PRRSV hijacks DDX3X protein and induces ferroptosis to facilitate viral replication.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a severe disease with substantial economic consequences for the swine industry. The DEAD-box helicase 3 (DDX3X) is an RNA helicase that plays a crucial role in regulating RNA metabolism, immunological response, and even RNA virus infection. However, it is unclear whether it contributes to PRRSV infection. Recent studies have found that the expression of DDX3X considerably increases in Marc-145 cells when infected with live PRRSV strains Ch-1R and SD16; however, it was observed that inactivated viruses did not lead to any changes. By using the RK-33 inhibitor or DDX3X-specific siRNAs to reduce DDX3X expression, there was a significant decrease in the production of PRRSV progenies. In contrast, the overexpression of DDX3X in host cells substantially increased the proliferation of PRRSV. A combination of transcriptomics and metabolomics investigations revealed that in PRRSV-infected cells, DDX3X gene silencing severely affected biological processes such as ferroptosis, the FoxO signalling pathway, and glutathione metabolism. The subsequent transmission electron microscopy (TEM) imaging displayed the typical ferroptosis features in PRRSV-infected cells, such as mitochondrial shrinkage, reduction or disappearance of mitochondrial cristae, and cytoplasmic membrane rupture. Conversely, the mitochondrial morphology was unchanged in DDX3X-inhibited cells. Furthermore, silencing of the DDX3X gene changed the expression of ferroptosis-related genes and inhibited the virus proliferation, while the drug-induced ferroptosis inversely promoted PRRSV replication. In summary, these results present an updated perspective of how PRRSV infection uses DDX3X for self-replication, potentially leading to ferroptosis via various mechanisms that promote PRRSV replication.

Synergistic collaboration between AMPs and non-direct antimicrobial cationic peptides.

Non-direct antimicrobial cationic peptides (NDACPs) are components of the animal innate immune system. But their functions and association with antimicrobial peptides (AMPs) are incompletely understood. Here, we reveal a synergistic interaction between the AMP AW1 and the NDACP AW2, which are co-expressed in the frog Amolops wuyiensis. AW2 enhances the antibacterial activity of AW1 both in vitro and in vivo, while mitigating the development of bacterial resistance and eradicating biofilms. AW1 and AW2 synergistically damage bacterial membranes, facilitating cellular uptake and interaction of AW2 with the intracellular target bacterial genomic DNA. Simultaneously, they trigger the generation of ROS in bacteria, contributing to cell death upon reaching a threshold level. Moreover, we demonstrate that this synergistic antibacterial effect between AMPs and NDACPs is prevalent across diverse animal species. These findings unveil a robust and previously unknown correlation between AMPs and NDACPs as a widespread antibacterial immune defense strategy in animals.

A nuclear RNA degradation code for eukaryotic transcriptome surveillance.

The RNA exosome plays critical roles in eukaryotic RNA degradation, but it remains unclear how the exosome specifically recognizes its targets. The PAXT connection is an adaptor that recruits the exosome to polyadenylated RNAs in the nucleus, especially transcripts polyadenylated at intronic poly(A) sites. Here we show that PAXT-mediated RNA degradation is induced by the combination of a 5' splice site and a poly(A) junction, but not by either sequence alone. These sequences are bound by U1 snRNP and cleavage/polyadenylation factors, which in turn cooperatively recruit PAXT. As the 5' splice site-poly(A) junction combination is typically not found on correctly processed full-length RNAs, we propose that it functions as a "nuclear RNA degradation code" (NRDC). Importantly, disease-associated single nucleotide polymorphisms that create novel 5' splice sites in 3' untranslated regions can induce aberrant mRNA degradation via the NRDC mechanism. Together our study identified the first NRDC, revealed its recognition mechanism, and characterized its role in human diseases.

Transcriptome and small RNA analysis unveils novel insights into the C4 gene regulation in sugarcane.

