The neuroprotective effect of pituitary adenylate cyclase-activating polypeptide on cerebellar granule cells is mediated through inhibition of the CED3-related cysteine protease caspase-3/CPP32

Caspase-3 knockout mice exhibit thickening of the internal granule cell layer of the cerebellum. Concurrently, it has been shown that intracerebral injection of pituitary adenylate cyclase-activating polypeptide (PACAP) induces a transient increase of the thickness of the cerebellar cortex. In the present study, we have investigated the possible effect of PACAP on caspase activity in cultured cerebellar granule cells from 8-day-old rat. Incubation of granule neurons with PACAP for 24 h promoted cell survival and prevented DNA fragmentation. Exposure of cerebellar granule cells to the specific caspase-3 inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethylketone (Z-DEVD-FMK) for 24 h markedly enhanced cell survival and inhibited apoptotic cell death. Time-course studies revealed that PACAP causes a prolonged inhibition of caspase-3 activity without affecting caspase-1. Administration of graded concentrations of PACAP for 3 h induced a dose-dependent inhibition of caspase-3 activity. Incubation of granule cells with both dibutyryl-cAMP (dbcAMP) and phorbol 12-myristate 13-acetate (PMA) mimicked the inhibitory effect of PACAP on caspase-3. Cotreatment of cultured neurons with the protein kinase A inhibitor H89 and the protein kinase C inhibitor chelerythrine abrogated the effect of PACAP on caspase-3 activity. In contrast, the ERK kinase inhibitor U0126 did not affect the action of PACAP on caspase-3 activity. These data demonstrate that PACAP prevents cerebellar granule neurons from apoptotic cell death through a protein kinase A- and protein kinase C-dependent inhibition of caspase-3 activity.

Segment-specific pattern of sympathetic preganglionic projections in the chicken embryo spinal cord is altered by retinoids

Sympathetic preganglionic neurons exhibit segment-specific projections. Preganglionic neurons located in rostral spinal segments project rostrally within the sympathetic chain, those located in caudal spinal segments project caudally, and those in midthoracic segments project either rostrally or caudally in segmentally graded proportions. Moreover, rostrally and caudally projecting preganglionic neurons are skewed toward the rostral and caudal regions, respectively, of each midthoracic segment. The mechanisms that establish these segment-specific projections are unknown. Here we show that experimental manipulation of retinoid signaling in the chicken embryo alters the segment-specific pattern of sympathetic preganglionic projections and that this effect is mediated by the somitic mesoderm. Application of exogenous retinoic acid to a single rostral thoracic somite decreases the number of rostrally projecting preganglionic neurons at that level. Conversely, disrupting endogenous synthesis of retinoic acid in a single caudal thoracic somite increases the number of rostrally projecting preganglionic neurons at that level. The number of caudally projecting neurons does not change in either case, indicating that the effect is specific for rostrally projecting preganglionic neurons. These results indicate that the sizes of the rostrally and caudally projecting populations may be independently regulated by different factors. Opposing gradients of such factors along the longitudinal axis of the thoracic region of the embryo could be sufficient, in combination, to determine the segment-specific identity of preganglionic projections.

Inositol 1,4,5-tris-phosphate activation of inositol tris-phosphate receptor Ca2+ channel by ligand tuning of Ca2+ inhibition

