Regional variations in ABC transporter expression along the mouse intestinal tract.

The ATP-binding cassette (ABC) family of proteins comprise a group of membrane transporters involved in the transport of a wide variety of compounds, such as xenobiotics, vitamins, lipids, amino acids, and carbohydrates. Determining their regional expression patterns along the intestinal tract will further characterize their transport functions in the gut. The mRNA expression levels of murine ABC transporters in the duodenum, jejunum, ileum, and colon were examined using the Affymetrix MuU74v2 GeneChip set. Eight ABC transporters (Abcb2, Abcb3, Abcb9, Abcc3, Abcc6, Abcd1, Abcg5, and Abcg8) displayed significant differential gene expression along the intestinal tract, as determined by two statistical models (a global error assessment model and a classic ANOVA, both with a P < 0.01). Concordance with semiquantitative real-time PCR was high. Analyzing the promoters of the differentially expressed ABC transporters did not identify common transcriptional motifs between family members or with other genes; however, the expression profile for Abcb9 was highly correlated with fibulin-1, and both genes share a common complex promoter model involving the NFkappaB, zinc binding protein factor (ZBPF), GC-box factors SP1/GC (SP1F), and early growth response factor (EGRF) transcription binding motifs. The cellular location of another of the differentially expressed ABC transporters, Abcc3, was examined by immunohistochemistry. Staining revealed that the protein is consistently expressed in the basolateral compartment of enterocytes along the anterior-posterior axis of the intestine. Furthermore, the intensity of the staining pattern is concordant with the expression profile. This agrees with previous findings in which the mRNA, protein, and transport function of Abcc3 were increased in the rat distal intestine. These data reveal regional differences in gene expression profiles along the intestinal tract and demonstrate that a complete understanding of intestinal ABC transporter function can only be achieved by examining the physiologically distinct regions of the gut.

Regional and cellular patterns of reelin mRNA expression in the forebrain of the developing and adult mouse

The reelin gene encodes an extracellular protein that is crucial for neuronal migration in laminated brain regions. To gain insights into the functions of Reelin, we performed high-resolution in situ hybridization analyses to determine the pattern of reelin expression in the developing forebrain of the mouse. We also performed double-labeling studies with several markers, including calcium-binding proteins, GAD65/67, and neuropeptides, to characterize the neuronal subsets that express reelin transcripts. reelinexpression was detected at embryonic day 10 and later in the forebrain, with a distribution that is consistent with the prosomeric model of forebrain regionalization. In the diencephalon, expression was restricted to transverse and longitudinal domains that delineated boundaries between neuromeres. During embryogenesis,reelin was detected in the cerebral cortex in Cajal-Retzius cells but not in the GABAergic neurons of layer I. At prenatal stages, reelin was also expressed in the olfactory bulb, and striatum and in restricted nuclei in the ventral telencephalon, hypothalamus, thalamus, and pretectum. At postnatal stages, reelin transcripts gradually disappeared from Cajal-Retzius cells, at the same time as they appeared in subsets of GABAergic neurons distributed throughout neocortical and hippocampal layers. In other telencephalic and diencephalic regions,reelin expression decreased steadily during the postnatal period. In the adult, there was prominent expression in the olfactory bulb and cerebral cortex, where it was restricted to subsets of GABAergic interneurons that co-expressed calbindin, calretinin, neuropeptide Y, and somatostatin. This complex pattern of cellular and regional expression is consistent with Reelin having multiple roles in brain development and adult brain function.

Effects of overinflation on procollagen type III expression in experimental acute lung injury