MAIN CONCLUSION: This study reveals miRNA indirect regulation of C4 genes in sugarcane through transcription factors, highlighting potential key regulators like SsHAM3a. C4 photosynthesis is crucial for the high productivity and biomass of sugarcane, however, the miRNA regulation of C4 genes in sugarcane remains elusive. We have identified 384 miRNAs along the leaf gradients, including 293 known miRNAs and 91 novel miRNAs. Among these, 86 unique miRNAs exhibited differential expression patterns, and we identified 3511 potential expressed targets of these differentially expressed miRNAs (DEmiRNAs). Analyses using Pearson correlation coefficient (PCC) and Gene Ontology (GO) enrichment revealed that targets of miRNAs with positive correlations are integral to chlorophyll-related photosynthetic processes. In contrast, negatively correlated pairs are primarily associated with metabolic functions. It is worth noting that no C4 genes were predicted as targets of DEmiRNAs. Our application of weighted gene co-expression network analysis (WGCNA) led to a gene regulatory network (GRN) suggesting miRNAs might indirectly regulate C4 genes via transcription factors (TFs). The GRAS TF SsHAM3a emerged as a potential regulator of C4 genes, targeted by miR171y and miR171am, and exhibiting a negative correlation with miRNA expression along the leaf gradient. This study sheds light on the complex involvement of miRNAs in regulating C4 genes, offering a foundation for future research into enhancing sugarcane's photosynthetic efficiency.

RNA Interference Reveals the Impacts of CYP6CY7 on Imidacloprid Resistance in Aphis glycines.

Cytochrome P450 (CYP) is a group of important detoxification enzymes found in insects related to their resistance to insecticides. To elucidate the CYP6 family genes of P450, which are potentially related to imidacloprid resistance in Aphis glycines, the CYP6 cDNA sequences of A. glycines were studied. The transcriptome of A. glycines was constructed, and the CYP6 cDNA sequences of A. glycines were screened. Their relative expression levels in response to imidacloprid induction were examined through qRT-PCR, and the CYP6s with higher expression levels were used to study the detoxification of imidacloprid through RNA interference and a bioassay. Twelve CYP6s were obtained from the A. glycines transcriptome. These samples were named by the International P450 Nomenclature Committee and registered in GenBank. After 3, 6, 12, 24 and 48 h of induction with LC50 concentrations of imidacloprid, the relative expression levels of these CYP6s increased; the expression level of CYP6CY7 experienced the highest increase, being more than 3-fold higher than that of those of the non-imidacloprid-induced CYP6s. After RNA interference for CYP6CY7, the relative expression level of CYP6CY7 significantly decreased after 3, 6 and 12 h, while the corresponding P450 enzyme activity decreased after 12 and 24 h. The mortality of A. glycines due to imidacloprid treatment increased by 14.71% at 24 h. CYP6CY7 might detoxify imidacloprid in A. glycines. This study provides a theoretical basis for the further study of the mechanism of action of CYP6s and potential new methods for improving insecticidal efficacy.

Renal ischaemia-reperfusion injury is promoted by transcription factor NF-kB p65, which inhibits TRPC6 expression by activating miR-150.

AIM: To investigate the mechanism by which NF-κB p65 activates miR-150 to suppress TRPC6 expression and promote renal ischemia-reperfusion injury. METHODS: To assess the transcription of miR-150, NF-B p65, and TRPC6 in HK-2 cells treated with hypoxia reperfusion and rat kidney tissue damaged by ischemia-reperfusion (I/R), qPCR was implemented. The protein production of NF-κB p65 and TRPC6 was assessed by Western blot (WB) analysis. The histological score of rat kidney tissue was assessed using H&E (hematoxylin and eosin) staining. To assess the rate of apoptosis of renal tissue cells following I/R injury, we used the TACS TdT In Situ Apoptosis Detection Kit. To find out the impairment of renal function, blood levels of creatinine (Cr) and blood urea nitrogen (BUN) were tested in rats. Concentrations of inflammatory cytokines, including IL-1ß, IL-10, and TNF-α, were detected in HK-2 cells and rat renal tissue cells utilizing ELISA kits. FITC and CCK-8 were employed to analyze the death rate and cellular proliferation of HK-2 cells. To analyse the mechanism of engagement between NF-κB p65 and the miR-150 promoter, coupled with the detrimental impact of miR-150 on TRPC6, we adopted the dual-luciferase reporter assay. To confirm the activating effect of NF-κB p65 on miR-150,we implemented the ChIP assay. RESULTS: NF-κB p65 expression was significantly upregulated in rat renal tissue following IRI. Applying the dual-luciferase reporter assay, we demonstrated that the specific attachment of NF-B p65 with the miR-150 promoter location is viable, resulting in the promotion of the activity of the promoter. When miR-150 was overexpressed, we observed a notable reduction in cell proliferation. And it notably increased the rate of cellular apoptosis rate and amounts of the proinflammatory cytokines IL-1ß, IL-10, and TNF-α. Employing the dual-luciferase reporter assay, we demonstrated that miR-150 transfection diminished the function of luciferase in the TRPC6-WT group, whereas luciferase activity in the TRPC6-MUT group remained unchanged, indicating that miR-150 is a targeted inhibitor of TRPC6. In the rat renal I/R model, when miR-150 was inhibited or TRPC6 was overexpressed in the rat kidney I/R model, the histological score of rat kidney tissue significantly decreased, so did the quantities of proinflammatory cytokines IL-1ß, IL-10, TNF-α, creatinine (Cr) and blood urea nitrogen (BUN) contents and the rate of cell apoptosis in kidney tissue. CONCLUSION: Activation of miR-150 by NF-κB p65 results in downregulation of TRPC6 expression and promotion of IRI in the kidney.

