Generation of human computational models with machine learning

Services in smart environments pursue to increase the quality of people?s lives. The most important issues when developing this kind of environments is testing and validating such services. These tasks usually imply high costs and annoying or unfeasible real-world testing. In such cases, artificial societies may be used to simulate the smart environment (i.e. physical environment, equipment and humans). With this aim, the CHROMUBE methodology guides test engineers when modeling human beings. Such models reproduce behaviors which are highly similar to the real ones. Originally, these models are based on automata whose transitions are governed by random variables. Automaton?s structure and the probability distribution functions of each random variable are determined by a manual test and error process. In this paper, it is presented an alternative extension of this methodology which avoids the said manual process. It is based on learning human behavior patterns automatically from sensor data by using machine learning techniques. The presented approach has been tested on a real scenario, where this extension has given highly accurate human behavior models,

Nagele: un nuevo asentamiento en un paisaje artificial

Nagele es un asentamiento urbano situado en el Noordoostpolder, territorio neerlandés ganado al mar. Fue diseñado por arquitectos de los grupos De 8 en Opbouw entre los que destacaron Rietveld, Van Eesteren, Van Eyck, Bakema, Stam y Ruys. El proyecto se desarrolló entre 1947 y 1956, un periodo de tiempo con formas de proyectar muy ricas en interpretaciones. Los arquitectos pusieron en crisis los planteamientos historicistas de las nuevas poblaciones de los pólderes. Propusieron un nuevo prototipo, una morfología compacta y concéntrica que transmitiría igualdad a una comunidad agrícola, entendida como una sociedad urbana del siglo XX. La administración apoyó la propuesta que convertiría el proyecto en un arriesgado reto por su falta de antecedentes. La vigencia de las formulaciones permanece hoy en día en la ciudad construida, aunque con alteraciones. En los dibujos del proceso se encuentran los principales enunciados teóricos que este trabajo pretende descubrir. El trabajo aborda aspectos no suficientemente explorados, como su relación con el pólder, la evolución de las estrategias proyectivas, la ordenación paisajista y los elementos urbanos. El Noordoostpolder es la culminación de una serie de experiencias multidisciplinares en el reclamo de tierras a gran escala. Se estudia su estructura urbana policéntrica, la parcelación agrícola que origina el proyecto urbano y la vinculación de la vegetación con la infraestructura, proporcionando orientación, protección climática y escala humana, conceptos que impregnan las estrategias del proyecto urbano. La primera fase de la ordenación configuró áreas monofuncionales que respondían a cada una de las cuatro necesidades básicas del método científico de la ciudad higienista. El acontecimiento que marcó el final de la primera fase fue su presentación en el séptimo CIAM de 1949, cuyo título fue Aplicación de la Carta de Atenas. El programa residencial se dividió en clusters organizados en torno a una pradera vecinal central, vinculándose el orden vecinal, urbano y territorial. La segunda fase fue un catalizador de nuevos planteamientos. El proyecto se transformó en un In-between Realm, un escenario teórico donde coexisten fenómenos tradicionalmente antagónicos que Van Eyck denominó Twin Phenomena, convirtiéndose la ciudad en una réplica formal de la ambivalencia de la mente humana. La indefinición espacial no programada en la propuesta anterior se transformó en un conjunto de espacios urbanos, con límites y dimensiones adaptados a la escala humana. El proyecto es anterior a la obra escrita de Van Eyck por lo que estimuló sus enunciados teóricos. Unas ideas también reconocidas en los tres CIAM posteriores en los que también se expuso el proyecto. El diseño paisajista se integra en el proyecto urbano desde sus orígenes. El límite se compone de una barrera boscosa que protege climáticamente, proporciona escala humana y control visual frente a las llanuras infinitas del pólder. Van Eyck sintetizó el proyecto como una habitación verde sin techo, afirmación que dilucida su equivalencia con el de un interior doméstico. Exteriormente la ciudad se convierte en una unidad autónoma del territorio. Interiormente, un sistema jerarquizado de vegetación vinculado a la arquitectura y la infraestructura constituye espacios urbanos de diferentes escalas. La propuesta fue transformada por Boer y Ruys en un nuevo espacio urbano único, no asimilando los conceptos anteriores. El proyecto y construcción de los primeros elementos urbanos consistió en un reparto de tareas a De 8 en Opbouw, hecho que estimula estudiar su relación con el proyecto urbano. La estructura policéntrica organiza las aulas de las escuelas de Van Eyck, las diferentes áreas confesionales del cementerio de Ruys y las unidades residenciales, diseñadas por Stam, Rietveld y Stam-Besse. Los Twin Phenomena alcanzan un acuerdo en el corredor comercial, diseñado por Bakema y Van der Broek. La generación de espacios dentro de otros aparece también en el cementerio, a través de una nueva barrera boscosa y en el sistema de pliegues del muro que configura la iglesia de Bakema y Van der Broek. El proyecto se vincula a un planteamiento holístico, mediante el que el diseño de cada uno de sus elementos