RESUMEN
A 75,000-dalton protein has been purified approximately 1,000-fold from rat liver, based on its capacity to organize poly(A) in a 27-residue repeating structure. This protein may be identified with the major polypeptide component of cytoplasmic poly(A)-ribonucleoprotein (RNP) previously described. The poly(A)-organizing activity of the protein is detected only in cytoplasmic fractions. Upon nuclease digestion of the 75,000-dalton protein-poly(A) complex, monomers, and higher multimers of RNP subunits can be resolved in a sucrose gradient. The sedimentation rate of the monomer is compatible with a composition of one 75,000-dalton protein molecule and one 27-residue segment of poly(A).
Asunto(s)
Citoplasma/análisis , Nucleoproteínas/análisis , Poli A , Ribonucleoproteínas/análisis , Animales , Núcleo Celular/análisis , Centrifugación por Gradiente de Densidad , Fenómenos Químicos , Química , Nucleasa Microcócica , Poli A/análisis , Ratas , Ribonucleoproteínas/aislamiento & purificaciónRESUMEN
Evolutionary ecologists are traditionally gene-focused, as genes propagate phenotypic traits across generations and mutations and recombination in the DNA generate genetic diversity required for evolutionary processes. As a consequence, the inheritance of changed DNA provides a molecular explanation for the functional changes associated with natural selection. A direct focus on proteins on the other hand, the actual molecular agents responsible for the expression of a phenotypic trait, receives far less interest from ecologists and evolutionary biologists. This is partially due to the central dogma of molecular biology that appears to define proteins as the 'dead-end of molecular information flow' as well as technical limitations in identifying and studying proteins and their diversity in the field and in many of the more exotic genera often favored in ecological studies. Here we provide an overview of a newly forming field of research that we refer to as 'Evolutionary Proteomics'. We point out that the origins of cellular function are related to the properties of polypeptide and RNA and their interactions with the environment, rather than DNA descent, and that the critical role of horizontal gene transfer in evolution is more about coopting new proteins to impact cellular processes than it is about modifying gene function. Furthermore, post-transcriptional and post-translational processes generate a remarkable diversity of mature proteins from a single gene, and the properties of these mature proteins can also influence inheritance through genetic and perhaps epigenetic mechanisms. The influence of post-transcriptional diversification on evolutionary processes could provide a novel mechanistic underpinning for elements of rapid, directed evolutionary changes and adaptations as observed for a variety of evolutionary processes. Modern state-of the art technologies based on mass spectrometry are now available to identify and quantify peptides, proteins, protein modifications and protein interactions of interest with high accuracy and assess protein diversity and function. Therefore, proteomic technologies can be viewed as providing evolutionary biologist with exciting novel opportunities to understand very early events in functional variation of cellular molecular machinery that are acting as part of evolutionary processes.
Asunto(s)
Ecología/métodos , Evolución Molecular , Proteómica/métodos , Animales , Ecología/tendencias , Humanos , Proteómica/tendenciasRESUMEN
BACKGROUND: Individual case review of spontaneous adverse event (AE) reports remains a cornerstone of medical product safety surveillance for industry and regulators. Previously we developed the Vaccine Adverse Event Text Miner (VaeTM) to offer automated information extraction and potentially accelerate the evaluation of large volumes of unstructured data and facilitate signal detection. OBJECTIVE: To assess how the information extraction performed by VaeTM impacts the accuracy of a medical expert's review of the vaccine adverse event report. METHODS: The "outcome of interest" (diagnosis, cause of death, second level diagnosis), "onset time," and "alternative explanations" (drug, medical and family history) for the adverse event were extracted from 1000 reports from the Vaccine Adverse Event Reporting System (VAERS) using the VaeTM system. We compared the human interpretation, by medical experts, of the VaeTM extracted data with their interpretation of the traditional full text reports for these three variables. Two experienced clinicians alternately reviewed text miner output and full text. A third clinician scored the match rate using a predefined algorithm; the proportion of matches and 95% confidence intervals (CI) were calculated. Review time per report was analyzed. RESULTS: Proportion of matches between the interpretation of the VaeTM extracted data, compared to the interpretation of the full text: 93% for outcome of interest (95% CI: 91-94%) and 78% for alternative explanation (95% CI: 75-81%). Extracted data on the time to onset was used in 14% of cases and was a match in 54% (95% CI: 46-63%) of those cases. When supported by structured time data from reports, the match for time to onset was 79% (95% CI: 76-81%). The extracted text averaged 136 (74%) fewer words, resulting in a mean reduction in review time of 50 (58%) seconds per report. CONCLUSION: Despite a 74% reduction in words, the clinical conclusion from VaeTM extracted data agreed with the full text in 93% and 78% of reports for the outcome of interest and alternative explanation, respectively. The limited amount of extracted time interval data indicates the need for further development of this feature. VaeTM may improve review efficiency, but further study is needed to determine if this level of agreement is sufficient for routine use.
