Characterizing embryonic gene expression patterns in the mouse using nonredundant sequence-based selection

This article investigates the expression patterns of 160 genes that are expressed during early mouse development. The cDNAs were isolated from 7.5 d postcoitum (dpc) encloderm, a region that comprises visceral encloderm (VE), definitive encloderm, and the node-tissues that are required for the initial steps of axial specification and tissue patterning in the mouse. To avoid examining the same gene more than once, and to exclude potentially ubiquitously expressed housekeeping genes, cDNA sequence was derived from 1978 clones of the Endoderm library. These yielded 1440 distinct cDNAs, of which 123 proved to be novel in the mouse. In situ hybridization analysis was carried out on 160 of the cDNAs, and of these, 29 (18%) proved to have restricted expression patterns.

Guideline to reference gene selection for quantitative real-time PCR

Today, quantitative real-time PCR is the method of choice for rapid and reliable quantification of mRNA transcription. However, for an exact comparison of mRNA transcription in different samples or tissues it is crucial to choose the appropriate reference gene. Recently glyceraldehyde 3-phosphate dehydrogenase and P-actin have been used for that purpose. However, it has been reported that these genes as well as alternatives, like rRNA genes, are unsuitable references, because their transcription is significantly regulated in various experimental settings and variable in different tissues. Therefore, quantitative real-time PCR was used to determine the mRNA transcription profiles of 13 putative reference genes, comparing their transcription in 16 different tissues and in CCRF-HSB-2 cells stimulated with 12-O-tetradecanoylphorbol-13-acetate and ionomycin. Our results show that Classical reference genes are indeed unsuitable, whereas the RNA polymerase II gene was the gene with the most constant expression in different tissues and following stimulation in CCRF-HSB-2 cells. (C) 2003 Elsevier Inc. All rights reserved.

Expressed sequence tag analysis of genes expressed during development of the tropical abalone Haliotis asinina

The tropical abalone. Haliotis asinina. is,in ideal species to investigate the molecular mechanisms that control development. growth, reproduction and shell formation in all cultured haliotids. Here we describe the analysis of 232 expressed sequence tags (EST) obtained front a developmental H. asinina cDNA library intended for future microarray studies. From this data set we identified 183 unique gene Clusters. Of these, 90 clusters showed significant homology with sequences lodged in GenBank, ranging in function from general housekeeping to signal transduction, gene regulation and cell-cell communication. Seventy-one clusters possessed completely novel ORFs greater than 50 codons in length, highlighting the paucity of sequence data from molluscs and other lophotrochozoans. This study of developmental gene expression in H. asinina provides the foundation for further detailed analyses of abalone growth, development and reproduction.

Protein kinases associated with the yeast phosphoproteome

Background: Protein phosphorylation is an extremely important mechanism of cellular regulation. A large-scale study of phosphoproteins in a whole-cell lysate of Saccharomyces cerevisiae has previously identified 383 phosphorylation sites in 216 peptide sequences. However, the protein kinases responsible for the phosphorylation of the identified proteins have not previously been assigned. Results: We used Predikin in combination with other bioinformatic tools, to predict which of 116 unique protein kinases in yeast phosphorylates each experimentally determined site in the phosphoproteome. The prediction was based on the match between the phosphorylated 7-residue sequence and the predicted substrate specificity of each kinase, with the highest weight applied to the residues or positions that contribute most to the substrate specificity. We estimated the reliability of the predictions by performing a parallel prediction on phosphopeptides for which the kinase has been experimentally determined. Conclusion: The results reveal that the functions of the protein kinases and their predicted phosphoprotein substrates are often correlated, for example in endocytosis, cytokinesis, transcription, replication, carbohydrate metabolism and stress response. The predictions link phosphoproteins of unknown function with protein kinases with known functions and vice versa, suggesting functions for the uncharacterized proteins. The study indicates that the phosphoproteins and the associated protein kinases represented in our dataset have housekeeping cellular roles; certain kinases are not represented because they may only be activated during specific cellular responses. Our results demonstrate the utility of our previously reported protein kinase substrate prediction approach (Predikin) as a tool for establishing links between kinases and phosphoproteins that can subsequently be tested experimentally.

