STAT3 and Endothelial Cell—Cardiomyocyte Dialog in Cardiac Remodeling

This article presents an overview of the central role of STAT3 in the crosstalk between endothelial cells and cardiac myocytes in the heart. Endothelial cell STAT3 has a key role in inflammation that underlies cardiovascular disease and impacts on cardiac structure and function. STAT3 in endothelial cells contributes to adverse cardiomyocyte genetic reprograming, for instance, during peripartum cardiomyopathy. Conversely, cardiomyocyte STAT3 is important for maintaining endothelial cell function and capillary integrity with aging and hypertension. In addition, STAT3 serves as a sentinel for stress in the heart. Recent evidence has revealed that the redox nature of STAT3 is regulated, and STAT3 is responsive to oxidative stress (ischemia-reperfusion) so as to induce protective genes. At the level of the mitochondrion, STAT3 is important in regulating reactive oxygen species (ROS) formation, metabolism, and mitochondrial integrity. STAT3 may also control calcium release from the ER so as to limit its subsequent uptake by mitochondria and the induction of cell death. Under normal conditions, some STAT3 localizes to intercalated discs of cardiomyocytes and serves to transmit pro-fibrotic gene induction signals in the nucleus with increased blood pressure. Further research is needed to understand how the sentinel role of STAT3 in both endothelial cells and cardiomyocytes is integrated in order to coordinate the response of the heart to both physiological and pathological demands

The Role of Transforming Growth Factorβ (TGF-β)-activated Kinase 1 (TAK1) in Retinal Development

poster abstractPurpose: The formation of the retina is dependent on multiple transcription factors being expressed in the correct time and place. Transforming growth factor-β-activated kinase1 (TAK1), a serine threonine kinase, has been increasingly associated with regulation of proliferation, differentiation and apoptosis of many cell types both within and outside of the central nervous system. However, little is known about its role in development of the retina. Previous results from our lab have indicated that TAK1 is expressed throughout the developing retina; however activated TAK1 is found predominantly in the dividing progenitors of the early developing chick retina. Retinas injected with TAK1 inhibitor appeared to have an increase in progenitor population and a decrease in differentiating retinal ganglion cells. The present study evaluated the potential role of TAK1 in inducing apoptosis in the developing chick retina. Methods: Embryonic day 3(E3) chick retina were injected with vehicle and 1.0 M or 2.0 M concentration (5Z)-7-Oxozeaenol, an irreversible inhibitor of TAK1. 24 hours post inhibition the tissue was harvested. Immunohistochemistry (IHC) was performed to analyze the levels of cleaved caspase 3 expression, a protein activated during apoptosis. Nuclei stained with DAPI were used to quantify the number of cells expressing the caspase3. Lipopolysaccharide (LPS) treated adult, postnatal 30 (P30), mouse retina was used as a positive control for our IHC. Results: No difference in the level of cleaved (activated) caspase 3 immunolabel was found in vehicle-, 1.0 and 2.0 M inhibitor-injected retinas. Conclusion: Lack of cleaved caspase 3 immunolabel in TAK1-inhibited retinas indicates that TAK1 may not be playing any role in inducing cell death through apoptosis in the developing chick retina used in our study. These preliminary results suggest further research should be done to better understand its role in retinal development. Mentors: Teri Belecky-Adams and Sarika Tiwari, Center for Regenerative Biology and Medicine, IUPUI, Department of Biology, IUPU

Bruton's Tyrosine Kinase Inhibition Attenuates the Cardiac Dysfunction Caused by Cecal Ligation and Puncture in Mice

