Numerical Simulation of a Physiological Mathematical Model of Energy Consumption in a Sarcomere / Simulación Numérica de un Modelo Matemático Fisiológico de Consumo de Energía en un Sarcómero

ABSTRACT The paradigm of biological systems provides a framework to quantify the behavior of biological processes. Mathematical modeling is one of the analytical tools of biological systems used to reproduce the variables of a system for prediction. This article presents the analysis of muscular contraction, the physiological process responsible of generating force in skeletal muscle, from the point of view of mathematical modeling. The aim is to provide numerical evidences about the force generated by the sarcomere, and the energy required to produce such a force. The proposed scheme includes a model to activate the contractile cycle, based on the action potential that reaches the neuromuscular junction, the calcium release into the sarcoplasm, the contraction response, and the quantification of the energy that the sarcomere requires to perform mechanical work. The results shows that the proposed scheme is acceptable because it reproduces experimental data of force, velocity, and energy reported in the literature. The results of the proposed scheme are encouraging to scale the model at the muscle or muscle group level, in such a way that the quantification of energy can be an alternative to the indirect estimation methods of energy consumption that currently exist.

Vitrification of Mouse Blastocysts by Open or Closed System and Warming in Sucrose-containing or Sucrose-free Diluent.

BACKGROUND: Cryopreservation of embryos is of considerable relevance for the implementation of embryo transfer programs and the establishment of embryo banks in several mammalian species. OBJECTIVE: The present investigation compares two different vitrification systems and two different warming solutions. MATERIALS AND METHODS: Vitrification was performed using Open Pulled Straw (OPS) or CVM RingFibre plug (CVM) devices. Warming was carried out either in a warming solution containing 0.33 M sucrose or in a solution devoid of sucrose. RESULTS: Differences between vitrification systems were not significant. Warming in sucrose-containing diluent resulted in an expansion rate of 64%, as compared to 86% in a solution devoid of sucrose; reported hatching rates were 45% vs. 9%, respectively (p<0.05). Upon transfer, implantation rates for OPS- and CVM were 50% and 27%, respectively, compared with 55% for freshly collected embryos. The implantation rate after warming was 43% for sucrose-containing and 33% for sucrose-free medium. CONCLUSION: a) both vitrification systems are suitable for vitrifying mouse blastocysts; b) warming in sucrose-free diluent yields better embryo survival rates than in diluent containing 0.33 M sucrose.

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon.

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Use of a virtual human performance laboratory to improve integration of mathematics and biology in sports science curricula in Sweden and the United States.

New fields such as bioengineering are exploring the role of the physical sciences in traditional biological approaches to problems, with exciting results in device innovation, medicine, and research biology. The integration of mathematics, biomechanics, and material sciences into the undergraduate biology curriculum will better prepare students for these opportunities and enhance cooperation among faculty and students at the university level. We propose the study of sports science as the basis for introduction of this interdisciplinary program. This novel integrated approach will require a virtual human performance laboratory dual-hosted in Sweden and the United States. We have designed a course model that involves cooperative learning between students at Göteborg University and Stanford University, utilizes new technologies, encourages development of original research and will rely on frequent self-assessment and reflective learning. We will compare outcomes between this course and a more traditional didactic format as well as assess the effectiveness of multiple web-hosted virtual environments. We anticipate the grant will result in a network of original faculty and student research in exercise science and pedagogy as well as provide the opportunity for implementation of the model in more advance training levels and K-12 programs.

A functional CD40 receptor is expressed in pancreatic beta cells.

AIMS/HYPOTHESIS: Despite differences in function and embryonic origin, pancreatic islet cells and neurons express proteins belonging to the tumour necrosis factor receptor superfamily. While neurons express the CD40 receptor, it is unknown whether islet cells also express it. We investigated CD40 expression in human and mouse pancreatic islets as well as in NIT-1 insulinoma cells. METHODS: CD40 expression was studied by reverse transcriptase polymerase chain reaction, flow cytometry, immunohistochemistry and western blot. Responses mediated by CD40 were assessed by a luciferase gene reporter assay following stimulation with a CD40 agonist antibody. RESULTS: We found that CD40 is expressed in mouse and human pancreatic islet cells. CD40 is expressed by beta cells, and its expression is upregulated by proinflammatory cytokines (IL-1beta, IFN-gamma and TNF-alpha). CD40 signalling in NIT-1 insulinoma cells activates nuclear factor kappa-B, demonstrating that CD40 is functional. CONCLUSIONS/INTERPRETATION: We present evidence that, in addition to immune cell types, mouse and human pancreatic beta cells express CD40. Its expression is upregulated by proinflammatory stimuli, and signalling through this receptor activates NF-kappaB. We suggest that the effects of inflammatory stimuli that affect beta cell function and survival may be also mediated by signalling through the CD40 receptor. Thus, CD40 may have a role in processes associated with islet autoimmunity and transplantation.

