Arabidopsis calcineurin B-like-interacting protein kinase 8 and its functional homolog in tomato negatively regulates ABA-mediated stomatal movement and drought tolerance.

Stomatal movement is critical for water transpiration, gas exchange, and responses to biotic stresses. Abscisic acid (ABA) induces stomatal closure to prevent water loss during drought. We report that Arabidopsis CIPK8 negatively regulates ABA-mediated stomatal closure and drought tolerance. CIPK8 is highly enriched in guard cells and transcriptionally induced by ABA. Functional loss of CIPK8 results in hypersensitive stomatal closure to ABA and enhanced drought tolerance. Guard cell-specific downregulation of CIPK8 mimics the phenotype of cipk8 whereas guard cell-specific expression of a constitutive active CIPK8 (CIPK8CA) has an opposite effect, suggesting a cell autonomous activity of CIPK8. CIPK8 physically interacts with CBL1 and CBL9. Functional loss of CBL1 and CBL9 mimics ABA-hypersensitive stomatal closure of cipk8 whereas abolishes the effect of CIPK8CA, indicating that CIPK8 and CBL1/CBL9 form a genetic module in ABA-responsive stomatal movement. SlCIPK7, the functional homolog of CIPK8 in tomato (Solanum lycopersicum), plays a similar role in ABA-responsive stomatal movement. Genomic editing of SlCIPK7 results in more drought-tolerant tomato, making it a good candidate for germplasm improvement.

RAE1 promotes gastric carcinogenesis and epithelial-mesenchymal transition.

AIMS: The purpose of this study was to explore the role of RAE1 in the invasion and metastasis of gastric cancer (GC) cells. MATERIALS AND METHODS: RAE1 expression in GC cells was determined by reverse-transcription polymerase chain reaction (qRT-PCR) and Western blotting (WB). Cell models featuring RAE1 gene silencing and overexpression were constructed by lentiviral transfection; The proliferation, migration, and invasion ability of cells were detected by cell counting, colony formation assay, would healing assay, and transwell invasion and migration test. WB analysis of ERK/MAPK signaling pathway (ERK1/2, p-ERK1/2, c-Myc) and EMT-related molecules (ZEB1, E-cadherin, N-cadherin, and Vimentin). RESULTS: The expression level of RAE1 in GC was notably higher than in adjacent tissues. Elevated RAE1 expression correlated with an unfavorable prognosis for GC patients. Knockdown of RAE1, as compared to the control group, resulted in a significant inhibition of proliferation, migration, and invasion abilities in GC cell lines. Furthermore, RAE1 knockdown led to a substantial decrease in the expression of N-cadherin, vimentin, ZEB1, p-ERK1/2, and c-Myc proteins, coupled with a marked increase in E-cadherin expression. The biological effects of RAE1 in GC cells were effectively reversed by the inhibition of the ERK/MAPK signaling pathway using SCH772984. Additionally, RAE1 knockdown demonstrated a suppressive effect on GC tumor size in vivo. Immunohistochemistry (IHC) results revealed significantly lower expression of Ki-67 in RAE1 knockout mice compared to the control group. CONCLUSIONS: RAE1 promotes GC cell migration and invasion through the ERK/MAPK pathway and is a potential therapeutic target for GC therapy.

Impact of chemotherapy delay on long-term prognosis of laparoscopic radical surgery for locally advanced gastric cancer: a pooled analysis of four randomized controlled trials.

BACKGROUND: Adjuvant chemotherapy following curative surgery for locally advanced gastric cancer (AGC) significantly improves long-term patient prognosis. However, delayed chemotherapy (DC), in which patients are unable to receive timely treatment, is a common phenomenon in clinical practice for various reasons. This study aimed to investigate the impact of DC on the prognosis of patients with stage II-III locally AGC and explore the associated risk factors. METHODS: Data from four prospective studies were included in the pooled analysis. The planned chemotherapy (PC) group was defined as the time interval between surgery and the first chemotherapy ≤ 49 d, while the DC group was defined as the time interval between surgery and chemotherapy > 49 d. The prognosis, recurrence, and risk factors were compared, and a nomogram for predicting DC was established. RESULTS: In total, 596 patients were included, of whom 531 (89.1%) had PC and 65 (10.9%) had DC. Survival analysis revealed that the 5-year overall survival (OS) and disease-free survival (DFS) were significantly lower in the DC group than those in the PC group (log-rank P < 0.001). Cox univariable and multivariable analyses showed that DC was an independent risk factor for OS and DFS in stage II-III patients (P < 0.05). Based on the significant factors for DC, a prediction model was established that had a good fit, high accuracy (AUC = 0.780), and clinical applicability in both the training and validation sets. CONCLUSION: Delayed chemotherapy after gastrectomy is associated with poor long-term prognosis in patients with locally advanced stage II-III GC disease. But standardized, full-cycle adjuvant chemotherapy after surgery may play a remedial role, and can to a certain extent compensate the poor effects caused by delayed chemotherapy.

