T-PHYLLOPLANIN AND \u3ci\u3eCIS\u3c/i\u3e-ABIENOL, TWO NATURAL PRODUCTS FROM TOBACCO HAVE BROAD SPECTRUM, ANTI-FUNGAL ACTIVITIES

Tobacco phylloplanins (T-phylloplanin) are a group of closely-related glycoproteins that are formed and disposed at the interface between the plant aerial surface (the phylloplane) and the atmosphere. They are synthesized in short procumbent trichomes and are secreted to aerial surfaces where they are thought to serve the plant as a first line of defense against fungal pathogens. Here it is shown using in vitro and in planta assays that tobacco and sunflower phylloplanins have broad-spectrum antifungal activities against spores - and also hyphae for two species - of several true fungi. Field tests show that T-phylloplanin reduces diseases caused by three important fungal pathogens of turf grasses. Tobacco phylloplanins are distinct proteins but they have properties in common with small, membrane-pore-forming, antimicrobial peptides formed by other organisms. To directly determine if T-phylloplanin has pore-forming activity we monitored conductivity change and specific ion leakage from spores and hyphae in suspension. Results indicate that phylloplanin causes fungal membrane disruption that leads to ion depletion and cell death. Having observed broad efficacy of T-phylloplanin against spores and/or hyphae of several true fungi, but no activity towards hyphae of the oomycetes, Pythium and Peronospora parasitica, we tested for possible effects on zoospores of the latter two pathogens. T-phylloplanin was shown to be effective against their zoospores, extending the efficacy of T-phylloplanin to include water molds. In the course of these experiments we also tested the effects of the diterpene cis-abienol that is secreted from tall trichomes of tobaccos and found this compound impacted zoospores and could prevent black shank disease caused by P. parasitica when applied to soil-grown tobaccos as a root drench. Thus, results of these studies with phylloplanins and cis-abienol, two different tobacco surface accumulated compounds are consistent with their serving the plant as first line of defense systems against a wide array of invading fungal pathogens. Phylloplanins and cis-abienol may be useful for controlling fungal diseases in tobacco float beds. The efficacy shown here for T-phylloplanin control of fungal pathogens of turf grasses in the field suggests that this natural product may find use in IPM of turf and other crops

A 3D-printed microfluidic-enabled hollow microneedle architecture for transdermal drug delivery.

Embedding microfluidic architectures with microneedles enables fluid management capabilities that present new degrees of freedom for transdermal drug delivery. To this end, fabrication schemes that can simultaneously create and integrate complex millimeter/centimeter-long microfluidic structures and micrometer-scale microneedle features are necessary. Accordingly, three-dimensional (3D) printing techniques are suitable candidates because they allow the rapid realization of customizable yet intricate microfluidic and microneedle features. However, previously reported 3D-printing approaches utilized costly instrumentation that lacked the desired versatility to print both features in a single step and the throughput to render components within distinct length-scales. Here, for the first time in literature, we devise a fabrication scheme to create hollow microneedles interfaced with microfluidic structures in a single step. Our method utilizes stereolithography 3D-printing and pushes its boundaries (achieving print resolutions below the full width half maximum laser spot size resolution) to create complex architectures with lower cost and higher print speed and throughput than previously reported methods. To demonstrate a potential application, a microfluidic-enabled microneedle architecture was printed to render hydrodynamic mixing and transdermal drug delivery within a single device. The presented architectures can be adopted in future biomedical devices to facilitate new modes of operations for transdermal drug delivery applications such as combinational therapy for preclinical testing of biologic treatments

Stable heterologous expression of biologically active terpenoids in green plant cells