Mutations in the yeast cyclin-dependent kinase Cdc28 reveal a role in the spindle assembly checkpoint.

Anaphase onset and mitotic exit are regulated by the spindle assembly or kinetochore checkpoint, which inhibits the anaphase-promoting complex (APC), preventing the degradation of anaphase inhibitors and mitotic cyclins. As a result, cells arrest with high cyclin-dependent kinase (CDK) activity due to the accumulation of cyclins. Aside from this, a clear-cut demonstration of a direct role for CDKs in the spindle checkpoint response has been elusive. Cdc28 is the main CDK driving the cell cycle in budding yeast. In this report, mutations in cdc28 are described that confer specific checkpoint defects, supersensitivity towards microtubule poisons and chromosome loss. Two alleles encode single mutations in the N and C terminal regions, respectively (R10G and R288G), and one allele specifies two mutations near the C terminus (F245L, I284T). These cdc28 mutants are unable to arrest or efficiently prevent sister chromatid separation during treatment with nocodazole. Genetic interactions with checkpoint and apc mutants suggest Cdc28 may regulate checkpoint arrest downstream of the MAD2 and BUB2 pathways. These studies identify a C-terminal domain of Cdc28 required for checkpoint arrest upon spindle damage that mediates chromosome stability during vegetative growth, suggesting that it has an essential surveillance function in the unperturbed cell cycle.

Differential splicing of the IA-2 mRNA in pancreas and lymphoid organs as a permissive genetic mechanism for autoimmunity against the IA-2 type 1 diabetes autoantigen.

Type 1 diabetes results from the autoimmune destruction of pancreatic beta-cells in genetically susceptible individuals. Growing evidence suggests that genetically determined variation in the expression of self-antigens in thymus may affect the shaping of the T-cell repertoire and susceptibility to autoimmunity. For example, both allelic variation and parent-of-origin effects influence the thymic expression of insulin (a known type 1 diabetes autoantigen), and insulin gene transcription levels in thymus inversely correlate with susceptibility in both humans and transgenic models. It is unclear why patients lose tolerance to IA-2 (insulinoma-associated tyrosine phosphatase-like protein, or islet cell antigen 512 [ICA512]), especially because IA-2 polymorphisms are not associated with type 1 diabetes. We report that alternative splicing determines differential IA-2 expression in islets compared with thymus and spleen. Islets express full-length mRNA and two alternatively spliced transcripts, whereas thymus and spleen exclusively express an alternatively spliced transcript lacking exon 13. This encodes for the transmembrane (TM) and juxta-membrane (JM) domains that comprise several type 1 diabetes target epitopes, supporting the concept that tolerance to IA-2 epitopes not expressed in lymphoid organs may not be achieved. We propose differential splicing as a regulatory mechanism of gene expression playing a permissive role in the development of autoimmune responses to IA-2. Our findings also show that candidate gene expression studies can help in dissecting the complex genetic determinants of a multifactorial disease such as type 1 diabetes.

Self-antigen-presenting cells expressing diabetes-associated autoantigens exist in both thymus and peripheral lymphoid organs.

Recent reports indicate that genes with tissue-restricted expression, including those encoding the type 1 diabetes autoantigens insulin, glutamic acid decarboxylase (GAD), and the tyrosine-phosphatase-like protein IA-2 (or ICA512), are transcribed in the thymus. The reported modulation of diabetes susceptibility by genetically determined differences in thymic insulin levels and studies in transgenic mice provide correlative and functional evidence that thymic expression of peripheral proteins is crucial for immunological self-tolerance. However, there are no specific data about the existence, tissue distribution, phenotype, and function of those cells that express insulin and other self-antigens in the human thymus. We find that the human thymus harbors specialized cells synthesizing (pro)insulin, GAD, and IA-2, mainly localized in the medulla, and we demonstrate such cells also in peripheral lymphoid organs (spleen and lymph nodes). Phenotypic analysis qualifies these cells as antigen-presenting cells (APCs), including both dendritic cells and macrophages. These cells often appear surrounded by apoptotic lymphocytes, both in thymus and spleen, and may therefore be involved in the deletion of autoreactive lymphocytes. Our findings demonstrate the existence of, and define the tissue distribution and phenotype of, a novel subset of APCs expressing self-antigens in human lymphoid organs that appear to be involved in the regulation of self-tolerance throughout life.

The 62E early-late puff of Drosophila contains D-spinophilin, an ecdysone-inducible PDZ-domain protein dynamically expressed during metamorphosis.