The 'Kidney' model for optimising feedback in undergraduate clinical communication: A meta-ethnographic systematic review.

OBJECTIVES: Feedback frameworks/models focus on certain aspects of the feedback process, but a coherent and systematic model is lacking. A meta-ethnography was conducted to identify and synthesise guidance for optimising feedback interactions in undergraduate clinical communication simulations. METHODS: A systematic search of 4 electronic databases and grey literature was conducted. Following Noblit and Hare's seven phases for conducting meta-ethnography, key themes and concepts were synthesised to provide new interpretations of components in effective feedback interactions. RESULTS: 373 publications were identified and 14 included for the final synthesis, which informed the development of a new Feedback Kidney Model. The Model illustrates the interconnections of various components that allow for effective feedback interactions. The main processes include preparation, proactivity, analysis and feedback information, reception and response, and influencing factors. CONCLUSIONS: This meta-ethnography moves beyond providing an up-to-date synthesis of feedback guidance to proposing the brand-new Feedback Kidney Model, which can guide medical education and future research into how feedback is co-constructed and utilised to promote learning. PRACTICE IMPLICATIONS: Clinical communication should incorporate meta-cognitive training and using this Model will help students better utilise on-site face-to-face feedback to enhance their learning and improve future communication with patients.

Quantitative Relaxometry Assessment of Brain Microstructural Abnormality of Preschool Children With Autism Spectrum Disorder With Synthetic Magnetic Resonance Imaging.

OBJECTIVE: This study aimed to perform an assessment of brain microstructure in children with autism aged 2 to 5 years using relaxation times acquired by synthetic magnetic resonance imaging. MATERIALS AND METHODS: Thirty-four children with autism spectrum disorder (ASD) (ASD group) and 17 children with global developmental delay (GDD) (GDD group) were enrolled, and synthetic magnetic resonance imaging was performed to obtain T1 and T2 relaxation times. The differences in brain relaxation times between the 2 groups of children were compared, and the correlation between significantly changed T1/T2 and clinical neuropsychological scores in the ASD group was analyzed. RESULTS: Compared with the GDD group, shortened T1 relaxation times in the ASD group were distributed in the genu of corpus callosum (GCC) ( P = 0.003), splenium of corpus callosum ( P = 0.002), and right thalamus (TH) ( P = 0.014), whereas shortened T2 relaxation times in the ASD group were distributed in GCC ( P = 0.011), left parietal white matter ( P = 0.035), and bilateral TH (right, P = 0.014; left, P = 0.016). In the ASD group, the T2 of the left parietal white matter is positively correlated with gross motor (developmental quotient [DQ] 2) and personal-social behavior (DQ5), respectively ( r = 0.377, P = 0.028; r = 0.392, P = 0.022); the T2 of the GCC was positively correlated with DQ5 ( r = 0.404, P = 0.018); and the T2 of the left TH is positively correlated with DQ2 and DQ5, respectively ( r = 0.433, P = 0.009; r = 0.377, P = 0.028). All significantly changed relaxation values were not significantly correlated with Childhood Autism Rating Scale scores. CONCLUSIONS: The shortened relaxometry times in the brain of children with ASD may be associated with the increased myelin content and decreased water content in the brain of children with ASD in comparison with GDD, contributing the understanding of the pathophysiology of ASD. Therefore, the T1 and T2 relaxometry may be used as promising imaging markers for ASD diagnosis.

A critical review of cultural competence frameworks and models in medical and health professional education: A meta-ethnographic synthesis: BEME Guide No. 79.