Sepsis is one of the most prevalent diseases in the world. The development of cardiac dysfunction in sepsis results in an increase of mortality. It is known that Bruton's tyrosine kinase (BTK) plays a role in toll-like receptor signaling and NLRP3 inflammasome activation, two key components in the pathophysiology of sepsis and sepsis-associated cardiac dysfunction. In this study we investigated whether pharmacological inhibition of BTK (ibrutinib 30 mg/kg and acalabrutinib 3 mg/kg) attenuates sepsis associated cardiac dysfunction in mice. 10-week old male C57BL/6 mice underwent CLP or sham surgery. One hour after surgery mice received either vehicle (5% DMSO + 30% cyclodextrin i.v.), ibrutinib (30 mg/kg i.v.), or acalabrutinib (3 mg/kg i.v.). Mice also received antibiotics and an analgesic at 6 and 18 h. After 24 h, cardiac function was assessed by echocardiography in vivo. Cardiac tissue underwent western blot analysis to determine the activation of BTK, NLRP3 inflammasome and NF-\u3baB pathway. Serum analysis of 33 cytokines was conducted by a multiplex assay. When compared to sham-operated animals, mice subjected to CLP demonstrated a significant reduction in ejection fraction (EF), fractional shortening (FS), and fractional area change (FAC). The cardiac tissue from CLP mice showed significant increases of BTK, NF-\u3baB, and NLRP3 inflammasome activation. CLP animals resulted in a significant increase of serum cytokines and chemokines (TNF-\u3b1, IL-6, IFN-\u3b3, KC, eotaxin-1, eotaxin-2, IL-10, IL-4, CXCL10, and CXCL11). Delayed administration of ibrutinib and acalabrutinib attenuated the decline of EF, FS, and FAC caused by CLP and also reduced the activation of BTK, NF-\u3baB, and NLRP3 inflammasome. Both ibrutinib and acalabrutinib significantly suppressed the release of cytokines and chemokines. Our study revealed that delayed intravenous administration of ibrutinib or acalabrutinib attenuated the cardiac dysfunction associated with sepsis by inhibiting BTK, reducing NF-\u3baB activation and the activation of the inflammasome. Cytokines associated with sepsis were significantly reduced by both BTK inhibitors. Acalabrutinib is found to be more potent than ibrutinib and could potentially prove to be a novel therapeutic in sepsis. Thus, the FDA-approved BTK inhibitors ibrutinib and acalabrutinib may be repurposed for the use in sepsis

The CXCL10/CXCR3 Axis and Cardiac Inflammation: Implications for Immunotherapy to Treat Infectious and Noninfectious Diseases of the Heart.

Accumulating evidence reveals involvement of T lymphocytes and adaptive immunity in the chronic inflammation associated with infectious and noninfectious diseases of the heart, including coronary artery disease, Kawasaki disease, myocarditis, dilated cardiomyopathies, Chagas, hypertensive left ventricular (LV) hypertrophy, and nonischemic heart failure. Chemokine CXCL10 is elevated in cardiovascular diseases, along with increased cardiac infiltration of proinflammatory Th1 and cytotoxic T cells. CXCL10 is a chemoattractant for these T cells and polarizing factor for the proinflammatory phenotype. Thus, targeting the CXCL10 receptor CXCR3 is a promising therapeutic approach to treating cardiac inflammation. Due to biased signaling CXCR3 also couples to anti-inflammatory signaling and immunosuppressive regulatory T cell formation when activated by CXCL11. Numbers and functionality of regulatory T cells are reduced in patients with cardiac inflammation, supporting the utility of biased agonists or biologicals to simultaneously block the pro-inflammatory and activate the anti-inflammatory actions of CXCR3. Other immunotherapy strategies to boost regulatory T cell actions include intravenous immunoglobulin (IVIG) therapy, adoptive transfer, immunoadsorption, and low-dose interleukin-2/interleukin-2 antibody complexes. Pharmacological approaches include sphingosine 1-phosphate receptor 1 agonists and vitamin D supplementation. A combined strategy of switching CXCR3 signaling from pro- to anti-inflammatory and improving Treg functionality is predicted to synergistically lessen adverse cardiac remodeling

Conflicting vascular and metabolic impact of the IL-33/sST2 axis.

Interleukin 33 (IL-33), which is expressed by several immune cell types, endothelial and epithelial cells, and fibroblasts, is a cytokine of the IL-1 family that acts both intra- and extracellularly to either enhance or resolve the inflammatory response. Intracellular IL-33 acts in the nucleus as a regulator of transcription. Once released from cells by mechanical stress, inflammatory cytokines, or necrosis, extracellular IL-33 is proteolytically processed to act in an autocrine/paracrine manner as an 'alarmin' on neighbouring or various immune cells expressing the ST2 receptor. Thus, IL-33 may serve an important role in tissue preservation and repair in response to injury; however, the actions of IL-33 are dampened by a soluble form of ST2 (sST2) that acts as a decoy receptor and is produced by endothelial and certain immune cells. Accumulating evidence supports the conclusion that sST2 is a biomarker of vascular health with diagnostic and/or prognostic value in various cardiovascular diseases, including coronary artery disease, myocardial infarction, atherosclerosis, giant-cell arteritis, acute aortic dissection, and ischaemic stroke, as well as obesity and diabetes. Although sST2 levels are positively associated with cardiovascular disease severity, the assumption that IL-33 is always beneficial is naïve. It is increasingly appreciated that the pathophysiological importance of IL-33 is highly dependent on cellular and temporal expression. Although IL-33 is atheroprotective and may prevent obesity and type 2 diabetes by regulating lipid metabolism, IL-33 appears to drive endothelial inflammation. Here, we review the current knowledge of the IL-33/ST2/sST2 signalling network and discuss its pathophysiological and translational implications in cardiovascular diseases