tiene en cuenta las estrategias proyectivas del todo del cual forma parte, convirtiéndose, al igual que las obras de De Stijl, en parte de una composición infinita que acerca arte y diseño en la vida cotidiana de la sociedad. La diversidad generacional e ideológica de estos arquitectos convirtió el proyecto en un tablero de juego sobre el que se aplicaron diferentes formas de proyectar la ciudad, ubicando a Nagele en un punto de inflexión del Movimiento Moderno. ABSTRACT The research focuses on the Nagele project, a Dutch urban settlement located in the Noordoostpolder, a territory which was entirely reclaimed from the IJsselmeer lake. It was designed by a group of architects from the De 8 and Opbouw teams, the leading protagonists being Rietveld, Van Eesteren, Van Eyck, Bakema, Stam y Ruys. It was designed from 1947 to 1956, a fruitful period in urban planning. These architects questioned the traditionalist urban design applied to the new populations in the IJsselmeer polders. Facing their principles, the work group proposed a new prototype; a compact and concentric urban pattern to foster equality in a new community of farm labourers, which was recognized by the architects as a twentieth century urban society. The government supported their new proposals. The lack of implementation of the innovatory conceptual statements subjected the project into a high-risk challenge. However, in spite of these difficulties, the basic concepts remain though partially transformed, in the actual city. The project drawings reflect the principle concepts that this work aims to discover. Some approaches that have not been sufficiently studied are tackled in this thesis. Firstly, the project´s relationship with the polder. Secondly, the evolution of projective strategies during the period of urban planning, the landscape design and the design of urban elements. The Noordoostpolder is the culmination of a series of multi-disciplinary experiences in large scale land reclamation, whose polycentric urban structure and agricultural subdivision provide the framework of Nagele. Linking the vegetation to infrastructure fostered orientation, climate protection and human scale; strategies which were repeated, though on a smaller scale, in the actual city. The first phase of the project was composed of mono-functional urban areas which responded to each of the four basic human needs indicated by the scientific method of the functional city. The presentation of the project at the seventh CIAM in 1949 was the event which marked the end of the first phase of the planning. This congress was entitled Implementation of the Athens Charter. The residential program was divided into housing clusters surrounding a central prairie, a pattern which was related to its urban and territorial whole. The second phase of the plan was subjected to a new theoretical approach. The urban planning became an In-between Realm, a theoretical scenario where traditionally antagonistic concepts coexist. Van Eyck named these concepts Twin Phenomena. The city thus conceived of as a counterform of the ambivalence of the human mind where spatial indefinition in the previous proposals was transformed into a Bunch of Places with defined boundaries and dimensions, all of which reflecting human scale. The landscape design was integrated into the urban project from its inception. The limits consist of a green wind-barrier which not only provides climate protection but also provides human scale and visual control towards the unlimited plains of the polder. Van Eyck summarised the project as a green room without a roof. This statement elucidates its equivalence to a domestic interior. Outwardly, the city becomes an autonomous unit on the territory. Inwardly a hierarchical vegetation system is linked to architecture and infrastructure. Together, they configure different scales of urban spaces. The proposal was transformed by Boer and Ruys into a unique urban space without assimilating Van Eyck´s concepts. The study of the Nagele landscape project of Nagele and the writings of Van Eyck verify the fact that many of his theoretical foundations (In-between Realm, Twin Phenomena, Bunch of Places, Right Scale) can be applied not only to architecture and city but also to landscape design. The application of these principles led the Nagele project to become a counterform of Van Eyck´s thinking. The design and construction of the first urban elements involved a distribution of tasks to De 8 en Opbouw, which stimulated their relationship with the urban project. The polycentric structure organised the school classrooms outlined by Van Eyck, the different areas of the cemetery planned by Ruys and the housing clusters designed by Stam, Rietveld and Stam-Besse. The Twin Phenomena concept can be applied in Van der Broek´s shopping corridor. The concept space within another space is also implemented in the cemetery surrounded by a new green barrier, and in the church built by Van der Broek and Bakema, whose spaces are configured by a folding wall. The project takes a holistic approach, which considers the design of each element within the strategies of the whole, where they become parts of an infinite composition, as in the art works of De Stijl fostering art and design to ordinary people´s daily lives. The generational and ideological diversity of these architects turned the project into a game board on which different ways of planning the city were played, obtaining Nagele the distinction of being a turning point of Modernism.

