Elastic fiber formation involves the secretion of tropoelastin which is converted to insoluble elastin by cross-linking, initiated by the oxidative deamination of lysine residues by lysyl oxidase. ...Five lysyl oxidase genes have been discovered. This study deals with the expression of two isoforms, LOX and LOX-like (LOXL), in human foreskin and in a human skin-equivalent (SE) model that allows the formation of elastic fibers. In this model, keratinocytes are added to a dermal equivalent made of fibroblasts grown on a chitosan-cross-linked collagen-GAG matrix. LOX and LOXL were detected by immunohistochemistry in the dermis and the epidermis of both normal skin and in a SE. This expression was confirmed by in situ hybridization on the SE. LOX and LOXL expression patterns were confirmed in human skin. The ultrastructural localization of LOXL was indicative of its association with elastin-positive materials within the SE and human skin, though interaction with collagen could not be discarded. LOX was found on collagen fibers and could be associated with elastin-positive materials in the SE and human skin. LOXL and LOX were detected in keratinocytes where LOX was mainly expressed by differentiating keratinocytes, in contrast to LOXL that can be found in both proliferating and differentiating fibroblasts. These data favor a role for LOXL in elastic fiber formation, together with LOX, and within the epidermis where both enzymes should play a role in post-translational modification of yet unknown substrates.
Small cell lung cancer (SCLC) is an aggressive tumour with an abysmal prognosis. These cancers are characteristically resistant to glucocorticoid (Gc) action, owing to impaired expression of the ...glucocorticoid receptor (GR). We identified reduced GR expression in human SCLC cell lines, compared to a non-SCLC cell line. The SCLC cells also showed no Gc inhibition of proliferation, in contrast to non-SCLC cells. Retroviral overexpression of GR resulted in significantly increased cell death, which was partially blocked by the GR antagonist, RU486. Indeed, in cells sorted for GR expression, there was rapid, near complete loss of live cells by 72 h, in contrast to control cells that proliferated as expected. Flow cytometry using Annexin V revealed that cell death was by apoptosis. In addition, confocal analysis of fixed cells showed that cells overexpressing GR displayed a significant increase in fragmenting apoptotic nuclei. Microarray studies showed that transgenic GR expression upregulated the proapoptotic genes, BAD and BAX. We have, therefore, identified a profound apoptotic effect of GR in SCLC cells, which may explain the low levels of endogenous GR in SCLC cells. Understanding how GR overexpression leads to apoptotic cell death in SCLC cells may uncover new therapeutic strategies.
The purpose of this study was to identify synergistic effects in the interaction of light with biosystems in the presence of chemical agents. Their systematic analysis promises therapeutic ...strategies.
Light intensities around 1000 Wm(2) potentially induce density variations in nanoscopic water layers adhering to surfaces in air or subaquatically. In permeable nanoscopic compartments in the interior of biosystems, this could result in powerful flow processes and bidirectional flows for repetitive applications of light. Consequently, external stimulation with light will force microorganisms and cells to incorporate a suitable antiinfective. Nanoscale biosystems, which respond to both light stimulation and antibiotics, are nanobacteria. Responses include growth, inhibition, and slime secretion. Slime secretion was provoked in vitro by gentamycin, an agent proposed for in vivo eradication, and blocked by light. Depending on the field of action, co-operative effects between light and an antiinfective can be exploited by considering two properties of the drug: transmission of light and resorption by the tissue. Antiinfectives can be administered in an active form or via drug delivery systems. In the latter case, a double action of the light could be exploited: stimulated release from the carrier and subsequent uptake by the targeted biosystem.
The attenuation of laser light (670 nm) by antiinfectives was measured in films of different thickness of a vaginal suppository. The effect of 670-nm laser light - not absorbed by water - on nanoscopic water layers was examined by comparing the evaporation time of irradiated drops of water-based nanosuspensions with non-irradiated controls.
The 6-microm-thick suppository films were virtually transparent to the laser light, and the 1-mm-thick films totally attenuated it. Nanosuspension drops irradiated with 670-nm light needed more time to evaporate than controls.
Low-level light (LLL) therapy is compatible with antiinfectives, and even capable of boosting effects of superficially applied and/or absorbed antiinfectives. Temporal coordination between light treatment and drug administration maximizes drug effects and minimizes possible adverse effects. Irradiation should start when the drug concentration has reached its maximum in the desired field of action. Light-induced flow in nanoscale cavities could represent one mechanism of LLL therapy.
In the next 15–30 years, manned space flight to Mars, our planetary neighbour, will become a reality and astronauts are likely to spend at least 2–3 years away from Earth. Time spent in such extreme ...environments will result in a diminution of immune status and profound changes in the human bacterial microflora. In microgravity, the efficacy of antibiotics is reduced and microbial mutation rates increase dramatically. These factors will impinge on the capacity to treat effectively the infections that will doubtless arise during such long and stressful endeavour. We highlight new rationales for the treatment of infectious disease that may be applicable to therapy in extreme environments such as deep space.