At the onset of metamorphosis in Drosophila melanogaster, the steroid hormone 20-OH ecdysone induces a small number of early and early-late puffs in the polytene chromosomes of the third-instar larval salivary gland whose activity is required for regulating the activity of a larger set of late puffs. Most of the corresponding early and early-late genes have been found to encode transcription factors that regulate a much larger set of late genes. In contrast, we describe here the identification of an ecdysone-regulated gene in the 62E early-late puff, denoted D-spinophilin, that encodes a protein similar to the mammalian protein spinophilin/neurabin II. The D-spinophilin protein is predicted to contain a highly conserved PP1-binding domain and adjacent PDZ domain, as well as a coiled-coil domain and SAM domain, and belongs to a family of related proteins from diverse organisms. Transcription of D-spinophilin is correlated with 62E puff activity during the early stages of metamorphosis and is ecdysone-dependent, making this the first member of this gene family shown to be regulated by a steroid hormone. Examination of the dynamic patterns of D-spinophilin expression during the early stages of metamorphosis are consistent with a role in central nervous system metamorphosis as well as a more general role in other tissues. As D-spinophilin appears to be the only member of this gene family in Drosophila, its study provides an excellent opportunity to elucidate the role of an important adaptor protein in a genetic model organism.

Alaska Natives assessing the health of their environment.

The changes in Alaska's ecosystems caused by pollution, contaminants and global climate change are negatively impacting Alaska Natives and rural residents who rely on natural resources for food, culture and community identity. While Alaska commerce has contributed little to these global changes and impacts, Alaska and its resources are nonetheless affected by the changes. While Alaska Natives have historically relied on Alaska's land, water and animals for survival and cultural identity, today their faith in the safety and quality of these resources has decreased. Alaska Natives no longer believe that these wild resources are the best and many are turning to alternative store-bought foods. Such a change in diet and activity may be contributing to a decline in traditional activities and a decline in general health. Contaminants are showing up in the animals, fish and waters that Alaska Natives use. Efforts need to be expanded to empower Alaska Native Tribes to collect and analyze local wild foods for various contaminants. In addition existing information on contaminants and pollution should be made readily available to Alaska residents. Armed with this type of information Alaska Native residents will be better prepared to make informed decisions on using wild foods and materials.

Benchmarking graduate health administration programs: stakeholders' perceptions.

The purpose of this study was to identify and compare among key stakeholders the factors in graduate health care administration education that are perceived to be important for ranking, or benchmarking, based on the opinions of those stakeholders, i.e., program directors, faculty, graduate students, and accrediting agency commissioners. We used an original survey to obtain stakeholders' perceptions and opinions about important process and outcome measures. We sent it to all ACEHSA-accredited graduate health care administration programs in the United States, Canada, and Puerto Rico; to full-time faculty members in each program; to three current graduate students in each program, and to all ACEHSA commissioners. We performed frequency of responses, Analysis of Variances (ANOVA) tests, and Dunnett T3 tests. A response rate of 32 percent (n = 156) was achieved for all stakeholders. A total of 67 percent of all respondents reported that benchmarking graduate health care administration programs was important. The study results revealed a significant difference between populations on the importance of evaluating certain process and outcome measures related to curriculum, research, student characteristics, and resources. However, most of the stakeholders reported that curriculum, faculty, and graduate student performance were the key quality indicators of a program. The results of this study provide preliminary information for health care administration programs to begin to develop an educational benchmarking effort.

Construction and transposition of a 100-kilobase extended P element in Drosophila.

We have used P element deletion derivatives at defined locations in the Drosophila genome to construct a 100-kb extended P element more than twice the size of any previously available. We demonstrate that this prototypical extended P element is capable of transposition to new sites in the genome. The structural and functional integrity of a transposed extended P element was confirmed using molecular, genetic, and cytogenetic criteria. This is the first method shown to be capable of producing large, unlinked transpositional duplications in Drosophila. The ability to produce functional transposable elements from half-elements is novel and has many potential applications for the functional analysis of complex genomes.

The E23 early gene of Drosophila encodes an ecdysone-inducible ATP-binding cassette transporter capable of repressing ecdysone-mediated gene activation.