Asunto(s)
Procesamiento de Lenguaje Natural , Informe de Investigación , Vacunas/efectos adversosRESUMEN
Multiple mating (and insemination) by females with different males, polyandry, is widespread across animals, due to material and/or genetic benefits for females. It reaches particularly high levels in some social insects, in which queens can produce significantly fitter colonies by being polyandrous. It is therefore a paradox that two thirds of eusocial hymenopteran insects appear to be exclusively monandrous, in spite of the fitness benefits that polyandry could provide. One possible cost of polyandry could be sexually transmitted parasites, but evidence for these in social insects is extremely limited. Here we show that two different species of Nosema microsporidian parasites can transmit sexually in the honey bee Apis mellifera. Honey bee males that are infected by the parasite have Nosema spores in their semen, and queens artificially inseminated with either Nosema spores or the semen of Nosema-infected males became infected by the parasite. The emergent and more virulent N. ceranae achieved much higher rates of infection following insemination than did N. apis. The results provide the first quantitative evidence of a sexually transmitted disease (STD) in social insects, indicating that STDs may represent a potential cost of polyandry in social insects.
Asunto(s)
Abejas/microbiología , Microsporidiosis/transmisión , Nosema/fisiología , Animales , ADN de Hongos/análisis , ADN de Hongos/metabolismo , Femenino , Intestinos/microbiología , Masculino , Microsporidiosis/veterinaria , Nosema/genética , Ovario/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Espermatozoides/microbiología , Esporas Fúngicas/aislamiento & purificación , Esporas Fúngicas/patogenicidadRESUMEN
Multiple mating by females (polyandry) is taxonomically widespread but the evolution of such behaviors is not clearly understood given potential costs of polyandry such as time, energy, or predation risk. The genetic variability versus parasites hypothesis predicts a reduction of parasitism due to increased genetic variability among offspring and an associated fitness gain. We tested this hypothesis with a field experiment in the bumblebee, Bombus terrestris L. Worker heterogeneity within the colony was experimentally altered by artificially inseminating queens with sperm from one male, four brothers, two males, or four unrelated males. We found genetic variability to be effective, because intensity and prevalence of the most common parasite, Crithidia bombi, a trypanosome, decreased with increasing levels of colony heterogeneity. Fitness differed between treatments but did not increase in a simple way, with increasing genetic heterogeneity among colony workers. Instead, fitness followed a U-shaped function with a minimum for small amounts of genetic heterogeneity. We therefore suggest that polyandry also induces a cost, perhaps as a result of the social structure within the colony. In evolutionary terms, singly mated females appear to be constrained by an adaptive valley that needs to be crossed before high degrees of mating frequency can be reached. This may help to explain why B. terrestris and most other social insects are often monandrous.
Asunto(s)
Abejas/fisiología , Abejas/parasitología , Variación Genética , Conducta Sexual Animal/fisiología , Animales , Abejas/genética , Crithidia/fisiología , Femenino , MasculinoRESUMEN
During copulation, males of Bombus terrestris fill the queen's sexual tract with a mating plug after transferring their sperm. The sticky secretion is produced by the male's accessory glands and disappears within a couple of days. Experiments now show that the primary function of the plug is to reduce the subsequent mating probability of the queen. The plug is not efficient in preventing sperm migration into the spermatheca. Due to its low energetic value, the plug is also unlikely to serve as a nuptial gift. This type of male interference with female mating propensity has so far not been found in social insects. This finding could, at least tentatively, explain why females of B. terrestris may not be able to take advantage of the demonstrated benefits of multiple mating. Furthermore, such male interference could be a more general phenomenon in social insects, with obvious ramifications for the evolution of polyandry in this group.
Asunto(s)
Abejas/fisiología , Conducta Sexual Animal/fisiología , Espermatozoides/fisiología , Animales , Copulación/fisiología , Metabolismo Energético , Femenino , Masculino , Transporte Espermático/fisiologíaRESUMEN
Recently, Conger, Garcia, Kauffman, Lust, Murakami and Passonneau, (1981) proposed the use of brain alanine-glutamate ratios (A:G) for the prediction of outcome after brain ischemia. This study evaluates this parameter and brain lactate concentration during and after asphyxial insults in rats. During the first 15 min of asphyxial death in rats (n = 37), lactate increased sharply from mean values of 1.48 to 18.06 mumol g-1 wet brain, and thereafter to 19.44 mumol g-1 wet brain at 180 min. During total body asphyxia (n = 38) and recovery after resuscitation, brain lactates increased to mean values of 15 and 17.5 mumol g-1 wet brain at 5 and 10 min, respectively, to recover after 30-60 min to baseline. The alanine-glutamate ratios did not rise during the insult; however, after restoration of circulation, the ratios rose to peak at about 15 min post-restoration of circulation and recovered slowly during the next 165 min to still slightly increased levels. During intermittent asphyxia (n = 15), lactate and alanine-glutamate ratios followed the same patterns as found before except at lower levels. The conclusions of this study are: (1) brain lactate concentrations had no value in predicting the potential of recovery; (2) increased lactate concentrations during recovery indicated secondary insult; (3) brain A:G's did not increase during asphyxiation; (4) brain alanine-glutamate ratios increased after restoration of circulation and may have reflected the quality of reflow; (5) increased ratios during recovery beyond 20 min indicated secondary insult. Brain alanine-glutamate ratios could not be used for prediction of outcome in asphyxial insults in rats.
