Endothelial Sp1/Sp3 are essential to the effect of captopril on blood pressure in male mice.

Endothelial dysfunction represents a major cardiovascular risk factor for hypertension. Sp1 and Sp3 belong to the specificity protein and Krüppel-like transcription factor families. They are ubiquitously expressed and closely associated with cardiovascular development. We investigate the role of Sp1 and Sp3 in endothelial cells in vivo and evaluate whether captopril, an angiotensin-converting enzyme inhibitor (ACEI), targets Sp1/Sp3 to exert its effects. Inducible endothelial-specific Sp1/Sp3 knockout mice are generated to elucidate their role in endothelial cells. Tamoxifen-induced deletion of endothelial Sp1 and Sp3 in male mice decreases the serum nitrite/nitrate level, impairs endothelium-dependent vasodilation, and causes hypertension and cardiac remodeling. The beneficial actions of captopril are abolished by endothelial-specific deletion of Sp1/Sp3, indicating that they may be targets for ACEIs. Captopril increases Sp1/Sp3 protein levels by recruiting histone deacetylase 1, which elevates deacetylation and suppressed degradation of Sp1/Sp3. Sp1/Sp3 represents innovative therapeutic target for captopril to prevent cardiovascular diseases.

The Multi-Step Chain Extension for Waterborne Polyurethane Binder of Para-Aramid Fabrics.

The comprehensive balance of the mechanical, interfacial, and environmental requirements of waterborne polyurethane (WPU) has proved challenging, but crucial in the specific application as the binder for high-performance polymer fiber composites. In this work, a multi-step chain extension (MCE) method was demonstrated using three kinds of small extenders and one kind of macro-chain extender (CE) for different chain extension steps. One dihydroxyl blocked small molecular urea (1,3-dimethylolurea, DMU) was applied as one of the CEs and, through the hybrid macrodiol/diamine systems of polyether, polyester, and polysiloxane, the WPU was developed by the step-by-step optimization on each chain extending reaction via the characterization on the H-bonding association, microphase separation, and mechanical properties. The best performance was achieved when the ratio of polyether/polyester was controlled at 6:4, while 2% of DMU and 1% of polysiloxane diamine was incorporated in the third and fourth chain extension steps, respectively. Under the condition, the WPU exhibited not only excellent tensile strength of 30 MPa, elongation of break of about 1300%, and hydrophobicity indicated by the water contact angle of 98°, but also effective interfacial adhesion to para-aramid fabrics. The peeling strength of the joint based on the polysiloxane incorporated WPU after four steps of chain extension was 430% higher than that prepared through only two steps of chain extension. Moreover, about 44% of the peeling strength was sustained after the joint had been boiling for 40 min in water, suggesting the potential application for high-performance fabric composites.

Endothelial G protein stimulatory α-subunit is a critical regulator of post-ischemic angiogenesis.

Post-ischemic angiogenesis is a vital pathophysiological process in diseases such as peripheral arterial disease (PAD), heart ischemia, and diabetic retinopathy. The molecular mechanisms of post-ischemic angiogenesis are complicated and not fully elucidated. The G protein stimulatory alpha subunit (Gsα) is essential for hormone-stimulated cyclic adenosine monophosphate (cAMP) production and is an important regulator for many physiological processes. In the present study, we investigated the role of endothelial Gsα in post-ischemic angiogenesis by generating adult mice with endothelial-specific Gsα deficiency (GsαECKO). GsαECKO mice had impaired blood flow recovery after hind limb ischemic injury, and reduced neovascularization in allograft transplanted tumors. Mechanically, Gsα could regulate the expression of angiogenic factor with G patch and FHA domains 1 (AGGF1) through cAMP/CREB pathway. AGGF1 plays a key role in angiogenesis and regulates endothelial cell proliferation as well as migration. Knockdown of CREB or mutation of the CRE site on the AGGF1 promoter led to reduced AGGF1 promoter activity. In addition, knockdown of AGGF1 reduced the proangiogenic effect of Gsα in endothelial cells, and overexpression of AGGF1 reversed the impaired angiogenesis in GsαECKO mice in vivo. The finding may prove useful in designing new therapeutic targets for treatments of post-ischemic angiogenesis-related diseases.

