PROX1 Inhibits PDGF-B Expression to Prevent Myxomatous Degeneration of Heart Valves.

BACKGROUND: Cardiac valve disease is observed in 2.5% of the general population and 10% of the elderly people. Effective pharmacological treatments are currently not available, and patients with severe cardiac valve disease require surgery. PROX1 (prospero-related homeobox transcription factor 1) and FOXC2 (Forkhead box C2 transcription factor) are transcription factors that are required for the development of lymphatic and venous valves. We found that PROX1 and FOXC2 are expressed in a subset of valvular endothelial cells (VECs) that are located on the downstream (fibrosa) side of cardiac valves. Whether PROX1 and FOXC2 regulate cardiac valve development and disease is not known. METHODS: We used histology, electron microscopy, and echocardiography to investigate the structure and functioning of heart valves from Prox1ΔVEC mice in which Prox1 was conditionally deleted from VECs. Isolated valve endothelial cells and valve interstitial cells were used to identify the molecular mechanisms in vitro, which were tested in vivo by RNAScope, additional mouse models, and pharmacological approaches. The significance of our findings was tested by evaluation of human samples of mitral valve prolapse and aortic valve insufficiency. RESULTS: Histological analysis revealed that the aortic and mitral valves of Prox1ΔVEC mice become progressively thick and myxomatous. Echocardiography revealed that the aortic valves of Prox1ΔVEC mice are stenotic. FOXC2 was downregulated and PDGF-B (platelet-derived growth factor-B) was upregulated in the VECs of Prox1ΔVEC mice. Conditional knockdown of FOXC2 and conditional overexpression of PDGF-B in VECs recapitulated the phenotype of Prox1ΔVEC mice. PDGF-B was also increased in mice lacking FOXC2 and in human mitral valve prolapse and insufficient aortic valve samples. Pharmacological inhibition of PDGF-B signaling with imatinib partially ameliorated the valve defects of Prox1ΔVEC mice. CONCLUSIONS: PROX1 antagonizes PDGF-B signaling partially via FOXC2 to maintain the extracellular matrix composition and prevent myxomatous degeneration of cardiac valves.

Promoting Lymphangiogenesis and Lymphatic Growth and Remodeling to Treat Cardiovascular and Metabolic Diseases.

Lymphatic vessels are low-pressure, blind-ended tubular structures that play a crucial role in the maintenance of tissue fluid homeostasis, immune cell trafficking, and dietary lipid uptake and transport. Emerging research has indicated that the promotion of lymphatic vascular growth, remodeling, and function can reduce inflammation and diminish disease severity in several pathophysiologic conditions. In particular, recent groundbreaking studies have shown that lymphangiogenesis, which describes the formation of new lymphatic vessels from the existing lymphatic vasculature, can be beneficial for the alleviation and resolution of metabolic and cardiovascular diseases. Therefore, promoting lymphangiogenesis represents a promising therapeutic approach. This brief review summarizes the most recent findings related to the modulation of lymphatic function to treat metabolic and cardiovascular diseases such as obesity, myocardial infarction, atherosclerosis, and hypertension. We also discuss experimental and therapeutic approaches to enforce lymphatic growth and remodeling as well as efforts to define the molecular and cellular mechanisms underlying these processes.

Prostate specific membrane antigen (PSMA) avid nonprostatic benign and malignant disease: a pictorial review.

Prostate specific membrane antigen (PSMA) positron emission tomography-computed tomography (PET-CT) is revolutionising the management of prostate cancer (PC) in primary staging and assessment of biochemical recurrence (BCR) through its higher diagnostic accuracy compared to both conventional imaging and previously available PET radiopharmaceuticals. PSMA is a transmembrane glycoprotein, highly expressed in prostate cancer, with its extracellular domain the target for PSMA PET radiopharmaceuticals. However, PSMA expression is not prostate specific and resultant PSMA uptake on PET-CT is not restricted to pathologies arising from the prostate gland. The increasing use of PSMA PET-CT has revealed PSMA uptake in a variety of non-prostatic benign and malignant diseases, which adds complexity to PET-CT interpretation and subsequent clinical management. This pictorial review will provide a thorough knowledge and understanding of the comprehensive range of PSMA avid non-prostatic benign and malignant diseases demonstrable on PSMA PET-CT, whilst highlighting the complimentary nature of other imaging modalities.