Appraisal of surgical outcomes and oncological efficiency of intraoperative adverse events in robotic radical gastrectomy for gastric cancer.

BACKGROUND: Surgical quality control is a crucial determinant of evaluating the tumor efficacy. OBJECTIVE: To assess the ClassIntra grade for quality control and oncological outcomes of robotic radical surgery for gastric cancer (GC). METHODS: Data of patients undergoing robotic radical surgery for GC at a high-volume center were retrospectively analyzed. Patients were categorized into two groups, the intraoperative adverse event (iAE) group and the non-iAE group, based on the occurrence of intraoperative adverse events. The iAEs were further classified into five sublevels (ranging from I to V according to severity) based on the ClassIntra grade. Surgical performance was assessed using the Objective Structured Assessment of Technical Skill (OSATS) and the General Error Reporting Tool. RESULTS: This study included 366 patients (iAE group: n = 72 [19.7%] and non-iAE group: n = 294 [80.3%]). The proportion of ClassIntra grade II patients was the highest in the iAE group (54.2%). In total and distal gastrectomies, iAEs occurred most frequently in the suprapancreatic area (50.0% and 54.8%, respectively). In total gastrectomy, grade IV iAEs were most common during lymph node dissection in the splenic hilum area (once for bleeding [grade IV] and once for injury [grade IV]). The overall survival (OS) and disease-free survival of the non-iAE group were significantly better than those of the iAE group (Log rank P < 0.001). Uni- and multi-variate analyses showed that iAEs were key prognostic indicators, independent of tumor stage and adjuvant chemotherapy (P < 0.001). CONCLUSION: iAEs in patients who underwent robotic radical gastrectomy significantly correlated with the occurrence of postoperative complications and a poor long-term prognosis. Therefore, utilization and inclusion of ClassIntra grading as a crucial surgical quality control and prognostic indicator in the routine surgical quality evaluation system are recommended.

Associations of the Serum KL-6 with Severity and Prognosis in Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease.

BACKGROUND: As a biomarker of alveolar-capillary basement membrane injury, Krebs von den Lungen-6 (KL-6) is involved in the occurrence and development of pulmonary diseases. However, the role of the KL-6 in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) has yet to be elucidated. This prospective study was designed to clarify the associations of the serum KL-6 with the severity and prognosis in patients with AECOPD. METHODS: This study enrolled 199 eligible AECOPD patients. Demographic data and clinical characteristics were recorded. Follow-up was tracked to evaluate acute exacerbation and death. The serum KL-6 concentration was measured via an enzyme-linked immunosorbent assay. RESULTS: Serum KL-6 level at admission was higher in AECOPD patients than in control subjects. The serum KL-6 concentration gradually elevated with increasing severity of AECOPD. Pearson and Spearman analyses revealed that the serum KL-6 concentration was positively correlated with the severity score, monocyte count and concentrations of C-reactive protein, interleukin-6, uric acid, and lactate dehydrogenase in AECOPD patients during hospitalization. A statistical analysis of long-term follow-up data showed that elevated KL-6 level at admission was associated with longer hospital stays, an increased risk of future frequent acute exacerbations, and increased severity of exacerbation in COPD patients. CONCLUSION: Serum KL-6 level at admission is positively correlated with increased disease severity, prolonged hospital stay and increased risk of future acute exacerbations in COPD patients. There are positive dose-response associations of elevated serum KL-6 with severity and poor prognosis in COPD patients. The serum KL-6 concentration could be a novel diagnostic and prognostic biomarker in AECOPD patients.

Situational simulation teaching effectively improves dental students' non-operational clinical competency and objective structured clinical examination performance.

