Caracterização citoarquitetônica e por imunoistoquímica para tirosina-hidroxilase da substância negra, área tegmentar ventral e zona retrorubral do Sagui (Callithrix jacchus)

It is known that the catecholamine group is constituted by dopamine, noradrenaline and adrenaline, in which the synthesis is regulated by an enzyme named tyrosine hydroxylase. Thus, 3-hydroxytyramine/dopamine (DA) is a precursor of the noradrenaline and adrenaline synthesis and acts as a neurotransmitter in the central nervous system. The three main nuclei, named the retrorubral field (A8 group), the substantia nigra pars compacta (A9 group) and the ventral tegmental area (A10 group), are arranged in the die-mesencephalic portion and are involved in three complexes circuitries - the mesostriatal, mesolimbic and mesocortical pathways. These pathways are related to behavioral manifestations, motricity, learning, reward and pathologies such as Parkinson’s Disease and Schizophrenia. Thus, the aim of this study was to perform de morphological analysis of the A8, A9 and A10 nuclei of the common marmoset (Callithrix jacchus). The marmoset is a neotropical primate, whose morphological and functional characteristics supports the suitability of use of this animal in biomedical research. Coronal sections of the marmoset brain were submitted to cytoarchitectonic characterization and TH-immunohistochemistry. Based on the morphology of the neurons, it was possible to subdivide the A10 group in seven regions: interfascicular nucleus, raphe rostral linear nucleus and raphe caudal linear nucleus, in the middle line; paranigral and parainterfascicular nucleus, in the middle zone; rostral portion of the ventral tegmental area nucleus and parabrachial pigmented nucleus, located in the dorsolateral portion of the mesencephalic tegmentum. A9 group was divided into four regions: substantia nigra compacta dorsal and ventral tiers; substantia nigra compacta lateral and medial clusters. No subdivisions were founded into A8 group. These results revealed that A8, A9 and A10 are phylogenetically conserved between species, but it’s necessary to expand the studies about this compartmentalization, investigating its occurrence in other primate species or investigating its functional relevance.

Particelle di cellulosa come nuclei di ghiacciamento in nube

La formazione del ghiaccio nelle nubi avviene prevalentemente per nucleazione eterogenea, grazie alla presenza di nuclei di ghiacciamento (Ice Nucleating Particles, INP), ovvero particelle di aerosol (prevalentemente polveri minerali) in grado di favorire la solidificazione di una gocciolina o il passaggio diretto dalla fase vapore alla fase ghiaccio. Recentemente, l'interesse si è esteso anche all'aerosol di tipo biologico (funghi, spore, etc.), in grado di agire come INP a temperature più elevate rispetto all'aerosol minerale. Il lavoro sperimentale di questa tesi, svolto presso il laboratorio del gruppo Nubi e Precipitazioni dell’ISAC-CNR di Bologna, ha avuto come obiettivo lo studio della capacità della cellulosa di agire come INP. Esso si inserisce in un’attività di ricerca internazionale dedicata allo studio degli INP (progetto Ice NUclei research unIT, INUIT, Germania). Il lavoro sperimentale ha riguardato la messa a punto di due diversi sistemi di generazione dell'aerosol di cellulosa, la caratterizzazione dimensionale delle particelle, la loro osservazione al microscopio elettronico e la preparazione dei filtri per le misure di INP. I risultati ottenuti hanno evidenziato che le proprietà nucleanti della cellulosa sono inferiori rispetto alle polveri minerali ma paragonabili ad altri materiali, come la cenere vulcania e l'aerosol marino. Quindi, in aree ricche di vegetazione o dove le polveri minerali non sono abbondanti, la cellulosa potrebbe costituire un importante materiale per la formazione del ghiaccio nelle nubi miste. Misure preliminari hanno evidenziato come le particelle di cellulosa con dimensioni inferiori a 0,5μm risultano meno attive nella nucleazione del ghiaccio rispetto a quelle di dimensioni maggiori. I risultati ottenuti sono stati presentati al Congresso PM2016 (Roma, 17-19 maggio 2016) e saranno presentati in un poster alla prossima European Aerosol Conference (Tours, Francia, 4-6 settembre 2016).

Are luminous radio-loud active galactic nuclei triggered by galaxy interactions?

