Isolation and characterization of Stenotrophomonas pavanii GXUN74707 with efficient flocculation performance and application in wastewater treatment.

The identification of microorganisms with excellent flocculants-producing capability and optimization of the fermentation process are necessary for the wide-scale application of bioflocculants. Therefore, we isolated and identified a highly efficient flocculation performance strain of Stenotrophomonas pavanii GXUN74707 from the sludge. The optimal fermentation and flocculation conditions of strain S. pavanii GXUN74707 was in fermentation medium with glucose and urea as the carbon and nitrogen sources, respectively, at pH 7.0 for 36 h, which treatment of kaolin suspension with 0.5 mL of the fermentation broth resulted in a flocculation rate of 99.0%. The bioflocculant synthesized by strain S. pavanii GXUN74707 was found mainly in the supernatant of the fermentation broth. Chemical analysis revealed that the pure bioflocculant consisted of 79.70% carbohydrates and 14.38% proteins. The monosaccharide components of MBF-GXUN74707 are mainly mannose (5.96 µg/mg), galactose (1.86 µg/mg), and glucose (1.73 µg/mg). Infrared spectrometric analysis showed the presence of carboxyl (COO-), hydroxyl (-OH) groups. The SEM images showed clumps of rod-shaped bacteria with adhesion of extracellular products. Furthermore, the strain decolored dye wastewater containing direct black, direct blue, and Congo red by 89.2%, 95.1%, 94.1%, respectively. The chemical oxygen demand (COD) and biological oxygen demand (BOD) removal rates after treatment of aquaculture wastewater with the fermentation broth were 68% and 23%, respectively. This study is the first to report the performance and application of strain Stenotrophomonas pavanii in wastewater flocculation. The results indicate that strain S. pavanii is a good candidate for the production novel bioflocculants and demonstrates its potential industrial practicality in biotechnology processes.

Colon-targeted oral nanoliposomes loaded with psoralen alleviate DSS-induced ulcerative colitis.

Oral administration, while convenient, but complex often faces challenges due to the complexity of the digestive environment. In this study, we developed a nanoliposome (NLP) encapsulating psoralen (P) and coated it with chitosan (CH) and pectin (PT) to formulate PT/CH-P-NLPs. PT/CH-P-NLPs exhibit good biocompatibility, superior to liposomes loaded with psoralen and free psoralen alone. After oral administration, PT/CH-P-NLPs remain stable in the stomach and small intestine, followed by a burst release of psoralen after reaching the slightly alkaline and gut microbiota-rich colon segment. In the DSS-induced ulcerative colitis of mice, PT/CH-P-NLPs showed significant effects on reducing inflammation, mitigating oxidative stress, protecting the integrity of the colon mucosal barrier, and modulating the gut microbiota. In conclusion, the designed nanoliposomes demonstrated the effective application of psoralen in treating ulcerative colitis.

pH- and Matrix Metalloproteinase-Responsive Multifunctional Bilayer Microneedles Platform for Treatment of Tinea Pedis.

Persistent foot odor and itchiness are common symptoms of tinea pedis, significantly disrupting the daily life of those affected. The cuticular barrier at the site of the tinea pedis is thickened, which impedes the effective penetration of antifungal agents. Additionally, fungi can migrate from the skin surface to deeper tissues, posing challenges in the current clinical treatment for tinea pedis. To effectively treat tinea pedis, we developed a platform of bilayer gelatin methacrylate (GelMA) microneedles (MNs) loaded with salicylic acid (SA) and FK13-a1 (SA/FK13-a1@GelMA MNs). SA/FK13-a1@GelMA MNs exhibit pH- and matrix metalloproteinase (MMP)-responsive properties for efficient drug delivery. The MNs are designed to deliver salicylic acid (SA) deep into the stratum corneum, softening the cuticle and creating microchannels. This process enables the antibacterial peptide FK13-a1 to penetrate through the stratum corneum barrier, facilitating intradermal diffusion and exerting antifungal and anti-inflammatory effects. In severe cases of tinea pedis, heightened local pH levels and MMP activity further accelerate drug release. Our research demonstrates that SA/FK13-a1@GelMA MNs are highly effective against Trichophyton mentagrophytes, Trichophyton rubrum, and Candida albicans. They also reduced stratum corneum thickness, fungal burden, and inflammation in a guinea pig model of tinea pedis induced by T. mentagrophytes. Furthermore, it was discovered that SA/FK13-a1@GelMA MNs exhibit excellent biocompatibility. These findings suggest that SA/FK13-a1@GelMA MNs have significant potential for the clinical treatment of tinea pedis as well as other fungal skin disorders.