BACKGROUND: Appropriate communication with dental patients enhances treatment outcomes and patient satisfaction. Implementing simulated patient interviews courses can improve patient-centered care and reduce conflict during clerkship training. Therefore, this study explored the relationship among student participation in a situational simulation course (SSC), academic performance, clerkship performance, and objective structured clinical examination (OSCE) performance. METHODS: This study was conducted with a sample of fifth-year dental students undergoing clerkship training. After implementing a situational simulation course to investigate the relationship among participation in SSC, academic performance, clerkship performance, and OSCE performance, a path analysis model was developed and tested. RESULTS: Eighty-seven fifth-year dental students were eligible for the SSC, and most (n = 70, 80.46%) volunteered to participate. The path analysis model revealed that academic performance had a direct effect on OSCE performance (ß = 0.281, P = 0.003) and clerkship performance (ß = 0.441, P < 0.001). In addition, SSC teaching had a direct effect on OSCE performance (ß = 0.356, P < 0.001). CONCLUSIONS: SSCs can enhance dental students' non-operational clinical competency and OSCE performance effectively. Simulated patient encounters with feedback, incorporated into the dental curricula, have led to improved communication. Based on our findings, we suggest implementing SSC teaching before the OSCE to improve communication and cognitive skills.

Patient perception of service quality to preanesthetic oral examination: a cross-sectional study using the SERVQUAL model.

BACKGROUND: A phase-III interdisciplinary quality improvement program, the preanesthetic oral examination (PAOE), was implemented as a new program in an academic medical center to prevent perioperative dental injuries. This study was aimed at surveying the perceived service quality and satisfaction of patients who had undergone PAOE based on the SERVQUAL model. METHODS: This cross-sectional survey was conducted at the Kaohsiung Medical University Hospital using convenience sampling. Patients referred for PAOE (PAOE group) and those who had voluntarily availed dental services (control group) were recruited. A modified SERVQUAL questionnaire was used to assess the perceived service quality and patient satisfaction with dental services. Cronbach's alpha for SERVQUAL was 0.861. RESULTS: We enrolled 286 (68.8%) and 130 (31.2%) participants in the PAOE and control groups, respectively. The path analysis revealed that the PAOE group scored lower in dimensions of reliability (ß = -0.074, P = 0.003), responsiveness (ß = -0.148, P = 0.006), and empathy (ß = -0.140, P = 0.011). Furthermore, reliability (ß = 0.655, P < 0.001) and responsiveness (ß = 0.147, P = 0.008) showed a direct effect on patient satisfaction. Overall, participants were highly satisfied with the dental services. CONCLUSIONS: The PAOE group showed lower satisfaction and perceived quality of dental services compared to the control group. Although implementing an interdisciplinary program reduces the perceived service quality, its influence is limited. Employing an interdisciplinary teamwork is a win-win strategy encouraged to improve patient safety and reduce malpractice claims. Future suggestions should focus on establishing waiting times that are considered reasonable by patients. Patient-centered education related to the risk of perioperative dental injuries should be provided, and awareness of oral conditions for patient safety should be improved. Moreover, interprofessional education in continuous and undergraduate programs is necessary to improve professional quality.

The impact of Lacticaseibacillus paracasei GMNL-143 toothpaste on gingivitis and oral microbiota in adults: a randomized, double-blind, crossover, placebo-controlled trial.

BACKGROUND: This study examines the oral health benefits of heat-killed Lacticaseibacillus paracasei GMNL-143, particularly its potential in oral microbiota alterations and gingivitis improvement. METHODS: We assessed GMNL-143's in vitro interactions with oral pathogens and its ability to prevent pathogen adherence to gingival cells. A randomized, double-blind, crossover clinical trial was performed on gingivitis patients using GMNL-143 toothpaste or placebo for four weeks, followed by a crossover after a washout. RESULTS: GMNL-143 showed coaggregation with oral pathogens in vitro, linked to its surface layer protein. In patients, GMNL-143 toothpaste lowered the gingival index and reduced Streptococcus mutans in crevicular fluid. A positive relationship was found between Aggregatibacter actinomycetemcomitans and gingival index changes, and a negative one between Campylobacter and gingival index changes in plaque. CONCLUSION: GMNL-143 toothpaste may shift oral bacterial composition towards a healthier state, suggesting its potential in managing mild to moderate gingivitis. TRIAL REGISTRATION: ID NCT04190485 ( https://clinicaltrials.gov/ ); 09/12/2019, retrospective registration.

Arabidopsis KETCH1 Is Critical for the Nuclear Accumulation of Ribosomal Proteins and Gametogenesis.