Inositol 1,4,5-tris-phosphate (IP3) binding to its receptors (IP3R) in the endoplasmic reticulum (ER) activates Ca2+ release from the ER lumen to the cytoplasm, generating complex cytoplasmic Ca2+ concentration signals including temporal oscillations and propagating waves. IP3-mediated Ca2+ release is also controlled by cytoplasmic Ca2+ concentration with both positive and negative feedback. Single-channel properties of the IP3R in its native ER membrane were investigated by patch clamp electrophysiology of isolated Xenopus oocyte nuclei to determine the dependencies of IP3R on cytoplasmic Ca2+ and IP3 concentrations under rigorously defined conditions. Instead of the expected narrow bell-shaped cytoplasmic free Ca2+ concentration ([Ca2+]i) response centered at ≈300 nM–1 μM, the open probability remained elevated (≈0.8) in the presence of saturating levels (10 μM) of IP3, even as [Ca2+]i was raised to high concentrations, displaying two distinct types of functional Ca2+ binding sites: activating sites with half-maximal activating [Ca2+]i (Kact) of 210 nM and Hill coefficient (Hact) ≈2; and inhibitory sites with half-maximal inhibitory [Ca2+]i (Kinh) of 54 μM and Hill coefficient (Hinh) ≈4. Lowering IP3 concentration was without effect on Ca2+ activation parameters or Hinh, but decreased Kinh with a functional half-maximal activating IP3 concentration (KIP3) of 50 nM and Hill coefficient (HIP3) of 4 for IP3. These results demonstrate that Ca2+ is a true receptor agonist, whereas the sole function of IP3 is to relieve Ca2+ inhibition of IP3R. Allosteric tuning of Ca2+ inhibition by IP3 enables the individual IP3R Ca2+ channel to respond in a graded fashion, which has implications for localized and global cytoplasmic Ca2+ concentration signaling and quantal Ca2+ release.

A Computer-Assisted Personalized Approach in an Undergraduate Plant Physiology Class1

We used Computer-Assisted Personalized Approach (CAPA), a networked teaching and learning tool that generates computer individualized homework problem sets, in our large-enrollment introductory plant physiology course. We saw significant improvement in student examination performance with regular homework assignments, with CAPA being an effective and efficient substitute for hand-graded homework. Using CAPA, each student received a printed set of similar but individualized problems of a conceptual (qualitative) and/or quantitative nature with quality graphics. Because each set of problems is unique, students were encouraged to work together to clarify concepts but were required to do their own work for credit. Students could enter answers multiple times without penalty, and they were able to obtain immediate feedback and hints until the due date. These features increased student time on task, allowing higher course standards and student achievement in a diverse student population. CAPA handles routine tasks such as grading, recording, summarizing, and posting grades. In anonymous surveys, students indicated an overwhelming preference for homework in CAPA format, citing several features such as immediate feedback, multiple tries, and on-line accessibility as reasons for their preference. We wrote and used more than 170 problems on 17 topics in introductory plant physiology, cataloging them in a computer library for general access. Representative problems are compared and discussed.

Structure and function of declarative and nondeclarative memory systems

This article reviews recent studies of memory systems in humans and nonhuman primates. Three major conclusions from recent work are that (i) the capacity for nondeclarative (nonconscious) learning can now be studied in a broad array of tasks that assess classification learning, perceptuomotor skill learning, artificial grammar learning, and prototype abstraction; (ii) cortical areas adjacent to the hippocampal formation, including entorhinal, perirhinal, and parahippocampal cortices, are an essential part of the medial temporal lobe memory system that supports declarative (conscious) memory; and (iii) in humans, bilateral damage limited to the hippocampal formation is nevertheless sufficient to produce severe anterograde amnesia and temporally graded retrograde amnesia covering as much as 25 years.

Signals and signs in the nervous system: The dynamic anatomy of electrical activity is probably information-rich

The dichotomy between two groups of workers on neuroelectrical activity is retarding progress. To study the interrelations between neuronal unit spike activity and compound field potentials of cell populations is both unfashionable and technically challenging. Neither of the mutual disparagements is justified: that spikes are to higher functions as the alphabet is to Shakespeare and that slow field potentials are irrelevant epiphenomena. Spikes are not the basis of the neural code but of multiple codes that coexist with nonspike codes. Field potentials are mainly information-rich signs of underlying processes, but sometimes they are also signals for neighboring cells, that is, they exert influence. This paper concerns opportunities for new research with many channels of wide-band (spike and slow wave) recording. A wealth of structure in time and three-dimensional space is different at each scale—micro-, meso-, and macroactivity. The depth of our ignorance is emphasized to underline the opportunities for uncovering new principles. We cannot currently estimate the relative importance of spikes and synaptic communication vs. extrasynaptic graded signals. In spite of a preponderance of literature on the former, we must consider the latter as probably important. We are in a primitive stage of looking at the time series of wide-band voltages in the compound, local field, potentials and of choosing descriptors that discriminate appropriately among brain loci, states (functions), stages (ontogeny, senescence), and taxa (evolution). This is not surprising, since the brains in higher species are surely the most complex systems known. They must be the greatest reservoir of new discoveries in nature. The complexity should not deter us, but a dose of humility can stimulate the flow of imaginative juices.