Cardiac STAT3 Deficiency Impairs Contractility and Metabolic Homeostasis in Hypertension

Signal transducer and activator of transcription 3 (STAT3) protects the heart from acute ischemic stress. However, the importance of STAT3 to the heart in chronic stress, such as hypertension, is not known. To study this, we used cardiomyocyte-targeted STAT3 knockout (KO) mice and ANG II infusion by osmotic minipumps. After 4 weeks, ANG II induced similar cardiac hypertrophy in wild type (WT) and cardiac Cre-expressing control (CTRL) mice with no impairment of cardiac function. In contrast, STAT3 KO mice exhibited reduced contractile function but similar hypertrophy to CTRL mice. Ejection fraction and fractional shortening decreased by 22.5% and 27.3%, respectively. Since STAT3 has direct protective effects on mitochondrial function, we examined rates of glucose and oleate oxidation by isolated perfused hearts using a Langendorff system. Hearts of ANG II-treated STAT3 KO and CTRL mice had similar rates of oleate oxidation as saline-infused WT mice. Rates of glucose oxidation were similar between hearts of WT plus saline and CTRL plus ANG II mice; however, glucose oxidation was increased by 66% in hearts of ANG II-treated STAT3 KO mice. The ratio of maximal ATP yield from glucose to fatty acid oxidation was 21.1 ± 3.1 in hearts of ANG II-treated STAT3 KO mice vs. 12.6 ± 2.2 in hearts of ANG II-treated CTRL mice. Lactate production was also elevated in hearts of ANG II-treated STAT3 KO mice by 162% compared to ANG II-treated CTRL mice. Our findings indicate that (1) STAT3 is important for maintaining contractile function and metabolic homeostasis in the hypertensive heart, and (2) STAT3 deficiency promotes a switch toward glucose utilization

Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample

BACKGROUND: Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. RESULTS: The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). CONCLUSIONS: The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget

PEMANFAATAN TEKNOLOGI MEMBRAN REVERSE OSMOSIS (RO) UNTUK PENGOLAHAN AIR BERSIH DI KAMPUNG NELAYAN, DESA KEDUNGPANDAN, KECAMATAN JABON, KABUPATEN SIDOARJO

Abstract. The Fishermen's Village in Tlocor Hamlet, Kedungpandan Village, Jabon District, is one of the coastal areas in Sidoarjo Regency, which has several islands. This has become an opportunity for most local residents to become fishermen and a tourist spot. Behind its natural beauty, the people of Dusun Tlocor still lack clean water, and the majority of their mothers are only housewives. They have to buy water for drinking and cooking because it is difficult to find clean water, even though it is close to the islands. The purpose of this Capacity Strengthening Program (PPK) activity is to find out the impact of implementing reverse osmosis membrane (MRO) technology on the costs incurred to buy clean water and increase the creativity of the surrounding community towards processed food as a special food for Tlocor Marine Tourism. The target of this activity is to help people get clean water more easily and reduce expenses for purchasing clean water. The implementation will be carried out from June to August 2022. The initial stage is a location survey by visiting the fishermen's chief's house to socialize the work program to be implemented. The next stage is procuring tools and installing WTP and MRO tools, then providing assistance to members of the fisherman group on how to use and maintain WTP and MRO tools. The result of this activity is an increase in the quality of clean water that can be utilized by the community. Communities can easily obtain clean water for drinking, sanitation, and cooking purposes.Abstrak. Kampung Nelayan di Dusun Tlocor, Desa Kedungpandan, Kecamatan Jabon merupakan salah satu daerah pesisir di Kabupaten Sidoarjo yang memiliki beberapa pulau hal itu menjadi peluang sebagian besar warga sekitar untuk menjadi nelayan dan sebagai tempat wisata. Dibalik keindahan alamnya, masyarakat Dusun Tlocor masih kekurangan air (bersih) mayoritas ibu-ibunya hanya sebagai IRT. Air untuk minum dan masak meraka harus membeli, karna sulit mencari air bersih walaupun dekat dengan pulau-pulau. Tujuan dari kegiataan Program Penguatan Kapasitas (PPK) ini adalah mengetahui dampak yang ditimbulkan setelah dilakukannya penerapan teknologi Membran Reverse Osmosis (MRO) tehadap biaya yang dikeluarkan untuk membeli air bersih serta meningkatkan kreativitas masyarakat sekitar terhadap olahan pangan sebagai makanan khas Wisata Bahari Tlocor. Target kegiatan ini adalah masyarakat dapat memperoleh air bersih dengan lebih mudah dan mengurangi biaya pengeluaran untuk pembelian air bersih. Pelaksanaan dilakukan pada bulan Juni sampai Agustus 2022, tahapan awal yaitu survei lokasi dengan mengunjungi rumah ketua nelayan untuk melakukan sosialisasi program kerja yang akan dilaksanakan, tahapan berikutnya pengadaan alat dan pemasangan Alat WTP & MRO, kemudian dilakukan pendampingan kepada anggota kelompok nelayan tentang cara penggunaan dan pemeliharaan alat WTP dan MRO. Hasil dari kegiatan ini adalah meningkatnya kualitas air bersih yang dapat dimanfaatkan oleh masyarakat. Masyarakat dapat memperoleh air bersih dengan mudah untuk keperluan air minum, air sanitasi, dan memasak.

