[Approach to cardiac function analysis: introduction (author's transl)].
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Heart↗
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[Statistical considerations on the evaluation of the Krogh-Poulsen clinical functional analysis].
361 patients, who five years previously were supplied with removable partial protheses, were submitted to Krogh-Poulson clinical function analyses. The test showed no suggestion of functional disturbances in only 12% of patients. Investigations showed that the test should be improved as to reliability, objectivity and validity before being used as a screening procedure in practice.
Denture Retention↗
Localization and functional analysis of structural and regulatory dehalogenase genes carried on DEH from Pseudomonas putida PP3.
Pseudomonas putida PP3 expressed two dehalogenases, DehI and DehII. The DehI gene (dehI) was located on a mobile DNA element (DEH) which inserted at high frequencies into target plasmids from its chromosomal location. From a recombinant TOL plasmid (pWW0) containing a 6.0-kb DEH element inserted into the plasmid's 5.6-kb EcoRI-G restriction endonuclease fragment, an 11.6-kb EcoRI fragment was cloned. Subcloning analysis and insertion mutagenesis produced a structural map of the DEH element and located the dehalogenase functions. The gene dehI was transcribed from a regulated promoter on DEH which was expressed in P. putida and Escherichia coli. The direction of transcription of dehI was determined, and it was also found to be under positive control, activated by an adjacent regulatory gene (dehRI). Expression of dehI in clones containing the intact DEH supported good growth on 2-monochloropropionate (2MCPA). Subclones lacking dehRI expressed dehI at levels which allowed only slow growth on 2MCPA, even when dehI expression was initiated from vector promoters. Expression of dehI in P. putida containing the intact DEH element required rpoN, suggesting that it was omega 54 dependent. The intact DEH element transferred to P. putida on a suicide plasmid donor pAWT34 (pBR325 replicon), and dehI was stably inherited, without vector DNA sequences, in transformants selected on 2MCPA. This indicated that the cloned DEH element contained functions associated with recombination.
Bacterial Proteins↗
Functional analysis of the homeodomain-related proteins of the A alpha locus of Schizophyllum commune.
DNA-mediated transformation was used to correlate function with putative genes from three alternative A alpha mating-type loci (A alpha 1, A alpha 3, and A alpha 4) of Schizophyllum commune. Each DNA was tested in at least nine haploid strains, one for each of the nine A alpha mating types found in the world-wide population of S. commune. The Y and Z genes (tentatively identified by sequence analysis elsewhere) individually activate A alpha-regulated development when transformed into any strain with a different A alpha mating type. The only exceptions are when the Y alleles of A alpha 3 or A alpha 4 (i.e., Y3 or Y4, respectively) are introduced into an A alpha 1 strain (the A alpha 1 locus encodes Y1 but lacks a Z gene). These observations indicate that A alpha-regulated development is activated by the interaction (direct or indirect) of products from different genes (e.g., Z3 and Y1) rather than from different alleles of the same gene (e.g., Y1 and Y3). Therefore, the activating interaction is of the form ZiYj where i not equal to j and i and j are the A alpha mating types from which the Z and Y polypeptides, respectively, are derived. Transformations with truncated or mutagenized genes begin to define essential regions of the genes and their products. Activity is in some cases dependent upon the particular A alpha mating type of the recipient. A working hypothesis for the activation of A alpha-regulated development is proposed.
Crosses, Genetic↗
Immunophenotyping and functional analysis of purified human uterine mast cells.
Human mast cells have been purified from uterine tissues, and their surface marker profile and function have been evaluated as part of ongoing studies of mast cell heterogeneity. Using a panel of antibodies, purified uterine mast cells (UMC; 81% +/- 7% purity, n = 10) were analyzed by immunofluorescence and flow cytometry for surface expression of various antigens. Consistent with previous analyses of mast cells from other tissues, UMC expressed HLA class I, IgE, c-kit receptor, CD9, CD33, CD43, CD45, and CD54, while CD11a, CD11b, CD14, CD16, CD23, and CD64 were not detected. Unlike other mast cells, UMC expressed CD11c/CD18 (p150,95) and CD32 (Fc gamma RII). Additional antigens not previously studied on mast cells included the selectin LECAM-1 (Leu-8) and several beta 1 and beta 3 integrins; expression of very late activation antigen-4 (VLA-4) (CD49d/CD29), VLA-5 (CD49e/CD29), and the vitronectin receptor (CD51/CD61) was seen. Functional studies showed that treatment of human umbilical vein endothelial cells with interleukin-1 (5 ng/mL for 4 hours) resulted in a twofold to threefold increase in adhesiveness for UMC. Purification procedures did not alter histamine release responses to anti-IgE or the calcium ionophore A23187, and treatment of UMC with an anti-CD32 monoclonal antibody (IV.3) did not induce histamine release or alter anti-IgE-induced release. These data suggest that UMC may possess unique phenotypic characteristics, and support the concept of mast cell heterogeneity.
