungi in mutualism. This symbiotic relationship plays an essential role in the overall life cycle of orchids from germination, growth, settlement, and reproduction. Among the 1000 species of the orchid, the Korean lady's slipper, Cypripedium japonicum, is known as an endangered species. Currently, only five natural habitats of the Korean lady's slipper remain in South Korea, and the population of Korean lady's slipper in their natural habitat is not increasing. To prevent extinction, this study was designed to understand the fungal community interacting in the rhizosphere of the Korean lady's slipper living in the native and artificial habitats. In-depth analyses were performed to discover the vital mycorrhizal fungi contributing to habitat expansion and cultivation of the endangered orchid species. Our results suggested that Lycoperdon nigrescens contributed most to the increase in natural habitats and Russula violeipes as a characteristic of successful cultivation. And the fungi that helped L. nigrescens and R. violeipes to fit into the rhizosphere community in Korean lady's slipper native place were Paraboeremia selaginellae and Metarhizium anisopliae, respectively. The findings will contribute to restoring and maintaining the endangered orchid population in natural habitats.

Cytological Study of Cypripedium japonicum Thunb. (Orchidaceae Juss.): An Endangered Species from Korea.

Changes in chromosome number and karyotype evolution are important to plant diversification, as they are both major drivers of speciation processes. Herein, chromosome number, karyotype, and genome size of the Korean lady's slipper orchid Cypripedium japonicum Thunb., an endangered species, were investigated in natural populations. Furthermore, all cytological data from this species are reported herein for the first time. The chromosome number of all investigated C. japonicum plants was diploid (2n = 2x = 22), with x = 11 as base chromosome number, whereby the species can now be clearly distinguished from the Japanese lady's slipper orchid. The karyotypes of all studied individuals were of similar length, symmetrical, and rather unimodal. Flow cytometry of the C. japonicum revealed that the genome size ranged from 28.38 to 30.14 pg/1C. Data on chromosome number and karyotypes were largely consistent with previous results indicating that Korean (x = 11) populations of C. japonicum are more closely related to Chinese populations (x = 11) compared to Japanese (x = 10) populations. These comprehensive cytological results will benefit the efforts to discriminate the geographically isolated and endangered Eastern Asian (China, Japan, and Korea) lady's slipper orchid species.

Microstructural evolution in self-catalyzed GaAs nanowires during in-situ TEM study.

The microstructural evolutions in self-catalyzed GaAs nanowires (NWs) were investigated by using in situ heating transmission electron microscopy (TEM). The morphological changes of the self-catalyst metal gallium (Ga) droplet, the GaAs NWs, and the atomic behavior at the interface between the self-catalyst metal gallium and GaAs NWs were carefully studied by analysis of high-resolution TEM images. The microstructural change of the Ga-droplet/GaAs-NWs started at a low temperature of ∼200 °C. Formation and destruction of atomic layers were observed at the Ga/GaAs interface and slow depletion of the Ga droplet was detected in the temperature range investigated. Above 300 °C, the evolution process dramatically changed with time: The Ga droplet depleted rapidly and fast growth of zinc-blende (ZB) GaAs structures were observed in the droplet. The Ga droplet was completely removed with time and temperature. When the temperature reached ∼600 °C, the decomposition of GaAs was detected. This process began in the wurtzite (WZ) structure and propagated to the ZB structure. The morphological and atomistic behaviors in self-catalyzed GaAs NWs were demonstrated based on thermodynamic considerations, in addition to the effect of the incident electron beam in TEM. Finally, GaAs decomposition was demonstrated in terms of congruent vaporization.

First Report of Gray Mold Disease on Endangered Species Cypripedium japonicum.

Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease symptoms relating to gray mold were observed on C. japonicum in May 2019. The suspected pathogen was successfully isolated from the symptomatic leaf tissue and conducted a pure culture of the fungi. The conidia formed consisted of a colorless or light brown single cell, which was either egg or oval-shaped with a size of 7.1 to 13.4 × 5.2 to 8.6 µm. Molecular phylogenetic relationship analysis was also confirmed that the pathogen concerned belonging to the family of Botrytis cinerea. Therefore, the findings confirmed that the pathogen isolated from C. japonicum was consistent with the unique properties of B. cinerea.