We present the results of a comparison between the optical morphologies of a complete sample of 46 southern 2 Jy radio galaxies at intermediate redshifts (0.05 < z < 0.7) and those of two control samples of quiescent early-type galaxies: 55 ellipticals at redshifts z ≤ 0.01 from the Observations of Bright Ellipticals at Yale (OBEY) survey, and 107 early-type galaxies at redshifts 0.2 < z < 0.7 in the Extended Groth Strip (EGS). Based on these comparisons, we discuss the role of galaxy interactions in the triggering of powerful radio galaxies (PRGs). We find that a significant fraction of quiescent ellipticals at low and intermediate redshifts show evidence for disturbed morphologies at relatively high surface brightness levels, which are likely the result of past or on-going galaxy interactions. However, the morphological features detected in the galaxy hosts of the PRGs (e.g. tidal tails, shells, bridges, etc.) are up to 2 mag brighter than those present in their quiescent counterparts. Indeed, if we consider the same surface brightness limits, the fraction of disturbed morphologies is considerably smaller in the quiescent population (53 per cent at z < 0.2 and 48 per cent at 0.2 ≤ z < 0.7) than in the PRGs (93 per cent at z < 0.2 and 95 per cent at 0.2 ≤ z < 0.7 considering strong-line radio galaxies only). This supports a scenario in which PRGs represent a fleeting active phase of a subset of the elliptical galaxies that have recently undergone mergers/interactions. However, we demonstrate that only a small proportion (≲20 per cent) of disturbed early-type galaxies are capable of hosting powerful radio sources.

Spatial organisation and behaviour of the parental chromosome sets in the nuclei of Saccharomyces cerevisiae × S. paradoxus hybrids

Peer reviewed

Spatial organisation and behaviour of the parental chromosome sets in the nuclei of Saccharomyces cerevisiae × S. paradoxus hybrids

Peer reviewed

QUADRUPOLE CENTRAL TRANSITION NMR SPECTROSCOPY OF QUADRUPOLAR NUCLEI IN SOLUTION

This thesis reports on 17O (I = 5/2) and 59Co (I = 7/2) quadrupole central transition (QCT) NMR studies of three classes of biologically important molecules: glucose, nicotinamide and Vitamin B12 derivatives. Extensive QCT NMR experiments were performed over a wide range of molecular motion by changing solvent viscosity and temperature. 17O-labels were introduced at the 5- and 6-positions respectively: D-[5-17O]-glucose and D-[6-17O]-glucose following the literature method. QCT NMR greatly increased the molecular size limit obtained by ordinary solution NMR. It requires much lower temperatures to get the optimal spectral resolution, which are preferable for biological molecules. In addition, quadrupolar product parameter (PQ) and shielding anisotropy product parameter (PSA) were obtained for hydroxide group and amide group for the first time. For conventional NMR studies of quadrupolar nuclei, only PQ is accessible while QCT NMR obtained both PQ and PSA simultaneously. Our experiments also suggest the resolution of QCT NMR can be even better than that obtained by conventional NMR. We observed for the first time that the second-order quadrupolar interaction becomes a dominant relaxation mechanism under ultraslow motion. All these observations suggest that QCT NMR can become a standard technique for studying quadrupolar nuclei in solution.

Nuclei of Nearby Active Galaxies: NIR and Sub-mm Views and Finalizing the LINC-NIRVANA Fringe and Flexure Tracking System: Flexure and Temperature Behavior