Antibodies, Monoclonal↗
The guinea pig I region. II. Functional analysis.
We have examined whether an association exists between specific Ia antigen genes and Ir genes which are encoded within the same haplotype. Functionally monospecific sera to the Ia antigens of the guinea pig MHC were selective in their ability to inhibit antigen-specific T-cell proliferation and we were thus able to demonstrate an association between individual Ia specificities and specific Ir genes. The results of these studies in inbred animals were confirmed by examining the association of Ir genes and Ia antigens in the outbred guinea pig population. Of great interest was the observation that antisera made against cross-reactive Ia antigens of strains lacking specific Ir genes would still inhibit immune responses of strains possessing the Ir gene, if the Ir gene was associated with that Ia antigen in the responder strain.
Albumins↗
Functional analysis of T cells expressing Ia antigens. I. Demonstration of helper T-cell heterogeneity.
We have examined the expression of I-region antigens on functional subpopulations of murine T cells. A.TH anti-A.TL (anti-Ik, Sk, Gk) alloantiserum was raised by immunization of recipients with concanavalin A (Con A) stimulated thymic and peripheral T-cell blasts. In contrast to similar antisera made by conventional methods, the anti-Ia blast serum was highly cytotoxic for purified T lymphocytes. Moreover, it reacted in a specific fashion with T cells having particular functions. Treatment of keyhole limpet hemocyanin (KLH)-primed B10.A (H-2 alpha) T cells with this antiserum plus complement resulted in the elimination of helper activity for B-cell responses to trinitrophenyl-KLH. Inhibition was shown to be a result of the selective killing of one type of helper T cell whose activity could be replaced by a factor(s) found in the supernate of Con A-activated spleen cells. A second type of helper cell required for responses to protein-bound antigens appeared to be Ia-. By absorption and analysis on H-2 recombinants, at least two specificities were detectable on helper T cells; one mapping in the I-A subregion and a second in a region(s) to the right of I-J. In addition, the helper T cell(s) involved in the generation of alloreactive cytotoxic lymphocytes was shown to be Ia+, whereas cytotoxic effector cells and their precursors were Ia- with this antiserum. These results provide strong evidence for the selective expression of I-region determinants on T-cell subsets and suggest that T-cell-associated Ia antigens may play an important role in T-lymphocyte function.
Animals↗
Structure-function analysis of the human papillomavirus type 16 E7 oncoprotein.
The E7 gene of human papillomavirus type 16 encodes a multifunctional nuclear phosphoprotein that is functionally and structurally similar to the adenovirus (Ad) E1A proteins and the T antigens of other papovaviruses. E7 can cooperate with an activated ras oncogene to transform primary rodent cells, trans activate the Ad E2 promoter, and abrogate transforming growth factor beta-mediated repression of c-myc. Recent studies suggest that these functions may in part be a consequence of the ability of E7 to associate with the product of the retinoblastoma tumor suppressor gene (pRB). In this study, a series of site-specific mutations of the human papillomavirus type 16 E7 gene product were constructed and assessed for their effects on intracellular protein stability, ras cooperativity, transcriptional trans activation, pRB association, and phosphorylation. The results of these studies indicate that the transforming and trans-activating domains extensively overlap within a region of the protein analogous to conserved region 2 of Ad E1A, suggesting that pRB binding is necessary for both activities. Deletion of sequences in conserved region 1 abrogates cellular transformation but has only a marginal effect on trans activation. These data suggest that E7 trans activation and cellular transformation are interrelated but separable functions.
