MicroRNA-132 is a physiological regulator of hematopoietic stem cell function and B-cell development

MicroRNAs are a class of small non-coding RNAs that negatively regulate gene expression. Several microRNAs have been implicated in altering hematopoietic cell fate decisions. Importantly, deregulation of many microRNAs can lead to deleterious consequences in the hematopoietic system, including the onset of cancer, autoimmunity, or a failure to respond effectively to infection. As such, microRNAs fine-tune the balance between normal hematopoietic output and pathologic consequences. In this work, we explore the role of two microRNAs, miR-132 and miR-125b, in regulating hematopoietic stem cell (HSC) function and B cell development. In particular, we uncover the role of miR-132 in maintaining the appropriate balance between self-renewal, differentiation, and survival in aging HSCs by buffering the expression of a critical transcription factor, FOXO3. By maintain this balance, miR-132 may play a critical role in preventing aging-associated hematopoietic conditions such as autoimmune disease and cancer. We also find that miR-132 plays a critical role in B cell development by targeting a key transcription factor, Sox4, that is responsible for the differentiation of pro-B cells into pre-B cells. We find that miR-132 regulates B cell apoptosis, and by delivering miR-132 to mice that are predisposed to developing B cell cancers, we can inhibit the formation of these cancers and improve the survival of these mice. In addition to miR-132, we uncovered the role of another critical microRNA, miR-125b, that potentiates hematopoietic stem cell function. We found that enforced expression of miR-125b causes an aggressive myeloid leukemia by downregulation of its target Lin28a. Importantly, miR-125b also plays a critical role in inhibiting the formation of pro-B cells. Thus, we have discovered two microRNAs with important roles in regulating normal hematopoiesis, and whose dregulation can lead to deleterious consequences such as cancer in the aging hematopoietic system. Both miR-132 and miR-125b may therefore be targeted for therapeutics to inhibit age-related immune diseases associated with the loss of HSC function and cancer progression.

On the population dynamics of Eudiaptomus gracilis Sars and Heterocope borealis Fischer in the Bodensee-Obersee. Part B. Eudiaptomus gracilis Sars. [Translation from: Trudy Instituta Biologii Vnutrennykh Vodnany 12(15) 170-174, 1966.]

Experimental research was conducted to study the development of eggs of Eudiaptomus gracilis Sars. The egg production was studied as well as the population dynamics. Factors like losses in the lake and through the effluent Rhine at Konstanz were considered.

A biologically active peptide in the skin of lampreys (Eudontomyzon danfordi vladykovi). [Translation from: Z.Naturforsch. (B), 26(10), 1021-1023, 1971. ]

In recent years interest in the production and description of kinin-type substances has been greatly intensified. So, for example, bradykinin, phyllokinin, physalaemin, ranatensin and caerulein could be extracted from the skin of amphibians as well as. eledoisin out of the salivary glands of Eledon moschata. An examination of lampreys seemed to us particularly profitable in the search for the incidence of further kinins. Ammocoetes of different sizes and also adults of both sexes of the species Eudontomyzon danfordi vladykovi were studied in this research. This species is found in many tributaries of the Danube. Skin extracts were tested on on isolated rat uterus, rat duodenum, guinea pig ileum and rabbit jejunum, further tests were done in order to determine a peptide character of the biologically active substance.

New Catalytic Methods for the Preparation of Tryptophans and Pyrroloindolines: Total Synthesis of (+)-Naseseazines A and B and (–)-Aspergilazine A

Tryptophan and unnatural tryptophan derivatives are important building blocks for the total synthesis of natural products, as well as the development of new drugs, biological probes, and chiral small molecule catalysts. This thesis describes various catalytic methods for the preparation of tryptophan derivatives as well as their functionalization and use in natural product total synthesis.

Herein, the tandem Friedel–Crafts conjugate addition/asymmetric protonation reaction between 2-substituted indoles and methyl 2-acetamidoacrylate to provide enantioenriched trytophans is reported. This method inspired further work in the area of transition metal catalyzed arylation reactions. We report the development of the coppercatalyzed arylation of tryptamine and tryptophan derivatives. The utility of these transformations is highlighted in the five-step syntheses of the natural products (+)-naseseazine A and B. Further work on the development of a mild and general Larock indolization protocol to access unnatural tryptophans is also discussed.