Piel artificial : metamorfosis arquitectónica del cuerpo a través de la superficie

El cuerpo, como conjunto organizado de partes que configuran el organismo, es una entidad metamórfica. El ser humano procura dar continuidad a esta condición mutante que le caracteriza, mediante diversas acciones de carácter arquitectónico. A partir de la observación de los procesos naturales, el individuo se autodefine artificialmente, transformando su realidad innata en una versión distorsionada de sí misma. Por adición, sustracción o modificación, la piel como última capa natural, se convierte en lienzo de manipulación plástica primordial para asegurar la existencia y controlar la identidad, individual y colectiva. La evolución experimental de estas intervenciones primarias, permite suplantar la piel natural por una reinterpretación construida; una piel exenta y desmontable con la que proyectar un yo diferente provisionalmente. El uso constante de esta prótesis removible e intercambiable, provoca que el cuerpo desnudo se transforme en un cuerpo vestido, en un entorno social en el que la desnudez deja de ser el estado natural del ser humano. La piel artificial se construye mediante una gran diversidad de procesos proyectuales, siendo la transformación de la superficie bidimensional en envolvente tridimensional el más utilizado a lo largo de la existencia de la vestimenta. El plano, concebido como principal formato de revestimiento humano, se adapta a su irregularidad topográfica por modelado, perforación, fragmentación, trazado, parametrización e interacción, transformándose en una envolvente cada vez más compleja y perfecta. Su diseño implica la consideración de variables como la dimensión y la escala, la función y la forma, la estructura, el material y la construcción, la técnica y los instrumentos. La vestimenta es una arquitectura habitacional individual, un límite corporal que relaciona el espacio entre el exterior e el interior, lo ajeno y lo propio, el tú y el yo; un filtro concreto y abstracto simultáneamente; una interfaz en donde el vestido es el continente y el cuerpo su contenido. ABSTRACT The body as a whole, organized of parts that make up the organism, is a metamorphic entity. The human being seeks to give continuity to this mutant condition which characterizes him through various actions of architectural character. From the observation of the natural processes, the individual defines itself artificially, transforming its innate reality into a distorted version of itself. By addition, subtraction or modification, the skin, as the last natural layer, becomes canvas of primary plastic handling in order to ensure the existence and to control the identity, both individual and collective. The experimental evolution of these primary interventions allows to impersonate the natural skin by a constructed reinterpretation; a free and detachable skin together with which to be able to project, temporarily, a different “I”. The constant use of this removable and interchangeable prosthesis causes the naked body to be transformed into a dressed body, in a social setting in which the nudity is no longer the natural state of the human being. The artificial skin is constructed by a variety of projectual processes; the most used throughout the existence of the outfit is transforming the two-dimensional surface into a three-dimensional covering. The plan, conceived as the main human lining format, adapts to its topographic irregularity by modeling, drilling, fragmentation, outline, parameters and interaction, thus becoming a type of increasingly more complex and perfect covering. Its design implies the consideration of different variables such as the dimension and the scale, the function and the shape, the structure, the material and the construction, the technique and the instruments. The clothing is an individual residential architecture, a body boundary which relates the space between outside and inside, between the external and the self, between “you” and “I”; at the same time a specific and abstract filter; an interface where the dress is the container and the body its content.