The work presented in my thesis addresses the two cornerstones of modern astronomy: Observation and Instrumentation. Part I deals with the observation of two nearby active galaxies, the Seyfert 2 galaxy NGC 1433 and the Seyfert 1 galaxy NGC 1566, both at a distance of $\sim10$ Mpc, which are part of the Nuclei of Galaxies (NUGA) sample. It is well established that every galaxy harbors a super massive black hole (SMBH) at its center. Furthermore, there seems to be a fundamental correlation between the stellar bulge and SMBH masses. Simulations show that massive feedback, e.g., powerful outflows, in Quasi Stellar Objects (QSOs) has an impact on the mutual growth of bulge and SMBH. Nearby galaxies follow this relation but accrete mass at much lower rates. This gives rise to the following questions: Which mechanisms allow feeding of nearby Active Galactic Nuclei (AGN)? Is this feeding triggered by events, e.g., star formation, nuclear spirals, outflows, on $\sim500$ pc scales around the AGN? Does feedback on these scales play a role in quenching the feeding process? Does it have an effect on the star formation close to the nucleus? To answer these questions I have carried out observations with the Spectrograph for INtegral Field Observation in the Near Infrared (SINFONI) at the Very Large Telescope (VLT) situated on Cerro Paranal in Chile. I have reduced and analyzed the recorded data, which contain spatial and spectral information in the H-band ($1.45 \mic-1.85 \mic$) and K-band ($1.95 \mic-2.45 \mic$) on the central $10\arcsec\times10\arcsec$ of the observed galaxies. Additionally, Atacama Large Millimeter/Sub-millimeter Array (ALMA) data at $350$ GHz ($\sim0.87$ mm) as well as optical high resolution Hubble Space Telescope (HST) images are used for the analysis. For NGC 1433 I deduce from comparison of the distributions of gas, dust, and intensity of highly ionized emission lines that the galaxy center lies $\sim70$ pc north-northwest of the prior estimate. A velocity gradient is observed at the new center, which I interpret as a bipolar outflow, a circum nuclear disk, or a combination of both. At least one dust and gas arm leads from a $r\sim200$ pc ring towards the nucleus and might feed the SMBH. Two bright warm H$_2$ gas spots are detected that indicate hidden star formation or a spiral arm-arm interaction. From the stellar velocity dispersion (SVD) I estimate a SMBH mass of $\sim1.74\times10^7$ \msol. For NGC 1566 I observe a nuclear gas disk of $\sim150$ pc in radius with a spiral structure. I estimate the total mass of this disk to be $\sim5.4\times10^7$ \msol. What mechanisms excite the gas in the disk is not clear. Neither can the existence of outflows be proven nor is star formation detected over the whole disk. On one side of the spiral structure I detect a star forming region with an estimated star formation rate of $\sim2.6\times10^{-3}$ \msol\ yr$^{-1}$. From broad Br$\gamma$ emission and SVD I estimate a mean SMBH mass of $\sim5.3\times10^6$ \msol\ with an Eddington ratio of $\sim2\times10^{-3}$. Part II deals with the final tests of the Fringe and Flexure Tracker (FFTS) for LBT INterferometric Camera and the NIR/Visible Adaptive iNterferometer for Astronomy (LINC-NIRVANA) at the Large Binocular Telescope (LBT) in Arizona, USA, which I conducted. The FFTS is the subsystem that combines the two separate beams of the LBT and enables near-infrared interferometry with a significantly large field of view. The FFTS has a cryogenic system and an ambient temperature system which are separated by the baffle system. I redesigned this baffle to guarantee the functionality of the system after the final tests in the Cologne cryostat. The redesign did not affect any scientific performance of LINC-NIRVANA. I show in the final cooldown tests that the baffle fulfills the temperature requirement and stays $<110$ K whereas the moving stages in the ambient system stay $>273$ K, which was not given for the old baffle design. Additionally, I test the tilting flexure of the whole FFTS and show that accurate positioning of the detector and the tracking during observation can be guaranteed.

Messenger ribonucleic acid of cerebral nuclei.

1. RNA was isolated from crude nuclear preparations and from ribosomes derived from rat brain and liver. Nuclear RNA was obtained by lysis of the nuclei with sodium dodecyl sulphate, followed by denaturation and removal of DNA and protein with hot phenol. 2. Base composition analyses indicated that the cerebral nuclear RNA preparation contained a higher proportion of non-ribosomal RNA than the analogous hepatic preparation. 3. Sucrose-density-gradient analyses revealed a heterogeneous profile for each nuclear RNA preparation, with two major peaks possessing the sedimentation properties of ribosomal RNA (18s and 28s). 4. Template activities of both preparations were widely distributed through the sucrose density gradients. 5. The cerebral nuclear RNA preparation was more active than the hepatic nuclear RNA preparation in promoting amino acid incorporation in cell-free systems from Escherichia coli and rat brain. 6. Cerebral nuclear RNA stimulated amino acid incorporation in a cerebral ribosomal system even in the presence of an excess of purified E. coli transfer RNA. 7. It is concluded that a significant proportion of cerebral nuclear RNA has the characteristics of messenger RNA.