BACKGROUND: Cultural competence resides at the core of undergraduate and postgraduate medical and health professional education. The evolution of studies on cultural competence has resulted in the existence of multiple theoretical frameworks and models, each emphasising certain elements of culturally appropriate care, but generally lacking in providing a coherent and systematic approach to teaching this subject. METHODS: Following a meta-ethnographic approach, a systematic search of five databases was undertaken to identify relevant articles published between 1990 and 2022. After citation searching and abstract and full article screening, a consensus was reached on 59 articles for final inclusion. Key constructs and concepts of cultural competence were synthesised and presented as themes, using the lens of critical theory. RESULTS: Three key themes were identified: competences; roles and identities; structural competency. Actionable concepts and themes were incorporated into a new transformative ACT cultural model that consists of three key domains: activate consciousness, connect relations, and transform to true cultural care. CONCLUSION: This critical review provides an up-to-date synthesis of studies that conceptualise cultural competence frameworks and models in international medical and healthcare settings. The ACT cultural model provides a set of guiding principles for culturally appropriate care, to support high-quality educational interventions.

Transcriptome dynamics provides insights into divergences of the photosynthesis pathway between Saccharum officinarum and Saccharum spontaneum.

Saccharum spontaneum and Saccharum officinarum contributed to the genetic background of modern sugarcane cultivars. Saccharum spontaneum has shown a higher net photosynthetic rate and lower soluble sugar than S. officinarum. Here, we analyzed 198 RNA-sequencing samples to investigate the molecular mechanisms for the divergences of photosynthesis and sugar accumulation between the two Saccharum species. We constructed gene co-expression networks based on differentially expressed genes (DEGs) both for leaf developmental gradients and diurnal rhythm. Our results suggested that the divergence of sugar accumulation may be attributed to the enrichment of major carbohydrate metabolism and the oxidative pentose phosphate pathway. Compared with S. officinarum, S. spontaneum DEGs showed a high enrichment of photosynthesis and contained more complex regulation of photosynthesis-related genes. Noticeably, S. spontaneum lacked gene interactions with sulfur assimilation stimulated by photorespiration. In S. spontaneum, core genes related to clock and photorespiration displayed a sensitive regulation by the diurnal rhythm and phase-shift. Small subunit of Rubisco (RBCS) displayed higher expression in the source tissues of S. spontaneum. Additionally, it was more sensitive under a diurnal rhythm, and had more complex gene networks than that in S. officinarum. This indicates that the differential regulation of RBCS Rubisco contributed to photosynthesis capacity divergence in both Saccharum species.

Running exercise improves spatial learning and memory ability and enhances angiogenesis in the cerebral cortex via endogenous nitric oxide.

BACKGROUND: The molecular mechanisms by which exercise improves brain function and capillaries in the cerebral cortex are unclear. Exercise can increase the expression of nitric oxide (NO) in the brain, and endogenous NO is thought to exert beneficial effects on proangiogenic factors, antiangiogenic factors and brain function. Therefore, we hypothesized that running exercise might improve brain function and enhance angiogenesis through endogenous NO. METHODS AND RESULTS: The following three groups of rats were administered intracerebroventricular (i.c.v.) injections before running exercise each day for 4 weeks: exercise+L-NAME group (i.c.v. L-NAME, an NO synthase blocker, dose: 1 µmol/µl and 5 µl/day; treadmill exercise, 20 min/day), exercise group (i.c.v. normal saline, 5 µl/day; treadmill exercise, 20 min/day), and sham group (i.c.v. normal saline, 5 µl/day; no treadmill exercise). Subsequently, the spatial learning and memory abilities were tested using a Morris water maze, and the nitric oxide synthase (NOS) activity in the cerebral cortex in each group of rats was measured using a method involving nitric acid reductase and metabolic chemistry. The parameters of the cortical capillaries were quantitatively investigated using an immunohistochemistry technique and stereological methods. The expression levels of proangiogenic factors (VEGF and FGF-2) and an antiangiogenic inhibitor (endostatin) in the cerebral cortex were tested using a Western blot analysis. Running exercise significantly improved the rats' spatial learning and memory abilities and increased NOS activity in the cortex. Running exercise also subsequently improved the expression of proangiogenic factors (VEGF and FGF-2) and the length, volume and surface area of capillaries and reduced the expression of antiangiogenic factors (endostatin) in the cortex. In contrast, the L-NAME treatment attenuated the effects of running exercise. CONCLUSIONS: Running exercise regulates proangiogenic factors, antiangiogenic factors and angiogenesis in the cerebral cortex via a partially NO-dependent mechanism, and influencing endogenous NO might potentially affect the exercise-related beneficial effects on cognitive ability and cortical capillaries.

