Children who appeared or remained overweight or obese predict a higher follow-up blood pressure and higher risk of hypertension: a 6-year longitudinal study in Yantai, China.

Few longitudinal studies have examined the effect of weight status change on blood pressure change over time and incidence of hypertension among Chinese children. The longitudinal study enrolled 17,702 Chinese children aged 7 years in Yantai in 2014 as baseline, with a continuous 5 years of follow-up to 2019. Generalized estimating equation model was fitted to examine the main and interaction effects of weight status change and time with blood pressure and the incidence of hypertension. Compared with the participants who remained normal weight, the participants who remained overweight or obese had higher systolic blood pressure (SBP) (ß = 2.89, p < 0.001) and diastolic blood pressure (DBP) (ß = 1.79, p < 0.001). Significant interactions were identified between weight status change and time with SBP (χ2interaction = 697.77, p < 0.001) and DBP (χ2interaction = 270.49, p < 0.001). The odds ratio (OR) and 95% confidence interval (CI) of hypertension were 1.70 (1.59-1.82) for participants who appeared overweight or obese, 2.26 (2.14-2.40) for participants who remained overweight or obese, compared with the participants who remained normal weight. Those who switched from overweight or obesity to normal weight had almost the same risk of developing hypertension (OR = 1.13, 95% CI: 1.02 to 1.26) as children who remained normal weight. Children who appeared or remained overweight or obese predict a higher follow-up blood pressure and higher risk of hypertension, whereas losing weight could reduce blood pressure and the risk of hypertension. Children who appeared or remained overweight or obese predict a higher follow-up blood pressure and higher risk of hypertension, whereas losing weight could reduce blood pressure and the risk of hypertension.

Fungal Pathogens Associated with Strawberry Crown Rot Disease in China.

Strawberry crown rot (SCR) is a serious disease that is generally referred to as seedling anthracnose due to its association with Colletotrichum spp. Presently, SCR is the main cause of death of strawberry seedlings. However, management strategies, including fungicides targeting Colletotrichum spp., have failed to obtain satisfactory results. Therefore, identifying the exact pathogen species causing SCR could guide its management. A total of 287 isolates were obtained from SCR-diseased plants. Based on the culture, morphology, and phylogenetic characteristics, the above 287 fungal isolates of SCR pathogens were identified as 12 different species, including Colletotrichum siamense, C. fructicola, Fusarium oxysporum, F. commune, F. equiseti, F. solani, F. tricinctum, Epicoccum sorghinum, Stemphylium lycopersici, Clonostachys rosea, Phoma herbarum, and Curvularia trifolii. Pathogenicity results showed that most isolates were pathogenic to strawberry seedlings and exhibited different degrees of virulence. In severe cases, poor growth on the ground, yellowing of the leaves, and even death of seedlings occurred. In mild cases, only black disease spots appeared on the stems of the strawberry seedlings, and a few withered leaves became necrotic. The inoculation experiments showed that the most virulent species were C. siamense and F. oxysporum, followed by F. equiseti, P. herbarum, Cl. rosea, S. lycopersici, and C. fructicola, which had disease incidences above 50%. E. sorghinum, S. lycopersici, Cl. rosea, P. herbarum and Cu. trifolii were reported to cause SCR for the first time herein. In conclusion, SCR is a sophisticated disease caused by a diversity of pathogenic fungi. This work provides new valuable data about the diversity and pathogenicity of SCR pathogens, which will help in formulating effective strategies to better control of the SCR disease.

Lateral Circumflex Femoral Vascular Chimeric Fascia Flap Reduces Pain and Promotes Wound Healing in Repairing Skin and Tendon Defects of Hand, Foot, and Ankle.

The purpose of this study was to investigate the clinical efficacy of lateral circumflex femoral artery embedded with fascia lata flap in the repair of skin and tendon defects in hand, foot, and ankle. From January 2020 to June 2021, 32 patients with skin and tendon defects of the hand, foot, and ankle admitted to our hospital were selected as the study subjects. According to the random number table method, they were divided into the observation group (16 cases, treated with rotational lateral femoral vascular inlay broad fascial flap repair) and the control group (16 cases, treated with conventional skin flap repair) and followed up for 6 months. The postoperative tendon midactivity measurement scale (TAM), ankle-hindfoot scoring system (AOFAS), and lower limb functional evaluation scale (LEFS) scores were all higher in the observer group than in the control group The number of people with infection, implant necrosis, and subcutaneous hematoma in the observation group (total incidence 6.25%) was less (lower) than that in the control group (50.00%), and the total number of people with significant and fair clinical efficacy in the observation group (total effective rate 100.00%) was more (higher) than that in the control group (68.75%). The application of early plastic surgery to the clinical treatment of patients with deep burns on the hand can reduce the patient's pain and promote the healing of the wound. It is of great significance to reduce the risk of complications such as necrosis of the skin graft and improve the efficacy of the surgery.