Male and female gametophytes are generated from micro- or megaspore mother cells through consecutive meiotic and mitotic cell divisions. Defects in these divisions often result in gametophytic lethality. Gametophytic lethality was also reported when genes encoding ribosome-related proteins were mutated. Although numerous ribosomal proteins (RPs) have been identified in plants based on homology with their yeast and metazoan counterparts, how RPs are regulated, e.g., through dynamic subcellular targeting, is unknown. We report here that an Arabidopsis (Arabidopsis thaliana) importin ß, KETCH1 (karyopherin enabling the transport of the cytoplasmic HYL1), is critical for gametogenesis. Karyopherins are molecular chaperones mediating nucleocytoplasmic protein transport. However, the role of KETCH1 during gametogenesis is independent of HYPONASTIC LEAVES 1 (HYL1), a previously reported KETCH1 cargo. Instead, KETCH1 interacts with several RPs and is critical for the nuclear accumulation of RPL27a, whose mutations caused similar gametophytic defects. We further showed that knocking down KETCH1 caused reduced ribosome biogenesis and translational capacity, which may trigger the arrest of mitotic cell cycle progression and lead to gametophytic lethality.

Transcriptional Regulation of PLETHORA1 in the Root Meristem Through an Importin and Its Two Antagonistic Cargos.

Plant roots are sustained through meristem activity at the root tip. Two transcriptional pathways, one mediated by PLETHORAs (PLTs) and the other by SHORTROOT and SCARECROW, play major roles in root meristem development. The role of PLTs during root meristem development requires a concentration gradient, which is not only contributed by posttranslational regulation such as growth dilution and intercellular movement but also likely by a largely unknown fine-tuned transcriptional regulatory mechanism. We report here that Arabidopsis (Arabidopsis thaliana) JANUS positively regulates PLT1 expression in the root meristem by recruiting RNA polymerase II (Pol II) to PLT1 and by interacting with PLT1. JANUS-dependent recruitment of Pol II is inhibited through the competitive binding of JANUS by GRF-INTERACTING FACTOR1 (GIF1)/ANGUSTIFOLIA3, a transcriptional cofactor that negatively regulates PLT1 expression. Finally, GIF1 and JANUS, the antagonistic regulators of PLT1, both depend on Arabidopsis IMPORTIN ß4 for their nuclear accumulation. The combination of an importin and its two antagonistic cargos in PLT1 transcription may have logistic benefits in fine-tuning the transcription of PLT1 in root meristem.

Association of high-risk comorbidity with overall survival among patients with gastric cancer and its sex-specific differences in China: a retrospective observational cohort study.

BACKGROUND: Concomitant diseases often occur in cancer patients and are important in decision-making regarding treatments. However, information regarding the prognostic relevance of comorbidities for mortality risk is still limited among Chinese gastric cancer (GC) patients. This study aimed to investigate the association between comorbidities and 3-year mortality risk. METHODS: This retrospective study enrolled 376 GC patients undergoing radical gastrectomy at the Affiliated Zhongshan Hospital of Dalian University from January 2011 to December 2019. Demographic and clinicopathological information and treatment outcomes were collected. Patients were divided into low-, moderate- and high-risk comorbidity groups based on their Charlson Comorbidity Index (CCI) and age-adjusted CCI (ACCI) scores. Kaplan-Meier survival and Cox regression analyses were used to examine 3-year overall survival (OS) and mortality risk for each group. RESULTS: The median follow-up time was 43.5 months, and 40.2% (151/376) of GC patients had died at the last follow-up. There were significant differences in OS rates between ACCI-based comorbidity groups (76.56; 64.51; 54.55%, log-rank P = 0.011) but not between CCI-based comorbidity groups (log-rank P = 0.16). The high-risk comorbidity group based on the ACCI remained a significant prognostic factor for 3-year OS in multivariate analysis, with an increased mortality risk (hazard ratio [HR], 1.99; 95% CI, 1.15-3.44). Subgroup analysis revealed that this pattern only held for male GC patients but not for female patients. CONCLUSION: The present study suggested that high-risk comorbidities were significantly associated with a higher mortality risk, particularly in Chinese male GC patients. Moreover, the ACCI score was an independent prognostic factor of long-term mortality.

Preoperative sarcopenia combined with prognostic nutritional index predicts long-term prognosis of radical gastrectomy with advanced gastric cancer: a comprehensive analysis of two-center study.