Visualization of unstained homo/heterogeneous DNA nanostructures by low-voltage scanning transmission electron microscopy.

Three-dimensional (3D) homo/heterogeneous DNA nanostructures were studied with low-voltage scanning transmission electron microscopy (LV-STEM). Four types of 3D DNA nanostructures were designed and fabricated by the origami method including newly proposed protocols. The low-energy electron probe and optimized dark-field STEM detector enabled individual unstained DNA nanostructures to be clearly imaged by the single acquisition without the averaging process. For the vertically stacked double structures, assembled through modified single-stranded domains, and the structures containing a square opening (i.e., a hole) in the center, the LV-STEM successfully reveals the vertical information of these 3D structures as the contrast differences compared to the reference. For the heterogeneous structures, the LV-STEM visualized both regions of the functionalized gold nanoparticles and the DNA base structure with distinct contrasts. This study introduces a straightforward method to fabricate stackable DNA nanostructures or nanoparticles by replacing a relatively small number of incumbent DNA strands, which could realize the simple and sophisticated fabrication of higher-order 3D DNA homo/hetero nanostructures. Together with these design techniques of DNA nanostructures, this study has demonstrated that the LV-STEM is the swift and simple method for visualizing the 3D DNA nanostructures and certifying the fabricated products as the specified design, which is applicable to various research fields on soft materials including DNA nanotechnology.

Effects of preanesthetic single administration of dexmedetomidine on the remifentanil and propofol requirement during laparoscopic cholecystectomy

BACKGROUND: Dexmedetomidine, an α2-adrenergic agonist, can be used for sedation and as an adjuvant to anesthetics. This study aimed to evaluate the effects of preanesthetic administration of dexmedetomidine on the propofol and remifentanil requirement during general anesthesia and postoperative pain in patients undergoing laparoscopic cholecystectomy. METHODS: Sixty patients were randomly assigned to group D or S (n = 30 each). Dexmedetomidine (0.5 µg/kg) and a comparable volume of saline were administered in groups D and S, respectively, over a 10 minutes period before induction. General anesthesia was induced and maintained with propofol and remifentanil; the bispectral index was maintained at 40–60. The intraoperative remifentanil and propofol dosages were recorded, and postoperative pain was assessed using a visual analog scale (VAS). RESULTS: In groups S and D, propofol dosage was 8.52 ± 1.64 and 6.83 ± 1.55 mg/kg/h, respectively (P < 0.001), while remifentanil dosage was 7.18 ± 2.42 and 4.84 ± 1.44 µg/ kg/h, respectively (P < 0.001). VAS scores for postoperative pain were 6.50 (6–7) and 6.0 (6–7), respectively, at 30 minutes (P = 0.569), 5 (4–5) and 4 (3–5), respectively, at 12 hours (P = 0.039), and 2 (2–3) and 2 (1.25–2), respectively, at 24 hours (P = 0.044). The Friedman test revealed that VAS scores changed over time in both groups (P < 0.001). CONCLUSIONS: Preanesthetic single administration of a low dose of dexmedetomidine (0.5 µg/kg) can significantly decrease the remifentanil and propofol requirement during short surgeries and alleviate postoperative pain.

Cardiac arrest caused by intramyometrial injection of vasopressin during a robotic-assisted laparoscopic myomectomy.

Vasopressin is a locally-injected vasoconstrictor used to reduce bleeding during gynaecological surgery. However, even in these cases, vasopressin can induce adverse effects, including bradycardia, myocardial infarction and cardiac arrest. Elevated blood concentrations of vasopressin may induce the sympathoinhibitory reflex by increasing blood pressure and augment the sympathoinhibitory reflex by activating the area postrema. In addition, pneumoperitoneum formation needed for laparoscopy as well as physiological changes caused by steep Trendelenburg positions used during robotic surgeries may cause bradycardia. Shoulder braces used to prevent slipping from a steep Trendelenburg position may also be hazardous. This case report describes a 31-year-old female patient who underwent a scheduled robotic-assisted laparoscopic myomectomy in a steep Trendelenburg position. The patient experienced a cardiac arrest 2 min after the vasopressin injection and was treated accordingly. There were no abnormal findings on the postoperative laboratory studies, chest X-ray and electrocardiogram. The patient also had clear consciousness with no other notable symptoms. The patient was discharged on postoperative day 2. The report discusses the potential adverse effects of local vasopressin injection during robotic-assisted laparoscopic myomectomy.