Bioactivity of mefentrifluconazole against different Fusarium spp.

Emergence and development of resistance to 14α-demethylase inhibitors (DMIs) have become a critical issue in both agriculture and medical fields. Mefentrifluconazole, the first isopropanol triazole fungicide belonging to a new subclass of DMIs, has been proposed to show high activity, minimal adverse side effects, and inconsistent cross resistance with other DMIs due to its high structural flexibility. In this study, mefentrifluconazole showed disparate inhibitory activity against the mycelium growth of seven tested Fusarium species. The most sensitive species included F. oxysporum, F. proliferatum, F. commuae, and F. fujikuroi, followed by F. equiseti and F. graminearum, while F. solani was most insensitive. Consistently, mefentrifluconazole presented the strongest inhibiting effects on conidium germination, cell membrane integrity, and ergosterol biosynthesis in F. fujikuroi, followed by F. graminearum, while F. solani ranked last. Further results indicated that all F. fujikuroi isolates causing rice bakanae disease (RBD) were sensitive to mefentrifluconazole regardless of their resistance to prochloraz, tebuconazole, carbendazim, and phenamacril. Additionally, the inoculation tests found that mefentrifluconazole presented a better protective efficacy on rice seedlings when applied 12 h before the F. fujikuroi inoculation, compared to applied 12 h post the inoculation. Overall, this study demonstrated the various bioactivity of mefentrifluconazole combating Fusarium spp. and put new insights into RBD management as well as the applications of DMIs.

Colletotrichum Spp. Diversity Between Leaf Anthracnose and Crown Rot From the Same Strawberry Plant.

Leaf anthracnose (LA) and anthracnose crown rot (ACR) represent serious fungal diseases that pose significant threats to strawberry production. To characterize the pathogen diversity associated with above diseases, 100 strawberry plants, including varieties of "Hongjia," "Zhangji," and "Tianxianzui," were sampled from Jiande and Zhoushan, the primary plantation regions of Zhejiang province, China. A total of 309 Colletotrichum isolates were isolated from crown (150 isolates) and leaves (159 isolates) of affected samples. Among these, 100 isolates obtained from the plants showing both LA and CR symptoms were selected randomly for further characterization. Based on the morphological observations combined with phylogenetic analysis of multiple genes (ACT, ITS, CAL, GAPDH, and CHS), all the 100 tested isolates were identified as C. gloeosporioides species complex, including 91 isolates of C. siamense, 8 isolates of C. fructicola causing both LA and ACR, and one isolate of C. aenigma causing ACR. The phenotypic characteristics of these isolated species were investigated using the BIOLOG phenotype MicroArray (PM) and a total of 950 different metabolic phenotype were tested, showing the characteristics among these isolates and providing the theoretical basis for pathogenic biochemistry and metabolism. The pathogenicity tests showed that even the same Colletotrichum species isolated from different diseased tissues (leaves or crowns) had significantly different pathogenicity toward strawberry leaves and crown. C. siamense isolated from diseased leaves (CSLA) was more aggressive than C. siamense isolated from rotted crown (CSCR) during the infection on "Zhangji" leaves. Additionally, C. fructicola isolated from affected leaf (CFLA) caused more severe symptoms on the leaves of four strawberry varieties compared to C. fructicola isolated from diseased crown (CFCR). For crown rot, the pathogenicity of CSCR was higher than that of CSLA.

Activity of a SDHI fungicide penflufen and the characterization of natural-resistance in Fusarium fujikuroi.

