Anhydrous volatile fatty acid extraction through omniphobic membranes by hydrophobic deep eutectic solvents: Mechanistic understanding and future perspective.

Volatile fatty acids (VFAs) derived from arrested anaerobic digestion (AD) can be recovered as a valuable commodity for value-added synthesis. However, separating VFAs from digestate with complex constituents and a high-water content is an energy-prohibitive process. This study developed an innovative technology to overcome this barrier by integrating deep eutectic solvents (DESs) with an omniphobic membrane into a membrane contactor for efficient extraction of anhydrous VFAs with low energy consumption. A kinetic model was developed to elucidate the mechanistic differences between this novel omniphobic membrane-enabled DES extraction and the previous hydrophobic membrane-enabled NaOH extraction. Experimental results and mechanistic modeling suggested that VFA extraction by the DES is a reversible adsorption process facilitating subsequent VFA separation via anhydrous distillation. High vapor pressure of shorter-chain VFAs and low Nernst distribution coefficients of longer-chain VFAs contributed to DES-driven extraction, which could enable continuous and in-situ recovery and conversion of VFAs from AD streams.

RNAi-mediated silencing of the neverland gene inhibits molting in the migratory locust, Locusta migratoria.

7-dehydrocholesterol (7-DHC) is a key intermediate product used for biosynthesis of molting hormone. This is achieved through a series of hydroxylation reactions catalyzed by the Halloween family of cytochrome P450s. Neverland is an enzyme catalyzes the first reaction of the ecdysteroidogenic pathway, which converts dietary cholesterol into 7-DHC. However, research on the physiological function of neverland in orthopteran insects is lacking. In this study, neverland from Locusta migratoria (LmNvd) was cloned and analyzed. LmNvd was mainly expressed in the prothoracic gland and highly expressed on days 6 and 7 of fifth instar nymphs. RNAi-mediated silencing of LmNvd resulted in serious molting delays and abnormal phenotypes, which could be rescued by 7-DHC and 20-hydroxyecdysone supplementation. Hematoxylin and eosin staining results showed that RNAi-mediated silencing of LmNvd disturbed the molting process by both promoting the synthesis of new cuticle and suppressing the degradation of the old cuticle. Quantitative real-time PCR results suggested that the mRNA expression of E75 early gene and chitinase 5 gene decreased and that of chitin synthase 1 gene was markedly upregulated after knockdown of LmNvd. Our results suggest that LmNvd participates in the biosynthesis process of molting hormone, which is involved in regulating chitin synthesis and degradation in molting cycles.

Defects in the cell wall and its deposition caused by loss-of-function of three RLKs alter root hydrotropism in Arabidopsis thaliana.

Root tips can sense moisture gradients and grow into environments with higher water potential. This process is called root hydrotropism. Here, we report three closely related receptor-like kinases (RLKs) that play critical roles in root hydrotropism: ALTERED ROOT HYDROTROPIC RESPONSE 1 (ARH1), FEI1, and FEI2. Overexpression of these RLKs strongly reduce root hydrotropism, but corresponding loss-of-function mutants exhibit an increased hydrotropic response in their roots. All these RLKs show polar localization at the plasma membrane regions in root tips. The biosynthesis of the cell wall, cutin, and wax (CCW) is significantly impaired in root tips of arh1-2 fei1-C fei2-C. A series of known CCW mutants also exhibit increased root hydrotropism and reduced osmotic tolerance, similar to the characteristics of the triple mutant. Our results demonstrat that the integrity of the cell wall, cutin, and root cap wax mediate a trade-off between root hydrotropism and osmotic tolerance.

Enhanced nutrient recovery from anaerobically digested poultry wastewater through struvite precipitation by organic acid pre-treatment and seeding in a bubble column electrolytic reactor.

