Glucocorticosteroids modify Langerhans cells to produce TGF-β and expand regulatory T cells

Although glucocorticosteroids (GCSs) have been used for many decades in transplantation and (auto)inflammatory diseases, the exact mechanisms responsible for their immunosuppressive properties are not fully understood. The purpose of this study was to characterize the effects of oral GCSs on the cutaneous immune response. We analyzed, by immunofluorescence staining and quantitative RT-PCR, residual skin biopsy material from a clinical study in which we had used oral GCS as positive control for determining the effects of candidate anti-inflammatory compounds on epicutaneous patch tests of Ni-allergic patients. Expectedly, oral GCS treatment led to a reduction of clinical symptoms and infiltrating leukocytes. Notably, we observed increased numbers of dermal FOXP3(+)CD25(+) T cells and epidermal Langerhans cells (LCs) that were associated with upregulated mRNA expression of TGF-β in lesions of GCS-treated Ni-allergic patients. To investigate this phenomenon further, we exposed purified LCs to GCS. They exhibited, in contrast to GCS-nonexposed LCs, 1) a more immature phenotype, 2) higher intracellular amounts of TGF-β, and 3) increased receptor activator for NF-κB expression, conditions that reportedly favor the expansion of regulatory T cells (Tregs). Indeed, we observed an enhancement of functionally suppressive FOXP3(+) T cells when CD3(+) cells were incubated with GCS-pretreated LCs. The expansion of Tregs was inhibited by TGF-β blockage alone, and their suppressive activity was neutralized by a combination of anti-TGF-β and anti-IL-10 Abs. Our data show that systemically applied GCSs endow LCs with Treg-promoting properties and thus shed new light on the mechanisms of GCS-mediated immunosuppression.     

IL-33 Induces IL-9 Production in Human CD4+ T Cells and Basophils

IL-33, an IL-1 family member and ligand for the IL-1 receptor-related protein ST2, has been associated with induction of Th2 cytokines such as IL-4, IL-5, and IL-13. Here, we report that IL-33 can initiate IL-9 protein secretion in vitro in human CD4+ T cells and basophils isolated from peripheral blood. TGF-β has been described as a critical factor for IL-9 induction in Th2 cells; however, we found that TGF-β also induces co-production of IL-9 in purified, naïve (>99%) CD4⁺CD45RA⁺CD45RO⁻CD25⁻ T cells differentiated towards a Th1 profile. Subsequently, it was demonstrated that TGF-β is important, although not an absolute requirement, for IL-9 production in CD4+ T cells. IL-9 production by purified (>95%) human basophils, cultured for 24 h with IL-3 or IL-33, was found, with a strong synergy between the two, likely to be explained by the IL-3 upregulated ST2 expression. Collectively, these data indicate that barrier functioning cells are important for the regulation of IL-9 production by immune cells in inflamed tissue.     

Vaginal irritation models: the current status of available alternative and in vitro tests

Mucosal surfaces, such as the vaginal epithelium, are natural barriers to infection that are constantly exposed to bacteria and viruses, and are therefore potential sites of entry for numerous pathogens. The vaginal epithelium can be damaged mechanically, e.g. by the incorrect use of objects such as tampons, and by chemicals that are irritating or corrosive. Consequently, this can lead to an increase in susceptibility to further damage or infection. Pharmaceutical, cosmetic and personal care products that are specifically formulated for application onto human external mucosae can occasionally induce undesirable local or systemic side-effects. Therefore, the compatibility of applied materials with this mucosal surface represents a key issue to be addressed by manufacturers. The most frequently used method for assessing vaginal mucosal irritation is the in vivo rabbit vaginal irritation test. However, the current emphasis in the field of toxicology is to use alternative in vitro methods that reduce, refine, and replace the use of animals, and which model and predict human, not animal, responses. Such an approach is of particular interest to the personal care and cosmetic industries in their effort to comply with European legislative measures, such as the 7th Amendment to the EU Cosmetics Directive that does not permit the marketing of cosmetic products if they, or their ingredients, have been tested for irritation responses in animals. The focus of this review is to provide an overview of the alternative and in vitro tests that are currently available for vaginal mucosal irritation assessment, and which are already used, or may become useful, to establish the safety of newly-designed products for human use.     