Mediation Effects of Coping Styles on Fear of Progression and Reproductive Concerns in Breast Cancer Patients of Reproductive Age.

PURPOSE: This study aimed to investigate reproductive concerns among breast cancer patients of reproductive age, analyze the influencing factors, explore the relationship between coping styles, fear of progression (FOP), and reproductive concerns, and identify the multiple effects of coping styles on the relationship between FOP and reproductive concerns among Chinese breast cancer patients. METHODS: A cross-sectional, descriptive study was conducted among breast cancer patients in four tertiary grade A hospitals in Fujian, China, from January 2022 to September 2022. A total of 210 patients were recruited to complete paper-based questionnaires, which included the general data questionnaires, the Reproductive Concerns After Cancer Scale (RCACS), the Fear of Progression Questionnaire-Short Form (FOP-Q-SF), and the Medical Coping Modes Questionnaire (MCMQ). Structural equation models were utilized to evaluate the multiple effects of coping styles on FOP and reproductive concerns. RESULTS: Reproductive concerns in breast cancer patients had a mean score of 53.02 (SD, 10.69), out of a total score of 90, and coping styles for cancer (confrontation, avoidance) were closely associated with FOP and reproductive concerns. FOP showed a significant positive correlation with reproductive concerns (r = .52, p < .01). At the same time, confrontation was significantly negatively correlated with both FOP (r = -.28, p < .01) and reproductive concerns (r = -.39, p < .01). Avoidance was positively correlated to both FOP (r = .25, p < .01) and reproductive concerns (r = .34, p < .01). The impact of FOP on reproductive concerns is partially mediated by confrontation and avoidance, with effect sizes of .07 and .04, respectively. These mediating factors account for 22.0% of the total effect. CONCLUSIONS: The FOP directly impacted reproductive concerns, while coping styles could partially mediate the association between FOP and reproductive concerns. This study illustrates the role of confrontation and avoidance in alleviating reproductive concerns, suggesting that it is necessary to focus on the changes in reproductive concerns among reproductive-age breast cancer patients. Healthcare professionals can improve disease awareness and reduce patients' FOP, thereby promoting positive psychological and coping behaviors and ultimately alleviating reproductive concerns.

Synergistic effects of oxidized konjac glucomannan on rheological, thermal and structural properties of gluten protein.

Oxidation is an effective way to prepare depolymerized konjac glucomannan (KGM). The oxidized KGM (OKGM) differed from native KGM in physicochemical properties due to different molecular structure. In this study, the effects of OKGM on the properties of gluten protein were investigated and compared with native KGM (NKGM) and enzymatic hydrolysis KGM (EKGM). Results showed that the OKGM with a low molecular weight and viscosity could improve rheological properties and enhance thermal stability. Compared to native gluten protein (NGP), OKGM stabilized the protein secondary structure by increasing the contents of ß-sheet and α-helix, and improved the tertiary structure through increasing the disulfide bonds. The compact holes with shrunk pore size confirmed a stronger interaction between OKGM and gluten protein through scanning electron microscopy, forming a highly networked gluten structure. Furthermore, OKGM depolymerized by the moderate ozone-microwave treatment of 40 min had a higher effect on gluten proteins than that by the 100 min treatment, demonstrating that the excessive degradation of KGM weakened the interaction between the gluten protein and OKGM. These findings demonstrated that incorporating moderately oxidized KGM into gluten protein was an effective strategy to improve the properties of gluten protein.

Phosphate-solubilizing bacteria abate cadmium absorption and restore the rhizospheric bacterial community composition of grafted watermelon plants.