Enantioselective Total Synthesis of Diketopiperazinecontaining Natural Products: (–)-Lansai B, (+)-Nocardioazines A and B, and (–)-Acetylapoaranotin

Diketopiperazine (DKP) motif is found in a wide range of biologically active natural products. This work details our efforts toward two classes of DKP-containing natural products.

Class one features the pyrroloindoline structure, derived from tryptophans. Our group developed a highly enantioselective (3 + 2) formal cycloaddition between indoles and acrylates to provide pyrroloindoline products possessing three stereocenters. Utilizing this methodology, we accomplished asymmetric total synthesis of three natural products: (–)-lansai B, (+)-nocardioazines A and B. Total synthesis of (–)-lansai B was realized in six steps, and featured an amino acid dimerization strategy. The total synthesis of (+)-nocardioazine B was also successfully completed in ten steps. Challenges were met in approaching (+)-nocardioazine A, where a seemingly easy last-step epoxidization did not prove successful. After re-examining our synthetic strategy, an early-stage epoxidation strategy was pursued, which eventually yielded a nine-step total synthesis of (+)-nocardioazine A.

Class two is the epidithiodiketopiperazine (ETP) natural products, which possesses an additional episulfide bridge in the DKP core. With the goal of accessing ETPs with different peripheral structures for structure-activity relationship studies, a highly divergent route was successfully developed, which was showcased in the formal synthesis of (–)-emethallicin E and (–)-haematocin, and the first asymmetric synthesis of (–)-acetylapoaranotin.

Viabilidad económico-financiera del proyecto de negocio de libre elección B

En este TFG trato de ver la viabilidad económico-financiera de un proyecto empresarial de nueva empresa basado en la innovación, las tendencias actuales de compra y la tecnología. Tras el análisis de la evolución de los supermercados y la apuesta por la tecnología, de estos, en otros países, he tratado de buscar la forma de adaptarla a nuestra sociedad, cultura y costumbres. Scanner Market es un nuevo modelo de supermercado supeditado a un gran almacén de distribución, en este caso El Corte Inglés, pero puede ser aplicado para otros muchos. Basa su comercialización en una tienda física en la que existe una gran variedad de productos en un espacio reducido. Se consigue comercializar tal variedad de productos gracias a la exposición de uno solo, de forma que actúe como “muestra” para los clientes. En la tienda no hay almacén, sino que se trata de optimizar el espacio para dar cabida a una amplia oferta de productos (en forma de muestras). Los clientes hacen la compra físicamente escaneando el producto a través de una app. para Smartphone o unas pistolas de compra con el mismo software y el pedido es enviado a su hogar por la distribución de El corte Inglés.

I. Studies on bacteriophage DNA's and minicircular DNA's in M. lysodeikticus and E. coli 15. II. Flow dichroism of DNA solutions

Part I

Chapter 1.....A physicochemical study of the DNA molecules from the three bacteriophages, N1, N5, and N6, which infect the bacterium, M. lysodeikticus, has been made. The molecular weights, as measured by both electron microscopy and sedimentation velocity, are 23 x 10⁶ for N5 DNA and 31 x 10⁶ for N1 and N6 DNA's. All three DNA's are capable of thermally reversible cyclization. N1 and N6 DNA's have identical or very similar base sequences as judged by membrane filter hybridization and by electron microscope heteroduplex studies. They have identical or similar cohesive ends. These results are in accord with the close biological relation between N1 and N6 phages. N5 DNA is not closely related to N1 or N6 DNA. The denaturation T_m of all three DNA's is the same and corresponds to a (GC) content of 70%. However, the buoyant densities in CsCl of Nl and N6 DNA's are lower than expected, corresponding to predicted GC contents of 64 and 67%. The buoyant densities in Cs₂SO₄ are also somewhat anomalous. The buoyant density anomalies are probably due to the presence of odd bases. However, direct base composition analysis of N1 DNA by anion exchange chromatography confirms a GC content of 70%, and, in the elution system used, no peaks due to odd bases are present.

Chapter 2.....A covalently closed circular DNA form has been observed as an intracellular form during both productive and abortive infection processes in M. lysodeikticus. This species has been isolated by the method of CsC1-ethidium bromide centrifugation and examined with an electron microscope.

Chapter 3.....A minute circular DNA has been discovered as a homogeneous population in M. lysodeikticus. Its length and molecular weight as determined by electron microscopy are 0.445 μ and 0.88 x 10⁶ daltons respectively. There is about one minicircle per bacterium.