Mechanotransduction activates canonical Wnt/ß-catenin signaling to promote lymphatic vascular patterning and the development of lymphatic and lymphovenous valves.

Lymphatic vasculature regulates fluid homeostasis by returning interstitial fluid to blood circulation. Lymphatic endothelial cells (LECs) are the building blocks of the entire lymphatic vasculature. LECs originate as a homogeneous population of cells predominantly from the embryonic veins and undergo stepwise morphogenesis to become the lymphatic capillaries, collecting vessels or valves. The molecular mechanisms underlying the morphogenesis of the lymphatic vasculature remain to be fully understood. Here we show that canonical Wnt/ß-catenin signaling is necessary for lymphatic vascular morphogenesis. Lymphatic vascular-specific ablation of ß-catenin in mice prevents the formation of lymphatic and lymphovenous valves. Additionally, lymphatic vessel patterning is defective in these mice, with abnormal recruitment of mural cells. We found that oscillatory shear stress (OSS), which promotes lymphatic vessel maturation, triggers Wnt/ß-catenin signaling in LECs. In turn, Wnt/ß-catenin signaling controls the expression of several molecules, including the lymphedema-associated transcription factor FOXC2. Importantly, FOXC2 completely rescues the lymphatic vessel patterning defects in mice lacking ß-catenin. Thus, our work reveals that mechanical stimulation is a critical regulator of lymphatic vascular development via activation of Wnt/ß-catenin signaling and, in turn, FOXC2.

RASA1-driven cellular export of collagen IV is required for the development of lymphovenous and venous valves in mice.

RASA1, a negative regulator of Ras-MAPK signaling, is essential for the development and maintenance of lymphatic vessel valves. However, whether RASA1 is required for the development and maintenance of lymphovenous valves (LVV) and venous valves (VV) is unknown. In this study, we show that induced disruption of Rasa1 in mouse embryos did not affect initial specification of LVV or central VV, but did affect their continued development. Similarly, a switch to expression of a catalytically inactive form of RASA1 resulted in impaired LVV and VV development. Blocked development of LVV was associated with accumulation of the basement membrane protein, collagen IV, in LVV-forming endothelial cells (EC), and could be partially or completely rescued by MAPK inhibitors and drugs that promote collagen IV folding. Disruption of Rasa1 in adult mice resulted in venous hypertension and impaired VV function that was associated with loss of EC from VV leaflets. In conclusion, RASA1 functions as a negative regulator of Ras signaling in EC that is necessary for EC export of collagen IV, thus permitting the development of LVV and the development and maintenance of VV.

YAP and TAZ maintain PROX1 expression in the developing lymphatic and lymphovenous valves in response to VEGF-C signaling.

Lymphatic vasculature is an integral part of digestive, immune and circulatory systems. The homeobox transcription factor PROX1 is necessary for the development of lymphatic vessels, lymphatic valves (LVs) and lymphovenous valves (LVVs). We and others previously reported a feedback loop between PROX1 and vascular endothelial growth factor-C (VEGF-C) signaling. PROX1 promotes the expression of the VEGF-C receptor VEGFR3 in lymphatic endothelial cells (LECs). In turn, VEGF-C signaling maintains PROX1 expression in LECs. However, the mechanisms of PROX1/VEGF-C feedback loop remain poorly understood. Whether VEGF-C signaling is necessary for LV and LVV development is also unknown. Here, we report for the first time that VEGF-C signaling is necessary for valve morphogenesis. We have also discovered that the transcriptional co-activators YAP and TAZ are required to maintain PROX1 expression in LVs and LVVs in response to VEGF-C signaling. Deletion of Yap and Taz in the lymphatic vasculature of mouse embryos did not affect the formation of LVs or LVVs, but resulted in the degeneration of these structures. Our results have identified VEGF-C, YAP and TAZ as a crucial molecular pathway in valve development.

GATA2 regulates blood/lymph separation in a platelet-dependent and lymphovenous valve-independent manner.