At the onset of Drosophila metamorphosis, the steroid hormone 20-OH ecdysone directly induces a small number of early puffs in the polytene chromosomes of the larval salivary gland. Proteins encoded by the early genes corresponding to these transcriptional puffs then regulate the activity of both the early puffs themselves and a much larger set of late puffs. Three of these early genes encode transcription factors that play critical regulatory roles during metamorphosis. Here we report the cloning, DNA sequence, genomic structure, ecdysone inducibility, and temporal expression of an early gene residing in the 23E early puff and denoted E23 (Early gene at 23). In contrast to other early genes, E23 encodes a protein with similarity to ATP-binding cassette transporters. Using heat shock-inducible transgenes, we found that E23 overexpression not only produces phenotypic abnormalities and lethality, but also interferes with ecdysone-mediated gene activation, demonstrating that E23 is capable of modulating the ecdysone response. Our results suggest the existence of a previously unrecognized regulatory mechanism for modulating steroid hormone signaling in Drosophila.

The L63 gene is necessary for the ecdysone-induced 63E late puff and encodes CDK proteins required for Drosophila development.

The pulse of ecdysone that triggers Drosophila metamorphosis activates six early genes in a primary response made visible by polytene chromosome puffs. The secondary response is detected by the induction of over 100 late puffs, only a few of which have been subject to molecular genetic analysis. We present a molecular and mutational analysis of the L63 gene responsible for the late puff at 63E. This gene contains overlapping L63A, B, and C transcription units of which the A unit encodes two isoforms and the B unit three. The C unit, which exhibits little activity, encodes one of the B isoforms. Evidence that L63B, but not L63A, transcription is ecdysone responsive derives from their developmental transcription profiles and from P-element mutagenesis showing that ecdysone induction of the 63E puff requires sequences adjacent to the 5' end of L63B but not those adjacent to the 5' end of L63A. L63-specific lethal mutations showed that L63 is required not only for metamorphosis, but also maternally and for embryonic and larval development. The L63 proteins contain a common C-terminal 294-aa sequence that is 71% identical to the CDK sequence of the murine PFTAIRE protein. In vivo tests of L63 proteins altered by site-directed mutagenesis showed that they exhibit CDK functions. L63 proteins are widely distributed among late larval and prepupal tissues and are unlikely to be involved in cell cycle functions.

The 82F late puff contains the L82 gene, an essential member of a novel gene family.

Metamorphosis in Drosophila results from a hierarchy of ecdysone-induced gene expression initiated at the end of the third larval instar. A now classical model of this hierarchy was proposed based on observations of the activity of polytene chromosome "puffs" which distinguished "early" puffs as those directly induced by ecdysone and "late" puffs as those which become active as a secondary response to the hormone. We report here the isolation and characterization of the L82 gene corresponding to the extensively characterized late puff at 82F. L82 is a complex gene that spans at least 50 kb of genomic DNA, produces at least seven different nested mRNAs, and has homology to a novel gene family. In contrast to most previously characterized puff genes, the broad developmental expression pattern of L82 suggests that it is controlled by both ecdysone-dependent and ecdysone-independent regulatory mechanisms. L82 mutations were identified by transgene rescue of developmental delay and eclosion lethal phenotypes.

Process mapping in screening mammography.

Successful screening mammography programs aim to screen large numbers of women efficiently and inexpensively. Development of an effective screening mammography program requires skilled personnel, solid infrastructure, and a robust computer system. A group of physicians, technologists, computer support personnel, and administrators carefully analyzed a growing screening mammography program as a series of steps, starting with the request for the examination and ending with the receipt of a hard-copy consultation. The analysis involved a detailed examination of every step and every possible outcome in the screening process. The information gained through process mapping may be used for identification of systemic and personnel problems, allocation of resources, modification of workplace architecture, and design of computer networks. Process mapping is helpful for those involved in designing and improving screening mammography programs. Viewing a process (i.e., obtaining a screening mammogram) as a series of steps may allow for the identification of inefficient components that may limit growth.

[Isolation and characterization of Cryptococcus neoformans var. gattii from samples of Eucalyptus camaldulensis in Mexico city.]. / Aislamiento y caracterización de Cryptococcus neoformans var. gattii a partir de muestras de Eucalyptus camaldulensis en la ciudad de México.

The habitat of Cryptococcus neoformans var. gattii is not fully known in Mexico. We investigated the relationship of the yeast with Eucalyptus camaldulensis soil in three main Avenues of the city. A total of 135 trees of the species E. camaldulensis, were selected. Samples were taken in duplicate from the ground containing vegetable debris, tree cortex, leaves and flowers. Isolation of the yeast was made on Guizotia abyssinica media, using Staib technique. The identification was accomplished by biochemical and morphologic tests, and caraterization of the variety was made by bromotimol canavanine-glycine-blue (CGB) and D-proline tests. Isolation of 87 strains of Cryptococcus spp. was acomplished and eight of them were identified as C. neoformans var. gattii. These findings confirmed the close relationship of C. neoformans var. gattii and E. camaldulensis. To our knowledge this is the first report concerning the isolation of this variety from E. camaldulensis trees in Mexico.