Stimulatory G-Protein α Subunit Modulates Endothelial Cell Permeability Through Regulation of Plasmalemma Vesicle-Associated Protein.

Endothelial cell leakage occurs in several diseases. Intracellular junctions and transcellular fashion are involved. The definite regulatory mechanism is complicated and not fully elucidated. The alpha subunit of the heterotrimeric G-stimulatory protein (Gsα) mediates receptor-stimulated production of cyclic adenosine monophosphate (cAMP). However, the role of Gsα in the endothelial barrier remains unclear. In this study, mice with knockout of endothelial-specific Gsα (GsαECKO) were generated by crossbreeding Gsαflox/flox mice with Cdh5-CreERT2 transgenic mice, induced in adult mice by tamoxifen treatment. GsαECKO mice displayed phenotypes of edema, anemia, hypoproteinemia and hyperlipoproteinemia, which indicates impaired microvascular permeability. Mechanistically, Gsα deficiency reduces the level of endothelial plasmalemma vesicle-associated protein (PLVAP). In addition, overexpression of Gsα increased phosphorylation of cAMP response element-binding protein (CREB) as well as the mRNA and protein levels of PLVAP. CREB could bind to the CRE site of PLVAP promoter and regulate its expression. Thus, Gsα might regulate endothelial permeability via cAMP/CREB-mediated PLVAP expression.

Topology Reliable LCST-Type Behavior of ABA Triblock Polymer and Influence on Water Condensation and Crystallization.

As a kind of smart material, thermoresponsive hydrogels are widely investigated and applied in many fields. Due to the limitation of the freezing temperature of the water, it is a challenge to further broaden their sol-gel transition temperature (Tgel ) range, especially below 0 °C. Herein, the lower critical solution temperature type of amphiphilic ABA triblock copolymers, synthesized via two-step reversible addition-fragmentation chain transfer (RAFT) polymerization is demonstrated. The hydrophilic A-block and the hydrophobic B-block are composed of poly(N,N-dimethylacrylamide) (PDMAA) and poly(diacetone acrylamide) (PDAAM), respectively. The degree of polymerization (DP) of both A-block and B-block shows a significant influence on the Tgel of triblock copolymer dispersion. By changing the length of these two blocks or physically blending these copolymers dispersions, the Tgel can be well adjusted in a temperature range from 45 to -10 °C. Moreover, When the Tgel is higher than 4 °C, the triblock copolymer coatings show a good anti-fogging property. And when the Tgel is around or lower than the freezing temperature of the water, aqueous dispersions of the triblock copolymer have an ice recrystallization inhibition activity, resulting in the decrease of average maximum grain size (MLGS) of ice crystal.

Achievement of Both Mechanical Properties and Intrinsic Self-Healing under Body Temperature in Polyurethane Elastomers: A Synthesis Strategy from Waterborne Polymers.

Inspired by the growing demand for smart and environmentally friendly polymer materials, we employed 2,2'-disulfanediyldianiline (22DTDA) as a chain extender to synthesize a waterborne polyurethane (WPUR). Due to the ortho-substituted structure of the aromatic disulfide, the urea moieties formed a unique microphase structure in the WPUR, its mechanical strength was enhanced more 180 times relative to that of the material prepared without 22DTDA, and excellent self-healing abilities at body temperature in air or under ultrasound in water were obtained. If the self-healing process was carried out at 37 °C, 50 °C or under ultrasound, the ultimate tensile strength and elongation at break of the healed film could reach 13.8 MPa and 1150%, 15.4 MPa and 1215%, or 16 MPa and 1056%, respectively. Moreover, the WPUR films could be re-healed at the same fracture location over three cutting-healing cycles, and the recovery rates of the tensile strength and elongation at break remained almost constant throughout these cycles.