PURPOSE: This study aims to investigate the predictive value of the combined index smni(skeletal muscle index (SMI)-prognostic nutrition index(PNI)) for the postoperative survival of patients with advanced gastric cancer(AGC). METHODS: 650 patients with AGC from two centers (290 cases from the First Affiliated Hospital of Dalian University and 360 points from the Fujian Medical University Union Hospital) were selected as the study subjects based on unified screening criteria. Clinical data, preoperative abdominal CT images, results of hematology-related examinations, tumor-related characteristics, and surgical and follow-up data of the patients were collected and organized. The L3 vertebral level muscle area was measured using computer-assisted measurement techniques, and the skeletal muscle index(SMI) was calculated based on this measurement. The prognostic nutrition index (PNI) was calculated based on serum albumin and lymphocyte count indicators. The Kaplan-Meier survival analysis of data from the First Affiliated Hospital was used to determine that SMI and PNI are significantly correlated with the postoperative survival rate of patients with advanced gastric cancer. Based on this, a novel combined index smni was fitted and stratified for risk. Cox proportional hazards regression analysis was used to determine that the index smni is an independent prognostic risk factor for patients with AGC after surgery. The ROC curve was used to describe the predictive ability of the new combined index and its importance and predictive power in predicting postoperative survival of patients with AGC, which was verified in the data of Fujian Medical University Union Hospital. RESULT: The Kaplan-Meier curve analysis of the combined indicator smni Is clearly associated with long-term survival(3-year OS (P < 0.001) and DSS (P < 0.001)), univariate analysis and multivariate analysis showed that smni was an independent prognostic risk factor, The ROC curve for the first center 3-year OS(AUC = 0.678), DSS(AUC = 0.662) show good predictive ability and were validated in the second center. CONCLUSION: The combined index smni has a good predictive ability for the postoperative survival rate of patients with AGC and is expected to provide a new reference basis and more accurate and scientific guidance for the postoperative management and treatment of patients with AGC.

Cp2Fe-Mediated Electrochemical Synthesis of Phosphorylated Oxindoles and Indolo[2,1-a]isoquinolin-6(5H)-ones.

An efficient and environmentally friendly electrochemical synthesis of phosphorylated oxindoles and indolo[2,1-a]isoquinolin-6(5H)-ones mediated by readily available Cp2Fe has been developed, which illustrated a broad substrate scope and diverse functional group compatibility. This protocol featured an external oxidant-free process and was at room temperature, which was proposed to be driven by the anodic oxidation of Cp2Fe.

Electrical performance of monolayer MoS2transistor with MoS2nanobelt metallic edges as electrodes.

The contact electrodes have great influence on the performance of monolayer MoS2devices. In this paper, monolayer MoS2and MoS2nanobelts were synthesized on SiO2/Si substrates via the chemical vapor deposition method. By using wet and dry transfer process, MoS2nanobelt metallic edges were designed as the source/drain contact electrodes of monolayer MoS2field effect transistor. The 'nanobelt metallic edges' refers to the top surface of the nanobelt being metallic. Because the base planes of MoS2nanobelt vertically stand on the substrate, which makes the layer edges form the top surface of the nanobelt. The nonlinearIds-Vdscharacteristics of the device indicates that the contact between the monolayer MoS2and MoS2metallic edges displays a Schottky-like behavior. The back-gated transfer characteristics indicate that monolayer MoS2device with MoS2nanobelt metallic edges as electrodes shows an n-type behavior with a mobility of ∼0.44 cm2V-1·s-1, a carrier concentration of ∼7.31 × 1011cm-2, and an on/off ratio of ∼103. The results enrich the electrode materials of two-dimensional material devices and exhibit potential for future application of MoS2metallic edges in electronic devices.

Manganese-Mediated Electrochemical Dearomatization of Indoles To Access 2-Azido Spirocyclic Indolines.

An efficient and environmentally friendly electrochemical protocol for dearomatization of indoles was developed, delivering a series of azido-containing spirocyclic indolines with good functional group tolerance. This dearomatization process is proposed to result from the oxidation of MnII-N3 species, supported by cyclic voltammetry experiments. Moreover, synthetic transformations can provide an alternative approach to a range of functionalized indolines.

First Report of Aureobasidium pullulans var. pullulans Causing Spot Blight of Pear (Pyrus pyrifolia) in Zhejiang Province of China.