An ethnographic investigation of medical students' cultural competence development in clinical placements.

As a result of an increased understanding of culture's impact on health and healthcare, cultural competence and diversity curricula have been incorporated into many medical programs. However, little is known about how students develop their cultural competence during their training. This ethnographic case study combined participant observation with interviews and focus group to understand students' views and experiences in developing their cultural competence during clinical placements. The results show that students' development of cultural competence is an individually varied process via four distinctive yet interrelated learning avenues. Immersion in a diverse healthcare environment contributes to students' development of cultural awareness and knowledge. Observation of culturally appropriate or inappropriate practices allows students to enhance their practical skills and critical reflection. Interaction with other clinical professionals, patients, and their family members, enables students' engagement within the busy clinical practice. Reflection helps students to actively think about culture's impact on health and internalize the importance of cultural competence. Students' learning via each avenue is interrelated and constantly interacting with their learning environment, which collectively contributes to their development. Integrating the results allowed the authors to generate a theoretical model that conceptualizes medical students' cultural competence development in clinical placements, which unearths students' cultural learning within the informal and hidden curriculum. This study provides a rare view of students' development of cultural competence in clinical placements, which may inform the pedagogic development of cultural competence and diversity education in medicine and healthcare.

Changing trends in ethnicity and academic performance: observational cohort data from a UK medical school.

OBJECTIVE: Quantify differential attainment by ethnicity in undergraduate medical assessments and evaluate whether institutional efforts to reduce the attainment gap have had impact. DESIGN: Observational cohort study. SETTING: A single UK MBBS medical programme. PARTICIPANTS: Pseudonymised data of adults aged ≥18 years enrolled in one of the UK MBBS medical programmes between 2012 and 2018. Ethnicity was self-declared during enrolment as White, Asian, Black, mixed and other. MAIN OUTCOME MEASURE: Module mark (distinction, merit, pass, fail) graded according to a variety of assessments, including single best answer examinations, objective structured clinical examinations and coursework submissions. All modular assessments are graded as a percentage. Logistic regression models were used to calculate relative risk ratio to study the association between ethnicity and attainment gap over a calendar and scholastic year. Models were adjusted for age, gender, social deprivation and scholastic year of study. RESULTS: 3714 student records were included. In the sample, 2134 students (57%) were non-white. The proportion of non-white students increased from 2007 (49%) to 2018 (70%). Mean age was 18 (IQR 18-21) and 56.6% were females. Higher proportion of non-white students 218 (24.8%) were from more deprived backgrounds versus white 76 (14.8%). Compared with non-white, there were no significant differences in the proportion of students failing assessments. However, white students were more likely to achieve merit (relative risk ratio 1.29 (95% CI 1.08 to 1.45)) or distinction (1.69 (95% CI 1.37 to 2.08)). Differences in attainment gap have remained unchanged over time, and for black students, attainment gap grew between their first and final year of study. CONCLUSION: A similar proportion (97%) of non-white and white students had a passing score, but attainment gap for higher grades persists over years despite widespread efforts in medical schools to diminish the attainment gap linked to ethnicity. Our findings are from a single institution, thus affecting generalisability.

Review detection of Newcastle disease virus.

Newcastle disease (ND) is an acute and highly contagious disease caused by the Newcastle disease virus (NDV) infecting poultry, which has caused great harm to the poultry industry around the world. Rapid diagnosis of NDV is important to early treatment and early institution of control measures. In this review, we comprehensively summarize the most recent research into NDV, including historical overview, molecular structure, and infection mechanism. We then focus on detection strategies for NDV, including virus isolation, serological assays (such as hemagglutination and hemagglutination-inhibition tests, enzyme linked immunosorbent assay, reporter virus neutralization test, Immunofluorescence assay, and Immune colloidal gold technique), molecular assays (such as reverse transcription polymerase chain reaction, real-time quantitative PCR, and loop-mediated isothermal amplification) and other assays. The performance of the different serological and molecular biology assays currently available was also analyzed. To conclude, we examine the limitations of currently available strategies for the detection of NDV to lay the groundwork for new detection assays.