INTRODUCTION: Lymphatic vessels collect interstitial fluid, immune cells, and digested lipids and return these bodily fluids to blood through two pairs of lymphovenous valves (LVVs). Like other cardiovascular valves LVVs prevent the backflow of blood into the lymphatic vessels. In addition to LVVs, platelets are necessary to prevent the entry of blood into the lymphatic vessels. Platelet thrombi are observed at LVVs suggesting that LVVs and platelets function in synergy to regulate blood/lymphatic separation. OBJECTIVES: The primary objective of this work is to determine whether platelets can regulate blood/lymph separation independently of LVVs. METHODS: The transcription factor GATA2 is necessary for the development of both LVVs and hematopoietic stem cells. Using various endothelial- and hematopoietic cell expressed Cre-lines, we conditionally deleted Gata2. We hypothesized that this strategy would identify the tissue- and time-specific roles of GATA2 and reveal whether platelets and LVVs can independently regulate blood/lymph separation. RESULTS: Lymphatic vasculature-specific deletion of Gata2 results in the absence of LVVs without compromising blood/lymph separation. In contrast, deletion of GATA2 from both lymphatic vasculature and hematopoietic cells results in the absence of LVVs, reduced number of platelets and blood-filled lymphatic vasculature. CONCLUSION: GATA2 promotes blood/lymph separation through platelets. Furthermore, LVVs are the only known sites of interaction between blood and lymphatic vessels. The fact that blood is able to enter the lymphatic vessels of mice lacking LVVs and platelets indicates that under these circumstances the lymphatic and blood vessels are connected at yet to be identified sites.

Exploring invasive weed biochar as soil amendment: A study on fodder oats productivity and soil biological properties.

Biochar is known for the improvement of soil health, fertility, crop productivity, and quality in many agro-ecosystems globally, but information regarding fodder yield, quality, and soil microbial activity responses to biochar application remains very limited. The objective of this study was to prepare biochar from invasive weeds, i.e., Parthenium hysterophorus L. and Lantana camara L., and use it as a soil amendment along with inorganic fertilizers for oats (Avena sativa L.) growth, fodder yield, quality, and soil microbial activities in a two-year pot experiment. Treatments were comprised of control, 100% RDF (Recommended dose of fertilizers), 75% RDF along with three doses (2.5, 5.0, and 10 t/ha) of Parthenium hysterophorus L. biochar (PB) and Lantana camara L. biochar (LB), PB (10 t/ha), and LB (10 t/ha). Results showed that application of 75% RDF along with 10 t/ha LB gave significantly higher green (∼8%) and dry (∼7.8%) fodder yield and crude protein (∼6%) and decreased acid detergent fibre (ADF) and neutral detergent fibre (NDF) by 5.70 and 6.04% as compared to the 100% RDF treatment. The same treatment had a significantly higher population of bacteria (7.33 × 108 colony forming unit (CFU)/g soil), alkaline phosphatase activity (19.56 µg pNP/g soil/h), microbial biomass carbon (156.67 µg/g soil) and dehydrogenase activity (12.59 µg TPF/g/24 h), whereas the maximum fungal population (13.33 × 104 CFU/g soil) and acid phosphatase activity (14.45 µg pNP/g soil/h) were found in 75% RDF along with 10 t/ha PB and control treatment, respectively. This study concluded that application of invasive weed biochar along with inorganic fertilizers can benefit fodder yield and quality of oats by increasing plant height and number of tillers directly and by improving nutrient availability and water holding capacity (WHC) of soil indirectly, besides improving soil health. The findings from this study will provide a potential strategy for invasive weed management.

Sentimental analysis from imbalanced code-mixed data using machine learning approaches.

Knowledge discovery from various perspectives has become a crucial asset in almost all fields. Sentimental analysis is a classification task used to classify the sentence based on the meaning of their context. This paper addresses class imbalance problem which is one of the important issues in sentimental analysis. Not much works focused on sentimental analysis with imbalanced class label distribution. The paper also focusses on another aspect of the problem which involves a concept called "Code Mixing". Code mixed data consists of text alternating between two or more languages. Class imbalance distribution is a commonly noted phenomenon in a code-mixed data. The existing works have focused more on analyzing the sentiments in a monolingual data but not in a code-mixed data. This paper addresses all these issues and comes up with a solution to analyze sentiments for a class imbalanced code-mixed data using sampling technique combined with levenshtein distance metrics. Furthermore, this paper compares the performances of various machine learning approaches namely, Random Forest Classifier, Logistic Regression, XGBoost classifier, Support Vector Machine and Naïve Bayes Classifier using F1- Score.