Abstract Introduction In acute lung injury (ALI), elevation of procollagen type III (PC III) occurs early and has an adverse impact on outcome. We examined whether different high-inflation strategies of mechanical ventilation (MV) in oleic acid (OA) ALI alter regional expression of PC III. Methods We designed an experimental, randomized, and controlled protocol in which rats were allocated to two control groups (no injury, recruited [alveolar recruitment maneuver after tracheotomy without MV; n = 4 rats] and control [n = 5 rats]) or four injured groups (one exposed to OA only [n = 10 rats] and three OA-injured and ventilated). The three OA-injured groups were ventilated for 1 hour according to the following strategies: LVHP-S (low volume-high positive end-expiratory pressure [PEEP], supine; n = 10 rats, tidal volume [VT] = 8 ml/kg, PEEP = 12 cm H2O), HVLP-S (high volume-low PEEP, supine; n = 10 rats, VT = 20 ml/kg, PEEP = 5 cm H2O), and HVLP-P (high volume-low PEEP, prone; n = 10 rats). Northern blot analysis for PC III and interleukin-1-beta (IL-1β) and polymorphonuclear infiltration index (PMI) counting were performed in nondependent and dependent regions. Regional differences between groups were assessed by two-way analysis of variance after logarithmic transformation and post hoc tests. Results A significant interaction for group and region effects was observed for PC III (p = 0.012) with higher expression in the nondependent region for HVLP-S and LVHP-S, intermediate for OA and HVLP-P, and lower for control (group effect, p < 0.00001, partial η2 = 0.767; region effect, p = 0.0007, partial η2 = 0.091). We found high expression of IL-1β (group effect, p < 0.00001, partial η2 = 0.944) in the OA, HVLP-S, and HVLP-P groups without regional differences (p = 0.16). PMI behaved similarly (group effect, p < 0.00001, partial η2 = 0.832). Conclusion PC III expression is higher in nondependent regions and in ventilatory strategies that caused overdistension. This response was partially attenuated by prone positioning.

Proprotein convertases in amphioxus: predicted structure and expression of proteases SPC2 and SPC3.

SPC2 and SPC3 are two members of a family of subtilisin-related proteases which play essential roles in the processing of prohormones into their mature forms in the pancreatic B cell and many other neuroendocrine cells. To investigate the phylogenetic origins and evolutionary functions of SPC2 and SPC3 we have identified and cloned cDNAs encoding these enzymes from amphioxus (Branchiostoma californiensis), a primitive chordate. The amino acid sequence of preproSPC2 contains 689 aa and is 71% identical to human SPC2. In contrast, amphioxus prproSPC3 consists of 774 aa and exhibits 55% identity to human SPC3. These results suggest that the primary structure of SPC2 has been more highly conserved during evolution than that of SPC3. To further investigate the function(s) of SPC2 and SPC3 in amphioxus, we have determined the regional expression of these genes by using a reverse transcriptase-linked polymerase chain reaction (RT-PCR) assay. Whole amphioxus was dissected longitudinally into four equal-length segments and RNA was extracted. Using RT-PCR to simultaneously amplify SPC2 and SPC3 DNA fragments, we found that the cranial region (section 1) expressed equal amounts of SPC2 and SPC3 mRNAs, whereas in the caudal region (section 4) the SPC2-to-SPC3 ratio was 5:1. In the mid-body sections 2 and 3 the SPC2-to-SPC3 ratio was 1:5. By RT-PCR we also determined that amphioxus ILP, a homologue of mammalian insulin/insulin-like growth factor, was expressed predominately in section 3. These results suggest that the relative levels of SPC2 and SPC3 mRNAs are specifically regulated in various amphioxus tissues. Furthermore, the ubiquitous expression of these mRNAs in the organism indicates that they are involved in the processing of other precursor proteins in addition to proILP.

Pax3 expression enhances PDGF-B-induced brainstem gliomagenesis and characterizes a subset of brainstem glioma.

High-grade Brainstem Glioma (BSG), also known as Diffuse Intrinsic Pontine Glioma (DIPG), is an incurable pediatric brain cancer. Increasing evidence supports the existence of regional differences in gliomagenesis such that BSG is considered a distinct disease from glioma of the cerebral cortex (CG). In an effort to elucidate unique characteristics of BSG, we conducted expression analysis of mouse PDGF-B-driven BSG and CG initiated in Nestin progenitor cells and identified a short list of expression changes specific to the brainstem gliomagenesis process, including abnormal upregulation of paired box 3 (Pax3). In the neonatal mouse brain, Pax3 expression marks a subset of brainstem progenitor cells, while it is absent from the cerebral cortex, mirroring its regional expression in glioma. Ectopic expression of Pax3 in normal brainstem progenitors in vitro shows that Pax3 inhibits apoptosis. Pax3-induced inhibition of apoptosis is p53-dependent, however, and in the absence of p53, Pax3 promotes proliferation of brainstem progenitors. In vivo, Pax3 enhances PDGF-B-driven gliomagenesis by shortening tumor latency and increasing tumor penetrance and grade, in a region-specific manner, while loss of Pax3 function extends survival of PDGF-B-driven;p53-deficient BSG-bearing mice by 33%. Importantly, Pax3 is regionally expressed in human glioma as well, with high PAX3 mRNA characterizing 40% of human BSG, revealing a subset of tumors that significantly associates with PDGFRA alterations, amplifications of cell cycle regulatory genes, and is exclusive of ACVR1 mutations. Collectively, these data suggest that regional Pax3 expression not only marks a novel subset of BSG but also contributes to PDGF-B-induced brainstem gliomagenesis.