Limited mineralization of organic phosphorus to phosphate during the anaerobic digestion process poses a significant challenge in the development of cost-effective nutrient recovery strategies from anaerobically digested poultry wastewater (ADPW). This study investigated the influence of organic acids on phosphorus solubilization from ADPW, followed by its recycling in the form of struvite using a bubble column electrolytic reactor (BCER) without adding chemicals. The impact of seeding on the efficiency of PO43- and NH3-N recovery as well as the size distribution of recovered precipitates from the acid pre-treated ADPW was also evaluated. Pre-treatment of the ADPW with oxalic acid achieved complete solubilization of phosphorus, reaching ∼100% extraction efficiency at pH 2.5. The maximum removal efficiency of phosphate and ammonia-nitrogen from the ADPW were 88.9% and 90.1%, respectively, while the addition of 5 and 10 g/L struvite seed to the BCER increased PO43- removal efficiency by 9.6% and 11.5%, respectively. The value of the kinetic rate constant, k, increased from 0.0176 min-1 (unseeded) to 0.0198 min-1, 0.0307 min-1, and 0.0375 min-1 with the seed loading rate of 2, 5, and 10 g/L, respectively. Concurrently, the average particle size rose from 75.3 µm (unseeded) to 82.1 µm, 125.7 µm, and 148.9 µm, respectively. Results from XRD, FTIR, EDS, and dissolved chemical analysis revealed that the solid product obtained from the recovery process was a multi-nutrient fertilizer consisting of 94.7% struvite with negligible levels of heavy metals.

Construction and validation of a nomogram to predict left ventricular hypertrophy in low-risk patients with hypertension.

Electrocardiography (ECG) is an accessible diagnostic tool for screening patients with hypertensive left ventricular hypertrophy (LVH). However, its diagnostic sensitivity is low, with a high probability of false-negatives. Thus, this study aimed to establish a clinically useful nomogram to supplement the assessment of LVH in patients with hypertension and without ECG-LVH based on Cornell product criteria (low-risk hypertensive population). A cross-sectional dataset was used for model construction and divided into development (n = 2906) and verification (n = 1447) datasets. A multivariable logistic regression risk model and nomogram were developed after screening for risk factors. Of the 4353 low-risk hypertensive patients, 673 (15.4%) had LVH diagnosed by echocardiography (Echo-LVH). Eleven risk factors were identified: hypertension awareness, duration of hypertension, age, sex, high waist-hip ratio, education level, tea consumption, hypochloremia, and other ECG-LVH diagnostic criteria (including Sokolow-Lyon, Sokolow-Lyon products, and Peguero-Lo Presti). For the development and validation datasets, the areas under the curve were 0.724 (sensitivity = 0.606) and 0.700 (sensitivity = 0.663), respectively. After including blood pressure, the areas under the curve were 0.735 (sensitivity = 0.734) and 0.716 (sensitivity = 0.718), respectively. This novel nomogram had a good predictive ability and may be used to assess the Echo-LVH risk in patients with hypertension and without ECG-LVH based on Cornell product criteria.

Transcription factor CncC regulates the expression of antennal CYP6MU1 gene responsible for trans-2-hexen-1-al and nonanal recognition in Locusta migratoria.

Cytochrome P450 monooxygenases (P450s) are a superfamily of multifunctional heme-containing proteins and could function as odorant-degrading enzymes (ODEs) in insect olfactory systems. In our previous study, we identified a P450 gene from the antennal transcriptome of Locusta migratoria, LmCYP6MU1, which could be induced by a variety of volatiles. However, the regulatory mechanisms of this gene in response to volatiles remain unknown. In current study, we investigated the tissues and development stages expression patterns of LmCYP6MU1 and determined its olfactory function in the recognition of the main host plant volatiles which induced LmCYP6MU1 expression. The results showed that LmCYP6MU1 was antenna-rich and highly expressed throughout the antennal developmental stages of locusts. LmCYP6MU1 played important roles in the recognition of trans-2-hexen-1-al and nonanal. Insect CncC regulates the expression of P450 genes. We tested whether LmCncC regulates LmCYP6MU1 expression. It was found that LmCncC knockdown in the antennae resulted in the downregulation of LmCYP6MU1 and repressed the volatiles-mediated induction of LmCYP6MU1. LmCncC knockdown reduced the electroantennogram (EAG) and behavioral responses of locusts to volatiles. These results suggested that LmCncC could regulate the basal and volatiles-mediated inducible expression of LmCYP6MU1 responsible for the recognition of trans-2-hexen-1-al and nonanal. These findings provide an original basis for understanding the regulation mechanisms of LmCncC on LmCYP6MU1 expression and help us better understand the LmCncC-mediated olfactory plasticity.

Transcription factors, cap 'n' collar isoform C regulates the expression of CYP450 genes involving in insecticides susceptibility in Locusta migratoria.