Plants biosynthesize a great diversity of biologically active small molecules of interest for fragrances, flavours, and pharmaceuticals. Among specialized metabolites, terpenoids represent the greatest molecular diversity. Many terpenoids are very complex, and total chemical synthesis often requires many steps and difficult chemical reactions, resulting in a low final yield or incorrect stereochemistry. Several drug candidates with terpene skeletons are difficult to obtain by chemical synthesis due to their large number of chiral centres. Thus, biological production remains the preferred method for industrial production for many of these compounds. However because these chemicals are often found in low abundance in the native plant, or are produced in plants which are difficult to cultivate, there is great interest in engineering increased production or expression of the biosynthetic pathways in heterologous hosts. Although there are many examples of successful engineering of microbes such as yeast or bacteria to produce these compounds, this often requires extensive changes to the host organism's metabolism. Optimization of plant gene expression, post-translational protein modifications, subcellular localization, and other factors often present challenges. To address the future demand for natural products used as drugs, new platforms are being established that are better suited for heterologous production of plant metabolites. Specifically, direct metabolic engineering of plants can provide effective heterologous expression for production of valuable plant-derived natural products. In this review, our primary focus is on small terpenoids and we discuss the benefits of plant expression platforms and provide several successful examples of stable production of small terpenoids in plants

Religious Identity, Religious Attendance, and Parental Control

Using a national sample of adolescents aged 10–18 years and their parents (N = 5,117), this article examines whether parental religious identity and religious participation are associated with the ways in which parents control their children. We hypothesize that both religious orthodoxy and weekly religious attendance are related to heightened levels of three elements of parental control: monitoring activities, normative regulations, and network closure. Results indicate that an orthodox religious identity for Catholic and Protestant parents and higher levels of religious attendance for parents as a whole are associated with increases in monitoring activities and normative regulations of American adolescents

Halo Structures of Gravitational Lens Galaxies

We explore the halo structure of four gravitational lenses with well-observed, thin Einstein rings. We find that the gravitational potentials are well described by ellipsoidal density distributions in the sense that the best-fit nonellipsoidal models have parameters consistent with their ellipsoidal counterparts. We find upper limits on the standard parameters for the deviation from an ellipse of |a_3/a_0|<0.023, 0.019, 0.037, and 0.035, and |a_4/a_0|<0.034, 0.041, 0.051, and 0.064 for SDSS J0924+0219, HE0435-1223, B1938+666, and PG1115+080, respectively. We find that the lens galaxies are at the centers of their dark matter halos, and obtain upper limits for the offset of each center of mass from the center of light of |Delta x|<0.004, 0.005, 0.009, and 0.005 arcsec, corresponding to 22, 29, 70, and 23 pc. These limits also exclude the possibility of any significant lopsidednessof the dark matter halos and set an upper limit of f_sat<sqrt(N)% on the mass fraction of massive substructures inside the Einstein ring if they are divided over N satellites. We also explore the properties of galaxies as substructures in groups for the lens PG1115+080, finding evidence for dark matter halos associated with the galaxies but no evidence for a clear distinction between satellite and central galaxies.Comment: 10 pages, 5 figures, accepted for publication in The Astrophysical Journa

Microglia regulate blood clearance in subarachnoid hemorrhage by heme oxygenase-1

Subarachnoid hemorrhage (SAH) carries a 50% mortality rate. The extravasated erythrocytes that surround the brain contain heme, which, when released from damaged red blood cells, functions as a potent danger molecule that induces sterile tissue injury and organ dysfunction. Free heme is metabolized by heme oxygenase (HO), resulting in the generation of carbon monoxide (CO), a bioactive gas with potent immunomodulatory capabilities. Here, using a murine model of SAH, we demonstrated that expression of the inducible HO isoform (HO-1, encoded by Hmox1) in microglia is necessary to attenuate neuronal cell death, vasospasm, impaired cognitive function, and clearance of cerebral blood burden. Initiation of CO inhalation after SAH rescued the absence of microglial HO-1 and reduced injury by enhancing erythrophagocytosis. Evaluation of correlative human data revealed that patients with SAH have markedly higher HO-1 activity in cerebrospinal fluid (CSF) compared with that in patients with unruptured cerebral aneurysms. Furthermore, cisternal hematoma volume correlated with HO-1 activity and cytokine expression in the CSF of these patients. Collectively, we found that microglial HO-1 and the generation of CO are essential for effective elimination of blood and heme after SAH that otherwise leads to neuronal injury and cognitive dysfunction. Administration of CO may have potential as a therapeutic modality in patients with ruptured cerebral aneurysms