The occurrence of bakanae disease of rice caused by the fungus Fusarium fujikuroi in Zhejiang Province has become increasingly aggravated in recent years, concomitant with the development of resistance to the widely applied fungicides, prochloraz and phenamacril. In this study, the activity of a novel succinate dehydrogenase inhibitor (SDHI) fungicide, penflufen, against different fungi was evaluated in addition to the potential of penflufen in controlling F. fujikuroi infections. Penflufen exhibited good bioactivity against F. fujikuroi, but weak activity against Fusarium spp. and other investigated plant-pathogenic fungi including Colletotrichum spp. In addition to inhibiting mycelial growth, penflufen effectively inhibited F. fujikuroi conidium production. For germination, penflufen could effectively inhibit that of small conidia, but only delay the germination of large conidia. In addition, the sensitivity to penflufen among 100 F. fujikuroi isolates that were collected in areas that were never exposed to SDHIs was determined based on mycelium growth. Sensitivities surprisingly exhibited bimodal distributions, indicating the presence of natural resistance. Cross-resistance was not observed between penflufen in F. fujikuroi and two fungicides that have been extensively applied in field including prochloraz (a DMI) and phenamacril (a 2-cyanoacrylate fungicide), nor with the three SDHIs, fluopyram, benzovindiflupyr, and pydiflumetofen. Additional analysis identified five different point mutations in SDH-A (i.e., at residues 46, 225, 283, 430, and 586) of naturally resistant isolates. These results inform the potential application of the new SDHI fungicide penflufen for managing crop diseases and understanding possible resistance mechanisms among pathogens.

Cobalt-Catalyzed C-H Activation/Annulation of Benzamides with Fluorine-Containing Alkynes: A Route to 3- and 4-Fluoroalkylated Isoquinolinones.

The C-H activation/annulation reaction of various benzamides with fluoroalkylated alkynes in the presence of a Co(acac)2·2H2O catalyst proceeded very smoothly to give the corresponding 3- and 4-fluoroalkylated isoquinolinones in excellent yields with approximately 70% regioselectivities. These regioisomers could be successfully separated and obtained in pure form. Major or minor regioisomers were determined as 4- or 3-fluoroalkylated isoquinolinones, respectively, based on X-ray crystallographic analyses.

Regioselective cobalt(II)-catalyzed [2 + 3] cycloaddition reaction of fluoroalkylated alkynes with 2-formylphenylboronic acids: easy access to 2-fluoroalkylated indenols.

[2 + 3] cycloaddition reactions of fluorinated alkynes with 2-formylphenylboronic acids under the influence of Co(acac)2·2H2O in two-component solvents of acetonitrile/2-propanol at reflux temperature for 18 h took place smoothly, affording the corresponding fluoroalkylated indenol derivatives in good yields. This reaction shows excellent regioselectivity, giving 2-fluoroalkylated indenols, together with a very small amount of 3-fluoroalkylated indanones as side products.

Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection.

Tomato spotted wilt virus (TSWV) is one of the most devastating plant viruses and often causes severe crop losses worldwide. Generally, mature plants become more resistant to pathogens, known as adult plant resistance. In this study, we demonstrated a new phenomenon involving developmentally regulated susceptibility of Arabidopsis thaliana to TSWV. We found that Arabidopsis plants become more susceptible to TSWV as plants mature. Most young 3-week-old Arabidopsis were not infected by TSWV. Infection of TSWV in 4-, 5-, and 6-week-old Arabidopsis increased from 9%, 21%, and 25%, respectively, to 100% in 7- to 8-week-old Arabidopsis plants. Different isolates of TSWV and different tospoviruses show a low rate of infection in young Arabidopsis but a high rate in mature plants. When Arabidopsis dcl2/3/4 or rdr1/2/6 mutant plants were inoculated with TSWV, similar results as observed for the wild-type Arabidopsis plants were obtained. A cell-to-cell movement assay showed that the intercellular movement efficiency of TSWV NSm:GFP fusion was significantly higher in 8-week-old Arabidopsis leaves compared with 4-week-old Arabidopsis leaves. Moreover, the expression levels of pectin methylesterase and ß-1,3-glucanase, which play critical roles in macromolecule cell-to-cell trafficking, were significantly up-regulated in 8-week-old Arabidopsis leaves compared with 4-week-old Arabidopsis leaves during TSWV infection. To date, this mature plant susceptibility to pathogen infections has rarely been investigated. Thus, the findings presented here should advance our knowledge on the developmentally regulated mature host susceptibility to plant virus infection.

Rescue of tomato spotted wilt virus entirely from complementary DNA clones.