Smooth muscle-specific Gsα deletion exaggerates angiotensin II-induced abdominal aortic aneurysm formation in mice in vivo.

OBJECTIVE: Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease without an effective pharmaceutical treatment. Genetic studies have proved the involvement of smooth muscle phenotype switch in the development of AAA. The alpha subunit of the heterotrimeric G stimulatory protein (Gsα) mediates receptor-stimulated production of cyclic adenosine monophosphate (cAMP). However, the role of smooth muscle Gsα in AAA formation remains unknown. APPROACH AND RESULTS: In this study, mice with knockout of smooth muscle-specific Gsα (GsαSMKO) were generated by cross-breeding Gsαflox/flox mice with SM22-CreERT2 transgenic mice, induced in adult mice by tamoxifen treatment. Gsα deficiency induced a smooth muscle phenotype switch from a contractile to a synthetic state. Mechanically, Gsα deletion reduced cAMP level and increased the level of human antigen R (HuR), which binds with the adenylate uridylate-rich elements of the 3' untranslated region of Krüppel-like factor 4 (KLF4) mRNA, thereby increasing the stability of KLF4. Moreover, genetic knockdown of HuR or KLF4 rescued the phenotype switch in Gsα-deficient smooth muscle cells. Furthermore, with acute infusion of angiotensin II, the incidence of AAA was markedly higher in ApoE-/-/GsαSMKO than ApoE-/-/Gsαflox/flox mice and induced increased elastic lamina degradation and aortic expansion. Finally, the levels of Gsα and SM α-actin were significantly lower while those of HuR and KLF4 were higher in human AAA samples than adjacent nonaneurysmal aortic sections. CONCLUSIONS: Gsα may play a protective role in AAA formation by regulating the smooth muscle phenotype switch and could be a potential therapeutic target for AAA disease.

ABA-type triblock copolymer micellar system with lower critical solution temperature-type sol-gel transition.

A temperature sensitive sol-gel transition induced by the self-assembly of amphiphilic copolymers and its application in industry have been the objects of increasing study. We demonstrate here a two-step, reversible addition-fragmentation chain transfer (RAFT) polymerization of an ABA-type copolymer consisting of poly(N,N-dimethylacrylamide)-b-poly(diacetone acrylamide)-b-poly(N,N-dimethylacrylamide) (PDMAA-b-PDAAM-b-PDMAA). This copolymer can be easily dispersed in water, and this dispersion is critical for its lower critical solution temperature (LCST)-type sol-gel transition, which was monitored using dynamic light scattering (DLS), transmission electron microscopy (TEM), and rheology analysis, in addition to temperature-dependent 1H nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FTIR). Results revealed an abnormal sphere-to-worm micellar transition of this ABA copolymer at the LCST point, which could be affected by the length of the PDAAM block (B-block), the length as well as the distribution of the PDMAA block (A-block), and the concentration of the copolymer dispersion. Thus, copolymer dispersion could be feasibly used for drug loading at a low temperature, which could then be transformed into a gel at an elevated temperature. The loading and controllable release of the model drug of paracetamol into and out of a copolymer gel was further determined. The sustained release behavior was also studied using the Rigter-Peppas model.

Inhibition of HSV-1 Replication by Gene Editing Strategy.