GATA2 controls lymphatic endothelial cell junctional integrity and lymphovenous valve morphogenesis through miR-126.

Mutations in the transcription factor GATA2 cause lymphedema. GATA2 is necessary for the development of lymphatic valves and lymphovenous valves, and for the patterning of lymphatic vessels. Here, we report that GATA2 is not necessary for valvular endothelial cell (VEC) differentiation. Instead, GATA2 is required for VEC maintenance and morphogenesis. GATA2 is also necessary for the expression of the cell junction molecules VE-cadherin and claudin 5 in lymphatic vessels. We identified miR-126 as a target of GATA2, and miR-126-/- embryos recapitulate the phenotypes of mice lacking GATA2. Primary human lymphatic endothelial cells (HLECs) lacking GATA2 (HLECΔGATA2) have altered expression of claudin 5 and VE-cadherin, and blocking miR-126 activity in HLECs phenocopies these changes in expression. Importantly, overexpression of miR-126 in HLECΔGATA2 significantly rescues the cell junction defects. Thus, our work defines a new mechanism of GATA2 activity and uncovers miR-126 as a novel regulator of mammalian lymphatic vascular development.

Effect of SARS-CoV-2 seropositivity on antigen - specific cytokine and chemokine responses in latent tuberculosis.

SARS-CoV-2 and latent Mycobacterium tuberculosis infection are both highly co-prevalent in many parts of the globe. Whether exposure to SARS-CoV-2 influences the antigen specific immune responses in latent tuberculosis has not been investigated. We examined the baseline, mycobacterial antigen and mitogen induced cytokine and chemokine responses in latent tuberculosis (LTBI) individuals with or without SARS-CoV-2 seropositivity, LTBI negative individuals with SARS-CoV-2 seropositivity and healthy control (both LTBI and SARS-CoV-2 negative) individuals. Our results demonstrated that LTBI individuals with SARS-CoV-2 seropositivity (LTBI+/IgG +) were associated with increased levels of unstimulated and TB-antigen stimulated IFNγ, IL-2, TNFα, IL-17, IL-1ß, IL-6, IL-12, IL-4, CXCL1, CXCL9 and CXCL10 when compared to those without seropositivity (LTBI+/IgG-). In contrast, LTBI+/IgG+ individuals were associated with decreased levels of IL-5 and IL-10. No significant difference in the levels of cytokines/chemokines was observed upon mitogen stimulation between the groups. SARS-CoV-2 seropositivity was associated with enhanced unstimulated and TB-antigen stimulated but not mitogen stimulated production of cytokines and chemokines in LTBI+ compared to LTBI negative individuals. Finally, most of these significant differences were not observed when LTBI negative individuals with SARS-CoV-2 seropositivity and controls were examined. Our data clearly demonstrate that both baseline and TB - antigen induced cytokine responses are augmented in the presence of SARS-CoV-2 seropositivity, suggesting an augmenting effect of prior SARS-CoV-2 infection on the immune responses of LTBI individuals.

Biochemical and mechanical signals in the lymphatic vasculature.

Lymphatic vasculature is an integral part of the cardiovascular system where it maintains interstitial fluid balance. Additionally, lymphatic vasculature regulates lipid assimilation and inflammatory response. Lymphatic vasculature is composed of lymphatic capillaries, collecting lymphatic vessels and valves that function in synergy to absorb and transport fluid against gravitational and pressure gradients. Defects in lymphatic vessels or valves leads to fluid accumulation in tissues (lymphedema), chylous ascites, chylothorax, metabolic disorders and inflammation. The past three decades of research has identified numerous molecules that are necessary for the stepwise development of lymphatic vasculature. However, approaches to treat lymphatic disorders are still limited to massages and compression bandages. Hence, better understanding of the mechanisms that regulate lymphatic vascular development and function is urgently needed to develop efficient therapies. Recent research has linked mechanical signals such as shear stress and matrix stiffness with biochemical pathways that regulate lymphatic vessel growth, patterning and maturation and valve formation. The goal of this review article is to highlight these innovative developments and speculate on unanswered questions.