BACKGROUND: The cap 'n' collar (Cnc) belongs to the Basic Leucine Zipper (bZIP) transcription factor super family. Cap 'n' collar isoform C (CncC) is highly conserved in the animal kingdom. CncC contributes to the regulation of growth, development, and aging and takes part in the maintenance of homeostasis and the defense against endogenous and environmental stress. Insect CncC participates in the regulation of various kinds of stress-responsive genes and is involved in the development of insecticide resistance. RESULTS: In this study, one full-length CncC sequence of Locusta migratoria was identified and characterized. Upon RNAi silencing of LmCncC, insecticide bioassays showed that LmCncC played an essential role in deltamethrin and imidacloprid susceptibility. To fully investigate the downstream genes regulated by LmCncC and further identify the LmCncC-regulated genes involved in deltamethrin and imidacloprid susceptibility, a comparative transcriptome was constructed. Thirty-five up-regulated genes and 73 down-regulated genes were screened from dsLmCncC-knockdown individuals. We selected 22 LmCncC-regulated genes and verified their gene expression levels using RT-qPCR. Finally, six LmCYP450 genes belonging to the CYP6 family were selected as candidate detoxification genes, and LmCYP6FD1 and LmCYP6FE1 were further validated as detoxification genes of insecticides via RNAi, insecticide bioassays, and metabolite identification. CONCLUSIONS: Our data suggest that the locust CncC gene is associated with deltamethrin and imidacloprid susceptibility via the regulation of LmCYP6FD1 and LmCYP6FE1, respectively.

Influence of hypertension and global or abdominal obesity on left ventricular hypertrophy: A cross-sectional study.

Although hypertension and obesity are both risk factors for left ventricular hypertrophy (LVH), the extent of their impact on LVH in the general population is still unclear, and the predictive value of obesity indicators for LVH remains to be elucidated. In this study, obesity-related indicators, including waist circumference (WC), waist-height ratio (WHTR), and waist-hip ratio (WHR), were used to define abdominal obesity (AO), whereas body mass index (BMI) was used to measure general obesity (GO). The effects of hypertension and obesity on LVH were estimated using logistic regression analysis, as was the relative risk of LVH based on the presence of obesity, hypertension, or both. Subgroup analyses were performed based on sex and age. Of the 9134 participants (≥35 years old), 915 (10.0%) developed LVH. After adjusting for covariates, the odds ratios (95% confidence intervals) for LVH were 3.94 (3.27-4.75) in patients with hypertension, 1.90 (1.60-2.26) in those with GO, and 1.45 (1.25-1.69), 1.69 (1.43-2.00), and 1.54 (1.33-4.75) in individuals with AO defined based on WC, WHTR, and WHR, respectively. Analysis by sex showed similar values in women, but AO based on WC and WHR were not significantly associated with LVH in men. Further, after adjusting for potential confounding factors, concomitant hypertension and obesity had an increased risk of developing LVH in all age ranges, particularly in patients aged 35-45 years (risk increased 14.14-fold, 10.84-fold, 7.97-fold, and 9.95-fold for BMI-based GO and WC-, WHTR-, and WHR-based AO, respectively), and in both men and women but particularly in men (risk increased 7.71-fold, 4.67-fold, 5.83-fold, and 5.58-fold, respectively). In summary, all obesity indicators (BMI, WC, WHTR, and WHR) had predictive value for LVH in women; however, only BMI and WHTR should be considered for men. Furthermore, monitoring for the occurrence and progression of LVH is imperative for rural Chinese patients with concomitant hypertension and obesity, especially men and those aged 35-45 years.

Associations of sleep duration, daytime napping, and snoring with depression in rural China: a cross-sectional study.

BACKGROUND: Most adult patients with depression complain about sleep symptoms, including insufficient and excessive sleep. However, previous studies investigating the impact of sleep duration on depression have yielded conflicting results. Therefore, this study aimed to analyse the link between depression and sleep duration, daytime napping, and snoring among rural Chinese adults. METHODS: A cross-sectional study was conducted with 9104 individuals. Interviews were conducted with the participants regarding their sleep patterns and their daytime napping routines. The individuals were then assessed for depression using the Patient Health Questionnaire-9. The risk of depression was assessed using a multifactor binary logistic regression analysis. A generalized additive model was used to evaluate the nonlinear relationship between depression and sleep duration/nap time. Additionally, subgroup analysis was conducted to investigate the correlation between sleep duration, daytime napping, snoring, and depression. RESULTS: Less than 6 h or more than 8 h of nighttime sleep, daytime napping for more than 1 h, and snoring were all significantly associated with an increased risk of depression. A U-shaped relationship was found between the duration of nighttime sleep and depression. In addition, we found that the nighttime duration of sleep, daytime naps, and snoring had a significant combined effect on the risk of depression. The subgroup analysis further revealed that lack of sleep at night significantly increased the risk of depression in all subgroups. However, snoring and excessive nighttime sleep and napping were only associated with the risk of depression in some subgroups. CONCLUSIONS: Lack of nighttime sleep (short sleep duration), excessive sleep, and napping for more than one hour during the day were associated with a high risk of depression and had a combined effect with snoring.