Intrusao alcalina de Jaboticabal, SP.

Five magnetic anomalies were found and mapped in the Faculdade de Ciencias Agrarias e Veterinarias, UNESP, campus of Jaboticabal, SP. The study showed that those anomalies represent intrusive alkaline bodies linked to magmatic manifestation with regional expression. -English summary

Luis Arana e i veterani di Euzkeldun Batzokija: la corrente ortodossa del nazionalismo basco

La tesi di Marco Perez intitolata “Luis Arana e i veterani di Euzkeldun Batzokija: la corrente ortodossa del nazionalismo basco”, può essere considerata come la biografia politica di uno dei personaggi più importanti del nazionalismo basco. Il lavoro di ricerca si centra fondamentalmente sull'ispiratore del nazionalismo euskaldun (e cofondatore del Partido Nacionalista Vasco) e della corrente che ne accompagnò e sostenne l'azione politica. Euzkeldun Batzokija fu il nome dato al primo circolo del PNV, fondato da Luis e Sabino Arana nel 1894. Successivamente, gli statuti del circolo e i suoi membri veterani furono presi come modello del nazionalismo primordiale (che si pretendeva definire sull'esempio dell'Ordine gesuita). Sul piano organizzativo la tesi si divide in sette capitoli che ricostruiscono il percorso politico di Luis Arana, dai primi documenti del 1879 fino alle ultime lettere inviate negli anni quaranta. Si tratta di un lungo periodo, che comprende momenti diversi della storia spagnola (dalle guerre carliste alla Guerra Civile spagnola) e del movimento aranista. In questo senso, sulla base di una generale e comparata riflessione sul nazionalismo, si analizza il movimento basco nei suoi rapporti con la modernità. Una realazione costruita attraverso concetti “diacronicamente” legati a un passato mitico e leggendario e comunque subalterna ai rapporti di forza tra le correnti del PNV. La corrente ortodossa fece sempre riferimento al nazionalismo “originario” (definito dai fratelli Arana nei primi anni del movimento) che fu un'espressione regionale del nazionalcattolicesimo spagnolo. Fu proprio Luis Arana a ricordare la finalità religiosa ed etnica del nazionalismo basco, respingendo qualsiasi aggiornamento teorico e organizzativo del PNV, intesi come una grave violazione dell'ortodossia aranista.

Continental-scale temperature variability during the past two millennia

Past global climate changes had strong regional expression. To elucidate their spatio-temporal pattern, we reconstructed past temperatures for seven continental-scale regions during the past one to two millennia. The most coherent feature in nearly all of the regional temperature reconstructions is a long-term cooling trend, which ended late in the nineteenth century. At multi-decadal to centennial scales, temperature variability shows distinctly different regional patterns, with more similarity within each hemisphere than between them. There were no globally synchronous multi-decadal warm or cold intervals that define a worldwide Medieval Warm Period or Little Ice Age, but all reconstructions show generally cold conditions between ad 1580 and 1880, punctuated in some regions by warm decades during the eighteenth century. The transition to these colder conditions occurred earlier in the Arctic, Europe and Asia than in North America or the Southern Hemisphere regions. Recent warming reversed the long-term cooling; during the period ad 1971–2000, the area-weighted average reconstructed temperature was higher than any other time in nearly 1,400 years.