The Prox1-Vegfr3 feedback loop maintains the identity and the number of lymphatic endothelial cell progenitors.

The mammalian lymphatic vasculature is important for returning fluids from the extracellular tissue milieu back to the blood circulation. We showed previously that Prox1 dosage is important for the development of the mammalian lymphatic vasculature. The lack of Prox1 activity results in the complete absence of lymphatic endothelial cells (LECs). In Prox1 heterozygous embryos, the number of LECs is reduced because of a decrease in the progenitor pool in the cardinal vein. This reduction is caused by some progenitor cells being unable to maintain Prox1 expression. In this study, we identified Vegfr3, the cognate receptor of the lymphangiogenic growth factor Vegfc, as a dosage-dependent, direct in vivo target of Prox1. Using various mouse models, we also determined that Vegfr3 regulates Prox1 by establishing a feedback loop necessary to maintain the identity of LEC progenitors and that Vegfc-mediated activation of Vegfr3 signaling is necessary to maintain Prox1 expression in LEC progenitors. We propose that this feedback loop is the main sensing mechanism controlling the number of LEC progenitors and, as a consequence, the number of budding LECs that will form the embryonic lymphatic vasculature.

Prevalence of early hip OA features on MRI in high-impact athletes. The femoroacetabular impingement and hip osteoarthritis cohort (FORCe) study.

OBJECTIVE: To compare early hip osteoarthritis (OA) features on magnetic resonance imaging (MRI) in high-impact athletes with and without hip and/or groin pain, and to evaluate associations between early hip OA features, the International Hip Outcome Tool (iHOT33) and Copenhagen Hip and Groin Outcome Score (HAGOS). DESIGN: This case-control study evaluated data of the femoroacetabular impingement and hip osteoarthritis cohort (FORCe). One hundred and eighty-two symptomatic (hip and/or groin pain >6 months and positive flexion-adduction-internal-rotation (FADIR) test) and 55 pain-free high-impact athletes (soccer or Australian football (AF)) without definite radiographic hip OA underwent hip MRI. The Scoring Hip Osteoarthritis with MRI (SHOMRI) method quantified and graded the severity of OA features. Each participant completed the iHOT33 and HAGOS. RESULTS: Hip and/or groin pain was associated with higher total SHOMRI (0-96) (mean difference 1.4, 95% CI: 0.7-2.2), labral score (adjusted incidence rate ratio (aIRR) 1.33, 95% CI: 1.1-1.6). Differences in prevalence of cartilage defects, labral tears and paralabral cysts between symptomatic and pain-free participants were inconclusive. There was a lower prevalence of effusion-synovitis in symptomatic participants when compared to pain-free participants (adjusted odds ratio (aOR) 0.46 (95% CI: 0.3-0.8). Early hip OA features were not associated with iHOT33 or HAGOS. CONCLUSIONS: A complex and poorly understood relationship exists between hip and/or groin pain and early hip OA features present on MRI in high-impact athletes without radiographic OA. Hip and/or groin pain was associated with higher SHOMRI and labral scores.

Dental pain, parental SARS-CoV-2 fear and distress on quality of life of 2 to 6 year-old children during COVID-19.

BACKGROUND: Coronavirus disease (COVID-19) has crippled life, families and oral healthcare delivery in India due to nationwide lockdown. AIM: Through cross-sectional design, we investigated the impact of child's dental pain, caregiver's fear of SARS-CoV-2 and parental distress on oral health-related quality of life (OHRQOL) of preschoolers during the nationwide COVID-19 pandemic lockdown. DESIGN: Preschool children self-reported their pain using Pieces of Hurt scale; caregiver SARS-CoV-2 fear was assessed using Fear of COVID-19 scale and parental distress evaluated using 4-item scale. Child's oral health was assessed using the dmft index and OHRQOL evaluated using early childhood oral health impact scale. Bivariate, multivariate regression analysis was conducted to identify predictors; statistical significance was set at 5%. RESULTS: Sample mean age was 4.58 years, and about 69% were boys. Children reporting higher pain scores (OR = 1.9) due to decayed teeth and having dmft > 5 (OR = 4.25), followed by greater parental distress (OR = 4.13) and fear of SARS-CoV-2 (OR = 3.84), were significantly associated with poor OHRQOL during the COVID-19 pandemic. CONCLUSIONS: Greater parental distress and fear of COVID-19 among caregivers, higher self-perceived dental pain among children and caries experience are associated with poor OHRQOL of preschool children during the COVID-19 pandemic.