FACS-based purification of Arabidopsis microspores, sperm cells and vegetative nuclei

Background: The male germline in flowering plants differentiates by asymmetric division of haploid uninucleated microspores, giving rise to a vegetative cell enclosing a smaller generative cell, which eventually undergoes a second mitosis to originate two sperm cells. The vegetative cell and the sperm cells activate distinct genetic and epigenetic mechanisms to control pollen tube growth and germ cell specification, respectively. Therefore, a comprehensive characterization of these processes relies on efficient methods to isolate each of the different cell types throughout male gametogenesis. Results: We developed stable transgenic Arabidopsis lines and reliable purification tools based on Fluorescence-Activated Cell Sorting (FACS) in order to isolate highly pure and viable fractions of each cell/nuclei type before and after pollen mitosis. In the case of mature pollen, this was accomplished by expressing GFP and RFP in the sperm and vegetative nuclei, respectively, resulting in 99% pure sorted populations. Microspores were also purified by FACS taking advantage of their characteristic small size and autofluorescent properties, and were confirmed to be 98% pure. Conclusions: We provide simple and efficient FACS-based purification protocols for Arabidopsis microspores, vegetative nuclei and sperm cells. This paves the way for subsequent molecular analysis such as transcriptomics, DNA methylation analysis and chromatin immunoprecipitation, in the developmental context of microgametogenesis in Arabidopsis.

A doorway to Borromean halo nuclei: the Samba configuration

We exploit the possibility of new configurations in three-body halo nuclei, Samba type (the neutron-core form a bound system) as a doorway to Borromean systems. The nuclei Be-12, B-15, N-23 and F-27 are of such nature, in particular N-23 with a half-life of 37.7 s and a halo radius of 6.07 fm is an excellent example of Samba-halo configuration. The fusion below the barrier of the Samba halo nuclei with heavy targets could reveal the so far elusive enhancement and a dominance of one-neutron over two-neutron transfers, in contrast to what was found recently for the Borromean halo nucleus He-6 + U-238.

Galanin n-terminal gragment (1-15) modifies the 5-HT1A receptor against [H3]-8-OH-DPAT binding in the dorsal raphe and hippocampus of the rat

We have described that Galanin N-terminal fragment (1-15) [GAL(1-15)] is associated with depressive effects and also modulates the antidepressant effects induced by the 5-HT1A receptor (5-HT1AR) agonist 8-OH-DPAT. The aim of this study is to analyze the ability of GAL(1-15) to modulate 5-HT1AR at the autoreceptor and postsynaptic receptor level in rats by using quantitative autoradiography. We analyzed the effect of intracerebroventricular GAL(1-15)-3nmol (n=6) or aCSF (n=6), 10 minutes, 2 and 5 hours after the injection, on the binding characteristics of the 5-HT1AR agonist [H3]-8-OH-DPAT in sections of the Dorsal Raphe (DR) and Dorsal Hippocampus, specifically CA1 and Dentate Gyrus (DG). Student’s t-test was used to compare the experimental groups. GAL(1-15) produced a time-dependent effect on the binding of [H3]-8-OH-DPAT. In CA1 and DG, a significant increase in the KD and Bmax was observed, by 90%(p<0.05), at 10 minutes and 2 hours after injection. However, 5 hours after GAL(1-15) the only significant change remaining was the increase in Bmax at the DG. The coinjection of the GALR2 antagonist M871 blocked significantly the effects induced by GAL(1-15) in both areas. In DR, 2 hours after injection GAL(1-15) only produced a decrease in the Bmax by 20%(p<0.05). These results indicate that GAL(1-15) interacts with 5-HT1AR at the receptor level in DR and Dorsal Hippocampus. Therapeutic strategies based on these results could be developed for the treatment of depression disorders. This work has been supported by Junta de Andalucia CVI646 and Spanish Ministry of Economy PSI2013-44901-P.