Expression of prostate-specific antigen variant MRNA transcripts in prostate cancer and benign prostatic hyperplasia

Prostate-specific antigen (PSA) is important in tumour detection, monitoring disease progression and tumour recurrence. however, PSA is not a cancerspecific marker as levels can also be elevated in benign prostatic disease. A number of different mRNA transcripts of PSA have also been identified in prostatic tissue, but have not been fully characterized (PSA 424, PSA 525, Schulz transcript). Tissue specimens from transurethral resection of the prostate (TURP) or radical prostatectomy were obtained from 17 men with BPH and 15 men with prostate cancer. Total RNA was extracted, and reverse-transcriptionpolymerase chain reaction (RT-PCR) and Southern analysis carried out using transcript-specific primers and probes to determine which mRNA PSA transcripts were expressed. Real-time PCR was performed to determine transcript levels between the two groups using transcript-specific primers and SYBR green fluorescence. Values obtained were normalized to a standard housekeeping gene, B2-microglobulin. Transcripts amplified by RT-PCR and real-time PCR were confirmed by DNA sequencing. Our results show that the transcripts were present in some, but not all, BPH and cancer samples indicating that they are not specific to either BPH or cancer. Analysis of real-time PCR normalized values using a Student’s t -test, shows that there is a significant difference between the two groups for PSA 424, but not wild-type PSA, PSA 525 or the Schulz transcript. Although a larger cohort of samples is needed to further confirm these results, these findings suggest that mRNA levels of PSA 424 may have some utility as a diagnostic or prognostic marker in prostate cancer detection.

Dynamic simulation of chemical plant

A CSSL- type modular FORTRAN package, called ACES, has been developed to assist in the simulation of the dynamic behaviour of chemical plant. ACES can be harnessed, for instance, to simulate the transients in startups or after a throughput change. ACES has benefited from two existing simulators. The structure was adapted from ICL SLAM and most plant models originate in DYFLO. The latter employs sequential modularisation which is not always applicable to chemical engineering problems. A novel device of twice- round execution enables ACES to achieve general simultaneous modularisation. During the FIRST ROUND, STATE-VARIABLES are retrieved from the integrator and local calculations performed. During the SECOND ROUND, fresh derivatives are estimated and stored for simultaneous integration. ACES further includes a version of DIFSUB, a variable-step integrator capable of handling stiff differential systems. ACES is highly formalised . It does not use pseudo steady- state approximations and excludes inconsistent and arbitrary features of DYFLO. Built- in debug traps make ACES robust. ACES shows generality, flexibility, versatility and portability, and is very convenient to use. It undertakes substantial housekeeping behind the scenes and thus minimises the detailed involvement of the user. ACES provides a working set of defaults for simulation to proceed as far as possible. Built- in interfaces allow for reactions and user supplied algorithms to be incorporated . New plant models can be easily appended. Boundary- value problems and optimisation may be tackled using the RERUN feature. ACES is file oriented; a STATE can be saved in a readable form and reactivated later. Thus piecewise simulation is possible. ACES has been illustrated and verified to a large extent using some literature-based examples. Actual plant tests are desirable however to complete the verification of the library. Interaction and graphics are recommended for future work.

A new phylogenetic group of Propionibacterium acnes

Immunofluorescence microscopy-based identification of presumptive Propionibacterium acnes isolates, using the P. acnes-specific mAb QUBPa3, revealed five organisms with an atypical cellular morphology. Unlike the coryneform morphology seen with P. acnes types I and II, these isolates exhibited long slender filaments (which formed large tangled aggregates) not previously described in P. acnes. No reaction with mAbs that label P. acnes types IA (QUBPa1) and II (QUBPa2) was observed. Nucleotide sequencing of the 16S rRNA gene (1484 bp) revealed the isolates to have between 99.8 and 99.9 % identity to the 16S rRNA gene of the P. acnes type IA, IB and II strains NCTC 737, KPA171202 and NCTC 10390, respectively. Analysis of the recA housekeeping gene (1047 bp) did reveal, however, a greater number of conserved nucleotide polymorphisms between the sequences from these isolates and those from NCTC 737 (98.9 % identity), KPA171202 (98.9 % identity) and NCTC 10390 (99.1 % identity). Phylogenetic investigations demonstrated that the isolates belong to a novel recA cluster or lineage distinct from P. acnes types I and II. We now propose this new grouping as P. acnes type III. The prevalence and clinical importance of this novel recA lineage amongst isolates of P. acnes remains to be determined.

Development of a computerised library system in the construction industry

This thesis examines the ways that libraries have employed computers to assist with housekeeping operations. It considers the relevance of such applications to company libraries in the construction industry, and describes more specifically the development of an integrated cataloguing and loan system. A review of the main features in the development of computerised ordering, cataloguing and circulation control systems shows that fully integrated packages are beginning to be completed, and that some libraries are introducing second generation programs. Cataloguing is the most common activity to be computerised, both at national and company level. Results from a sample of libraries in the construction industry suggest that the only computerised housekeeping system is at Taylor Woodrow. Most of the firms have access to an in-house computer, and some of the libraries, particularly those in firms of consulting engineers, might benefit from computerisation, but there are differing attitudes amongst the librarians towards the computer. A detailed study of the library at Taylor Woodrow resulted in a feasibility report covering all the areas of its activities. One of the main suggestions was the possible use of a computerised loans and cataloguing system. An integrated system to cover these two areas was programmed in Fortran and implemented. This new system provides certain benefits and saves staff time, but at the cost of time on the computer. Some improvements could be made by reprogramming, but it provides a general system for small technical libraries. A general equation comparing costs for manual and computerised operations is progressively simplified to a form where the annual saving from the computerised system is expressed in terms of staff and computer costs and the size of the library. This equation gives any library an indication of the savings or extra cost which would result from using the computerised system.