Distribution of Rhizosphere and Endosphere Fungi on the First-Class Endangered Plant Cypripedium japonicum.

Endangered native plant habitats and populations are rapidly disappearing because of climate and environmental changes. As a representative, the abundance of the first-class endangered wild plant, Cypripedium japonicum, has been rapidly decreasing in Korea. The purpose of this study was to evaluate the distribution of rhizosphere and endophytic fungi on C. japonicum in its native habitat. A total of 440 rhizosphere and 79 endosphere fungi isolates were isolated and identified on the basis of their molecular characteristics. Sixty-five genera and 119 fungi species were identified in this study. The genus Trichoderma showed the highest abundance among both rhizosphere and endosphere fungi. Mortierella, Hypocrea, and Penicillium spp. were also relatively dominant species on C. japonicum. The community structures of rhizosphere and endosphere fungi were similar, but endosphere fungi showed greater diversity.

Graphene-supporting films and low-voltage STEM in SEM toward imaging nanobio materials without staining: Observation of insulin amyloid fibrils.

Utilization of graphene-supporting films and low-voltage scanning transmission electron microscopy (LV-STEM) in scanning electron microscopy (SEM) is shown to be an effective means of observing unstained nanobio materials. Insulin amyloid fibrils, which are implicated as a cause of type II diabetes, are formed in vitro and observed without staining at room temperature. An in-lens cold field-emission SEM, equipped with an additional homemade STEM detector, provides dark field (DF)-STEM images in the low energy range of 5-30keV, together with secondary electron (SE) images. Analysis based on Lenz's theory is used to interpret the experimental results. Graphene films, where the fibrils are deposited, reduce the background level of the STEM images compared with instances when conventional amorphous carbon films are used. Using 30keV, which is lower than that for conventional TEM (100-300keV), together with low detection angles (15-55mrad) enhances the signals from the fibrils. These factors improve image quality, which enables observation of thin fibrils with widths of 7-8nm. STEM imaging clearly reveals a twisted-ribbon structure of a fibril, and SE imaging shows an emphasized striped pattern of the fibril. The LV-STEM in SEM enables acquisition of two types of images of an identical fibril in a single instrument, which is useful for understanding the structure. This study expands the application of SEM to other systems of interest, which is beneficial to a large number of users. The method in this study can be applied to the observation of various nanobio materials and analysis of their native structures, thus contributing to research in materials and life sciences.

Comparison of 0.075% and 0.1% ropivacaine in terms of motor dysfunction after piriformis muscle injection

BACKGROUND: Motor weakness occurs frequently after piriformis injection and it could put patients at risk of falls. We investigated the appropriate concentration and volume of ropivacaine required to minimize motor dysfunction. METHODS: A total of 120 patients who received piriformis injection were included in this study. Piriformis injections of triamcinolone 10 mg in various concentrations (0.1%, 0.075%) and volumes (8, 10, 12 ml) of ropivacaine were administered in 20 patients, respectively. One hour after the injection, we compared motor function according to the concentrations and volumes of ropivacaine. RESULTS: There were significant differences (P < 0.05) in the occurrence of motor dysfunction according to body mass index (BMI) and the concentration of ropivacaine. No significant differences were found in terms of gender, age, weight, height, or the volume of ropivacaine. Logistic regression analysis showed that the likelihood of motor dysfunction with administration of 0.1% ropivacaine was 58.249 times greater than that with administration of 0.075% concentration (P < 0.001), while BMI did not have a significant effect on motor dysfunction. CONCLUSIONS: According to the results of this study, 0.075% ropivacaine rather than 0.1% ropivacaine is appropriate in terms of reducing motor dysfunction after piriformis injection.