Asunto(s)
Alanina/metabolismo , Asfixia/metabolismo , Encéfalo/metabolismo , Glutamatos/metabolismo , Lactatos/metabolismo , Animales , Circulación Cerebrovascular , Ácido Glutámico , Ácido Láctico , Masculino , Ratas , Ratas Endogámicas , Respiración Artificial , Resucitación , Factores de TiempoRESUMEN
The genetic architecture of fitness-relevant traits in natural populations is a topic that has remained almost untouched by quantitative genetics. Given the importance of parasitism for the host's fitness, we used QTL mapping to study the genetic architecture of traits relevant for host-parasite interactions in the trypanosome parasite, Crithidia bombi and its host, Bombus terrestris. The three traits analysed were the parasite's infection intensity, the strength of the general immune response (measured as the encapsulation of a novel antigen) and body size. The genetic architecture of these traits was examined in three natural, unmanipulated mapping populations of B. terrestris. Our results indicate that the intracolonial phenotypic variation of all three traits is based on a network of QTLs and epistatic interactions. While these networks are similar between mapping populations in complexity and number of QTLs, as well as in their epistatic interactions, the variability in the position of QTL and the interacting loci was high. Only one QTL for body size was plausibly found in at least two populations. QTLs for encapsulation and Crithidia infection intensity were located on the same linkage groups.
Asunto(s)
Abejas/genética , Abejas/parasitología , Crithidia/genética , Variación Genética , Genética de Población , Fenotipo , Animales , Abejas/inmunología , Tamaño Corporal/genética , Crithidia/patogenicidad , Epistasis Genética , Femenino , Interacciones Huésped-Parásitos , Masculino , Sitios de Carácter CuantitativoRESUMEN
The interlinking of patient monitors, which allows observation of current measurement data from several patients in a treatment unit not only at the bedside, but also at more distant monitors attached to the unit, has become generally established in intensive care medicine. In several years of use in the anesthesia department of a surgical unit, such an interconnection of information has also proved to be useful.
Asunto(s)
Servicio de Anestesia en Hospital , Anestesiología/instrumentación , Departamentos de Hospitales , Monitoreo Fisiológico/instrumentación , Servicio de Cirugía en Hospital , HumanosRESUMEN
In 47 gas mixtures particularly made for that purpose the concentration of halothane, enflurane, or isoflurane was estimated with Normac; the estimated and measured values were found to be identical on the average. To verify precision, 40 measurements of one specimen were performed; the standard deviation was 0.03 vol-% (ca. 1% relative error). Any influence of the carrier gas components O2, N2, N2O, and H2O proved to be negligible. The practical clinical application of the apparatus was easy and without malfunctions.
Asunto(s)
Anestesia por Inhalación/instrumentación , Enflurano/análisis , Halotano/análisis , Isoflurano/análisis , Administración por Inhalación , HumanosRESUMEN
A repeating structure of cytoplasmic poly(A)-ribonucleoprotein is revealed by digestion with T2 RNase. A pattern of fragments that are multiples of about 27 residues is obtained. The repeating structure is readily reconstituted from purified poly(A) and cytoplasmic factors. Reconstitution is specific for poly(A), as shown by the lack of competition by poly(G), poly(C), poly(dA), and tRNA. The repeating structure is absent from the nucleus, and so appears to be formed upon transport to the cytoplasm.
Asunto(s)
Nucleoproteínas/metabolismo , Poli A/metabolismo , Ribonucleoproteínas/metabolismo , Animales , Secuencia de Bases , Sitios de Unión , Células Cultivadas , Citoplasma/metabolismo , Ratones , Peso Molecular , Ribonucleasas/metabolismo , Fagos T/enzimologíaRESUMEN
Purified RNA polymerase II from calf thymus can bind to about 15% of the nucleosome core particles prepared from mouse myeloma cells, forming a discrete complex having a sedimentation coefficient of 18S. These bound nucleosome cores are heavily enriched in transcribed DNA sequences, are deficient in histones H2A and H2B, and undergo a reversible change in structure on RNA polymerase II binding.