Chapter 4.....Several strains of E. coli 15 harbor a prophage. Viral growth can be induced by exposing the host to mitomycin C or to uv irradiation. The coliphage 15 particles from E. coli 15 and E, coli 15 T^- appear as normal phage with head and tail structure; the particles from E. coli 15 TAU are tailless. The complete particles exert a colicinogenic activity on E.coli 15 and 15 T^-, the tailless particles do not. No host for a productive viral infection has been found and the phage may be defective. The properties of the DNA of the virus have been studied, mainly by electron microscopy. After induction but before lysis, a closed circular DNA with a contour length of about 11.9 μ is found in the bacterium; the mature phage DNA is a linear duplex and 7.5% longer than the intracellular circular form. This suggests the hypothesis that the mature phage DNA is terminally repetitious and circularly permuted. The hypothesis was confirmed by observing that denaturation and renaturation of the mature phage DNA produce circular duplexes with two single-stranded branches corresponding to the terminal repetition. The contour length of the mature phage DNA was measured relative to φX RFII DNA and λ DNA; the calculated molecular weight is 27 x 10⁶. The length of the single-stranded terminal repetition was compared to the length of φX 174 DNA under conditions where single-stranded DNA is seen in an extended form in electron micrographs. The length of the terminal repetition is found to be 7.4% of the length of the nonrepetitious part of the coliphage 15 DNA. The number of base pairs in the terminal repetition is variable in different molecules, with a fractional standard deviation of 0.18 of the average number in the terminal repetition. A new phenomenon termed "branch migration" has been discovered in renatured circular molecules; it results in forked branches, with two emerging single strands, at the position of the terminal repetition. The distribution of branch separations between the two terminal repetitions in the population of renatured circular molecules was studied. The observed distribution suggests that there is an excluded volume effect in the renaturation of a population of circularly permuted molecules such that strands with close beginning points preferentially renature with each other. This selective renaturation and the phenomenon of branch migration both affect the distribution of branch separations; the observed distribution does not contradict the hypothesis of a random distribution of beginning points around the chromosome.

Chapter 5....Some physicochemical studies on the minicircular DNA species in E. coli 15 (0.670 μ, 1.47 x 10⁶ daltons) have been made. Electron microscopic observations showed multimeric forms of the minicircle which amount to 5% of total DNA species and also showed presumably replicating forms of the minicircle. A renaturation kinetic study showed that the minicircle is a unique DNA species in its size and base sequence. A study on the minicircle replication has been made under condition in which host DNA synthesis is synchronized. Despite experimental uncertainties involved, it seems that the minicircle replication is random and the number of the minicircles increases continuously throughout a generation of the host, regardless of host DNA synchronization.

Part II

The flow dichroism of dilute DNA solutions (A₂₆₀≈0.1) has been studied in a Couette-type apparatus with the outer cylinder rotating and with the light path parallel to the cylinder axis. Shear gradients in the range of 5-160 sec.^-1 were studied. The DNA samples were whole, "half," and "quarter" molecules of T4 bacteriophage DNA, and linear and circular λb₂b₅c DNA. For the linear molecules, the fractional flow dichroism is a linear function of molecular weight. The dichroism for linear A DNA is about 1.8 that of the circular molecule. For a given DNA, the dichroism is an approximately linear function of shear gradient, but with a slight upward curvature at low values of G, and some trend toward saturation at larger values of G. The fractional dichroism increases as the supporting electrolyte concentration decreases.

Rearing cyprinid fry in cages [conclusions only] [Translation from: Gospodarka rybna, Warszawa 27(8/9) 3-5, 1975]

This short translation presents the conclusions only of a study of the proposed method of rearing cyprinid fry in cages and how this method can increase the amount of material produced of the stocked fry.

The effect of the group delay ripple of chirped fiber grating on composite second-order in optical fiber CATV system

The effect of group delay ripple of chirped fiber gratings on composite second-order (CSO) performance in optical fiber CATV system is investigated. We analyze the system CSO performances for different ripple amplitudes, periods and residual dispersion amounts in detail. It is found that the large ripple amplitude and small ripple period will deteriorate the system CSO performance seriously. Additionally, the residual dispersion amount has considerable effect on CSO performance in the case of small ripple amplitude and large ripple period. (c) 2004 Elsevier B.V. All rights reserved.