Modeling back propagating action potential in weakly excitable dendrites of neocortical pyramidal cells.

Simultaneous recordings from the soma and apical dendrite of layer V neocortical pyramidal cells of young rats show that, for any location of current input, an evoked action potential (AP) always starts at the axon and then propagates actively, but decrementally, backward into the dendrites. This back-propagating AP is supported by a low density (-gNa = approximately 4 mS/cm2) of rapidly inactivating voltage-dependent Na+ channels in the soma and the apical dendrite. Investigation of detailed, biophysically constrained, models of reconstructed pyramidal cells shows the following. (i) The initiation of the AP first in the axon cannot be explained solely by morphological considerations; the axon must be more excitable than the soma and dendrites. (ii) The minimal Na+ channel density in the axon that fully accounts for the experimental results is about 20-times that of the soma. If -gNa in the axon hillock and initial segment is the same as in the soma [as recently suggested by Colbert and Johnston [Colbert, C. M. & Johnston, D. (1995) Soc. Neurosci. Abstr. 21, 684.2]], then -gNa in the more distal axonal regions is required to be about 40-times that of the soma. (iii) A backward propagating AP in weakly excitable dendrites can be modulated in a graded manner by background synaptic activity. The functional role of weakly excitable dendrites and a more excitable axon for forward synaptic integration and for backward, global, communication between the axon and the dendrites is discussed.

Interleukin 7 independent development of human B cells.

Mammalian hematopoietic stem cell (HSC) commitment and differentiation into lymphoid lineage cells proceed through a series of developmentally restricted progenitor compartments. A complete understanding of this process, and how it differs from HSC commitment and differentiation into cells of the myeloid/erythroid lineages, requires the development of model systems that support HSC commitment to the lymphoid lineages. We now describe a human bone marrow stromal cell culture that preferentially supports commitment and differentiation of human HSC to CD19+ B-lineage cells. Fluorescence activated cell sorterpurified CD34++/lineage-cells were isolated from fetal bone marrow and cultured on human fetal bone marrow stromal cells in serum-free conditions containing no exogenous cytokines. Over a period of 3 weeks, CD34++/lineage- cells underwent commitment, differentiation, and expansion into the B lineage. Progressive changes included: loss of CD34, acquisition of and graded increases in the level of cell surface CD19, and appearance of immature B cells expressing mu/kappa or mu/lambda cell surface Ig receptors. The tempo and phenotype of B-cell development was not influenced by the addition of IL-7 (10 ng/ml), or by the addition of goat anti-IL-7 neutralizing antibody. These results indicate a profound difference between mouse and human in the requirement for IL-7 in normal B-cell development, and provide an experimental system to identify and characterize human bone marrow stromal cell-derived molecules crucial for human B lymphopoiesis.

Potentiation of the bioavailability of daidzin by an extract of Radix puerariae.

The dose effect of pure daidzin on the suppression of ethanol intake in Syrian golden hamsters was compared with that of crude daidzin contained in a methanol extract of Radix puerariae (RP). EC50 values estimated from the graded dose-response curves for pure daidzin and RP extract daidzin are 23 and 2.3 mg per hamster per day, respectively. Apparently the antidipsotropic activity of the RP extract cannot be accounted for solely by its daidzin content (22 mg/g). In addition to daidzin, six other isoflavones were identified in the RP extract and quantified--namely, puerarin (160 mg per g of extract), genistin (3.7 mg/g), daidzein (2.6 mg/g), daidzein-4',7-diglucoside (1.2 mg/g), genistein (0.2 mg/g), and formononetin (0.16 mg/g). None of these, administered either alone or combined, contributes in any significant way to the antidipsotropic activity of the extract. Plasma daidzin concentration-time curves determined in hamsters administered various doses of pure daidzin or RP extract by i.p.injection indicate that the crude extract daidzin has approximately 10 times greater bioavailability than the pure compound. Reconstruction of the dose-response effects for pure and crude daidzin using bioavailable daidzin rather than administered dose gives a single curve. Synthetic daidzin added to the RP extract acquires the bioavailability of the endogenous daidzin that exists naturally in the extract. These results show that (i) daidzin is the major active principle in methanol extracts of RP, and (ii) additional constituents in the methanol extract of RP assist uptake of daidzin in golden hamsters.