Late memory-related genes in the hippocampus revealed by RNA fingerprinting

Although long-term memory is thought to require a cellular program of gene expression and increased protein synthesis, the identity of proteins critical for associative memory is largely unknown. We used RNA fingerprinting to identify candidate memory-related genes (MRGs), which were up-regulated in the hippocampus of water maze-trained rats, a brain area that is critically involved in spatial learning. Two of the original 10 candidate genes implicated by RNA fingerprinting, the rat homolog of the ryanodine receptor type-2 and glutamate dehydrogenase (EC 1.4.1.3), were further investigated by Northern blot analysis, reverse transcription–PCR, and in situ hybridization and confirmed as MRGs with distinct temporal and regional expression. Successive RNA screening as illustrated here may help to reveal a spectrum of MRGs as they appear in distinct domains of memory storage.

Calcium sensing receptor: molecular cloning in rat and localization to nerve terminals.

We have molecularly cloned a calcium sensing receptor (CaSR) from a rat striatal cDNA library. Rat CaSR displays 92% overall homology to its bovine counterpart with seven putative transmembrane domains characteristic of the superfamily of guanine nucleotide-binding proteins and significant homology with the metabotropic glutamate receptors. Northern blot analysis reveals two transcripts in thyroid, kidney, lung, ileum, and pituitary. In brain highest regional expression of the RNA occurs in the hypothalamus and the corpus striatum. Immunohistochemistry reveals discrete punctate localizations throughout the brain that appear to be associated with nerve terminals. No staining is evident in cell bodies of neurons or glia. Cerebral arteries display an intense network of CaSR immunoreactive fibers associated with vessel innervation. CaSR on nerve terminal membranes may regulate neurotransmitter disposition in response to Ca2+ levels in the synaptic space.

Selective loss of NMDA receptor NR1 subunit isoforms in Alzheimer's disease

Previous work had shown that the ratio of NMDA receptor NR1 subunit mRNA transcripts containing an N-terminal splice cassette to those that do not is markedly lower in regions of the Alzheimer's disease (AD) brain that are susceptible to pathological damage, compared with spared regions in the same cases or homotropic regions in controls. To elucidate the origins of this difference in proportionate expression, we measured the absolute levels of each of the eight NR1 transcripts by quantitative internally standardized RT-PCR assay. Expression of transcripts with the cassette was strongly attenuated in susceptible regions of Alzheimer's brain, whereas expression of non-cassette transcripts differed little from that in controls. The expression of other NR1 splice variants was not associated with pathology relevant to disease status, although some combinations of splice cassettes were well maintained in AD cases. The population profile of NR1 transcripts in occipital cortex differed from the profiles in other brain regions studied. Western analysis confirmed that the expression of protein isoforms containing the N-terminal peptide was very low in susceptible areas of the Alzheimer's brain. Cells that express NR1 subunits with the N-terminal cassette may be selectively vulnerable to toxicity in AD.

Sea surface temperature and primary productivity reconstruction of sediment core SO130-275KL from the northeastern Arabian Sea during the late Holocene

Variability in the oceanic environment of the Arabian Sea region is strongly influenced by the seasonal monsoon cycle of alternating wind directions. Prominent and well studied is the summer monsoon, but much less is known about late Holocene changes in winter monsoon strength with winds from the northeast that drive convective mixing and high surface ocean productivity in the northeastern Arabian Sea. To establish a high-resolution record of winter monsoon variability for the late Holocene, we analyzed alkenone-derived sea surface temperature (SST) variations and proxies of primary productivity (organic carbon and d15N) in a well-laminated sediment core from the Pakistan continental margin. Weak winter monsoon intensities off Pakistan are indicated from 400 B.C. to 250 A.D. by reduced productivity and relatively high SST. At about 250 A.D., the intensity of the winter monsoon increased off Pakistan as indicated by a trend to lower SST. We infer that monsoon conditions were relatively unstable from ~500 to 1300 A.D., because primary production and SST were highly variable. Declining SST and elevated biological production from 1400 to 1900 A.D. suggest invigorated convective winter mixing by strengthening winter monsoon circulation, most likely a regional expression of colder climate conditions during the Little Ice Age on the Northern Hemisphere. The comparison of winter monsoon intensity with records of summer monsoon intensity suggests that an inverse relationship between summer and winter monsoon strength exists in the Asian monsoon system during the late Holocene, effected by shifts in the Intertropical Convergence Zone.