Telmisartan attenuates aortic hypertrophy in hypertensive rats by the modulation of ACE2 and profilin-1 expression

Profilin-1 has recently been linked to vascular hypertrophy and remodeling. Here, we assessed the hypothesis that angiotensin (Ang) II type I receptor antagonist telmisartan improves vascular hypertrophy by modulation of expression of profilin-1 and angiotensin-converting enzyme 2 (ACE2). Ten-week-old male spontaneously hypertensive rats (SHR) were received oral administration of telmisartan (5 or 10 mg/kg; daily) or saline for 10 weeks. Compared with Wistar–Kyoto (WKY) rats, there were marked increases in systolic blood pressure and profilin-1 expression and reduced ACE2 and peroxisome proliferator activated receptor-γ (PPARγ) levels in aorta of SHR, associated with elevated extracellular-signal regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK) phosphorylation signaling and aortic hypertrophy characterized with increased media thickness, which were strikingly reversed by telmisartan. In cultured human umbilical artery smooth muscle cells (HUASMCs), Ang II induced a dose-dependent increase in profilin-1 expression, along with decreased ACE2 protein expression and elevated ERK1/2 and JNK phosphorylation. In addition, blockade of ERK1/2 or JNK by either specific inhibitor was able to abolish Ang II-induced ACE2 downregulation and profilin-1 upregulation in HUASMCs. Importantly, treatment with telmisartan (1 or 10 μM) or recombinant human ACE2 (2 mg/ml) largely ameliorated Ang II-induced profilin-1 expression and ERK1/2 and JNK phosphorylation and augmented PPARγ ?expression in the cultured HUASMCs. In conclusion, telmisartan treatment attenuates vascular hypertrophy in SHR by the modulation of ACE2 and profilin-1 expression with a marked reversal of ERK1/2 and JNK phosphorylation signaling pathways.     

Breast tumor cells with PI3K mutation or HER2 amplification are selectively addicted to Akt signaling

Background

Dysregulated PI3K/Akt signaling occurs commonly in breast cancers and is due to HER2 amplification, PI3K mutation or PTEN inactivation. The objective of this study was to determine the role of Akt activation in breast cancer as a function of mechanism of activation and whether inhibition of Akt signaling is a feasible approach to therapy.

Methodology/Principal Findings

A selective allosteric inhibitor of Akt kinase was used to interrogate a panel of breast cancer cell lines characterized for genetic lesions that activate PI3K/Akt signaling: HER2 amplification or PI3K or PTEN mutations in order to determine the biochemical and biologic consequences of inhibition of this pathway. A variety of molecular techniques and tissue culture and in vivo xenograft models revealed that tumors with mutational activation of Akt signaling were selectively dependent on the pathway. In sensitive cells, pathway inhibition resulted in D-cyclin loss, G1 arrest and induction of apoptosis, whereas cells without pathway activation were unaffected. Most importantly, the drug effectively inhibited Akt kinase and its downstream effectors in vivo and caused complete suppression of the growth of breast cancer xenografts with PI3K mutation or HER2 amplification, including models of the latter selected for resistance to Herceptin. Furthermore, chronic administration of the drug was well-tolerated, causing only transient hyperglycemia without gross toxicity to the host despite the pleiotropic normal functions of Akt.

Conclusions/Significance

These data demonstrate that breast cancers with PI3K mutation or HER2 amplification are selectively dependent on Akt signaling, and that effective inhibition of Akt in tumors is feasible and effective in vivo. These findings suggest that direct inhibition of Akt may represent a therapeutic strategy for breast and other cancers that are addicted to the pathway including tumors with resistant to Herceptin.     