Coupling physical selection with biological selection for the startup of a pilot-scale, continuous flow, aerobic granular sludge reactor without treatment interruption.

This study removes two technical constraints for transitioning full-scale activated sludge infrastructure to continuous flow, aerobic granular sludge (AGS) facilities. The first of these is the loss of treatment capacity as a result of the rapid washout of flocculent sludge inventory and in turn the potential loss of nitrification during initial AGS reactor startup. The second is the physical selector design which currently is limited to either the complex sequencing batch reactor selection or sidestream hydrocyclones. Briefly, real wastewater data collected from this study suggested that by increasing the surface overflow rate (SOR) of an upflow clarifier to 10 m h - 1, the clarifier can be taken advantage of as a physical selector to separate flocculant sludge from AGS. Redirecting the physical selector underflow and overflow sludge to the feast and famine zones of a treatment train, respectively, can create a biological selection that not only promotes AGS formation but also safeguards the effluent quality throughout the AGS reactor startup period. This study provides a novel concept for economically implementing continuous flow AGS within existing full-scale, continuous flow treatment trains.

Comparison of conventional and unconventional obesity indices associated with new-onset hypertension in different sex and age populations.

We aimed to compare the relationship between hypertension and obesity-related anthropometric indices (waist circumference [WC], waist-height ratio, waist-hip ratio [WHR], and body mass index; unconventional: new body shape index [ABSI] and body roundness index [BRI]) to identify best predictors of new-onset hypertension. The study included 4123 adult participants (2377 women). Hazard ratios (HRs) and 95% confidence intervals (CIs) were determined using a Cox regression model to estimate the risk of new-onset hypertension with respect to each obesity index. In addition, we assessed the predictive value of each obesity index for new-onset hypertension using area under the receiver operating characteristic curve (AUC) after adjusting for common risk factors. During the median follow-up of 2.59 years, 818 (19.8%) new hypertension cases were diagnosed. The non-traditional obesity indices BRI and ABSI had predictive value for new-onset hypertension; however, they were not better than the traditional indexes. WHR was the best predictor of new-onset hypertension in women aged ≤ 60 and > 60 years, with HRs of 2.38 and 2.51 and AUCs of 0.793 and 0.716. However, WHR (HR 2.28, AUC = 0.759) and WC (HR 3.24, AUC = 0.788) were the best indexes for predicting new-onset hypertension in men aged ≤ 60 and > 60 years, respectively.

CYP6FD5, an antenna-specific P450 gene, is potentially involved in the host plant recognition in Locusta migratoria.

Cytochrome P450 monooxygenases (P450s) are a large superfamily of heme-thiolate proteins and play a vital role in the biosynthesis and inactivation of endogenous substances as well as the detoxification of exogenous substances. They also function as odor-degrading enzymes (ODEs) in insect olfactory sensory systems. In the present study, a P450 gene was obtained from the antennae of Locusta migratoria and named as CYP6FD5. Multiple alignment of P450 proteins revealed that LmCYP6FD5 contained five conserved motifs, including the helix C motif, an oxygen-binding site, helix K motif, a meander region, and the haem-binding motif. The expression of LmCYP6FD5 in various tissues and antennal development stages was determined by using RT-qPCR. Our results showed that LmCYP6FD5 was antenna-specific and highly expressed throughout the antennal developmental stages of female and male locusts. Furthermore, the role of LmCYP6FD5 in the perception of host plant volatiles was assessed using RNAi in combination with electroantennogram (EAG) and behavioral responses. Our findings showed that after silencing LmCYP6FD5, the EAG responses of female and male locusts to the main volatiles of gramineous plants, including trans-2-Hexen-1-al, cis-3-Hexenyl acetate, and decanal, were significantly diminished. Moreover, a significant decrease in EAG response of male antennae to benzaldehyde was also observed. In addition, behavioral assay showed that the locust response to single volatile from host plant or wheat remained unchanged after the silencing of LmCYP6FD5. Antenna-specific expression and EAG responses of locusts to host plant volatiles still suggested that LmCYP6FD5 was potentially involved in host plant recognition, although no behavioral changes were observed.