A cluster of oppositely imprinted transcripts at the Gnas locus in the distal imprinting region of mouse chromosome 2

Imprinted genes tend to occur in clusters. We have identified a cluster in distal mouse chromosome (Chr) 2, known from early genetic studies to contain both maternally and paternally imprinted, but unspecified, genes. Subsequently, one was identified as Gnas, which encodes a G protein α subunit, and there is clinical and biochemical evidence that the human homologue GNAS1, mutated in patients with Albright hereditary osteodystrophy, is also imprinted. We have used representational difference analysis, based on parent-of-origin methylation differences, to isolate candidate imprinted genes in distal Chr 2 and found two oppositely imprinted genes, Gnasxl and Nesp. Gnasxl determines a variant G protein α subunit associated with the trans-Golgi network and Nesp encodes a secreted protein of neuroendocrine tissues. Gnasxl is maternally methylated in genomic DNA and encodes a paternal-specific transcript, whereas Nesp is paternally methylated with maternal-specific expression. Their reciprocal imprinting may offer insight into the distal Chr 2 imprinting phenotypes. Remarkably, Gnasxl, Nesp, and Gnas are all part of the same transcription unit; transcripts for Gnasxl and Nesp are alternatively spliced onto exon 2 of Gnas. This demonstrates an imprinting mechanism in which two oppositely imprinted genes share the same downstream exons.

Segregating the functions of human hippocampus

It is now accepted that hippocampal lesions impair episodic memory. However, the precise functional role of the hippocampus in episodic memory remains elusive. Recent functional imaging data implicate the hippocampus in processing novelty, a finding supported by human in vivo recordings and event-related potential studies. Here we measure hippocampal responses to novelty, using functional MRI (fMRI), during an item-learning paradigm generated from an artificial grammar system. During learning, two distinct types of novelty were periodically introduced: perceptual novelty, pertaining to the physical characteristics of stimuli (in this case visual characteristics), and exemplar novelty, reflecting semantic characteristics of stimuli (in this case grammatical status within a rule system). We demonstrate a left anterior hippocampal response to both types of novelty and adaptation of these responses with stimulus familiarity. By contrast to these novelty effects, we also show bilateral posterior hippocampal responses with increasing exemplar familiarity. These results suggest a functional dissociation within the hippocampus with respect to the relative familiarity of study items. Neural responses in anterior hippocampus index generic novelty, whereas posterior hippocampal responses index familiarity to stimuli that have behavioral relevance (i.e., only exemplar familiarity). These findings add to recent evidence for functional segregation within the human hippocampus during learning.