Identificación de minerales mediante difracción de rayos X utilizando bases de datos online de libre acceso

El artículo proporciona una metodología sencilla para la identificación de minerales mediante la técnica de difracción de rayos X de polvo utilizando bases de datos mineralógicos de libre acceso y online. Las bases de datos utilizadas son la base de datos mineralógicas webmineral y la base de datos de estructuras cristalinas de la American Mineralogist Crystal Structure Database, AMS. En el presente trabajo se han elaborado 3 actividades resueltas de estudios reales y en orden creciente de dificultad. Se ha pretendido hacer hincapié en puntos donde el profesor puede interactuar con el alumno y promover la capacidad de análisis, síntesis y razonamiento crítico del alumno ante un problema de investigación en geología. Finalmente se ha elaborado un Anexo donde se recogen recomendaciones para que el profesor desarrolle sus propias actividades.

Normas de orientação clínica portuguesas : abordagem terapêutica de doenças e factores de risco cardiovasculares : que evidência, que método e que impacto?

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Efavirenz versus Nevirapina : comparação da efectividade e do perfil de segurança, a longo prazo, no tratamento da infecção por vírus da imunodeficiência humana tipo 1

Tese de mestrado, Doenças Infecciosas Emergentes, Universidade de Lisboa, Faculdade de Medicina, 2016

Caratterizzazione dinamica di miscele bituminose SMA contenenti polverino di gomma da Pneumatici Fuori Uso

Lo Stone Matrix Asphalt (SMA) è un tipo di miscela chiusa costituita da uno scheletro litico di aggregato grosso, assortito in modo tale da ottenere una distribuzione granulometrica indicata con il termine gap-graded, e da un mastice, con funzione riempitiva, ottenuto dalla miscelazione di bitume, filler ed additivi stabilizzanti. In ambito di progettazione delle miscele per conglomerati bituminosi sta assumendo sempre più importanza l’utilizzo di materiali derivanti dalla frantumazione degli Pneumatici Fuori Uso, quali granulato e polverino di gomma. Quest’ultimo può essere impiegato come valida alternativa alla modifica polimerica del bitume garantendo maggiori prestazioni in termini di resistenza all’ormaiamento, a fatica e durabilità, con un conseguente contenimento dei costi di manutenzione della sovrastruttura nel medio e lungo periodo. Il presente studio è stato condotto con lo scopo di valutare le prestazioni meccaniche che una miscela di conglomerato bituminoso può esplicare a seguito della sua mescolazione con il polverino di gomma. In particolare, è stata impiegata una miscela bituminosa di tipo SMA che, data la sua composizione interna, conferisce allo strato di usura della pavimentazione ottime qualità soprattutto in termini di resistenza alle sollecitazioni, durabilità, fonoassorbenza e macrotessitura superficiale. Al fine di rendere più esaustiva la fase sperimentale, sono state messe a confronto due miscele di tipo SMA differenti tra loro per l’aggiunta del polverino di gomma. I dati ottenuti e le considerazioni effettuate al termine della fase sperimentale hanno permesso di affermare che la miscela indagata possiede proprietà meccaniche idonee per essere impiegata nella realizzazione di nuove infrastrutture o nella manutenzione delle pavimentazioni esistenti.