Negative-stranded/ambisense RNA viruses (NSVs) include not only dangerous pathogens of medical importance but also serious plant pathogens of agronomic importance. Tomato spotted wilt virus (TSWV) is one of the most important plant NSVs, infecting more than 1,000 plant species, and poses major threats to global food security. The segmented negative-stranded/ambisense RNA genomes of TSWV, however, have been a major obstacle to molecular genetic manipulation. In this study, we report the complete recovery of infectious TSWV entirely from complementary DNA (cDNA) clones. First, a replication- and transcription-competent minigenome replication system was established based on 35S-driven constructs of the S(-)-genomic (g) or S(+)-antigenomic (ag) RNA template, flanked by the 5' hammerhead and 3' ribozyme sequence of hepatitis delta virus, a nucleocapsid (N) protein gene and codon-optimized viral RNA-dependent RNA polymerase (RdRp) gene. Next, a movement-competent minigenome replication system was developed based on M(-)-gRNA, which was able to complement cell-to-cell and systemic movement of reconstituted ribonucleoprotein complexes (RNPs) of S RNA replicon. Finally, infectious TSWV and derivatives carrying eGFP reporters were rescued in planta via simultaneous expression of full-length cDNA constructs coding for S(+)-agRNA, M(-)-gRNA, and L(+)-agRNA in which the glycoprotein gene sequence of M(-)-gRNA was optimized. Viral rescue occurred with the addition of various RNAi suppressors including P19, HcPro, and γb, but TSWV NSs interfered with the rescue of genomic RNA. This reverse genetics system for TSWV now allows detailed molecular genetic analysis of all aspects of viral infection cycle and pathogenicity.

Alternative Oxidase Is Involved in the Pathogenicity, Development, and Oxygen Stress Response of Botrytis cinerea.

Alternative oxidase (AOX) is a ubiquinol terminal oxidase that is involved in fungal mitochondrial oxidative phosphorylation. In this study, we analyzed the roles of AOX in Botrytis cinerea by generating BcAOX deletion mutants. The mutants exhibited defects in mycelial growth, sporulation, spore germination, and virulence. Furthermore, the sensitivity of the mutants to quinone outside inhibitor fungicides and oxidative stress were increased. All phenotypic variations could be restored in the complemented strain. In summary, these results showed that BcAOX is involved in the regulation for vegetative development, adaptation to environmental stress, and virulence of B. cinerea.

A multilayered regulatory mechanism for the autoinhibition and activation of a plant CC-NB-LRR resistance protein with an extra N-terminal domain.

The tomato resistance protein Sw-5b differs from the classical coiled-coil nucleotide-binding leucine-rich repeat (CC-NB-LRR) resistance proteins by having an extra N-terminal domain (NTD). To understand how NTD, CC and NB-LRR regulate autoinhibition and activation of Sw-5b, we dissected the function(s) of each domain. When viral elicitor was absent, Sw-5b LRR suppressed the central NB-ARC to maintain autoinhibition of the NB-LRR segment. The CC and NTD domains independently and additively enhanced the autoinhibition of NB-LRR. When viral elicitor was present, the NB-LRR segment of Sw-5b was specifically activated to trigger a hypersensitive response. Surprisingly, Sw-5b CC suppressed the activation of NB-LRR, whereas the extra NTD of Sw-5b became a positive regulator and fully activated the resistance protein, probably by relieving the inhibitory effects of the CC. In infection assays of transgenic plants, the NB-LRR segment alone was insufficient to confer resistance against Tomato spotted wilt tospovirus; the layers of NTD and CC regulation on NB-LRR were required for Sw-5b to confer resistance. Based on these findings, we propose that, to counter the negative regulation of the CC on NB-LRR, Sw-5b evolved an extra NTD to coordinate with the CC, thus developing a multilayered regulatory mechanism to control autoinhibition and activation.

Structure and function analysis of nucleocapsid protein of tomato spotted wilt virus interacting with RNA using homology modeling.

The nucleocapsid (N) protein of tomato spotted wilt virus (TSWV) plays key roles in assembling genomic RNA into ribonucleoprotein (RNP), which serves as a template for both viral gene transcription and genome replication. However, little is known about the molecular mechanism of how TSWV N interacts with genomic RNA. In this study, we demonstrated that TSWV N protein forms a range of higher ordered oligomers. Analysis of the RNA binding behavior of N protein revealed that no specific oligomer binds to RNA preferentially, instead each type of N oligomer is able to bind RNA. To better characterize the structure and function of N protein interacting with RNA, we constructed homology models of TSWV N and N-RNA complexes. Based on these homology models, we demonstrated that the positively charged and polar amino acids in its predicted surface cleft of TSWV N are critical for RNA binding. Moreover, by N-RNA homology modeling, we found that the RNA component is deeply embedded in the predicted protein cleft; consistently, TSWV N-RNA complexes are relatively resistant to digestion by RNase. Collectively, using homology modeling, we determined the RNA binding sites on N and found a new protective feature for N protein. Our findings also provide novel insights into the molecular details of the interaction of TSWV N with RNA components.

