HLA-DR alpha 2 mediates negative signalling via binding to Tirc7 leading to anti-inflammatory and apoptotic effects in lymphocytes in vitro and in vivo

Classically, HLA-DR expressed on antigen presenting cells (APC) initiates lymphocyte activation via presentation of peptides to TCR bearing CD4+ T-Cells. Here we demonstrate that HLA-DR alpha 2 domain (sHLA-DRalpha2) also induces negative signals by engaging TIRC7 on lymphocytes. This interaction inhibits proliferation and induces apoptosis in CD4+ and CD8+ T-cells via activation of the intrinsic pathway. Proliferation inhibition is associated with SHP-1 recruitment by TIRC7, decreased phosphorylation of STAT4, TCR-zeta chain & ZAP70, and inhibition of IFN-gamma and FasL expression. HLA-DRalpha2 and TIRC7 co-localize at the APC-T cell interaction site. Triggering HLA-DR - TIRC7 pathway demonstrates that sHLA-DRalpha2 treatment inhibits proinflammatory-inflammatory cytokine expression in APC & T cells after lipopolysaccaride (LPS) stimulation in vitro and induces apoptosis in vivo. These results suggest a novel antiproliferative role for HLA-DR mediated via TIRC7, revise the notion of an exclusive stimulatory interaction of HLA-DR with CD4+ T cells and highlights a novel physiologically relevant regulatory pathway.     

Intravital microscopy of the murine pulmonary microcirculation.

Intravital microscopy (IVM) is considered as the gold standard for in vivo investigations of dynamic microvascular regulation. The availability of transgenic and knockout animals has propelled the development of murine IVM models for various organs, but technical approaches to the pulmonary microcirculation are still scarce. In anesthetized and ventilated BALB/c mice, we established a microscopic access to the surface of the right upper lung lobe by surgical excision of a window of 7- to 10-mm diameter from the right thoracic wall. The window was covered by a transparent polyvinylidene membrane and sealed with alpha-cyanoacrylate. Removal of intrathoracic air via a trans-diaphragmal intrapleural catheter coupled the lung surface to the window membrane. IVM preparations were hemodynamically stable for at least 120 min, with mean arterial blood pressure above 70 mmHg, and mean arterial Po(2) and arterial Pco(2) in the range of 90-100 Torr and 30-40 Torr, respectively. Imaged lungs did not show any signs of acute lung injury or edema. Following infusion of FITC dextran, subpleural pulmonary arterioles and venules of up to 50-microm diameter and alveolar capillary networks could be visualized during successive expiratory plateau phases over a period of at least 2 h. Vasoconstrictive responses to hypoxia (11% O(2)) or infusion of the thromboxane analog U-46619 were prominent in medium-sized arterioles (30- to 50-microm diameter), minor in small arterioles <30 microm, and absent in venules. The presented IVM model may constitute a powerful new tool for investigations of pulmonary microvascular responses in mice.     

Nonflowering plants possess a unique folate-dependent phenylalanine hydroxylase that is localized in chloroplasts

Tetrahydropterin-dependent aromatic amino acid hydroxylases (AAHs) are known from animals and microbes but not plants. A survey of genomes and ESTs revealed AAH-like sequences in gymnosperms, mosses, and algae. Analysis of full-length AAH cDNAs from Pinus taeda, Physcomitrella patens, and Chlamydomonas reinhardtii indicated that the encoded proteins form a distinct clade within the AAH family. These proteins were shown to have Phe hydroxylase activity by functional complementation of an Escherichia coli Tyr auxotroph and by enzyme assays. The P. taeda and P. patens AAHs were specific for Phe, required iron, showed Michaelian kinetics, and were active as monomers. Uniquely, they preferred 10-formyltetrahydrofolate to any physiological tetrahydropterin as cofactor and, consistent with preferring a folate cofactor, retained activity in complementation tests with tetrahydropterin-depleted E. coli host strains. Targeting assays in Arabidopsis thaliana mesophyll protoplasts using green fluorescent protein fusions, and import assays with purified Pisum sativum chloroplasts, indicated chloroplastic localization. Targeting assays further indicated that pterin-4a-carbinolamine dehydratase, which regenerates the AAH cofactor, is also chloroplastic. Ablating the single AAH gene in P. patens caused accumulation of Phe and caffeic acid esters. These data show that nonflowering plants have functional plastidial AAHs, establish an unprecedented electron donor role for a folate, and uncover a novel link between folate and aromatic metabolism.     