Cloning and characterization of human protease-activated receptor 4

Protease-activated receptors 1–3 (PAR1, PAR2, and PAR3) are members of a unique G protein-coupled receptor family. They are characterized by a tethered peptide ligand at the extracellular amino terminus that is generated by minor proteolysis. A partial cDNA sequence of a fourth member of this family (PAR4) was identified in an expressed sequence tag database, and the full-length cDNA clone has been isolated from a lymphoma Daudi cell cDNA library. The ORF codes for a seven transmembrane domain protein of 385 amino acids with 33% amino acid sequence identity with PAR1, PAR2, and PAR3. A putative protease cleavage site (Arg-47/Gly-48) was identified within the extracellular amino terminus. COS cells transiently transfected with PAR4 resulted in the formation of intracellular inositol triphosphate when treated with either thrombin or trypsin. A PAR4 mutant in which the Arg-47 was replaced with Ala did not respond to thrombin or trypsin. A hexapeptide (GYPGQV) representing the newly exposed tethered ligand from the amino terminus of PAR4 after proteolysis by thrombin activated COS cells transfected with either wild-type or the mutant PAR4. Northern blot showed that PAR4 mRNA was expressed in a number of human tissues, with high levels being present in lung, pancreas, thyroid, testis, and small intestine. By fluorescence in situ hybridization, the human PAR4 gene was mapped to chromosome 19p12.

Computerized polymorphic marker identification: Experimental validation and a predicted human polymorphism catalog

A computational system for the prediction of polymorphic loci directly and efficiently from human genomic sequence was developed and verified. A suite of programs, collectively called pompous (polymorphic marker prediction of ubiquitous simple sequences) detects tandem repeats ranging from dinucleotides up to 250 mers, scores them according to predicted level of polymorphism, and designs appropriate flanking primers for PCR amplification. This approach was validated on an approximately 750-kilobase region of human chromosome 3p21.3, involved in lung and breast carcinoma homozygous deletions. Target DNA from 36 paired B lymphoblastoid and lung cancer lines was amplified and allelotyped for 33 loci predicted by pompous to be variable in repeat size. We found that among those 36 predominately Caucasian individuals 22 of the 33 (67%) predicted loci were polymorphic with an average heterozygosity of 0.42. Allele loss in this region was found in 27/36 (75%) of the tumor lines using these markers. pompous provides the genetic researcher with an additional tool for the rapid and efficient identification of polymorphic markers, and through a World Wide Web site, investigators can use pompous to identify polymorphic markers for their research. A catalog of 13,261 potential polymorphic markers and associated primer sets has been created from the analysis of 141,779,504 base pairs of human genomic sequence in GenBank. This data is available on our Web site (pompous.swmed.edu) and will be updated periodically as GenBank is expanded and algorithm accuracy is improved.

Peptide design by artificial neural networks and computer-based evolutionary search

A technique for systematic peptide variation by a combination of rational and evolutionary approaches is presented. The design scheme consists of five consecutive steps: (i) identification of a “seed peptide” with a desired activity, (ii) generation of variants selected from a physicochemical space around the seed peptide, (iii) synthesis and testing of this biased library, (iv) modeling of a quantitative sequence-activity relationship by an artificial neural network, and (v) de novo design by a computer-based evolutionary search in sequence space using the trained neural network as the fitness function. This strategy was successfully applied to the identification of novel peptides that fully prevent the positive chronotropic effect of anti-β1-adrenoreceptor autoantibodies from the serum of patients with dilated cardiomyopathy. The seed peptide, comprising 10 residues, was derived by epitope mapping from an extracellular loop of human β1-adrenoreceptor. A set of 90 peptides was synthesized and tested to provide training data for neural network development. De novo design revealed peptides with desired activities that do not match the seed peptide sequence. These results demonstrate that computer-based evolutionary searches can generate novel peptides with substantial biological activity.

Natural genetic exchange between Haemophilus and Neisseria: Intergeneric transfer of chromosomal genes between major human pathogens