The grafting of watermelon plants to rootstocks is common due to the strong capacity of rootstocks to adapt to abiotic and biotic stresses. However, the effect of phosphate-solubilizing bacteria (PSB) on grafted watermelon plant growth and bacterial structures in root soil is unclear. In this study, the growth and hormone levels of grafted plants were measured, and the bacterial communities under cadmium (Cd) stress and inoculation with PSB were sequenced in three treatments (S1, control; S2, 50 µmol Cd [CdCl2]; and S3, 50 µmol Cd plus inoculation with the Cd-resistant PSB strain 'N3'). The results showed that inoculation with PSB significantly (P < 0.05) improved the total dry weight of the grafted plants. Typically, inoculation with PSB significantly (P < 0.05) reduced Cd content in scions and roots. The level of the phytohormone jasmonic acid increased in treatment S2, but decreased in treatment S3 under inoculation with PSB. The functional annotation of prokaryotic taxa showed that Cd decreased the abundance of nitrogen respiration and chloroplast functional groups. Nevertheless, inoculation with PSB helped restore bacterial community structures. These findings provide a new understanding of the effect of PSB on the promotion of seedling growth and bacterial communities in grafted watermelon plants under Cd stress.

Effect of 4-chloro-2-methylphenoxy acetic acid on tomato gene expression and rhizosphere bacterial communities under inoculation with phosphate-solubilizing bacteria.

The herbicide 4-chloro-2-methylphenoxy acetic acid (MCPA) is widely used to control the spread of broad-leaved weeds in agricultural soils, though it remains unclear how tomato plants cope with the phytotoxic effects of MCPA at the molecular level. In this study, RNA-seq and Illumina MiSeq were used to sequence bacterial communities in tomato rhizosphere soils treated with MCPA and the phosphate-solubilizing bacterial strain N3. The results showed that MCPA induced abnormal growth of lateral roots in tomato seedlings and reduced uptake of the nutrients N, P, and K as well as the hormone (ABA and GA3) levels. Inoculation with strain N3 increased nutrient uptake by roots and increased levels of the hormones ABA, ZEA, and JA in tomato seedlings and also increased the abundance of the phyla Proteobacteria and Gemmatimonadetes in soil under MCPA treatment. GO functional groups in which differentially expressed genes (DEGs) are involved included DNA binding transcription factor activity, transcriptional regulator activity, enzyme inhibitor activity, and cell wall biogenesis. The highest numbers of DEGs are annotated to ribosome, photosynthesis, and carbon metabolism categories. Our findings provide valuable information for the application of strain N3, which is beneficial for reducing the toxic effect of MCPA on vegetable plants.

Establishment of the technology of cambial meristematic cells (CMCs) culture from shoots and high expression of FmPHV (PHAVOLUTA) functions in identification and differentiation of CMCs and promoting the shoot regeneration by hypocotyl in Fraxinus mandshurica.

In Fraxinus mandshurica, we successfully isolated and identified the loose, uniform and creamy-white cambial meristematic cells (CMCs) from newborn shoots, and established a culture technology for induction, proliferation and differentiation of CMCs. In this technology, higher induction rate (83.0%, 0.57-fold to the control) was obtained by an effective pretreatment after 28-day induction culture, CMCs can be better proliferation cultured than common calli and maintain same growth states after several times of cultures and 3.3% CMCs primarily realized differentiation. Gene expressions in the differentiated CMCs revealed that, low expression of FmWOX5 (regulator in establishment of competence for shoot formation, 0.09-fold to the control) and high expressions of FmWOX4 (cambium stem cell regulator, 16.7-fold to the control) and 9 key genes in shoot regeneration (2.4-fold-72.1-fold to the control) function in CMCs differentiation. In addition to the function of high expression of PHAVOLUTA (FmPHV) in CMCs differentiation (5.4-fold-157.3-fold to undifferentiated CMCs), functions of high expression of FmPHV in CMCs identification (22.4-fold to common calli) and generating more shoots (2.3-fold to the control) by significantly changing expressions of key regulators in HD-Zip Class III related shoot regeneration networks in positive transgenic plants through the hypocotyl transforming system in F. mandshurica, were further revealed. These works were of profound significance in providing the culture technology of CMCs from newborn shoots in F. mandshurica for the first time and revealing the positive functions of FmPHV in CMCs identification and differentiation in F. mandshurica and promoting the shoot regeneration by hypocotyls.