Detrital zircon geochronology data, petrographic point count data and paleocurrent indicator data of Triassic turbidites in the Nanpanjiang Basin, South China

Triassic turbidites of the Nanpanjiang basin of south China represent the most expansive and voluminous siliciclastic turbidite accumulation in south China. The Nanpanjiang basin occurs at a critical junction between the southern margin of the south China plate and the Indochina, Siamo and Sibumasu plates to the south and southwest. The Triassic Yangtze carbonate shelf and isolated carbonated platforms in the basin have been extensively studied, but silicilastic turbidites in the basin have received relatively little attention. Deciphering the facies, paleocurrent indicators and provenance of the Triassic turbidites is important for several reasons: it promises to help resolve the timing of plate collisions along suture zones bordering the basin to the south and southwest, it will enable evaluation of which suture zones and Precambrian massifs were source areas, and it will allow an evaluation of the impact of the siliciclastic flux on carbonate platform evolution within the basin. Turbidites in the basin include the Early Triassic Shipao Formation and the Middle-Late Triassic Baifeng, Xinyuan, Lanmu Bianyang and Laishike formations. Each ranges upward of 700 m and the thickest is nearly 3 km. The turbidites contain very-fine sand in the northern part of the basin whereas the central and southern parts of the basin also commonly contain fine and rarely medium sand size. Coarser sand sizes occur where paleocurrents are from the south, and in this area some turbidites exhibit complete bouma sequences with graded A divisions. Successions contain numerous alternations between mud-rich and sand-rich intervals with thickness trends corresponding to proximal/ distal fan components. Spectacularly preserved sedimentary structures enable robust evaluation of turbidite systems and paleocurrent analyses. Analysis of paleocurrent measurements indicates two major directions of sediment fill. The northern part of the basin was sourced primarily by the Jiangnan massif in the northeast, and the central and southern parts of the basin were sourced primarily from suture zones and the Yunkai massif to the south and southeast respectively. Sandstones of the Lower Triassic Shipao Fm. have volcaniclastic composition including embayed quartz and glass shards. Middle Triassic sandstones are moderately mature, matrix-rich, lithic wackes. The average QFL ratio from all point count samples is 54.1/18.1/27.8% and the QmFLt ratio is 37.8/ 18.1/ 44.1%. Lithic fragments are dominantly claystone and siltstone clasts and metasedimentary clasts such as quartz mica tectonite. Volcanic lithics are rare. Most samples fall in the recycled orogen field of QmFLt plots, indicating a relatively quartz and lithic rich composition consistent with derivation from Precambrian massifs such as the Jiangnan, and Yunkai. A few samples from the southwest part of the basin fall into the dissected arc field, indicating a somewhat more lithic and feldspar-rich composition consistent with derivation from a suture zone Analysis of detrial zircon populations from 17 samples collected across the basin indicate: (1) Several samples contain zircons with concordant ages greater than 3000 Ma, (2) there are widespread peaks across the basin at 1800 Ma and 2500, (3) a widespread 900 Ma population, (3) a widespread population of zircons at 440 Ma, and (5) a larger population of younger zircons about 250 Ma in the southwestern part which is replaced to the north and northwest by a somewhat older population around 260-290 Ma. The 900 Ma provenance fits derivation from the Jiangnan Massif, the 2500, 1800, and 440 Ma provenance fits the Yunkai massif, and the 250 Ma is consistent with convergence and arc development in suture zones bordering the basin on the south or southwest. Early siliciclastic turbidite flux, proximal to source areas impacted carbonate platform evolution by infilling the basin, reducing accommodation space, stabilizing carbonate platform margins and promoting margin progradation. Late arrival, in areas far from source areas caused margin aggradation over a starved basin, development of high relief aggradational escarpments and unstable scalloped margins.

(Table 1) Summary of maximum clast size and bed thickness of ODP Leg 126 samples

Subaerial debris flows, with water contents ranging from as little as 10 wt% up to no more than about 25 wt% (Pierson, 1986; Pierson and Costa, 1987), are non-Newtonian fluids that move as fairly coherent masses with yield strength (owing to bulk densities and viscosity that are much greater than those of clear water), which enables them to suspend and transport large clasts. Their flow behavior is thought to be predominantly laminar, although the relative importance of laminar and turbulent flow has not been established and is debatable. They leave deposits (debrites) that are characteristically poorly sorted with large clasts in their middle portions and commonly protruding from their tops. Although generally ungraded or normally graded in their upper portions, many have centimeter- to decimeter-thick inversely graded basal zones (Arguden and Rodolfo, 1990, doi:10.1130/0016-7606(1990)102<0865:SADDBH>2.3.CO;2).