Palynomorphs from the Lateglacial and Holocene of the Mt-Athos Basin, Aegean Sea

To unravel the climatic and environmental dynamics in the borderlands of the Aegean Sea during the early and middle Holocene, and notably for the interval of sapropel S1 (S1) formation, we have analysed terrestrial palynomorphs from a marine core in the northern Aegean Sea. The qualitative results were complemented by quantitative pollen-based climate reconstructions. A land-sea correlation was established based on pollen data and sediment lightness measurements from the same core, and previously published benthic foraminifer data from a nearby core. The borderlands of the Aegean Sea underwent a transition from an open vegetation to oak-dominated woodlands between ~10.4 and ~9.5 ka cal BP. A coeval increase in winter precipitation suggests that moisture availability was the main factor controlling Holocene reforestation. The ~50% higher winter precipitation during S1 formation relative to "pre-sapropelic" conditions suggests a strong contribution from the borderlands of the Aegean Sea to the freshwater surplus during S1 formation. The humid and mild winter conditions during S1 formation were repeatedly punctuated by short-term climatic events that caused a partial deforestation and a reorganisation within the broad-leaved arboreal vegetation. In the marine realm, these events are documented by improved benthic oxygenation. The strongest event represents the regional expression of the 8.2 ka cold event and led to an interruption in S1 formation. Except for the interval of S1 formation, the pollen-derived winter temperatures correlate with the smoothed GISP2 K+ series. They support the previously published, marine-based concept that the intensity of the Siberian High strongly controlled the winter climate in the Aegean region. During S1 formation in the Aegean Sea, however, climate conditions in the borderlands were more strongly affected by the monsoonally influenced climate system of the lower latitudes.

Summer sea surface temperatures calculated from radiolaria at ODP Site 177-1089 in the subantarctic Atlantic

A submillennial resolution, radiolarian-based record of summer sea surface temperature (SST) documents the last five glacial to interglacial transitions at the subtropical front, southern Atlantic Ocean. Rapid fluctuations occur both during glacial and interglacial intervals, and sudden cooling episodes at glacial terminations are recurrent. Surface hydrography and global ice volume proxies from the same core suggest that summer SST increases prior to terminations lead global ice-volume decreases by 4.7 ± 3.7 ka (in the eccentricity band), 6.9 ± 2.5 ka (obliquity), and 2.7 ± 0.9 ka (precession). A comparison between SST and benthic delta13C suggests a decoupling in the response of northern subantarctic surface, intermediate, and deep water masses to cold events in the North Atlantic. The matching features between our SST record and the one from core MD97-2120 (southwest Pacific) suggests that the super-regional expression of climatic events is substantially affected by a single climatic agent: the Subtropical Front, amplifier and vehicle for the transfer of climatic change. The direct correlation between warmer DeltaTsite at Vostok and warmer SST at ODP Site 1089 suggests that warmer oceanic/atmospheric conditions imply a more southward placed frontal system, weaker gradients, and therefore stronger Agulhas input to the Atlantic Ocean.

Regional and cellular gene expression changes in human Huntington's disease brain.

Huntington's disease (HD) pathology is well understood at a histological level but a comprehensive molecular analysis of the effect of the disease in the human brain has not previously been available. To elucidate the molecular phenotype of HD on a genome-wide scale, we compared mRNA profiles from 44 human HD brains with those from 36 unaffected controls using microarray analysis. Four brain regions were analyzed: caudate nucleus, cerebellum, prefrontal association cortex [Brodmann's area 9 (BA9)] and motor cortex [Brodmann's area 4 (BA4)]. The greatest number and magnitude of differentially expressed mRNAs were detected in the caudate nucleus, followed by motor cortex, then cerebellum. Thus, the molecular phenotype of HD generally parallels established neuropathology. Surprisingly, no mRNA changes were detected in prefrontal association cortex, thereby revealing subtleties of pathology not previously disclosed by histological methods. To establish that the observed changes were not simply the result of cell loss, we examined mRNA levels in laser-capture microdissected neurons from Grade 1 HD caudate compared to control. These analyses confirmed changes in expression seen in tissue homogenates; we thus conclude that mRNA changes are not attributable to cell loss alone. These data from bona fide HD brains comprise an important reference for hypotheses related to HD and other neurodegenerative diseases.