Transcriptome profiling of genes involved in nutrient uptake regulated by phosphate-solubilizing bacteria in pepper (Capsicum annuum L.).

Improving nutrient absorption in pepper has become a vital prerequisite for growth to produce a sustainable yield. In this study, transcriptome gene expression in pepper inoculated with two types of phosphate-solubilizing bacteria (PSB) and grown under low and high nutrient levels (LN and HN) was analyzed. Results showed that the root length increased when pepper was grown under LN; however, the root structure was intensively tight under HN. Our data revealed that the roots preferred horizontal growth than longitudinal growth under HN. PSB strains 'M01' and 'N3' significantly (P < 0.01) increased the P uptake by 70.44% and 98.20%, respectively, but decreased the Ca2+ content by 8.96% and 9.13%, respectively, compared with the control (L1). Although no remarkable difference was detected in the chlorophyll content, inoculation with the two PSB strains decreased the Fe3+ content in pepper under HN. The total clean sequenced data from samples ranged between 5,923,659,118 and 9,955,045,953 bp. Transcriptome profiling revealed 320 upregulated and 449 downregulated genes in L3 versus L1 and 468 upregulated and 532 downregulated genes in L4 versus L1. Gene ontology analysis revealed that the biological processes, including response to stress and secondary metabolic process, were involved. Several pathways were subordinate to glycosphingolipid biosynthesis and linoleic acid and nitrogen metabolisms. Analysis of the eukaryotic orthologous group function revealed that most differential genes were attributed to RNA processing and modification, transcription, and signal transduction. Our results provided new insights into the molecular mechanism related to nutrient uptake in pepper inoculated with PSBs.

Expression characteristics and function of CAS and a new beta-amyrin synthase in triterpenoid synthesis in birch (Betula platyphylla Suk.).

Triterpenoids produced by the secondary metabolism of Betula platyphylla Suk. exhibit important pharmacological activities, such as tumor inhibition, anti-HIV, and defense against pathogens, but the yield of natural synthesis is low, which is insufficient to meet people's needs. In this study, we identified two OSC genes of birch, named as BpCAS and Bpß-AS, respectively. The expression of BpCAS and Bpß-AS were higher levels in roots and in stems, respectively, and they induced expression in response to methyl jasmonate (MeJA), gibberellin (GA3), abscisic acid (ABA), ethylene and mechanical damage. The function of the two genes in the triterpene synthesis of birch was identified by reverse genetics. The inhibition of Bpß-AS gene positively regulates synthesis of betulinic acid. BpCAS interference can significantly promote the upregulation of lupeol synthase gene (BPW) and ß-amyrin synthase gene(BPY), and conversion of 2,3-oxidosqualene to the downstream products betulinic acid and oleanolic acid. This study provided a basis for the genetic improvement of triterpenoid synthesis in birch through genetic engineering. The obtained transgenic birch and suspension cells served as material resources for birch triterpenoid applications in further.

Cardioprotective effect of isorhamnetin against myocardial ischemia reperfusion (I/R) injury in isolated rat heart through attenuation of apoptosis.

In this study, we investigated the effects of isorhamnetin on myocardial ischaemia reperfusion (I/R) injury in Langendorff-perfused rat hearts. Isorhamnetin treatment (5, 10 and 20 µg/mL) significantly alleviated cardiac morphological injury, reduced myocardial infarct size, decreased the levels of marker enzymes (LDH and CK) and improved the haemodynamic parameters, reflected by the elevated levels of the left ventricular developed pressure (LVDP), coronary flow (CF) and the maximum up/down velocity of left ventricular pressure (+dp/dtmax ). Moreover, isorhamnetin reperfusion inhibited apoptosis of cardiomyocytes in the rats subjected to cardiac I/R in a dose-dependent manner concomitant with decreased protein expression of Bax and cleaved-caspase-3, as well as increased protein expression of Bcl-2. In addition, I/R-induced oxidative stress was manifestly mitigated by isorhamnetin treatment, as showed by the decreased malondialdehyde (MDA) level and increased antioxidant enzymes activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px). These results indicated that isorhamnetin exerts a protective effect against I/R-induced myocardial injury through the attenuation of apoptosis and oxidative stress.