A study of T = 2 states in ^(12)B, ^(12)C, ^(20)F and ^(28)Al

The lowest T = 2 states have been identified and studied in the nuclei ¹²C, ¹²B, ²⁰F and and ²⁸Al. The first two of these were produced in the reactions ¹⁴C(p,t)¹²C and ¹⁴C (p,³He)¹²B, at 50.5 and 63.4 MeV incident proton energy respectively, at the Oak Ridge National Laboratory. The T = 2 states in ²⁰F and ²⁸Al were observed in (³He,p) reactions at 12-MeV incident energy, with the Caltech Tandem accelerator.

The results for the four nuclei studied are summarized below:

(1) ¹²C: the lowest T = 2 state was located at an excitation energy of 27595 ± 20 keV, and has a width less than 35 keV.

(2) ¹²B: the lowest T = 2 state was found at an excitation energy of 12710 ± 20 keV. The width was determined to be less than 54 keV and the spin and parity were confirmed to be 0⁺. A second ¹²B state (or doublet) was observed at an excitation energy of 14860 ± 30 keV with a width (if the group corresponds to a single state) of 226 ± 30 keV.

(3) ²⁰F: the lowest T = 2 state was observed at an excitation of 6513 ± 5 keV; the spin and parity were confirmed to be 0⁺. A second state, tentatively identified as T = 2 from the level spacing, was located at 8210 ± 6 keV.

(4) ²⁸Al: the lowest T = 2 state was identified at an excitation of 5997 ± 6 keV; the spin and parity were confirmed to be 0⁺. A second state at an excitation energy of 7491 ± 11 keV is tentatively identified as T = 2, with a corresponding (tentative) spin and parity assignment J^π = 2⁺.

The results of the present work and the other known masses of T = 2 states and nuclei for 8 ≤ A ≤ 28 are summarized, and massequation coefficients have been extracted for these multiplets. These coefficients were compared with those from T = 1 multiplets, and then used to predict the mass and stability of each of the unobserved members of the T = 2 multiplets.

Desenvolvimento de técnica de análise de imagens digitais para avaliação do estágio da fibrose em biópsias hepáticas de pacientes portadores de hepatite crônica por vírus da hepatite B e C

As hepatites crônicas por vírus são as mais frequentes, destacando-se os vírus das hepatites B (VHB) e C (VHC). O estudo anatomopatológico da biópsia hepática é considerado o padrão ouro para avaliar com precisão a distorção arquitetural e o grau de fibrose do parênquima do fígado, importantes fatores prognósticos para os pacientes portadores de hepatites crônicas virais. Na avaliação histopatológica atual, em adição aos relatos subjetivos das alterações histológicas, escores semiquantitativos que correlacionam achados morfológicos com graus numéricos são usados, tais como os reconhecidos escores de Ishak e METAVIR. Entretanto, em todos estes sistemas há a desvantagem da subjetividade do examinador e da incorporação de alterações categóricas, sem referências às mudanças quantitativas do colágeno hepático. Técnicas de análise de imagens digitais (AID) que fornecem quantificação objetiva dos graus de fibrose em amostras histológicas têm sido desenvolvidas. Todavia, o alto custo e dificuldade ao acesso das tecnologias descritas restringem seu uso a poucos centros especializados. Este estudo visa o desenvolvimento de uma técnica de custo acessível para a análise de imagens digitais da fibrose hepática em hepatites crônicas virais. Foram estudadas 304 biópsias de pacientes com hepatite crônica por vírus B e C, obtidas através de agulhas Menghini. Todas as amostras tinham pelo menos 15 mm de comprimento ou cinco espaços-porta completos e foram coradas pelo método Tricrômico de Masson. O estadiamento foi feito por um único hepatopatologista experiente, sem o conhecimento dos dados clínicos dos pacientes. Os escores de Ishak e METAVIR foram aplicados. As imagens microscópicas foram digitalizadas. Os índices de fibrose foram determinados de forma automatizada, em técnica desenvolvida no programa Adobe Photoshop. Para o escore de Ishak, observamos os seguintes índices de Fibrose (IF) médios: 0,8% 0,0 (estágio 0), 2.4% 0,6 (estágio 1), 4,7% 1,6 (estágio 2), 7,4% 1,4 (estágio 3), 14,9% 3,7 (estágio 4), 23,4% 2,9 (estágio 5) e 34,5% 1,5 (estágio 6). Para a classificação METAVIR: 0,8% 0,1 (estágio F0), 3,8% 1,8 (estágio F1), 7,4% 1,4 (estágio F2), 20,4% 5,2 (estágio F3) e 34,5% 1,5 (estágio F4). Observamos uma excelente correlação entre os índices de fibrose da AID e os escores de Ishak (r=0,94; p<0,001) e METAVIR (r=0,92; p<0,001). Em relação à indicação de tratamento antiviral, foi observado IF médio de 16,4%. Em relação ao diagnóstico de cirrose, foi observado IF médio de 26,9%, para o escore de Ishak, e 34,5% para a classificação METAVIR. A reprodutibilidade intra-observador foi excelente. Este novo método de análise de imagens digitais para a quantificação de fibrose hepática tem custo acessível e foi desenvolvido com tecnologia que está disponível em todo o mundo, permitindo identificar com precisão todos os estágios de fibrose, com excelente reprodutibilidade intra-observador.