Viroplasm protein P9-1 of Rice black-streaked dwarf virus preferentially binds to single-stranded RNA in its octamer form, and the central interior structure formed by this octamer constitutes the major RNA binding site.

The P9-1 protein of Rice black-streaked dwarf virus (RBSDV) is an essential part of the viroplasm. However, little is known about its nature or biological function in the viroplasm. In this study, the structure and function of P9-1 were analyzed for in vitro binding to nucleic acids. We found that the P9-1 protein preferentially bound to single-stranded versus double-stranded nucleic acids; however, the protein displayed no preference for RBSDV versus non-RBSDV single-stranded ssRNA (ssRNA). A gel mobility shift assay revealed that the RNA gradually shifted as increasing amounts of P9-1 were added, suggesting that multiple subunits of P9-1 bind to ssRNA. By using discontinuous blue native gel and chromatography analysis, we found that the P9-1 protein was capable of forming dimers, tetramers, and octamers. Strikingly, we demonstrated that P9-1 preferentially bound to ssRNA in the octamer, rather than the dimer, form. Deletion of the C-terminal arm resulted in P9-1 no longer forming octamers; consequently, the deletion mutant protein bound to ssRNA with significantly lower affinity and with fewer copies bound per ssRNA. Alanine substitution analysis revealed that electropositive amino acids among residues 25 to 44 are important for RNA binding and map to the central interior structure that was formed only by P9-1 octamers. Collectively, our findings provide novel insights into the structure and function of RBSDV viroplasm protein P9-1 binding to RNA.

One-step reverse transcription loop-mediated isothermal amplification for the rapid detection of cucumber green mottle mosaic virus.

Cucumber green mottle mosaic virus (CGMMV) has caused serious damage to Cucurbitaceae crops worldwide. The virus is considered one of the most serious Cucurbitaceae quarantine causes in many countries. In this study, a highly efficient and practical one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for the detection of CGMMV. The total RNA or crude RNA extracted from watermelon plants or seeds could be detected easily by this RT-LAMP assay. The RT-LAMP assay was conducted in isothermal (63°C) conditions within 1h. The amplified products of CGMMV could be detected as ladder-like bands using agarose gel electrophoresis or visualized in-tube under UV light with the addition of a fluorescent dye. The RT-LAMP amplification was specific to CGMMV, as no cross-reaction was observed with other viruses. The RT-LAMP assay was 100-fold more sensitive than that of reverse-transcription polymerase chain reaction (RT-PCR). This is the first report of the application of the RT-LAMP assay to detect CGMMV. The sensitive, specific and rapid RT-LAMP assay developed in this study can be applied widely in laboratories, the field and quarantine surveillance of CGMMV.

The 2a protein of Cucumber mosaic virus induces a hypersensitive response in cowpea independently of its replicase activity.

Resistance in cowpea to infection with strains of Cucumber mosaic virus (CMV) involves a local hypersensitive response (HR), and previous studies indicated that the 2a replicase of CMV is involved in HR induction. In this study, we confirmed and extended this observation by demonstrating that the nonviral expression of the 2a protein encoded by CMV is able to induce a cell death response in cowpea plants, whereas no other CMV-encoded proteins elicits such response. The 2a single-amino acid mutant, F631Y, no longer induces the necrosis response, yet the A641S mutant still induces cell death. The 2a double mutant, F631Y and A641S, does not induce HR. However, the three 2a mutants have comparable replicase activities in a fluorescence reporter assay. The 2a(D610A) mutant that alters the highly conserved GDD motif abolishes the replicase activity, however it does not affect HR induction in cowpea. The 2a(301-778aa) fragment introduced with the same D610A mutation in the GDD motif is also capable of inducing HR in cowpea. Collectively, these findings suggest that the 2a protein of CMV is sufficient to induce HR in cowpea independently of its replicase activity.