Transcriptome Analysis of Ice Plant Growth-Promoting Endophytic Bacterium Halomonas sp. Strain MC1 to Identify the Genes Involved in Salt Tolerance.

Salt stress is an important adverse condition encountered during plant and microbe growth in terrestrial soil ecosystems. Currently, how ice plant (Mesembryanthemum crystallinum) growth-promoting endophytic bacteria (EB) cope with salt stress and regulate growth and the genes responsible for salt tolerance remain unknown. We applied RNA-Seq technology to determine the growth mechanism of the EB Halomonas sp. MC1 strain and the genes involved in salt tolerance. A total of 893 genes were significantly regulated after salt treatment. These genes included 401 upregulated and 492 downregulated genes. Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes analysis revealed that the most enriched genes included those related to the outer membrane-bounded periplasmic space, ATPase activity, catabolic process, and proton transmembrane transport. The quantitative real-time polymerase chain reaction data were similar to those obtained from RNA-Seq. The MC1 strain maintained survival under salt stress by regulating cellular and metabolic processes and pyruvate metabolism pathways such as organic and carboxylic acid catabolic pathways. We highlighted the response mechanism of Halomonas sp. MC1 to fully understand the dynamics of complex salt-microbe interactions.

Variations in pH significantly affect cadmium uptake in grafted muskmelon (Cucumis melo L.) plants and drive the diversity of bacterial communities in a seedling substrate.

Substrates are fundamental prerequisites for growing grafted seedlings. In this study, substrates with different pH levels (5.0, 5.5, 6.0, 6.5, 7.0, and 8.0) were set up to elucidate the effect of pH on cadmium (Cd) uptake in grafted muskmelon (Cucumis melo L.) plants. Bacterial diversity was also investigated. Results showed that pH and high Cd concentration greatly affected the growth of grafted plants. The chlorophyll content of the muskmelon leaves decreased at 100 µM Cd. The majority of the Cd ions accumulated in the rootstock rather than in the shoot tissue in all of the treatments. The shoots and roots showed the highest Cd content at pH 5.5 and the lowest Cd content at pH 8.0 regardless of the Cd concentration. The operational taxonomic units belonging to Proteobacteria and Bacteroidetes were significantly (p < 0.05) enriched at different substrate pH levels compared with those at pH 5.0. The operational taxonomic units belonging to the phyla Firmicutes, Acidobacteria, and Chloroflexi were significantly decreased. The available nitrogen, phosphorus, Cd, and pH were strongly linked to bacterial community compositions. On the contrary, the available potassium was weakly correlated with the bacterial structure. This study demonstrates that pH greatly affects Cd uptake in grafted muskmelon plants and predicts microbial community structures in breeding substrates with different pH levels. Our results suggest that Cd accumulation in grafted plants can be reduced by setting the appropriate substrate pH. This work can serve as a reference for growing high-quality grafted plants and ensuring food safety in the presence of Cd contamination.

Prevalence and determinants of depression in caregivers of cancer patients: A systematic review and meta-analysis.