Carta enviada por W.I.B. Beveridge a Justo Garate en 1955

1 carta (mecanografiada) ; 190x250mm

Propiedades catalíticas de la lipasa B de Candida antarctica inmovilizada en soportes magnéticos. Actividad hidrolítica en medios acuosos

Los enzimas son piezas fundamentales en el correcto funcionamiento de cualquier sistema biológico. Gracias a su naturaleza proteica y a las estructuras tridimensionales complejas que son capaces de adoptar, estas moléculas actúan como catalizadores de reacciones químicas. L a función de los enz imas es disminuir la energía de activación de la reacción, aumentando de este modo la velocidad de reacción. L o s enzimas no alteran el balance e nergético de las reacciones en que intervienen, ni modifican, por lo tanto, el equilibrio de la reacción . Por este motivo, en las reacciones catalizadas por enzimas se observa una mayor rapidez a la hora de alcanzar el equilibrio. La ciencia que estudia l a velocidad de las reacciones químicas que son catalizadas por enzimas es la cinética enzimática , e n la cual , las moléculas sobre las que actúan los enzimas se denominan sustratos y las moléculas resultantes de la conversión productos. El estudio de la cin ética de un enzima permite explicar los detalles de su mecanismo catalítico, su papel en el metabolismo o incluso cómo se controla su actividad en la célula. Las dos propiedades más importantes a la hora de trabajar con enzimas son: el tiempo que tarda en saturarse con un sustrato en particular y la velocidad máxima de reacción que puede alcanzar. Para el estudio de estas propiedades en el laboratorio se realizan los ensayos enzimáticos. El procedimiento a seguir en estos casos es medir la aparición de un producto o la desaparición de un sustrato frente al tiempo.

Propiedades catalíticas de la lipasa B de Candida antarctica inmovilizada en soportes magnéticos. Actividad sintética en disolventes orgánicos

Este trabajo se centra en la inmovilización de la lipasa B de Candida antarctica en nanopartículas magnéticas y la posterior caracterización cinética de su actividad sintética en medios orgánicos para la pr oducción de biodiesel. La historia del biodiesel comienza en 1893, cuando Rudolph Diesel, el padre del motor diésel, puso en marcha el primer motor de este tipo. Más tarde, en 1900, Diesel ganó el Grand Prix en la Feria Mundial de París con su m otor impulsado por un biodiesel de aceite de cacahuete. En 1903, además, comenzó la producción del Modelo T de Henry Ford, diseñado para utilizar etanol como combustible. Diesel creía que la utilización de biodiesel era el futuro de la automoción: “ el uso de aceites vegetales como combustibles para motor puede parecer insignificante hoy en día, pero estos aceites puede n convertirse, con el transcurso del tiempo, en combustibles tan importantes como el petróleo y el carbón lo son hoy en día ” Sin embargo, a p artir de 1920, los fueles basados en petróleo comenzaron a ganar terreno, debido a su mayor eficiencia, menor precio y mejor disponibilidad. De esta forma, el mercado de los biofueles quedó relegado hasta que las distintas crisis del petróleo (1973, 1979, 19 90) unidas a la creciente preocupación por la polución y la c onservación del medio ambiente, además de al aumento de la población y por tanto de la demanda de combustibles , llevaron a devolve r la mirada a estos fueles . Fue en esta época cuando comenzó, p rincipalmente en EEUU y Brasil, la producción a gran escala de biocombustibles de primera generación, basados en la utilización de excedentes agrícolas como el maíz y la caña de azúcar para la producción de bioetanol y aceites de maíz y grasas animales par a la producción de biodiesel .