LmCht5-1 and LmCht5-2 Promote the Degradation of Serosal and Pro-Nymphal Cuticles during Locust Embryonic Development.

The success of the degradation of the extraembryonic serosal cuticle and the second embryonic cuticle (pro-nymphal cuticle) is essential for the development and molting of nymph from egg in Orthoptera Locusta migratoria. Chitinase 5 is an important gene for chitin degradation in nymphs and in the egg stage. In this study, we investigated the important roles of chitinase 5-1 (LmCht5-1) and chitinase 5-2 (LmCht5-2) in the degradation of the serosal and pro-nymphal cuticles during locust embryonic development. The serosal cuticle degrades from 7-day-old embryos (E7) to E13, along with the degradation of the pro-nymphal cuticle, which begins at E12 to E14. The mRNA and protein of LmCht5-1 and LmCht5-2 are expressed during the degradation process of the serosal cuticle and the pro-nymphal cuticle. RNAi experiments at the embryonic stage show that both dsLmCht5-1 and dsLmCht5-2 contribute to the failure of development in early and late embryogenesis. Further, during the serosal cuticle molting process, ultra-structure analysis indicated that dsLmCht5-1 prevented the loss of the coarse chitin layer in the upper part in both early and late embryogenesis. Meanwhile, dsLmCht5-2 blocked the degradation of the lower fine chitin layer at the early stage and blocked the chitin degradation of loose coarse chitin in the late molting process. During the degradation of the pro-nymphal cuticle, dsLmCht5-1 suppresses chitin degradation between layers in the procuticle, while dsLmCht5-2 suppresses chitin degradation into filaments inside of the layer. In summary, our results suggest that both LmCht5-1 and LmCht5-2 contribute to the degradation of the serosal and pro-nymphal cuticles during the locust embryonic stage.

Combined superposition effect of hypertension and dyslipidemia on left ventricular hypertrophy.

BACKGROUND: Hypertension and dyslipidemia are considered reversible risk factors for cardiovascular disease. The purpose of this study was to explore the impact of traditional and nontraditional blood lipid profiles on the risk of left ventricular hypertrophy (LVH) and to explore the superposition effect of dyslipidemia combined with hypertension. METHODS: Data on 9134 participants (53.5 ± 10.3 years old) from the Northeast China Rural Cardiovascular Health Study (NCRCHS) were statistically analyzed. The blood lipid profile was measured by total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), total glyceride (TG), and calculated nontraditional blood lipid indices including non-HDL-C, atherosclerosis index (AI), TC/HDL-C, and residual cholesterol (RC). RESULTS: After the adjustment of age and gender, the odds ratios (ORs) of LVH in patients with hypertension, high LDL-C, high non-HDL-C, high AI, and high TC/HDL-C were 3.97 (3.31-4.76), 1.27 (1.02-1.59), 1.21 (1.04-1.39), 1.33 (1.15-1.53), and 1.42 (1.22-1.65), respectively. After full adjustment of potential confounding factors, high AI and TC/HDL-C were associated with LVH rather than traditional blood lipid indices. The combination of hypertension and nontraditional dyslipidemia (defined by high AI and TC/HDL-C) was associated with the highest risk of LVH, especially in participants under 45 years of age. The risk was more significant in men, 5.09-fold and 6.24-fold, respectively, compared with 3.66-fold and 4.01-fold in women. CONCLUSIONS: People with dyslipidemia defined by nontraditional blood lipid indices (high AI and high TC/HDL-C) and hypertension were more likely to develop LVH.

Nomogram Including Serum Ion Concentrations to Screen for New-Onset Hypertension in Rural Chinese Populations Over a Short-Term Follow-up.