BACKGROUND: Aim of this study was to estimate the prevalence rate of depression in cancer patient caregivers and to identify factors affecting depression and quality of life of cancer caregivers. METHODS: Relevant research articles were retrieved after literature search in several electronic databases. Random effects meta-analyses were performed to obtain pooled estimates of the prevalence rates of depression and anxiety; their respective scores, and quality of life scores. Significant relationships between depression and factors related to depression and quality of life reported in individual studies were identified. RESULTS: Thirty studies were included. Overall, 21,149 caregivers were appraised in these studies (age 52.65 years [95% CI: 49.65, 55.65]; 31.14% [28.40, 33.89] men). The prevalence of depression and anxiety were 42.30% [33.31, 51.29] % and 46.55% [35.59, 57.52], respectively. Quality of life score, as measured with Caregiver Quality of Life-Cancer scale was 64.55 [47.44, 81.66]. Patient's condition, caregiving burden, duration of caregiving, spouse caregiver, caregiver being unemployed, caregiver with chronic disease, caregiver's sleep quality, caregiver's avoidance, financial problems, and female sex were positively associated with depression whereas overall quality of life of caregiver, pre-loss grief, caregiver's education level, caregiver's age, caregiver's sense of coherence, and caregiver's bondage with patient were negatively associated with depression in caregivers. CONCLUSION: A considerably high prevalence of depression is found in cancer patient caregivers. Several factors may affect depression and their quality of life of cancer patient caregivers.

Oleanane-Type Saponins from the Roots of Ligulariopsis shichuana and Their α-Glucosidase Inhibitory Activities.

Five new oleanane-type saponins, named ligushicosides A-E, and three known oleanane-type saponins were isolated from the roots of Ligulariopsis shichuana. Their structures were established by a combination of spectroscopic techniques, including 1D and 2D NMR and high resolution electrospray ionization mass spectroscopy (HR-ESI-MS). Furthermore, all isolates were evaluated for their yeast α-glucosidase inhibitory effects and exhibited potent inhibition against α-glucosidase, while compounds 1 and 2 showed excellent inhibitory activities. The 3-O-glycoside moiety in oleanane-type saponin is important for the α-glucosidase inhibitory effects.

Endothelial Kruppel-like factor 4 regulates angiogenesis and the Notch signaling pathway.

Regulation of endothelial cell biology by the Notch signaling pathway (Notch) is essential to vascular development, homeostasis, and sprouting angiogenesis. Although Notch determines cell fate and differentiation in a wide variety of cells, the molecular basis of upstream regulation of Notch remains poorly understood. Our group and others have implicated the Krüppel-like factor family of transcription factors as critical regulators of endothelial function. Here, we show that Krüppel-like factor 4 (KLF4) is a central regulator of sprouting angiogenesis via regulating Notch. Using a murine model in which KLF4 is overexpressed exclusively in the endothelium, we found that sustained expression of KLF4 promotes ineffective angiogenesis leading to diminished tumor growth independent of endothelial cell proliferation or cell cycling effects. These tumors feature increased vessel density yet are hypoperfused, leading to tumor hypoxia. Mechanistically, we show that KLF4 differentially regulates expression of Notch receptors, ligands, and target genes. We also demonstrate that KLF4 limits cleavage-mediated activation of Notch1. Finally, we rescue Notch target gene expression and the KLF4 sprouting angiogenesis phenotype by supplementation of DLL4 recombinant protein. Identification of this hitherto undiscovered role of KLF4 implicates this transcription factor as a critical regulator of Notch, tumor angiogenesis, and sprouting angiogenesis.

Endothelial Krüppel-like factor 4 modulates pulmonary arterial hypertension.

Krüppel-like factor 4 (KLF4) is a transcription factor expressed in the vascular endothelium, where it promotes anti-inflammatory and anticoagulant states, and increases endothelial nitric oxide synthase expression. We examined the role of endothelial KLF4 in pulmonary arterial (PA) hypertension (PAH). Mice with endothelial KLF4 knockdown were exposed to hypoxia for 3 weeks, followed by measurement of right ventricular and PA pressures, pulmonary vascular muscularization, and right ventricular hypertrophy. The effect of KLF4 on target gene expression was assessed in lungs from these mice, verified in vitro by small interfering RNA (siRNA) knockdown of KLF4, and further studied at the promoter level with cotransfection experiments. KLF4 expression was measured in lung tissue from patients with PAH and normal control subjects. We found that, after hypoxia, right ventricular and PA pressures were significantly higher in KLF4 knockdown animals than controls. Knockdown animals also had more severe pulmonary vascular muscularization and right ventricular hypertrophy. KLF4 knockdown resulted in increased pulmonary expression of endothelin-1 and decreased expression of endothelial nitric oxide synthase, endothelin receptor subtype B, and prostacyclin synthase. Concordant findings were observed in vitro, both with siRNA knockdown of KLF4 and promoter activity assays. Finally, KLF4 expression was reduced in lungs from patients with PAH. In conclusion, endothelial KLF4 regulates the transcription of genes involved in key pathways implicated in PAH, and its loss exacerbates pulmonary hypertension in response to chronic hypoxia in mice. These results introduce a novel transcriptional modulator of PAH, with the potential of becoming a new therapeutic target.