Asian pear (Pyrus pyrifolia Nakai cv. cuiguan), is widely grown in Zhejiang province of China. In April 2019, symptoms consisting of small black, round leaf spots and blight flower petals were observed on over 30 % of 'Cuiguan' pear trees in an orchard (ca. 0.8 ha) near Cixi city, Zhejiang province, China. Initially, leaf spots were observed on leaf petioles, which, with time, enlarged and coalesced into necrotic streaks (1-2 cm) along the length of the petioles. Irregularly, reddish brown spots developed on flower petals, which hastened their senescence. Additional symptoms included round, black spots on leaves (2-3 mm in diameter) and necrosis of shoot tips. Symptomatic tissues from petals, petioles and leaves were plated onto potato dextrose agar (PDA). After five days of incubation at 26 °C, slimy fungal colonies (48×48 mm) with pinkish to orange-colored mycelia and with regular annulations were isolated from all tissues. After 10 days, cultured were shiny and dark brown in the center. The color of conidia ranged from hyaline to dark brown. Hyaline conidia were blastic, unicellular, ellipsoidal, smooth, with lengths that ranged from 11.03 to 27.14 (avg. 18.38) µm, and widths that ranged from 3.45 to 8.86 (avg. 6.04) µm (n = 50). Dark brown conidia were 1 to 2 celled, 10.89 to 26.03 (avg. 17.41) µm in length and 4.26 to 12.15 (avg. 6.94) µm in width (n = 50), and a slight septal constriction. Conidiogenous cells were clavate, hyaline, eseptate and top smoothly with 3-11 spores. Indistinct scars remained when the conidia dislodge from the conidiogenous cells. Single spore isolation was used to obtain pure cultures. Mycelia and conidia were scraped from cultures and DNA was extracted using Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech). Amplified PCR products from the internal transcribed spacer (ITS) region ITS1F/ITS4 (White et al. 1990), the partial 28S rDNA (LSU) NL1/NL4 (Boekhout et al. 1995), the ß-tubulin (TUB) gene Bt2a/Bt2b (Glass & Donaldson 1995) and the partial elongase gene (ELO) ELO2-F/ELO2-R (Zalar et al. 2008) were sequenced (Tsingke Biotechnology Co., Hangzhou, Zhejiang). A blast search (GenBank Accession No. MT107050, OK485685, OK631951, OK631950) showed 99% Aureobasidium pullulans reference isolate CBS584.75 and EXF-150, which was consistent with the morphological data (Cene et al. 2014). Three-yr-old seedlings from 'Cuiguan' pears were spray with conidial suspension (106 conidia/ml) on the both sides of leaves without wounding. In a greenhouse (26 °C, natural light), six inoculated plants and three noninoculated plants (sprayed with sterile distilled water) enclosed in plastic bags to maintain humidity for 72 h. At 5 days after inoculation, shoot tips blackened and began to wilt. At 15 days after inoculation, symptoms similar to those on the original sample developed on inoculated petioles and leaves, whereas the control plants remained healthy. No symptoms were observed on leaves that were mature at the time of inoculation. Aureobasidium pullulans var. pullulans was reisolated from all inoculated plant. Overall, this disease shortened the life of pear flowers and reduced fruit set. To our knowledge, A. pullulans var. pullulans has not previously been reported as a pathogen of P. pyrifolia.

First report of Colletotrichum scovillei causing anthracnose fruit rot on Eriobotrya japonicain in Zhejiang province, China.

Loquat (Eriobotrya japonica (Thunb.) Lindl.) is a fruit tree of high economic impotance in China. In May 2021, fruit rot on cv. "Baozhu" was observed in Yuhang district (119°40'E, 30°09'N), Zhejiang province, at an incidence of more than 30% within five orchards (the total affected area were about 121, 000 m2). Early symptoms of naturally affected fruits were dark brown, necrotic lesion (Fig. 1 A). Lesions subsequently expanded, and orange conidia were observed (Fig. 1 B). Conidia were transferred using a sterilized needle into sterile water, diluted to several conidia (approximately 10 conidia/µL) in the field of light microscope, and spread onto potato dextrose agar (PDA). After 24 h, single colonies were transferred, and six single-spore strain isolated from different fruits or locations were obtained. Six-day old colonies grown on PDA at 27°C had gray-green centers with white edges, and on the reverse side, the centers were brown (Fig. 1 C). Isolate PPGS2 was selected for further characterization. Conidia were unicellular, smooth-walled, hyaline, cylindrical with one rounded and one acute end, or with both ends rounded and they (n = 30) measured 11.1 to 16.0 µm (mean = 13.3 µm) × 3.0 to 4.6 µm (mean = 4.1) (Fig. 1 D). Appressoria were single or in smalln groups, light brown to dark brown, ovoid or elliptical with a smooth or undulate outline, and measured (n = 30) 4.6 to 8.7 µm (mean = 6.5 µm) × 4.0 to 5.6 µm (mean = 4.9 µm) (Fig. 1 H). These features of PPGS2 were consistent with species of the Colletotrichum acutatum species complex (Damm et al. 2012). For species identification, the internal transcribed spacer (ITS), beta-tubulin (TUB), chitin synthase (CHS-1), calmodulin (CAL), and actin (ACT) genes were sequenced (O'Donnell et al. 2000; Weir et al. 2012). The five sequences were deposited in GenBank (OK054581, OK077960, OK077987, OK077988 and OK077989, respectively). A BLAST search of these sequences showed 99% identity with sequences of the ex-holotype C. scovillei CBS126529 (Damm et al. 2012). The evolutionary tree shown that PPGS2 and C. scovillei CBS126529 were clustered in a branch (SH-aLRT/approximate Bayes test/bootstrap support = 88.6/0.999/96). Combining morphological characteristics with phylogenetic analysis, PPGS2 was identified as C. scovillei. To fulfill Koch's postulates, E. japonica fruit were disinfected with 75% alcohol and air-dried for 5 mins to allow the alcohol to volatilize. A conidial suspension (10 mL of 1 × 106 conidia/ml) of PPGS2 was sprayed onto six fruits, and six fruit sprayed with sterilized water served as controls. The experiment was repeated three times. Fruits were kept on a mist bench at 27°C and 80% relative humidity for 5 days. Inoculated fruit developed dark brown necrotic lesions that later fused and expanded (Fig. 1, E-G), whereas control fruit remained symptomless. C. scovillei was re-isolated from lesions of inoculated fruit. C. scovillei has been reported from fruit of Capsicum sp., Musa sp., Mangifera indica, and Clausena lansium in China (Farr and Rossman 2021). To our knowledge, this is the first report of C. scovillei causing anthracnose fruit rot of E. japonica in China. This disease results in severe economic losses both in the field and after harvest, and it is necessary to develop more effective prevention and control strategies.