Taxonomic effect on plant base concentrations and stoichiometry at the tips of the phylogeny prevails over environmental effect along a large scale gradient

Despite the well‐known importance of all elements to plant growth and nutrient fluxes in ecosystems, most studies to date have been restricted to the roles of foliar nitrogen (N) and phosphorus (P). Much less is known about cycling and pools of base cations in ecosystems and the drivers of variation in cation concentrations among plant species, even though these cations are paramount for plant and ecosystem function. In particular, little is known about the contributions of taxonomic position and environmental variation on base cation concentrations. The extent to which concentrations of elements in plants are determined by phenotypic response to their availability in current environments versus by inherent species‐specific uptake and processing adaptations, should be most directly evident at the tips of the phylogeny, where inherent variation among species should reflect relatively recent adaptation to environmental variation since their common ancestry. To test this hypothesis, we explored the geographic pattern and the effects of taxonomy, climate and soil on concentrations and stoichiometry of the base cations potassium (K), sodium (Na), calcium (Ca) and magnesium (Mg) across a lineage of Artemisia species and their close relatives across northern China. We found that species identity explained the largest proportion of the total variance for all four base cations (38.3–53.8%) and their stoichiometry (35.2–59.6%). K, Na and Ca concentrations increased significantly with climate seasonality, while Ca concentration decreased with annual temperature and precipitation. Plant K concentration, K:Ca and K:Mg were negatively correlated with soil organic carbon concentrations, but positively with soil pH. Our results suggest that taxonomy still needs to be fully considered for interpreting variation in vegetation nutrition and stoichiometry along broad geographical gradients even for species at the tips of the phylogeny.     

Über die Bildung von Bakterienhemmstoffen durch Cosmarium impressulum

1. An already published test method for detecting bactericidal substances in paper chromatograms was further improved. 2. In cultures of Cosmarium impressulum free from bacteria, two bactericidal substances were found in the ether extracts from the algae and two others in the extracts of the culture medium. The are active against some or all bacteria testes (Serratia marcescens, Pseudomonas fluorescens; Aerobacter aerogenes or Bacillus pumilus). 3. If the culture medium of Cosmarium or another desmidiale was inoculated with the test bacteria, a clear bactericidal effect was never observed. 4. Because the activity of the bactericidal substances of Cosmarium is only small and the formation is not constant, it is concluded that the water-soluble bactericidal substances of the alga are not the cause that epiphytic bacteria do not grow normally on Cosmarium.     

Physiological ecology meets climate change

In this article, we pointed out that understanding the physiology of differential climate change effects on organisms is one of the many urgent challenges faced in ecology and evolutionary biology. We explore how physiological ecology can contribute to a holistic view of climate change impacts on organisms and ecosystems and their evolutionary responses. We suggest that theoretical and experimental efforts not only need to improve our understanding of thermal limits to organisms, but also to consider multiple stressors both on land and in the oceans. As an example, we discuss recent efforts to understand the effects of various global change drivers on aquatic ectotherms in the field that led to the development of the concept of oxygen and capacity limited thermal tolerance (OCLTT) as a framework integrating various drivers and linking organisational levels from ecosystem to organism, tissue, cell, and molecules. We suggest seven core objectives of a comprehensive research program comprising the interplay among physiological, ecological, and evolutionary approaches for both aquatic and terrestrial organisms. While studies of individual aspects are already underway in many laboratories worldwide, integration of these findings into conceptual frameworks is needed not only within one organism group such as animals but also across organism domains such as Archaea, Bacteria, and Eukarya. Indeed, development of unifying concepts is relevant for interpreting existing and future findings in a coherent way and for projecting the future ecological and evolutionary effects of climate change on functional biodiversity. We also suggest that OCLTT may in the end and from an evolutionary point of view, be able to explain the limited thermal tolerance of metazoans when compared to other organisms.     