A novel multilocus sequence typing scheme for the opportunistic pathogen Propionibacterium acnes and characterisation of type 1 cell surface-associated antigens

We have developed a novel multilocus sequence typing (MLST) scheme and database (http://pubmlst.org/pacnes/) for Propionibacterium acnes based on the analysis of seven core housekeeping genes. The scheme, which was validated against previously described antibody, single locus and random amplification of polymorphic DNA typing methods, displayed excellent resolution and differentiated 123 isolates into 37 sequence types (STs). An overall clonal population structure was detected with six eBURST groups representing the major clades I, II and III, along with two singletons. Two highly successful and global clonal lineages, ST6 (type IA) and ST10 (type IB1), representing 64?% of this current MLST isolate collection were identified. The ST6 clone and closely related single locus variants, which comprise a large clonal complex CC6, dominated isolates from patients with acne, and were also significantly associated with ophthalmic infections. Our data therefore support an association between acne and P. acnes strains from the type IA cluster and highlight the role of a widely disseminated clonal genotype in this condition. Characterization of type I cell surface-associated antigens that are not detected in ST10 or strains of type II and III identified two dermatan-sulphate-binding proteins with putative phase/antigenic variation signatures. We propose that the expression of these proteins by type IA organisms contributes to their role in the pathophysiology of acne and helps explain the recurrent nature of the disease. The MLST scheme and database described in this study should provide a valuable platform for future epidemiological and evolutionary studies of P. acnes.

Relatório de estágio no The Vintage House Lisboa Hotel

O presente relatório refere-se a um estágio curricular realizado no âmbito do Mestrado em Gestão e Direção Hoteleira. O estágio decorreu no hotel The Vintage House Lisboa, com inicio a 18 de agosto de 2014 e término a 31 de maio de 2015, consistindo num cross-training pelos departamentos: Housekeeping, F&B, Front-Office, “Célula”, Economato e Direção. O principal objetivo deste relatório é descrever as tarefas realizadas ao longo do estágio, bem como as competências adquiridas, as contribuições dadas a cada departamento, e sugestões de melhoria; que juntamente com a caracterização do hotel possibilitam um melhor entendimento de todo o trabalho. Foi através da observação e participação nas várias funções da unidade hoteleira que foi possível realizar uma intensa análise para, posteriormente, descrever os vários departamentos e, por fim, propor a melhoria da análise da satisfação dos clientes do hotel. A análise à satisfação do cliente é realizada através de três questionários, cada um dirigido a um público diferente: clientes externos dos outlets de F&B, clientes MICE, e hóspedes. Através desta análise, a unidade hoteleira poderá obter bastantes vantagens como o acréscimo da qualidade do serviço, o aumento da satisfação dos seus clientes, e a sua retenção e lealdade. Ao terminar este relatório, constatou-se a forte importância que o conhecimento das necessidades, perceções e expectativas dos clientes tem para o desenvolvimento e distinção de uma empresa ou unidade hoteleira; traduzindo-se numa extrema necessidade de uma continua adaptação às transformações do sector hoteleiro e dos seus consumidores, para conseguir um maior destacamento da feroz concorrência.

Relatório de Estágio no Hotel Villa Batalha

O presente relatório diz respeito ao estágio realizado no âmbito do Mestrado em Gestão e Direção Hoteleira. Este foi realizado no Hotel Villa Batalha e teve o seu início a 1 de agosto de 2013 e terminou a 17 de maio de 2014. O objetivo deste relatório é descrever o estágio, o que foi feito e quais os conhecimentos e competências adquiridos. Durante o estágio foi realizado um cross training pelos vários departamentos do hotel, incluindo o departamento de Front Office, o departamento de Food & Beverage (F&B), o departamento de Manutenção e o departamento de Housekeeping. No final deste relatório encontra-se uma análise dos segmentos de mercado em termos de ocupação e receita, que foi realizada através da observação ao longo do estágio e da recolha de dados do programa Protel. Esta análise traduz um desafio proposto durante o estágio, com o objetivo de complementar e colmatar conhecimentos

Un legado beauvoiriano : El trabajo doméstico en la perspectiva del feminismo materialista de Christine Delphy