Acceptance and experience of digital dental technology, burnout, job satisfaction, and turnover intention for Taiwanese dental technicians.

BACKGROUND: Digital dental technology (DDT) has progressed and been introduced to Taiwan in the recent years, gradually changing the industry ecology. Many studies have demonstrated that DDT is more accurate and faster than conventional dental technology. However, there is a paucity of research exploring dental technicians' perspectives on digital dental techniques, and their burnout, job satisfaction, and turnover intention. METHODS: This cross-sectional survey with convenience sampling was conducted at the conference venue of the Taiwan Association of Dental Technology to investigate the perspectives of dental technicians. We used the snowballing method in this study; two sampling methods were adopted, a convenience sampling of dental technicians to complete a survey, followed by asking the survey participants of the convenience sample to invite their colleagues to participate in the online survey. The survey questionnaire included questions on demographics, work-related information, acceptance and experiences of dental technicians toward DDT, occupational burnout, job satisfaction, and turnover intention. Regression models were used to determine the predictors of job satisfaction and determinants of turnover intention. RESULTS: In total, 341 valid questionnaires were obtained. Overall, the participants reported long working hours (95.5%), positive score on the DDT acceptance scale, moderate job satisfaction, higher personal burnout, and work burnout, along with lower over-commitment. Among them, 32.9% and 28.2% reported the intention to leave their organization and profession, respectively. The stepwise multiple regression model revealed that higher work burnout decreased job satisfaction, while higher DDT acceptance and position as employer increased job satisfaction. The binary logistic regression models revealed that geographical area of workplace, work burnout, and job satisfaction were significant predictors of turnover intentions. CONCLUSIONS: Many Taiwanese dental technicians reported turnover intentions and higher burnout. With the trend of digitalization in the dental industry, even though most dental technicians had a positive outlook toward DDT, its influence on job satisfaction appears limited. Retaining good and professional talents required of a dental technician is crucial, especially as Taiwan's dental care becomes increasingly specialized. Strategies for improving the work environment and occupational health of dental technicians should thus be the focus of future studies.

Effects of Different Component Contents of Colistin Methanesulfonate on the Pharmacokinetics of Prodrug and Formed Colistin in Human.

PURPOSES: To evaluate the effects of component contents in different colistin methanesulfonate (CMS) formulas on their clinical pharmacokinetics of the prodrug CMS and the formed colistin. METHODS: Two CMS formulas (CTTQ and Parkedale) were investigated in a single dose, randomized, open-label, crossover study conducted in 18 healthy Chinese subjects. Both CMS formulas met the requirements of European Pharmacopoeia 9.2 with 12.1% difference in the two major active components (CMS A and CMS B). The PK parameters after a single intravenous infusion of CMS at 2.5 mg/kg were calculated and the steady-state plasma colistin concentrations (Css,avg) following multiple dosing, once every 12 h for 7 days, were simulated with the non-compartment model. RESULTS: The systemic exposure (AUC0-inf) of CMS were 59.49 ± 5.90 h·µg/mL and 51.09 ± 4.70 h·µg/mL, and the AUC0-inf of colistin were 15.39 ± 2.63 h·µg/mL and 12.36 ± 2.10 h·µg/mL for CTTQ and Parkedale, respectively. The ratios (90% CI) of geometric mean of AUC0-inf of CTTQ to Parkedale were 116.38% (112.95%, 119.91%) and 124.49% (120.76%, 128.35%) for CMS and colistin, respectively. The predicted Css,avg (95% CI) were 0.92 (0.85, 0.99) µg/mL and 0.74 (0.69, 0.79) µg/mL for CTTQ and Parkedale, respectively. CONCLUSION: The difference in component content in the two CMS formulas had a significant (P < 0.001) impact on the systemic exposure of colistin in human, thus, warranted essential considerations in clinical applications.