Members of the bacterial families Haemophilus and Neisseria, important human pathogens that commonly colonize the nasopharynx, are naturally competent for DNA uptake from their environment. In each genus this process is discriminant in favor of its own and against foreign DNA through sequence specificity of DNA receptors. The Haemophilus DNA uptake apparatus binds a 29-bp oligonucleotide domain containing a highly conserved 9-bp core sequence, whereas the neisserial apparatus binds a 10-bp motif. Each motif (“uptake sequence”, US) is highly over-represented in the chromosome of the corresponding genus, particularly concentrated with core sequences in inverted pairs forming gene terminators. Two Haemophilus core USs were unexpectedly found forming the terminator of sodC in Neisseria meningitidis (meningococcus), and sequence analysis strongly suggests that this virulence gene, located next to IS1106, arose through horizontal transfer from Haemophilus. By using USs as search strings in a computer-based analysis of genome sequence, it was established that while USs of the “wrong” genus do not occur commonly in Neisseria or Haemophilus, where they do they are highly likely to flag domains of chromosomal DNA that have been transferred from Haemophilus. Three independent domains of Haemophilus-like DNA were found in the meningococcal chromosome, associated respectively with the virulence gene sodC, the bio gene cluster, and an unidentified orf. This report identifies intergenerically transferred DNA and its source in bacteria, and further identifies transformation with heterologous chromosomal DNA as a way of establishing potentially important chromosomal mosaicism in these pathogenic bacteria.

Mouse mammary tumor virus/v-Ha-ras transgene-induced mammary tumors exhibit strain-specific allelic loss on mouse chromosome 4

Hybrid mice carrying oncogenic transgenes afford powerful systems for investigating loss of heterozygosity (LOH) in tumors. Here, we apply this approach to a neoplasm of key importance in human medicine: mammary carcinoma. We performed a whole genome search for LOH using the mouse mammary tumor virus/v-Ha-ras mammary carcinoma model in female (FVB/N × Mus musculus castaneus)F1 mice. Mammary tumors developed as expected, as well as a few tumors of a second type (uterine leiomyosarcoma) not previously associated with this transgene. Genotyping of 94 anatomically independent tumors revealed high-frequency LOH (≈38%) for markers on chromosome 4. A marked allelic bias was observed, with M. musculus castaneus alleles almost exclusively being lost. No evidence of genomic imprinting effects was noted. These data point to the presence of a tumor suppressor gene(s) on mouse chromosome 4 involved in mammary carcinogenesis induced by mutant H-ras expression, and for which a significant functional difference may exist between the M. musculus castaneus and FVB/N alleles. Provisional subchromosomal localization of this gene, designated Loh-3, can be made to a distal segment having syntenic correspondence to human chromosome 1p; LOH in this latter region is observed in several human malignancies, including breast cancers. Evidence was also obtained for a possible second locus associated with LOH with less marked allele bias on proximal chromosome 4.

Molecular cloning and functional analysis of SUT-1, a sulfate transporter from human high endothelial venules

High endothelial venules (HEV) are specialized postcapillary venules found in lymphoid organs and chronically inflamed tissues that support high levels of lymphocyte extravasation from the blood. One of the major characteristics of HEV endothelial cells (HEVEC) is their capacity to incorporate large amounts of sulfate into sialomucin-type counter-receptors for the lymphocyte homing receptor L-selectin. Here, we show that HEVEC express two functional classes of sulfate transporters defined by their differential sensitivity to the anion-exchanger inhibitor 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), and we report the molecular characterization of a DIDS-resistant sulfate transporter from human HEVEC, designated SUT-1. SUT-1 belongs to the family of Na+-coupled anion transporters and exhibits 40–50% amino acid identity with the rat renal Na+/sulfate cotransporter, NaSi-1, as well as with the human and rat Na+/dicarboxylate cotransporters, NaDC-1/SDCT1 and NaDC-3/SDCT2. Functional expression studies in cRNA-injected Xenopus laevis oocytes showed that SUT-1 mediates high levels of Na+-dependent sulfate transport, which is resistant to DIDS inhibition. The SUT-1 gene mapped to human chromosome 7q33. Northern blotting analysis revealed that SUT-1 exhibits a highly restricted tissue distribution, with abundant expression in placenta. Reverse transcription–PCR analysis indicated that SUT-1 and the diastrophic dysplasia sulfate transporter (DTD), one of the two known human DIDS-sensitive sulfate transporters, are coexpressed in HEVEC. SUT-1 and DTD could correspond, respectively, to the DIDS-resistant and DIDS-sensitive components of sulfate uptake in HEVEC. Together, these results demonstrate that SUT-1 is a distinct human Na+-coupled sulfate transporter, likely to play a major role in sulfate incorporation in HEV.