Organic-inorganic hybrid ferrocene/AC as cathodes for wide temperature range aqueous Zn-ion supercapacitors.

Organic and inorganic materials have their own advantages and limitations, and new properties can be displayed in organic-inorganic hybrid materials by uniformly combining the two categories of materials at small scale. The objective of this study is to hybridize activated carbon (AC) with ferrocene to obtain a new material, ferrocene/AC, as the cathode for Zn-ion hybrid supercapacitors (ZHSCs). The optimized ferrocene/AC material owns fast charge transfer kinetics and can obtain pseudo-capacitance through redox reaction. Due to the introduction of ferrocene/AC, the ZHSCs exhibit remarkable electrochemical performances relative to that using ferrocene cathode, including high discharge specific capacity of 125.1 F g-1, high energy density (up to 44.8 Wh kg-1 at 0.1 A g-1) and large power density (up to 1839 W kg-1 at 5 A g-1). Meanwhile, the capacity retention rate remains 73.8% after 10 000 charge and discharge cycles. In particular, this cathode material can be used at low temperatures (up to -30 °C) with 60% capacity remained, which enlarges the application temperature range of ZHSCs. These results of this study can help understand new properties of organic-inorganic hybrid materials.

[Molecular analysis of 23 cases of B subgroup].

OBJECTIVE: To explore the molecular reasons of weak expression of B antigen on the red cell. METHODS: Serological test for blood group was carried out, including red cell and plasma grouping, and anti-A1 and anti-H testing, and confirming weak A or B antigens by adsorption and elution. Exons 1-7 were sequenced directly, and one of them was cloned and sequenced. RESULTS: All of the 23 samples showed the weak B antigen by serological method. The alleles of the subgroups were identified by DNA sequencing, including 2 Bel subgroup, 4 B3 subgroup, 14 Bw subgroup, 2 CisAB subgroup and a novel allele. The novel allele showed a nucleotide substitution 662G>A in the exon 7, and the sequence was submitted to Blood Group Antigen Gene Mutation Database, and the novel allele was named Bel10. CONCLUSION: Nucleotide substitution in exon results in blood subgroup, which showed that the antigens were weakened, and Bw phenotype was the most frequently subgroup.

Combined effectiveness of extracorporeal radial shockwave therapy and ultrasound-guided trigger point injection of lidocaine in upper trapezius myofascial pain syndrome.

OBJECTIVE: Myofascial pain syndrome (MPS) is a major musculoskeletal problem and a leading cause of disability worldwide. Extracorporeal shockwave therapy (ESWT) and trigger point injection (TPI) have shown positive results for MPS but no previous study has investigated the combined effects of radial shockwave and trigger point injection of lidocaine for upper trapezius myofascial pain syndrome. METHOD: For this purpose, forty-five participants were randomly divided into shockwave (n = 15), shockwave with ultrasound-guided trigger point injection (combined; n = 15), and control (standard care; n = 15) groups. Participants were assessed at baseline, one week and four weeks by using the visual analog scale, neck disability index, electromyography, infrared thermography, and sonoelastography. RESULTS: Compared with control group, both shockwave and combined groups showed a statistically significant reduction in pain (P<0.01), functional disability (P<0.01), skin temperature (P<0.01), and elastic stiffness, with greater reduction in the combined group (P<0.01) than shockwave group (P<0.05) at four weeks. However, no significant difference was found in electrical activity between the groups (P>0.05). The combined group also showed significant differences in pain (P<0.05) and elastic stiffness (P<0.01) compared with shockwave group at four weeks. CONCLUSION: Our study revealed that extracorporeal radial shockwave therapy combined with trigger point injection of lidocaine was more effective for decreasing pain and elastic stiffness in upper trapezius myofascial pain syndrome at four weeks.

Comparative genomic analysis of plasmids harboring bla OXA-48-like genes in Klebsiella pneumoniae.