BACKGROUND: This study aimed to establish a clinically useful nomogram to evaluate the probability of hypertension onset in the Chinese population.Methods and Results: A prospective cohort study was conducted in 2012-2013 and followed up in 2015 to identify new-onset hypertension in 4,123 participants. The dataset was divided into development (n=2,748) and verification (n=1,375) cohorts. After screening risk factors by lasso regression, a multivariate Cox regression risk model and nomogram were established. Among the 4,123 participants, 818 (19.8%) developed hypertension. The model identified 10 risk factors: age, waist-to-hip ratio, systolic blood pressure, diastolic blood pressure, high pulse rate, history of diabetes, family history of hypertension and stroke, intake frequency of bean products, and intensity of physical labor. The C-indices of the model in the development and validation cohorts were 0.744 and 0.768, respectively. After the inclusion of serum calcium and magnesium concentrations, the C-indices in the development and validation cohorts were 0.764 and 0.791, respectively, with areas under the curve for the updated model of 0.907 and 0.917, respectively. The calibration curve showed that the nomogram accurately predicted the probability of hypertension. The updated nomogram was clinically beneficial across thresholds of 10-60%. CONCLUSIONS: The newly developed nomogram has good predictive ability and may effectively assess hypertension risk in high-risk rural areas in China.

Evaluations of two glutathione S-transferase epsilon genes for their contributions to metabolism of three selected insecticides in Locusta migratoria.

The insect-specific epsilon class of glutathione S-transferases (GSTEs) plays important roles in insecticide detoxification in insects. In our previous work, five GSTEs were identified in Locusta migratoria, and two recombinant GSTEs, rLmGSTE1 and rLmGSTE4, showed high catalytic activity when 1-chloro-2,4-dinitrobenzene (CDNB) was used as a substrate. In this work, we further investigated whether these two GSTEs could metabolize three insecticides including malathion, deltamethrin and DDT. Using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC/MS) method, we found that rLmGSTE4, but not rLmGSTE1, can metabolize malathion and DDT. Malathion bioassays of L.migratoria after the expression of LmGSTE4 was suppressed by RNA interference (RNAi) showed increased insect mortality from 33.8% to 68.9%. However, no changes in mortality were observed in deltamethrin- or DDT-treated L.migratoria after the expression of LmGSTE4 was suppressed by RNAi. Our results provided direct evidences that LmGSTE4 participates in malathion detoxification in L.migratoria. These findings are important for understanding the mechanisms of insecticide resistance in L.migratoria and developing new strategies for managing the insect populations in the field.

Chromodomain Helicase DNA Binding Protein 1-like, a negative regulator of Forkhead box O3a, promotes the proliferation and migration of Angiotensin II-induced vascular smooth muscle cells.

Essential hypertension (EH) represents a major risk factor for stroke, myocardial infarction, and heart failure. Dysregulated proliferation and migration of vascular smooth muscle cells (VSMCs) play an important role in pathogenesis of EH. This study aims to investigate the effect of Chromodomain Helicase DNA Binding Protein 1-Like (CHD1L) on Angiotensin II (AngII)-induced VSMCs injury and reveal the underlying mechanism. The expression of CHD1L in EH patients was determined by bioinformatics analysis, and then it was silenced in AngII-induced VSMCs to detect the changes in cellular functions including proliferation, migration, invasion and phenotypic switching via CCK-8, EDU staining, wound healing, transwell and Western blot assays, respectively. Inflammation and oxidative stress were also measured by detecting related markers via commercial kits. After confirming the binding sites between forkhead box O3A (FOXO3a) and CHD1L and their negative association by bioinformatics analysis, FOXO3a was further silenced, and the cellular functions were assessed again to reveal the underlying mechanism. Results showed that CHD1L was highly expressed in EH, and interference of CHD1L suppressed the proliferation, migration, invasion and phenotypic switching in VSMCs. Inflammation and oxidative stress were also restrained by CHD1L knockdown. After validating the negative role of FOXO3a in regulating CHD1L, it was found that FOXO3a abrogated the effect of CHD1L knockdown on the cellular functions of AngII-induced VSMCs. In conclusion, FOXO3a suppresses the proliferation and migration of AngII-induced VSMCs by down-regulating CHD1L.

Silencing of transcription factor E93 inhibits adult morphogenesis and disrupts cuticle, wing and ovary development in Locusta migratoria.