First report of Colletotrichum fructicola and Colletotrichum nymphaeae causing leaf spot on Rubus corchorifolius in Zhejiang province, China.

Rubus corchorifolius is one of the most economically important fruit trees, (Tian et al. 2021). A severe leaf spot disease on leaves of R. corchorifolius was observed in Longquan county, Zhejiang province (118°42'E, 27°42'N) in 2019, with disease incidence of more than 20% on affected plants. The symptoms on leaves of the naturally affected plants were early necrotic lesion with white centers, surrounded by yellow halos (< 5 mm). Later, lesions were expanded with yellowish-brown centers, surrounded by yellow halos (< 5 mm). Putative pathogenic fungi were isolated as described by Fang (1998) and two pure single-colony fungal strains (FPZ1 and FPZ2) were selected for further analysis. The fungi was cultured on potato dextrose agar (PDA) medium for 6 days, at 25°C. The colonies had gray-green centers, white aerial mycelium and gelatinous orange conidial masses. The conidia were unicellular, smooth-walled, hyaline, cylindrical with obtuse to rounded ends, the size 10.15 to 14.09 µm (mean = 12.95 µm, n = 50) × 4.36 to 6.19 µm (mean = 5.19 µm, n = 50) were single, brown to dark brown, ovoid or irregular in shape, and 5.59 to 12.99 µm (mean = 8.77 µm, n = 50) × 4.68 to 10.36 µm (mean = 6.50 µm, n = 50). The characteristics of FPZ1 were consistent with the description of species in the Colletotrichum gloeosporioides complex (Weir et al. 2012). The conidia of FPZ2 were hyaline, smooth-walled, one-celled, fusiform, the size 9.34 to 14.09 µm (mean = 11.92 µm, n = 50) × 3.26 to 6.15 µm (mean = 4.89 µm, n = 50). Appressoria were single, darker brown, elliptical or irregular in outline, and 4.49 to 15.06 µm (mean = 9.88 µm, n = 50) × 3.23 to 7.42 µm (mean = 5.72 µm, n = 50) in size. The characteristics of FPZ2 were consistent with species of the Colletotrichum acutatum complex (Damn et al. 2012). For molecular identification of strains, the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-tubulin (TUB), chitin synthase (CHS-1), and actin (ACT) genes were sequenced (Weir et al. 2012). For the strain FPZ1, the five sequences obtain were deposited in GenBank (MT846907, MT849313, MT849317, MT849315 and MT849319, respectively). A BLAST search of FPZ1 sequences showed 99% identity with the five loci sequences of type strain C. fructicola ICMP 18581 (JX010165, JX010033, JX010405, JX009866 and FJ907426) (Jayawardena et al. 2016). Similarly, for the strain FPZ2, the five sequences (MT846885, MT849314, MT849318, MT849316 and MT849320, respectively) had 99% identity with the five loci sequences of type strain C. nymphaeae CBS 515.78 (JQ948197, JQ948527, JQ949848, JQ948858 and JQ949518, respectively) (Jayawardena et al. 2016). Based on morphological characteristics and phylogenetic analysis, FPZ1 was identified as C. fructicola and FPZ2 as C. nymphaeae, respestively. For pathogenicity tests, 10 µL conidial suspension (1 × 106 conidia per ml) of FPZ1 was used to inoculate five healthy, non-wounded detached leaves, while five leaves inoculated with sterilized water served as control. The experiment was repeated three times, and all leaves were kept on a mist bench at 27°C and relative humidity 80% for 6 days. The inoculation sites of both FPZ1 and FPZ2 became brown and necrotic, while control leaves developed no symptoms. C. fructicola and C. nymphaeae were re-isolated from the lesions of inoculated leaves, fulfilling Koch's postulates. To our knowledge, this is the first report of C. fructicola and C. nymphaeae causing leaf spot on Rubus corchorifolius in China, and reports on the prevalence of C. gloeosporioides and C. acutatum species complexes will be beneficial to management of anthracnose in R. corchorifolius.