The lichens <Emphasis Type="Italic">Xanthoria elegans</Emphasis> and <Emphasis Type="Italic">Cetraria islandica</Emphasis> maintain a high protection against UV-B radiation in Arctic habitats

This study reports UV screening pigments in the upper cortices of two widespread lichens collected in three sun-exposed locations along a latitudinal gradient from the Arctic lowland to alpine sites of the Central European Alps. Populations from the Alps receive 3–5 times higher UV-B irradiance than their Arctic counterparts from Svalbard because of latitudinal and altitudinal gradients in UV-B irradiance. In Cetraria islandica, the screening capacity of melanin in the upper cortices was assessed by direct measurements of cortical transmittance (250–1,000 nm). A comparison of cortical transmittances in brown sun-exposed and pale shade-adapted forest C. islandica thalli showed that fungal melanins strongly absorb both UV-B and photosynthetically active radiation (PAR). For Xanthoria elegans cortical UV-B absorbing pigments, mainly the orange parietin, were extracted and quantified. Field experiments with extracted, parietin-deficient X. elegans thalli cultivated under various filters showed that UV-B was essential for the induction of parietin synthesis. The parietin resynthesis in these parietin-deficient samples increased with decreasing latitude of their location in which they had been sampled, which may imply that the synthesis of pigments is habitat specific. However, no latitudinal gradient in cortical screening capacity was detected for any of the two species investigated in the field. This implies that Arctic populations maintain a high level of screening pigments in spite of low ambient UV-B, and that the studied lichen species presumably may tolerate an increase in UV-B radiation due to the predicted thinning of the ozone layer over polar areas     

Increased activin levels in cerebrospinal fluid of rabbits with bacterial meningitis are associated with activation of microglia

Activin, a member of the transforming growth factor superfamily, is upregulated in a number of inflammatory episodes such as septicemia and rheumatoid arthritis. In the CNS, activin has been predominantly assessed in terms of a neuroprotective role. In this report we characterized the activin response in the CNS in a rabbit model of meningitis. In normal animals, cerebrospinal fluid (CSF) activin levels were higher than those in serum, indicating an intracranial secretion of this cytokine. Following intracisternal inoculation with Streptococcus pneumoniae, activin in CSF was unchanged for the first 12 h and then rose progressively; levels were increased approximately 15-fold within 24 h. Activin levels were correlated positively with CSF protein content and with the number of apoptotic neurons in the dentate gyrus. No apparent correlation was observed between CSF activin concentrations and bacterial titer, lactate concentrations or leukocyte density. Using immunohistochemistry, activin staining was localized to epithelial cells of the choroid plexus, cortical neurons and the CA3 region of the hippocampus, with similar staining intensities in both normal and meningitic brains. However, in meningitic brains there was also strong staining in activated microglia and infiltrating macrophages. Taken together, these results demonstrate that activin forms part of the CNS response to immune challenge and may be an important mediator to modulate inflammatory processes in the brain.     

The design and discovery of novel amide CCR5 antagonists

The synthesis of a range of novel amine-containing structures and their primary potency as inhibitors of HIV-1 fusion via blocking of the CCR5 receptor is described. The development of the medicinal chemistry strategy and SAR’s which led to the identification of the piperidine amide compounds 33 and 36 as excellent leads for further evaluation is described, along with key physicochemical data which highlighted their lead potential.     

Weichselian chironomid and cladoceran assemblages from maar lakes

Sediments from maar lakes in the periglacial area were analysed to obtain information about the limnetic fauna of Pleniglacial lakes. On the basis of abundance and species composition of the subfossil chironomid and cladoceran assemblages, three major zones were biostratigraphically separated: the Upper Pleniglacial, the Middle Pleniglacial, and the Eem Interglacial.     

Evidence that humidity influences snake activity patterns: a field study of the Malayan pit viper Calloselasma rhodostoma

Multivariate statistical methods were used to elucidate which environmental factors influence the activity patterns of free-living Malayan pit vipers, Calloselasma rhodostoma. Fourteen adult snakes were implanted with miniature radiotransmitters and located a total of 887 times in 5 months. The pit vipers usually remained coiled on the ground for several consecutive days before moving at night to a new site. Partial correlation tests revealed that the frequency and distance of movements to new sites by tagged snakes were highly positively correlated with ambient relative humidity, but not with rainfall, ambient temperature or the lunar cycle. This finding was corroborated by the frequency with which active non-tagged C. rhodostoma were encountered at night, In each site, the proportion of the snakes' bodies exposed to view was positively correlated with ambient humidity, and the snakes retreated to areas with deeper undergrowth when ambient humidity was low. Overt thermoregulatory behaviour was not observed, and implanted thermosensitive transmitters revealed that the snakes were passive thermoconformers.
These findings seem lo contradict much of the current literature which shows temperature to be the dominant abiotic factor affecting reptilian activity, but most herpetologists have considered only temperate forms. Ambient temperature in our tropical study site was warm and relatively constant throughout the year (mean daily range = 24- 33°C), so the pit vipers could passively maintain body temperature within a fairly narrow range, with a daytime mean of 29.4°C. Ambient relative humidity, on the other hand, was very variable, and confining exposure and activity to periods of high ambient humidity may be necessary to avoid dehydration