The emergence and spread of carbapenem-resistant Klebsiella pneumoniae (CRKP) is a serious medical problem worldwide. Acquired OXA-48-like carbapenemases encoded by plasmids are important causes of carbapenem resistance in K. pneumoniae. To explore the links between plasmids and bla OXA-48-like genes in K. pneumoniae, we systematically analyzed the variants of bla OXA-48-like plasmid replicon types, phylogenetic patterns, geographic distribution, conjugative transfer regions, and the genetic environments surrounding bla OXA-48-like of 191 bla OXA-48-like-harboring plasmids, which were identified from 4451 plasmids of K. pneumoniae downloaded from GenBank. Our results showed that seven different variants of bla OXA-48-like genes were identified from the 191 bla OXA-48-like-harboring plasmids in K. pneumoniae, with bla OXA-48, bla OXA-232, and bla OXA-181 being highly prevalent. In K. pneumoniae, bla OXA-48 was mainly carried by the composite transposon Tn1999.2 located on IncL/M-type conjugative plasmids, which were mainly geographically distributed in Switzerland, Germany, and China. In K. pneumoniae, the blaOXA-232 gene was mainly carried by 6.1-kb ColKP3-type mobilizable plasmids, which were mainly isolated in India. In K. pneumoniae, bla OXA-181 was mainly carried by a group of 50-kb ColKP3-IncX3 hybrid conjugative plasmids and a group of small ColKP3-type mobilizable plasmids with lengths of 5.9-9.3 kb, the former was sporadically discovered in China, South Korea, India, and Czech Republic, while the latter was almost all isolated in India. In addition, five bla OXA-245-harboring 65.9-kb IncL plasmids of K. pneumoniae isolated in Spain were found to have the genetic context of bla OXA-245 more complicated than that of bla OXA-48-harboring IncL/M-type plasmids, with two copies of IS1R inserted both upstream and downstream of bla OXA-245-lysR. These findings enhance our understanding of the genetic diversity of bla OXA-48-like-harboring plasmids in K. pneumoniae.

Decreased carboxylesterase expression associated with increased susceptibility to insecticide in Mythimna separata.

Carboxylesterases are one of the three major types of detoxification enzyme in insects. In this study, we screened 12 full-length carboxylesterase cDNA sequences from the oriental armyworm Mythimna separata; they were named MsCarE1-MsCarE12 and registered in GenBank with accession numbers MK440541-MK440552. Treatment of fourth instar larvae of M. separata with the LD50 of the insecticide chlorantraniliprole increased the expression levels of MsCarE3 and MsCarE4, while treatment with the LD50 of lambda-cyhalothrin significantly increased the expression levels of MsCarE5 and MsCarE10. Spatiotemporal expression detection showed that MsCarE3, MsCarE4, MsCarE5, and MsCarE10 were expressed at different developmental stages and in different tissues of M. separata and their expression levels were different. Induction using a high dose of chlorantraniliprole resulted in lower expression of MsCarE3 and MsCarE4. LD50 of lambda-cyhalothrin induced higher expression of MsCarE5 and MsCarE10, while LD70 induced higher MsCarE10 expression at 3, 6, and 12 h after treatment. RNA interference successfully inhibited the expression of MsCarE3, MsCarE4, MsCarE5, and MsCarE10, to different degrees at different time points. Silencing of MsCarE5, or MsCarE5 and MsCarE10 simultaneously changed carboxylesterase activity and increased the susceptibility of M. separata larvae to lambda-cyhalothrin. This study provides a new method to increase the insect susceptibility to insecticide.

Revisiting cultural competence.

BACKGROUND: Although the importance of developing health care trainees and trainers' cultural competence is widely acknowledged, limited information is available on what elements of cultural competence should be taught. A lack of theoretical clarity and consensus over what constitutes the competence attributes is a key hurdle in faculty development (FD). This results in patchy cultural education across health schools globally. Based on a literature review, we propose a new framework that may inform clinical cultural education and the development of clinical educators in this area. METHODS: A critical interpretive synthesis was carried out on articles extracted from Medline, CINAHL and Web of Science. Sixty-nine articles were included for analysis, through which we engaged with the reconceptualisation of cultural competence in order to provide theoretical clarity and pedagogical guidance for FD in clinical cultural education. RESULTS: The synthesis illustrates that an ecological system is required to achieve comprehensive cultural competence development. We therefore composed a discursive multi-level framework to highlight the needs of achieving cultural competence at the individual, team, organisational and systemic levels. Affective, cognitive and behavioural domains are to be achieved at the individual level. Leadership skills training is a key component in achieving higher level competences. Successful FD needs to cover these developmental areas. DISCUSSION: This review offers a multi-level approach to developing cultural competence, which can be useful for clinical educators to improve their clinical practice and education in diverse cultural contexts. More research is needed to evaluate the effectiveness of the proposed framework and the methods.