En la década del 70, Christine Delphy presenta la perspectiva del feminismo materialista focalizándose en el trabajo doméstico no remunerado. El análisis de Delphy pone de manifiesto el legado del pensamiento de Beauvoir, puesto que retoma las críticas al marxismo clásico, en tanto que dicha teoría no explicita las condiciones de opresión y de explotación de las mujeres. Estos temas y su tratamiento desde el feminismo materialista los situamos en algunos estudios latinoamericanos. Nos referimos a la ocupación en el "servicio" y las tareas domésticas, y a la segregación que se produce entre las mujeres. Por último, ampliamos el marco de la recepción del pensamiento de Beauvoir, en particular la cuestión de las mujeres trabajadoras, a partir del análisis de Marcela Nari

Un legado beauvoiriano : El trabajo doméstico en la perspectiva del feminismo materialista de Christine Delphy

En la década del 70, Christine Delphy presenta la perspectiva del feminismo materialista focalizándose en el trabajo doméstico no remunerado. El análisis de Delphy pone de manifiesto el legado del pensamiento de Beauvoir, puesto que retoma las críticas al marxismo clásico, en tanto que dicha teoría no explicita las condiciones de opresión y de explotación de las mujeres. Estos temas y su tratamiento desde el feminismo materialista los situamos en algunos estudios latinoamericanos. Nos referimos a la ocupación en el "servicio" y las tareas domésticas, y a la segregación que se produce entre las mujeres. Por último, ampliamos el marco de la recepción del pensamiento de Beauvoir, en particular la cuestión de las mujeres trabajadoras, a partir del análisis de Marcela Nari

Un legado beauvoiriano : El trabajo doméstico en la perspectiva del feminismo materialista de Christine Delphy

En la década del 70, Christine Delphy presenta la perspectiva del feminismo materialista focalizándose en el trabajo doméstico no remunerado. El análisis de Delphy pone de manifiesto el legado del pensamiento de Beauvoir, puesto que retoma las críticas al marxismo clásico, en tanto que dicha teoría no explicita las condiciones de opresión y de explotación de las mujeres. Estos temas y su tratamiento desde el feminismo materialista los situamos en algunos estudios latinoamericanos. Nos referimos a la ocupación en el "servicio" y las tareas domésticas, y a la segregación que se produce entre las mujeres. Por último, ampliamos el marco de la recepción del pensamiento de Beauvoir, en particular la cuestión de las mujeres trabajadoras, a partir del análisis de Marcela Nari

Unraveling the function of bacterial-type phosphoenolpyruvate carboxylase in vascular plants

Two distinct phosphoenolpyruvate carboxylase (PEPC) isozymes occur in vascular plants and green algae: plant-type PEPC (PTPC) and bacterial-type PEPC (BTPC). PTPC polypeptides typically form a tightly regulated cytosolic Class-1 PEPC homotetramer. BTPCs, however, appear to be less widely expressed and to exist only as catalytic and regulatory subunits that physically interact with co-expressed PTPC subunits to form hetero-octameric Class-2 PEPC complexes that are highly desensitized to Class-1 PEPC allosteric effectors. Yeast two-hybrid studies indicated that castor plant BTPC (RcPPC4) interacts with all three Arabidopsis thaliana PTPC isozymes, and that it forms stronger interactions with AtPPC2 and AtPPC3, suggesting that specific PTPCs are preferred for Class-2 PEPC formation. In contrast, Arabidopsis BTPC (AtPPC4) appeared to interact very weakly with AtPPC2 and AtPPC3, suggesting that BTPCs from different species may have different physical properties, hypothesized to be due to sequence dissimilarities within their ~10 kDa intrinsically disordered region. Recent RNA-seq and microarray data were analyzed to obtain a better understanding of BTPC expression patterns in different tissues of various monocot and dicot species. High levels of BTPC transcripts, polypeptides and Class-2 PEPC complexes were originally discovered in developing castor seeds, but the analysis revealed a broad range of diverse tissues where abundant BTPC transcripts are also expressed, such as the developing fruits of cucumber, grape, and tomato. Marked BTPC expression correlated well with the presence of ~116 kDa immunoreactive BTPC polypeptides, as well as Class-2 PEPC complexes in the immature fruit of cucumbers and tomatoes. It is therefore hypothesized that in vascular plants BTPC and thus Class-2 PEPC complexes maintain anaplerotic PEP flux in tissues with elevated malate levels that would potently inhibit ‘housekeeping��� Class-1 PEPCs. Elevated levels of malate can be used by biosynthetically active sink tissues such as immature tomatoes and cucumbers for rapid cell expansion, drought or salt stressed roots for osmoregulation, and developing seeds and pollen as a precursor for storage lipid and protein biosynthesis.