Development of metabotropic glutamate receptors from trigeminal nuclei to barrel cortex in postnatal mouse

Expression patterns of group I (mGluR1α and mGluR5)and group II (mGluR2/3) metabotropic glutamate receptor subtypes were examined immunocytochemically in the trigeminal system of mice during the first 3 weeks of postnatal development, when somatotopic whisker representations are sequentially established from brainstem through thalamus to cerebral cortex. Immunostaining for all three epitopes formed whisker-related patterns in the trigeminal nuclei from postnatal day (P) 0, in the ventral posterior thalamic nucleus from P2, and in the posteromedial barrel subfield of somatosensory cortex (SI) from P4. The appearance of whisker-related patterns was preceded by increased levels of immunostaining of the neuropil, which subsequently declined from the trigeminal nuclei upward. In SI, mGluR1α-positive neurons were observed in all cortical layers from P2. mGluR5 was localized in neurons, glial cells, and neuropil from P2. mGluR2/3 immunostaining was distributed only in the neuropil at all ages. The three receptor subtypes showed moderate to high expression in deep layer V throughout development. Transient expression peaked in the hollows of layer IV barrels from P4 to P9, and then fell off as expression increased in supragranular layers from P14 to P21. The deep aspect of the cortical subplate (layer VIb) showed dense mGluR5 and less dense mGluR1α immunostaining throughout development. Up-regulation of expression of group I and II mGluRs is correlated with the growth and refinement of connectivity and the establishment of somatotopic patterns in the three main relay stations of the trigeminal system. This finding suggests roles for mGluRs in the early processing of sensory information and in developmental plasticity.

Estimating total cross sections from neutron-nucleus collisions using a simple functional form

Total cross sections for neutron scattering from nuclei, with energies ranging from 10 to 600 MeV and from many nuclei spanning the mass range 6Li to 238U, have been analyzed using a simple, three-parameter, functional form. The calculated cross sections are compared with results obtained by using microscopic (g-folding) optical potentials as well as with experimental data. The functional form reproduces those total cross sections very well. When allowance is made for Ramsauer-like effects in the scattering, the parameters of the functional form required vary smoothly with energy and target mass. They too can be represented by functions of energy and mass.

In vivo confocal microscopy of the palpebral conjunctiva and tarsal plate

Purpose The aim of this work is to develop a more complete understanding of the in vivo histology of the human palpebral conjunctiva and tarsal plate. Methods. The upper eyelids of 11 healthy human volunteer subjects were everted, and laser scanning confocal microscopy was used to examine the various tissue layers of the palpebral conjunctiva and tarsal plate. Results The superficial and basal epithelial layers are composed of cells with gray cytoplasm and thick, light gray borders.Nuclei can not be seen. The stroma has a varied appearance; fibrous tissue is sometimes observed, interspersed with dark,amorphous lacunae, and crevases. Numerous single white or gray cells populate this tissue, and fine blood vessels are seen traversing the field. Occasional conjunctival microcysts and Langerhans cells are observed. The tarsal plate is dark and amorphous, and meibomian gland acini with convoluted borders are clearly observed. Acini are composed of an outer lining of large cuboidal cells, and differentiated secretory cells can be seen within the acini lumen. Conclusions Laser scanning confocal microscopy is capable of studying the human palpebral conjunctiva, tarsal plate, and acini of meibomian glands in vivo. The observations presented here may provide useful supplementary anatomical information relating to the morphology of this tissue.

In vivo confocal microscopy of the bulbar conjunctiva

Background: The aim of this work is to develop a more complete qualitative and quantitative understanding of the in vivo histology of the human bulbar conjunctiva. Methods: Laser scanning confocal microscopy (LSCM) was used to observe and measure morphological characteristics of the bulbar conjunctiva of 11 healthy human volunteer subjects. Results: The superficial epithelial layer of the bulbar conjunctiva is seen as a mass of small cell nuclei. Cell borders are sometimes visible. The light grey borders of basal epithelial cells are clearly visible, but nuclei can not be seen. The conjunctival stroma is comprised of a dense meshwork of white fibres, through which traverse blood vessels containing cellular elements. Orifices at the epithelial surface may represent goblet cells that have opened and expelled their contents. Goblet cells are also observed in the deeper epithelial layers, as well as conjunctival microcysts and mature forms of Langerhans cells. The bulbar conjunctiva has a mean thickness of 32.9 1.1 mm, and a superficial and basal epithelial cell density of 2212 782 and 2368 741 cells/ mm2, respectively. Overall goblet and mature Langerhans cell densities are 111 58 and 23 25 cells/mm2, respectively. Conclusions: LSCM is a powerful technique for studying the human bulbar conjunctiva in vivo and quantifying key aspects of cell morphology. The observations presented here may serve as a useful marker against which changes in conjunctival morphology due to disease, surgery, drug therapy or contact lens wear can be assessed.