Endothelial Kruppel-like factor 4 protects against atherothrombosis in mice.

The endothelium regulates vascular homeostasis, and endothelial dysfunction is a proximate event in the pathogenesis of atherothrombosis. Stimulation of the endothelium with proinflammatory cytokines or exposure to hemodynamic-induced disturbed flow leads to a proadhesive and prothrombotic phenotype that promotes atherothrombosis. In contrast, exposure to arterial laminar flow induces a gene program that confers a largely antiadhesive, antithrombotic effect. The molecular basis for this differential effect on endothelial function remains poorly understood. While recent insights implicate Kruppel-like factors (KLFs) as important regulators of vascular homeostasis, the in vivo role of these factors in endothelial biology remains unproven. Here, we show that endothelial KLF4 is an essential determinant of atherogenesis and thrombosis. Using in vivo EC-specific KLF4 overexpression and knockdown murine models, we found that KLF4 induced an antiadhesive, antithrombotic state. Mechanistically, we demonstrated that KLF4 differentially regulated pertinent endothelial targets via competition for the coactivator p300. These observations provide cogent evidence implicating endothelial KLFs as essential in vivo regulators of vascular function in the adult animal.

Antisense repression of sucrose phosphate synthase in transgenic muskmelon alters plant growth and fruit development.

To unravel the roles of sucrose phosphate synthase (SPS) in muskmelon (Cucumis melo L.), we reduced its activity in transgenic muskmelon plants by an antisense approach. For this purpose, an 830 bp cDNA fragment of muskmelon sucrose phosphate synthase was expressed in antisense orientation behind the 35S promoter of the cauliflower mosaic virus. The phenotype of the antisense plants clearly differed from that of control plants. The transgenic plant leaves were markedly smaller, and the plant height and stem diameter were obviously shorter and thinner. Transmission electron microscope observation revealed that the membrane degradation of chloroplast happened in transgenic leaves and the numbers of grana and grana lamella in the chloroplast were significantly less, suggesting that the slow growth and weaker phenotype of transgenic plants may be due to the damage of the chloroplast ultrastructure, which in turn results in the decrease of the net photosynthetic rate. The sucrose concentration and levels of sucrose phosphate synthase decreased in transgenic mature fruit, and the fruit size was smaller than the control fruit. Together, our results suggest that sucrose phosphate synthase may play an important role in regulating the muskmelon plant growth and fruit development.

Cloning and characterization of a soluble acid invertase-encoding gene from muskmelon.

Soluble acid invertase (S-AIV; EC 3.2.1.26) is thought to play a critical role in sucrose hydrolysis in muskmelon (Cucumis melo L.) fruit. A full-length cDNA clone encoding S-AIV was isolated from muskmelon by RT-PCR and RACE. The clone, designated as CmS-AIV1, contains 2178 nucleotides with an open reading frame of 1908 nucleotides. The deduced 636 amino acid sequence showed high identities with other plant soluble acid invertases. Northern blot analysis indicated that CmS-AIV1 was expressed in flowers and fruit, but was not detected in roots, stems or leaves. Moreover, the mRNA accumulation of CmS-AIV1 showed its maximum level at 10 days after pollination (DAP) and decreased gradually during fruit development until its minimum level at mature fruit. Interestingly, the sucrose content was very low in fruit before 20 DAP but increased dramatically between 20 and 30 DAP during fruit development. In contrast to sucrose content, the activities of S-AIV was very high in fruit before 20 DAP and decreased apparently between 20 and 30 DAP, suggesting that sucrose metabolism may be linked to the CmS-AIV1 transcript level in muskmelon fruit.