The human XRCC9 gene corrects chromosomal instability and mutagen sensitivities in CHO UV40 cells

The Chinese hamster ovary (CHO) mutant UV40 cell line is hypersensitive to UV and ionizing radiation, simple alkylating agents, and DNA cross-linking agents. The mutant cells also have a high level of spontaneous chromosomal aberrations and 3-fold elevated sister chromatid exchange. We cloned and sequenced a human cDNA, designated XRCC9, that partially corrected the hypersensitivity of UV40 to mitomycin C, cisplatin, ethyl methanesulfonate, UV, and γ-radiation. The spontaneous chromosomal aberrations in XRCC9 cDNA transformants were almost fully corrected whereas sister chromatid exchanges were unchanged. The XRCC9 genomic sequence was cloned and mapped to chromosome 9p13. The translated XRCC9 sequence of 622 amino acids has no similarity with known proteins. The 2.5-kb XRCC9 mRNA seen in the parental cells was undetectable in UV40 cells. The mRNA levels in testis were up to 10-fold higher compared with other human tissues and up to 100-fold higher compared with other baboon tissues. XRCC9 is a candidate tumor suppressor gene that might operate in a postreplication repair or a cell cycle checkpoint function.

Inactivation of the survival motor neuron gene, a candidate gene for human spinal muscular atrophy, leads to massive cell death in early mouse embryos

Proximal spinal muscular atrophy is an autosomal recessive human disease of spinal motor neurons leading to muscular weakness with onset predominantly in infancy and childhood. With an estimated heterozygote frequency of 1/40 it is the most common monogenic disorder lethal to infants; milder forms represent the second most common pediatric neuromuscular disorder. Two candidate genes—survival motor neuron (SMN) and neuronal apoptosis inhibitory protein have been identified on chromosome 5q13 by positional cloning. However, the functional impact of these genes and the mechanism leading to a degeneration of motor neurons remain to be defined. To analyze the role of the SMN gene product in vivo we generated SMN-deficient mice. In contrast to the human genome, which contains two copies, the mouse genome contains only one SMN gene. Mice with homozygous SMN disruption display massive cell death during early embryonic development, indicating that the SMN gene product is necessary for cellular survival and function.

Direct isolation of human BRCA2 gene by transformation-associated recombination in yeast

Mutant forms of the BRCA2 gene contribute significantly to hereditary breast cancer. Isolation of the normal and mutant forms of the BRCA2 gene with its natural promoter would greatly facilitate analysis of the gene and its contribution to breast cancer. We have accomplished the direct isolation of the 90-kb gene from total human DNA by transformation-associated recombination in yeast using a small amount of 5′ and 3′ BRCA2 sequence information. Because the entire isolation procedure of a single chromosomal gene could be accomplished in approximately 2 weeks, the transformation-associated recombination cloning approach is readily applicable to studies of chromosome alterations and human genetic diseases.

Isolation of human and mouse genes based on homology to REC2, a recombinational repair gene from the fungus Ustilago maydis

A human and a mouse gene have been isolated based on homology to a recombinational repair gene from the corn smut Ustilago maydis. The new human (h) gene, termed hREC2, bears striking resemblance to several others, including hRAD51 and hLIM15. hREC2 is located on human chromosome 14 at q23–24. The overall amino acid sequence reveals characteristic elements of a RECA-like gene yet harbors an src-like phosphorylation site curiously absent from hRAD51 and hLIM15. Unlike these two relatives, hREC2 is expressed in a wide range of tissues including lung, liver, placenta, pancreas, leukocytes, colon, small intestine, brain, and heart, as well as thymus, prostate, spleen, and uterus. Of greatest interest is that hREC2 is undetectable by reverse transcription-coupled PCR in tissue culture unless the cells are treated by ionizing radiation.