HSV-1 induced illness affects greater than 85% of adults worldwide with no permanent curative therapy. We used RNA-guided CRISPR/Cas9 gene editing to specifically target for deletion of DNA sequences of the HSV-1 genome that span the region directing expression of ICP0, a key viral protein that stimulates HSV-1 gene expression and replication. We found that CRISPR/Cas9 introduced InDel mutations into exon 2 of the ICP0 gene profoundly reduced HSV-1 infectivity in permissive human cell culture models and protected permissive cells against HSV-1 infection. CRISPR/Cas9 mediated targeting ICP0 prevented HSV-1-induced disintegration of promonocytic leukemia (PML) nuclear bodies, an intracellular event critical to productive HSV-1 infection that is initiated by interaction of the ICP0 N-terminus with PML. Combined treatment of cells with CRISPR targeting ICP0 plus the immediate early viral proteins, ICP4 or ICP27, completely abrogated HSV-1 infection. We conclude that RNA-guided CRISPR/Cas9 can be used to develop a novel, specific and efficacious therapeutic and prophylactic platform for targeted viral genomic ablation to treat HSV-1 diseases.

Superior Versus Inferior Vestibular Neuritis: Are There Intrinsic Differences in Infection, Reactivation, or Production of Infectious Particles Between the Vestibular Ganglia?

HYPOTHESIS: Intrinsic differences in neurons of the vestibular ganglia result in the increased likelihood of superior vestibular ganglion involvement in vestibular neuritis. BACKGROUND: Vestibular neuritis is hypothesized to result from herpes simplex type I (HSV1) infection or reactivation in vestibular ganglia. Involvement of the inferior vestibular ganglion is extremely rare in patients with vestibular neuritis. METHODS: Primary cultures of rat superior and inferior vestibular ganglion neurons (VGNs) were cultivated separately. Neurons were lytically and latently infected with HSV1 with a US11-green fluorescent protein (GFP) chimera. Percentage lytic infection and baseline reactivation was assessed by microscopy for GFP fluorescence. Trichostatin-A (TSA) was used to stimulate HSV1 reactivation. Virion production was assessed by viral titers. Relative numbers of latency-associated (LAT) transcripts were determined by real-time reverse-transcription polymerase chain reaction (real-time RT-PCR). RESULTS: Lytic infection rates were equivalent between the two ganglia (p > 0.05). Lytic infections yielded similar amounts of plaque-forming units (p > 0.05). Relative amounts of LAT transcripts did not differ between latently infected superior and inferior VGNs. Latently infected cultures showed no differences in rates of baseline and TSA-induced HSV1 reactivation (p > 0.05). Production of virions was not significantly different between reactivated, latently infected superior versus inferior VGNs (p = 0.45). CONCLUSION: Differences in prevalence of superior and inferior vestibular neuritis do not result from intrinsic differences in HSV1 infection or virion production of these neurons. Other factors, such as the length and width of the bony canal containing the ganglia and nerves, account for the greater involvement of the superior vestibular ganglion in vestibular neuritis.

Preparation and electromagnetic properties of core/shell polystyrene@polypyrrole@nickel composite microspheres.

Through a novel method, we successfully synthesized electromagnetic (EM) functional polystyrene@polypyrrole@nickel (PS@PPy@Ni) composite microspheres. The PS@PPy spheres with well-defined core/shell structure have been synthesized via an in situ chemical oxidative copolymerization of pyrrole (Py) and N-2-carboxyethylpyrrole (PyCOOH) templated by PS microspheres. The reaction was carried out under heterophase conditions using the mixture of ethanol and water as the continuous phase. Tailored by the carboxyl groups on the surface of microspheres, magnetic nickel layer has been steady deposited onto the P(Py-PyCOOH) layer of the microspheres through an activation-electroless plating technology. The fine PS@P(Py-PyCOOH)@Ni core/shell structures could be obtained with the PyCOOH content up to 50 wt % in the P(Py-PyCOOH) layer. Moreover, the as-prepared PS@P(Py-PyCOOH)@Ni composites are ferromagnetic materials and behave as a good electromagnetic (EM) absorption material due to the coating of Ni layer around the PS@P(Py-PyCOOH) spheres. The PS@P(Py-PyCOOH)@Ni composite spheres show the remarkable EM wave absorption property with the maximum reflection loss (around -20.06 dB) at 10.69 GHz. The EM wave absorption can retained lower than -10 dB within a broad frequency range from 9.16 to 13.75 GHz.