Ecdysone-induced protein 93F (E93) plays important roles during the metamorphosis process in insects. In this study, a cDNA of the LmE93 gene was identified from the transcriptome of Locusta migratoria, which consists of the 3378-nucleotide open-reading frame (ORF) and encodes 1125 amino acids with helix-turn-helix (HTH) motifs. Reverse transcription quantitative polymerase chain reaction analysis revealed that LmE93 was highest expressed in ovary. The LmE93 expression level was markedly low from the 3rd to 4th instar nymphs, and greatly increased in 1-day-old 5th instar nymphs with a peak on middle nymphal days, then declined in the late nymphal days. Moreover, injected dsLmE93 into 4th and 5th instar nymphs greatly reduced LmE93 transcripts, respectively, and prevented the process of metamorphosis, causing supernumerary nymphal stages. Hematoxylin-eosin staining of the integument showed that the apolysis occurred in advance in 4th instar nymphs, and old cuticle degradation was decreased in dsLmE93-injected locusts of 5th instar nymphs. Smaller and no fully developed wings with reduced columns between the anterior and posterior regions were found in N6 and N7 supernumerary nymphs. In addition, the development of the ovary in dsLmE93-injected locusts was severely blocked, the yolk was almost not formed and there was no development of ovarioles. The results indicated that LmE93 play key roles in the metamorphosis, cuticle, wing and ovarian development of locusts.

Characteristics of Halloween genes and RNA interference-mediated functional analysis of LmCYP307a2 in Locusta migratoria.

Halloween genes are involved in the biosynthesis of the molting hormone, which plays a key role in insect ecdysis, development, metamorphosis, and reproduction. Our previous work identified five Halloween genes from Locusta migratoria, but their functions are currently unknown. In this study, the sequences of these five Halloween genes were analyzed and characterized. LmCYP307a2, LmCYP306a1, LmCYP302a1, and LmCYP315a1 were primarily expressed in the prothoracic glands, while LmCYP314a1 was universally expressed in peripheral tissues, especially in the ovaries and Malpighian tubules. All five Halloween genes were mainly expressed from the 5th to the 7th d in 5th-instar nymphs. RNA interference (RNAi) silencing of LmCYP307a2 resulted in severe molting delays and molting failure, which could be rescued by supplementary 20-hydroxyecdysone. A hematoxylin and eosin staining analysis suggested that the RNAi of LmCYP307a2 inhibited the ecdysis process by inhibiting the apolysis and degradation of the old cuticle, and by promoting the synthesis of a new cuticle. Quantitative reverse transcription polymerase chain reaction results showed that the expressions of LmE74, LmCht5, and LmCht10 were dramatically down-regulated, while that of LmChsI was substantially up-regulated, after knockdown of LmCYP307a2. The results suggest that LmCYP307a2 is related to the molt process via regulation of chitin synthesis and degradation.

Four alternative splicing transcripts of intracellular copper/zinc superoxide dismutase 1 in Oxya chinensis.

In this study, we obtained four alternative splicing transcripts of intracellular copper/zinc superoxide dismutase 1 (icCuZnSOD1) in Oxya chinensis. OcicCuZnSOD1a has all common characteristics of CuZnSOD family and is a canonical CuZnSOD. OcicCuZnSOD1b is missing a Zn binding site. OcicCuZnSOD1c lacks Zn ion and is a Cu-only SOD. OcicCuZnSOD1d is missing a CuZnSOD conserved sequence and lacks the E-loop, a conserved disulfide bond, and an active site arginine. OcicCuZnSOD1a was the most heat-resistant and OcicCuZnSOD1c was the most unstable at high temperatures above 55 °C. They were stable at a wide pH range, especially in alkaline conditions. The four variants expressed at the throughout developmental stages and had various tissue expression patterns. OcicCuZnSOD1a and OcicCuZnSOD1d were significantly induced by 8.79 mM CuCl2 and OcicCuZnSOD1b was significantly up-regulated by 14.67 mM CuCl2. OcicCuZnSOD1a was significantly inhibited by 19.13 mM ZnSO4 while OcicCuZnSOD1d were significantly induced by 22.61 mM ZnSO4. Disc diffusion assay showed that the four isoforms of OcicCuZnSOD1 made the killing zones smaller surrounding the CdCl2-soaked filter discs. However, the reduction ratios of OcicCuZnSOD1a were the highest. These results implied that the four transcripts played roles in defense against CdCl2-induced oxidative stress while OcicCuZnSOD1a had stronger antioxidant capacity.