Penentuan Karakteristik Tanah Desa Ngasem dengan Metode ASTM

Abstract: Soil is an important part of a sub-building structure, soil characteristics need to be considered in the initial planning of a construction, in order to achieve stability and security for the building above it. With different soil conditions, as is the case with the soil in Ngasem village which has low carrying capacity, it is necessary to know which structure meets the requirements for the soil conditions in Ngasem. To find out the characteristics of the soil is done by testing in the laboratory, namely testing the physical and mechanical properties. Testing the physical properties of the soil is testing the water content (Wc), specific gravity (Gs), Atterberg limits and soil unit weight (γ). Mechanical testing with proctor test and direct shear strength. The method used is an experimental method by taking soil samples at a depth of ± 80 cm and conducting research tests in the Kadiri University laboratory. So that the results of soil classification with the SW-SM code are obtained which are classified as well-graded sandy soils with silt mixtures based on USCS and classified as (A-1-b) according to the AASHTO classification system. The Atterberg Limit of Low Plasticity (clay low plasticity) with a Plastic Index Value of 3.243 which is classified as Slightly Plastic, a shear angle value of 38.748° with a cohesion value of 1.914, a maximum dry unit weight value of 1.77 gr/cm3 , and an optimum moisture content of 14.44 . So that with this study, the community is expected to choose and adjust the type and quality of the structure of building materials in accordance with the state of the land. Keywords: Soil properties, Compaction, Shear Test, Liquid Limit, Plastic LimitAbstrak: Tanah adalah bagian penting dari sebuah struktur bangunan bawah, karakteristik tanah perlu diperhatikan dalam perencanaan awal sebuah konstruksi, agar tercapai suatu kestabilan dan keamanan untuk bangunan diatasnya. Dengan kondisi tanah yang berbeda, seperti halnya tanah di Desa Ngasem yang berdaya dukung rendah sehingga perlu diketahui struktur yang memenuhi syarat untuk kondisi tanah di Ngasem. Untuk mengetahui karakteristik tanah dilakukan dengan pengujian di laboratorium yaitu pengujian sifat fisik dan mekanik. Pengujian sifat fisik tanah yaitu pengujian kadar air (Wc), berat jenis (Gs), batas-batas Atterberg dan berat volume tanah (γ). Pengujian mekanik dengan uji proctor dan kuat geser langsung. Metode yang digunakan adalah metode eksperimental dengan cara mengambil sampel tanah di kedalaman ±80 cm dan melakukan uji penelitian di laboratorium universitas kadiri. Sehingga didapatkan hasil klasifikasi tanah dengan kode SW-SM yang dikelompokkan sebagai tanah pasir bergradasi baik dengan campuran lanau berdasarkan USCS dan diklasifikasikan sebagai (A-1-b) menurut sistem klasifikasi AASHTO. Batas Atterberg Plastisitas Rendah (clay low plasticity) dengan nilai plasticity index adalah 3,243 yang tergolong Slightly Plastic, nilai sudut geser sebesar 38,748° dengan nilai kohesi 1,914, nilai berat volume kering maksimum  sebesar 1.77 gr/cm3 , dan  kadar air optimum sebesar 14,44. Sehingga dengan adanya penelitian ini, masyarakat diharapkan dapat memilih dan menyesuaikan jenis serta kualitas struktur bahan bangunan yang sesuai dengan keadaan tanah tersebut. Kata Kunci: Propertis tanah,Pemadatan,Uji Geser, Batas Cair, Batas Plasti