Simplified method to quantify valve back-leak uncovers severe mesenteric lymphatic valve dysfunction in mice deficient in connexins 43 and 37.

KEY POINTS: Lymphatic valve defects are one of the major causes of lymph transport dysfunction; however, there are no accessible methods for quantitatively assessing valve function. This report describes a novel technique for quantifying lymphatic valve back-leak. Postnatal endothelial-specific deletion of connexin 43 (Cx43) in connexin 37 null (Cx37-/- ) mice results in rapid regression of valve leaflets and severe valve dysfunction. This method can also be used for assessing the function of venous and lymphatic valves from various species, including humans. ABSTRACT: The lymphatic system relies on robust, spontaneous contractions of collecting lymphatic vessels and one-way secondary lymphatic valves to efficiently move lymph forward. Secondary valves prevent reflux and allow for the generation of propulsive pressure during each contraction cycle. Lymphatic valve defects are one of the major causes of lymph transport dysfunction. Genetic mutations in multiple genes have been associated with the development of primary lymphoedema in humans; and many of the same mutations in mice result in valve defects that subsequently lead to chylous ascites or chylothorax. At present the only experimental technique for the quantitative assessment of lymphatic valve function utilizes the servo-null micropressure system, which is highly accurate and precise, but relatively inaccessible and difficult to use. We developed a novel, simplified alternative method for quantifying valve function and determining the degree of pressure back-leak through an intact valve in pressurized, single-valve segments of isolated lymphatic vessels. With this diameter-based method, the competence of each lymphatic valve is challenged over a physiological range of pressures (e.g. 0.5-10cmH2 O) and pressure back-leak is extrapolated from calibrated, pressure-driven changes in diameter upstream from the valve. Using mesenteric lymphatic vessels from C57BL/6J, Ub-CreERT2 ;Rasa1fx/fx , Foxc2Cre/+ , Lyve1-Cre;Cx43fx/fx , and Prox1-CreERT2 ;Cx43fx/fx ;Cx37-/- mice, we tested our method on lymphatic valves displaying a wide range of dysfunction, from fully competent to completely incompetent. Our results were validated by simultaneous direct measurement of pressure back-leak using a servo-null micropressure system. Our diameter-based technique can be used to quantify valve function in isolated lymphatic valves from a variety of species. This method also revealed that haplodeficiency in Foxc2 (Foxc2Cre/+ ) is not sufficient to cause significant valve dysfunction; however, postnatal endothelial-specific deletion of Cx43 in Cx37-/- mice results in rapid regression of valve leaflets and severe valve dysfunction.

Mechanisms of Connexin-Related Lymphedema.

RATIONALE: Mutations in GJC2 and GJA1, encoding Cxs (connexins) 47 and 43, respectively, are linked to lymphedema, but the underlying mechanisms are unknown. Because efficient lymph transport relies on the coordinated contractions of lymphatic muscle cells (LMCs) and their electrical coupling through Cxs, Cx-related lymphedema is proposed to result from dyssynchronous contractions of lymphatic vessels. OBJECTIVE: To determine which Cx isoforms in LMCs and lymphatic endothelial cells are required for the entrainment of lymphatic contraction waves and efficient lymph transport. METHODS AND RESULTS: We developed novel methods to quantify the spatiotemporal entrainment of lymphatic contraction waves and used optogenetic techniques to analyze calcium signaling within and between the LMC and the lymphatic endothelial cell layers. Genetic deletion of the major lymphatic endothelial cell Cxs (Cx43, Cx47, or Cx37) revealed that none were necessary for the synchronization of the global calcium events that triggered propagating contraction waves. We identified Cx45 in human and mouse LMCs as the critical Cx mediating the conduction of pacemaking signals and entrained contractions. Smooth muscle-specific Cx45 deficiency resulted in 10- to 18-fold reduction in conduction speed, partial-to-severe loss of contractile coordination, and impaired lymph pump function ex vivo and in vivo. Cx45 deficiency resulted in profound inhibition of lymph transport in vivo, but only under an imposed gravitational load. CONCLUSIONS: Our results (1) identify Cx45 as the Cx isoform mediating the entrainment of the contraction waves in LMCs; (2) show that major endothelial Cxs are dispensable for the entrainment of contractions; (3) reveal a lack of coupling between lymphatic endothelial cells and LMCs, in contrast to arterioles; (4) point to lymphatic valve defects, rather than contraction dyssynchrony, as the mechanism underlying GJC2- or GJA1-related lymphedema; and (5) show that a gravitational load exacerbates lymphatic contractile defects in the intact mouse hindlimb, which is likely critical for the development of lymphedema in the adult mouse.