The effect of intrathecal baclofen single injection on neuropathic pain

BACKGROUND: Baclofen is a gamma-aminobutyric acid B-receptor agonist, which is usually used for patients with spasticity or patients with nerve injury inducing both spasticity and neuropathic pain. Both oral administration and intrathecal injection via a continuous infusion pump are common treatment methods. The aim of this study was to evaluate the effectiveness of a series of three individual injections of intrathecal baclofen for neuropathic pain without spasticity. METHODS: Thirty-one patients with neuropathic pain were treated with a series of three monthly individual injections of intrathecal baclofen without pump implantation A dose of 50 µg of baclofen was used. 10-cm visual analog scale (VAS) scores of spontaneous pain, allodynia, and hyperalgesia were recorded a week after each injection. Vital signs were monitored to detect any hemodynamic changes, and a myelogram was performed to detect any undesirable cerebrospinal fluid leakage. All patients were hospitalized for at least one day following each injection for close observation and to control any adverse effects. RESULTS: VAS scores of spontaneous pain, allodynia, and hyperalgesia decreased significantly (P < 0.001). The major complications were general weakness, sleepiness, and urinary retention; most of these resolved within one day without any further serious symptoms. CONCLUSIONS: A series of three individual intrathecal baclofen injections was effective for those patients who suffered from neuropathic pain without spasticity or dystonia; no serious complications were observed. However, the average satisfaction score recorded for spontaneous pain was lower than those for allodynia and hyperalgesia.

Analysis of Fungicide Sensitivity and Genetic Diversity among Colletotrichum Species in Sweet Persimmon.

Anthracnose, caused by Colletotrichum gloeosporioides (C. gloeosporioides; Teleomorph: Glomerella cingulata), is the most destructive disease that affects sweet persimmon production worldwide. However, the biology, ecology, and genetic variations of C. gloeosporioides remain largely unknown. Therefore, in this study, the development of fungicide resistance and genetic diversity among an anthracnose pathogen population with different geographical origins and the exposure of this population to different cultivation strategies were investigated. A total of 150 pathogen isolates were tested in fungicide sensitivity assays. Five of the tested fungicides suppressed mycelial pathogen growth effectively. However, there were significant differences in the sensitivities exhibited by the pathogen isolates examined. Interestingly, the isolates obtained from practical management orchards versus organic cultivation orchards showed no differences in sensitivity to the same fungicide. PCR-restriction fragment length polymorphism (RFLP) analyses were performed to detect internal transcribed spacer regions and the ß-tubulin and glutamine synthetase genes of the pathogens examined. Both the glutamine synthetase and ß-tubulin genes contained a complex set of polymorphisms. Based on these results, the pathogens isolated from organic cultivation orchards were found to have more diversity than the isolates obtained from the practical management orchards.

Effect of behavioral intervention using smartphone application for preoperative anxiety in pediatric patients.

BACKGROUND: Children and parents experience significant anxiety and distress during the preoperative period. This is important because preoperative anxiety in children is associated with adverse postoperative outcome. So we suggest behaviorally oriented preoperative anxiety intervention program based on the anesthesia and psychology with smartphone application, world-widely used. METHODS: A total 120 patients (aged 1-10 years old) who were scheduled for elective surgery under general anesthesia was included in this randomized controlled trial. We randomized the patients into three groups, with using intravenous (IV) midazolam sedation (M group), with using smartphone application program (S group), and with using low dose IV midazolam plus smartphone application program (SM group). And the child anxiety was assessed using the modified Yale Preoperative Anxiety Scale (mYPAS) at holding area, 5 min after intervention, entrance to operating room. RESULTS: In all three groups, mYPAS after intervention were lower than the preoperative holding area (M group 52.8 ± 11.8 vs 41.0 ± 7.0, S group 59.2 ± 17.6 vs 36.4 ± 7.3, SM group 58.3 ± 17.5 vs 26.0 ± 3.4). A comparison of mYPAS scores between each group showed that the S group reduced anxiety lower than M group (P < 0.01), and the SM group exhibited significantly lower anxiety than the two other groups (P < 0.01). CONCLUSIONS: The preoperative preparation program using smartphone application is simple and customized by individual development that effective in the reduction of preoperative anxiety.