Phytochrom-induzierte Enzymbildung (Phenylalanindesaminase), ein schnell ablaufender Prozeß

Zusammenfassung Bei Erstbelichtung des Senfkeimlings mit Dauer-Dunkelrot tritt der P₇₃₀-abhängige Anstieg der Phenylalanindesaminase-Aktivität erst mit einer lag-Phase von 1,5 Std ein (Abb. 3, 4 unten). Bei einer Zweitbelichtung (Programm: Erstbelichtung — längere Dunkelperiode — Zweitbelichtung) fehlt die lag-Phase (Abb. 4, oben). Die Enzymaktivität steigt sofort linear an. Da der Anstieg der Enzymaktivität wahrscheinlich auf eine de novo Synthese von RNS und Enzymprotein zurückzuführen ist (Tabelle), so erscheint der Schluß berechtigt, daß P₇₃₀ sehr rasch eine differentielle Genaktivierung mit anschließender Enzymsynthese bewirken kann, falls die Gene der Aktivierung durch P₇₃₀ zugänglich sind. Die relativ lange lag-Phase nach Einsetzen der Erstbelichtung benötigt das P₇₃₀ offenbar dazu, die potentiell aktiven Gene (P₇₃₀) für das P₇₃₀ zugänglich zu machen. Das Problem der primären lag-Phase ist in einer vorangegangenen Arbeit zur P₇₃₀-abhängigen Anthocyansynthese ausführlich diskutiert worden (vgl. Lange, Bienger und Mohr, 1967).

---

Phytochrome-mediated enzyme formation (Phenylalanine deaminase) as a rapid process

Summary In previous papers we have reported (Mohr and Durst, 1966a, b) that synthesis of phenylalanine deaminase (EC 4.3.1.5), an important enzyme of phenolic metabolism, can be stimulated by the physiologically active phytochrome (=P₇₃₀) in the mustard seedling. The data of the present paper suggest that induction of this enzyme is a rapid process if the gene in question is easily accessible for the activating action of P₇₃₀.The seedlings were irradiated with continuous standard far-red light. Longtime irradiation with far-red will maintain a low but virtually constant level of P₇₃₀ in the seedling over an extended period of time. At the moment when the far-red light is turned off the action of P₇₃₀ will virtually cease. — Fig. 3 and Fig. 4, lower part, show the kinetics of enzyme induction by P₇₃₀ in an etiolated seedling. The initial (or primary) lag-phase after the onset of far-red is 1.5 hours. If, however, a seedling which has been pre-irradiated with 12 hours of far-red is kept in darkness for 6 hours and is then re-irradiated with far-red no lag-phase for the action of the second irradiation can be found. Enzyme activity increases immediately after the onset of far-red. Since the action of the second irradiation as measured by increase of enzyme activity can be inhibited by relatively low doses of Puromycin and Cycloheximide (table) we conclude that the re-appearance of P₇₃₀ leads to de novo synthesis of enzyme protein. — Application of Actinomycin D (10 g/ml) only partially inhibits the action of the second irradiation as measured by increase of enzyme activity. This finding was to be expected. In preceding papers (e.g. Mohr and Bienger, 1967) it has been concluded that genes which have once been activated by P₇₃₀ remain less sensitive towards Actinomycin D even when P₇₃₀ has disappeared. Taking into account all available data the conclusion seems to be justified that the induction of enzyme synthesis by P₇₃₀ (i.e. differential gene activation followed by enzyme synthesis) is a rapid process if the genes are accessible for the action of P₇₃₀. The relatively long initial lag-phase (1.5 hours) is needed to make the potentially active genes (P₇₃₀) accessible for the action of P₇₃₀. The problem of how the initial lag-phase can be understood has been dealt with more in detail in a previous paper on phytochrome-mediated anthocyanin synthesis (Lange, Bienger and Mohr, 1967).