Asunto(s)
ARN Polimerasas Dirigidas por ADN/genética , Nucleosomas/fisiología , ARN Polimerasa II/genética , Transcripción Genética , Animales , Bovinos , Genes , Histonas/genética , Ratones , Nucleosomas/ultraestructura , Unión ProteicaRESUMEN
The location of nucleosomes on genes for 5 S rRNA in rat liver was determined by the preparation of nucleosome core DNA fragments, hybridization with 5 S rRNA, RNase digestion, and gel electrophoresis. The resulting size distribution of RNA fragments was essentially the same as that found when the experiment was carried out with random DNA fragments.
Asunto(s)
Hígado/metabolismo , Nucleosomas/metabolismo , ARN Ribosómico/biosíntesis , Transcripción Genética , Animales , Cinética , Peso Molecular , Desnaturalización de Ácido Nucleico , Hibridación de Ácido Nucleico , Ratas , RibonucleasasRESUMEN
The best mating strategy for males differs from that of females, because females gain from mating with several males (polyandry), but males gain from monopolizing the females. As a consequence, males have evolved a variety of methods, such as the transfer of inhibitory substances from their accessory glands, to ensure exclusive paternity of the female's offspring, generally with detrimental effects on female fitness. Inhibitory substances have been identified as peptides or other specific molecules. Unfortunately, in social insects male-mating traits are investigated only poorly, although male social insects might have the same fundamental influence on female-mating behavior as found in other species. A recently developed technique for the artificial insemination of bumblebee queens allowed us to investigate which chemical compound in the mating plug of male bumblebees, Bombus terrestris L., prevents females (queens) from further mating. Surprisingly, we found that the active substance is linoleic acid, a ubiquitous and rather unspecific fatty acid. Contrary to mating plugs in other insect species, the bumblebee mating plug is highly efficient and allows the males to determine queen-mating frequencies.
Asunto(s)
Ácidos Grasos/fisiología , Conducta Sexual Animal/fisiología , Animales , Abejas , Femenino , MasculinoRESUMEN
We investigated the dynamics of sperm transfer and the potential conflict between sexes over mating opportunities in the bumblebee, Bombus terrestris L. We recorded copulation duration in flight cages and manipulated copulation time by separating mating pairs. Sperm transfer and filling of the spermatheca were measured by dissecting queens at different time intervals after the onset of mating. On average, copulations lasted 37 min but most sperm were transferred into the female genital tract within the first 2 min. Sperm reached the spermatheca after 30-80 min. Males transferred a sticky gelatinous product of their accessory glands, the 'mating plug', to the female within 10-30 min of the onset of copulation. Hence the duration of the copulation matched the time required to deposit the plug fully. A likely function of the plug is to prevent backflow of sperm, but by artificially transferring mating plugs we showed that the plug also decreased sperm transfer into the female's spermatheca for at least 4 h and possibly for much longer (8 h or more). Increased mating costs arising from male selfishness may prevent females from seeking further matings. This may help to explain why females of B. terrestris seem to be mostly singly mated in the field, even though, in experiments with artificially inseminated queens, higher sperm diversity increases reproductive output. This is the first report of a mating plug for the Bombini but similar devices are known from two other tribes of the Apidae, the Apini and Meliponini. Copyright 1999 The Association for the Study of Animal Behaviour.
RESUMEN
cDNA and genomic cloning has been used to determine the mRNA and amino acid sequence of human plasma lecithin-cholesterol acyltransferase (LCATase; EC 2.3.1.43). The mature protein was found to contain 416 amino acid residues with a hydrophobic leader sequence of 24 amino acids. An unusual feature of the message is that the poly(A) signal AATAAA overlaps the COOH-terminal glutamic acid and stop codons, and the 3' untranslated region is only 23 bases. The protein itself is distinguished by a number of extended sequences of hydrophobic amino acids, one of which contains a hexapeptide identical with the interfacial binding segment of the active site of pancreatic lipase and is similar to the same site of lingual lipase. The cloned cDNA allows the expression of active LCATase by transfected tissue culture cells.
Asunto(s)
Esterol O-Aciltransferasa/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , ADN/genética , Regulación de la Expresión Génica , Humanos , Fragmentos de Péptidos/análisis , Proteínas Recombinantes/genéticaRESUMEN
Angle-resolved x-ray diffraction patterns of Xe to 127 GPa indicate that the fcc-to-hcp transition occurs martensitically between 3 and 70 GPa in diamond-anvil cells without an intermediate phase. These data also reveal that the transition occurs by the introduction of stacking disorder in the fcc lattice at low pressure, which grows into hcp domains with increasing pressure. The small energy difference between the hcp and the fcc structures may allow the two phases to coexist over a wide pressure range. Evidence of similar stacking disorder and incipient growth of an hcp phase are also observed in solid Kr.