Effect of behavioral intervention using smartphone application for preoperative anxiety in pediatric patients / 대한마취과학회지

BACKGROUND: Children and parents experience significant anxiety and distress during the preoperative period. This is important because preoperative anxiety in children is associated with adverse postoperative outcome. So we suggest behaviorally oriented preoperative anxiety intervention program based on the anesthesia and psychology with smartphone application, world-widely used. METHODS: A total 120 patients (aged 1-10 years old) who were scheduled for elective surgery under general anesthesia was included in this randomized controlled trial. We randomized the patients into three groups, with using intravenous (IV) midazolam sedation (M group), with using smartphone application program (S group), and with using low dose IV midazolam plus smartphone application program (SM group). And the child anxiety was assessed using the modified Yale Preoperative Anxiety Scale (mYPAS) at holding area, 5 min after intervention, entrance to operating room. RESULTS: In all three groups, mYPAS after intervention were lower than the preoperative holding area (M group 52.8 +/- 11.8 vs 41.0 +/- 7.0, S group 59.2 +/- 17.6 vs 36.4 +/- 7.3, SM group 58.3 +/- 17.5 vs 26.0 +/- 3.4). A comparison of mYPAS scores between each group showed that the S group reduced anxiety lower than M group (P < 0.01), and the SM group exhibited significantly lower anxiety than the two other groups (P < 0.01). CONCLUSIONS: The preoperative preparation program using smartphone application is simple and customized by individual development that effective in the reduction of preoperative anxiety.

Optimizing dose infusion of 0.125% bupivacaine for continuous femoral nerve block after total knee replacement.

BACKGROUND: The optimal dose infusion of 0.125% bupivacaine via a femoral catheter after total knee replacement (TKR) has not been defined. This study examined various dose infusions of bupivacaine to determine the analgesic quality in patients receiving a continuous femoral nerve block (CFNB). METHODS: Patients were randomized to receive a single-injection femoral nerve block (SFNB) or CFNB performed with 20 ml of 0.125% bupivacaine, followed by a continuous infusion of 0.125% bupivacaine in four groups (n = 20 per group): 1) 0 ml/h (SFNB), 2) 2 ml/h, 3) 4 ml/h, and 4) 6 ml/h. The pain intensity at rest and on knee movement was assessed using a visual analog scale (VAS) for the first 2 postoperative days. The cumulative bolus use of IV patientcontrolled analgesia (PCA) with a morphine-ketorolac combination was evaluated. RESULTS: A lower cumulative bolus of IV PCA was noted in all CFNB groups compared to SFNB on postoperative days (PODs) 1 and 2, respectively (P < 0.05). Lower VAS scores at rest were observed in the 4 ml/h and 6 ml/h groups than in the SFNB group on PODs 1 and 2, respectively, but only on POD 2 in the 2 ml/h group (P < 0.05). Lower VAS scores on movement were noted in the 4 ml/h than the SFNB group on PODs 1 and 2, but only on POD 1 in 6 ml/h (P < 0.05). CONCLUSIONS: The minimum effective infusion rate of 0.125% bupivacaine for CFNB after TKR appears to be 4 ml/h according to the VAS pain scores.

Optimizing dose infusion of 0.125% bupivacaine for continuous femoral nerve block after total knee replacement / 대한마취과학회지

BACKGROUND: The optimal dose infusion of 0.125% bupivacaine via a femoral catheter after total knee replacement (TKR) has not been defined. This study examined various dose infusions of bupivacaine to determine the analgesic quality in patients receiving a continuous femoral nerve block (CFNB). METHODS: Patients were randomized to receive a single-injection femoral nerve block (SFNB) or CFNB performed with 20 ml of 0.125% bupivacaine, followed by a continuous infusion of 0.125% bupivacaine in four groups (n = 20 per group): 1) 0 ml/h (SFNB), 2) 2 ml/h, 3) 4 ml/h, and 4) 6 ml/h. The pain intensity at rest and on knee movement was assessed using a visual analog scale (VAS) for the first 2 postoperative days. The cumulative bolus use of IV patientcontrolled analgesia (PCA) with a morphine-ketorolac combination was evaluated. RESULTS: A lower cumulative bolus of IV PCA was noted in all CFNB groups compared to SFNB on postoperative days (PODs) 1 and 2, respectively (P < 0.05). Lower VAS scores at rest were observed in the 4 ml/h and 6 ml/h groups than in the SFNB group on PODs 1 and 2, respectively, but only on POD 2 in the 2 ml/h group (P < 0.05). Lower VAS scores on movement were noted in the 4 ml/h than the SFNB group on PODs 1 and 2, but only on POD 1 in 6 ml/h (P < 0.05). CONCLUSIONS: The minimum effective infusion rate of 0.125% bupivacaine for CFNB after TKR appears to be 4 ml/h according to the VAS pain scores.