Results on reactive atom plasma etching performed on ULE® (Corning Ultra Low Expansion) glass samples at atmospheric pressure are presented for the first time. A reactive atomic plasma technology ...(RAPT®), has been developed by RAPT Industries and employed for the finishing of optical surfaces. An atmospheric pressure argon inductively coupled plasma (ICP) excites a reactive gas injected through its centre. The plume of hot neutral excited species reacts at the substrate yielding controlled and repeatable trenches. In the case of ULE a material removal (up to 0.55 mm3/s) is obtained without pre‐heating the samples. Among the factors influencing the results, an increase in gas concentration at the same power does not change the sample temperature, indicating that thermo‐chemical effects do not influence the removal rates. Due to the plasma constructive constrains, increasing the gas concentration is more practical and of wider effect than increasing the power. The benefits of the process are illustrated and the extension of the technology to large optical surfaces discussed.
Reactive atom plasma etching (RAPT®, developed by RAPT Industries) performed on ULE® (Corning Ultra Low Expansion) glass samples at atmospheric pressure with an atmospheric pressure inductively coupled plasma (ICP). The argon plasma excites a reactive gas injected through its centre. The excited species reacts at the substrate yielding controlled and repeatable pseudo‐gaussian trenches. For ULE , material removal rates up to 0.55 mm3/s have been obtained.
To analyze the impact of gravity on the self-organization of nanospheres on substrates, drops of an aqueous suspension containing 60-nm polystyrene nanospheres were evaporated on different ...biomaterials. By reducing the speed of evaporation and by selecting different substrate positions, the interplay between gravitational force and pattern formation could be studied in detail. Results reveal limits for the gravitational force: for rings formed by nanosuspension droplets with diameters below 1 mm, the effect of normal gravitational force can be neglected.
Lysyl oxidase initiates the enzymatic stage of collagen and elastin cross-linking. Among five isoforms comprising the lysyl oxidase family, LOX is the better studied. LOX is associated to an ...antitumor activity in ras-transformed fibroblasts, and its expression is down-regulated in many carcinomas. The aim of this work was to shed light on LOX functions within the epidermis by studying its expression in human basal and squamous cell carcinomas and analyzing the effect of its enzymatic activity inhibition and protein absence on human keratinocytes behavior in a skin equivalent. In both carcinomas, LOX expression by epidermal tumor cells was lacking, while it was up-regulated around invading tumor cells in association with the stromal reaction. Lysyl oxidase activity inhibition using beta-aminoproprionitrile in a skin equivalent model prepared with both primary human keratinocytes and HaCaT cell line affected keratin 10 and filaggrin expression and disorganized the collagen network and the basement membrane. In spite of all these changes, no invasion phenotype was observed. Modelization of the invasive phenotype was only noticed in the skin equivalent developed with LOX antisense HaCaT cell line, where the protein LOX is specifically absent. Our results clearly indicate that lysyl oxidase enzymatic activity is essential not only for the integrity maintenance of the dermis but also for the homeostasis of the epidermis. Moreover, LOX protein plays a role in the skin carcinomas and invasion but not through its enzymatic activity.
Aims: The effect of oxygen on the survival of Torulaspora delbrueckii and Kluyveromyces thermotolerans during mixed culture fermentations in grape juice with Saccharomyces cerevisiae was ...investigated. Methods and Results: Fermentations were carried out in two simple fermentation systems differing in the availability of oxygen. At low available oxygen conditions, T. delbrueckii and K. thermotolerans began to die off after two days of mixed culture fermentation. In filtrates from 2-day-old mixed cultures, single cultures of T. delbrueckii and K. thermotolerans survived and actively produced ethanol to concentrations of approx. 65 and 70 g l(-1), respectively, at low available oxygen conditions. Oxygen clearly increased the survival time and decreased the death rate of T. delbrueckii and K. thermotolerans in mixed cultures, whereas it did not affect the growth and survival of S. cerevisiae. Conclusions: Our results show that the deaths of T. delbrueckii and K. thermotolerans in mixed cultures at low available oxygen conditions are not due to toxic metabolites produced by the yeasts but rather to the lack of oxygen. Furthermore, they indicate that T. delbrueckii and K. thermotolerans are less tolerant to low available oxygen conditions than S. cerevisiae. Significance and Impact of the Study: Our study reveals new knowledge on the mechanisms underlying the succession of yeasts during wine fermentations. This knowledge may be of importance when creating defined, mixed starter cultures for the controlled production of wines with a wide range of flavour compositions.
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