Enhanced Biosynthesis Synthesis of Copper Oxide Nanoparticles (CuO-NPs) for their Antifungal Activity Toxicity against Major Phyto-Pathogens of Apple Orchards.

PURPOSE: The present study made an attempt to develop copper nanoparticles (Cu-NP) with antifungal property using green synthesis method. Copper oxide nanoparticles (CuO-NPs) botanically synthesized using Neem leaf extract (Azadirachta indica A. Juss) were characterized by using different techniques like; UV-visible spectrophotometry, FTIR, XRD, SEM and TEM. METHODS: Materials were chosen the disease free and fresh Azadirachta indica A. Juss were collected and identified at Center of Biodiversity and Taxonomy. The plant samples were vigorously washed with distilled water then shade dried followed by sterilization with 0.1% mercuric chloride for 20 s and again it was washed with distilled water. 15 g powder form of plant material was added to 200 ml double distilled, CO2 free and deionized water and kept in shaker at 80°C and 1500 rpm for six hours. After agitation, the extract was separated by regular centrifugation at 10,000 rpm followed by filtration by using whatmann filter paper. The final volume of 100 ml of supernatant was collected as pure extract and stored in cool place for further use. RESULTS: The final results confirm a significant inhibition of CuO-NPs for the test fungi. Additionally, CuO-NPs demonstrated an enhanced effect when combined with Neem leaf extract. A total of 20-30% improvement in activity was noticed after combination, which correlates with commonly used synthetic fungicides. The toxicity results reveal that A. indica extract and their combined fractions with CuO-NP were less toxic to the test seeds of experimental plant while as bulk Cu followed by biosynthesized CuO-NPs influenced the germination rate as compared to control pots. CONCLUSIONS: The study drops a concern of research and offers a promising route of developing Copper based green fungicides that can help to combat with modern issues of synthetic fungicides. An average size of 80 ± 15 nm monoclinic cupric oxide (CuO) and cubic cuprous oxides (Cu2O) nanocrystals that existed in mixed form were successfully developed.

SWAT ungauged: Water quality modeling in the Upper Mississippi River Basin.

Improving model performance in ungauged basins has been a chronic challenge in watershed model application to understand and assess water quality impacts of agricultural conservation practices, land use change, and climate adaptation measures in large river basins. Here, we evaluate a modified version of SWAT2012 (referred to as SWAT-EC hereafter), which integrates an energy balanced soil temperature module (STM) and the CENTRUY-based soil organic matter algorithm, for simulating water quality parameters in the Upper Mississippi River Basin (UMRB), and compare it against the original SWAT2012. Model evaluation was performed for simulating streamflow, sediment, and nitrate-N (NO3-N) and total nitrogen (TN) loadings at three stations near the outlets of UMRB. The model comparison was conducted without parameter calibration in order to assess their performance under ungauged conditions. The results indicate that SWAT-EC outperformed SWAT2012 for stream flow and NO3-N and TN loading simulation on both monthly and annual scales. For sediment, SWAT-EC performed better than SWAT2012 on a monthly time step basis, but no noticeable improvement was found at the annual scale. In addition, the performance of the uncalibrated SWAT-EC was comparable to other calibrated SWAT models reported in previous publications with respect to sediment and NO3-N loadings. These findings highlight the importance of advancing process representation in physically-based models to improve model credibility, particularly in ungauged basins.