Amino Acid Sequence↗
[On-line data processing in lung functions analysis (author's transl)].
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Diagnosis, Computer-Assisted↗
A chimeric EGFR/neu receptor in functional analysis of the neu oncoprotein.
As the factor binding to the neu protein has been unknown, it has not been possible to confirm experimentally the proposed growth-factor receptor like functions of the neu protein. To approach this problem we constructed a recombinant receptor which enabled ligand regulation of the neu tyrosine kinase. The hybrid receptor consisted of the extracellular ligand binding, transmembrane and protein kinase C-substrate domains joined to the intracellular tyrosine kinase and carboxyl-terminal domains of the neu protein. Several properties of NIH3T3 cells carrying this construct were tested. We obtained the first experimental evidence that the neu proto-oncogene has mitogenic and transforming activities only in the presence of a ligand stimulating its tyrosine kinase activity. Various cellular and molecular biological parameters indicated that the chimeric receptor behaved very similarly to the EGFR. Also, this chimeric receptor has allowed us to compare the constitutive oncogenic and the ligand-activated non-oncogenic activities of the neu tyrosine kinase. In the future we plan to focus on characterization of possible differences between EGFR and neu signalling in more differentiated cellular backgrounds.
3T3 Cells↗
Long-term effects of activator (Andresen appliance) treatment. A clinical, biometric, cephalometric roentgenographic and functional analysis.
The purpose of this investigation was to analyse the long-term effects of activator treatment on the dental arches and arch relationships, the dento-facial skeleton and the function of the masticatory system. A follow-up study was performed on 112 patients treated 10-20 years previously with activators as the sole orthodontic appliance. In connection with activator treatment extractions of permanent teeth were performed on 66 patients while 46 patients were treated without tooth extractions. The investigation consisted of four separate portions (Parts I-IV).
Activator Appliances↗
[Scintigraphic functional analysis with the gamma camera during renal perfusion (author's transl)].
The controlled progress of transplanted kidneys has already been useful. We have succeeded in finding a simple method to study the function of the renal parenchyma and the collecting system. This method was also applied to determine a flow volume relation during perfusion in comparison to impulse rates. In contrast the conventional parameters, which only describe a partial function of the organ, this simple method will deliver comprehensive information before transplantation.
Animals↗
A comparative study of infantile autism and specific developmental receptive language disorders. III. Discriminant function analysis.
A psychometric, observational, and interview study was undertaken with 47 boys, aged 4 1/2 to 10 years, with nonverbal IQs of 70+ and a severe developmental disorder of language comprehension. Separate discriminant function analyses, based on behavioral, language, or cognitive features, showed little overlap between clinically defined autistic and dysphasic subgroups. Moreover, the discrimination could be made as clearly on language or cognitive characteristics as on social or behavioral critera. Language abnormalities and behavioral features also intercorrelated within the autistic subgroup. It is concluded that autism and dysphasia differ in important ways and that a cognitive deficit is an essential part of the syndrome of autism.
Autistic Disorder↗
Functional analysis of the replicator structure of lambdoid bacteriophage DNAs.
In our hybrid-plasmid reconstruction analysis of lambda (lambdoid) DNA signal structures involved in phage DNA replication, we have detected a dual system alternatingly able to initiate a first primer-RNA synthesis. Both of them--the major, primase-dependent ori system and the minor and usually suppressed, RNA-polymerase-dependent oop system--act in conjunction with a common signal structure for inception of DNA synthesis. It appears that in situations such as this, where one has to deal with the existence of regular as well as backup systems serving the same function, straightforward conclusions are no longer possible in their genetic analysis. For example, even though the oop-DNA segment can be deleted entirely from bacteriophage lambda DNA without disturbing its ability to replicate, it may not be valid to conclude that the oop system has no function in DNA replication. Dual systems of this type or organization in general have also been observed previously for some other replicons such as the R-factors R6-5 and R6K (Timmis et al. 1978; Crosa et al., this volume) or the F factor (Helinski et al., this volume), and they may be more common than presently expected.
Bacteriophage lambda↗
Genome-wide screening and functional analysis of protein glycosylation-related genes involved in tomato fruit ripening.
Protein glycosylation, an essential co- and post-translational modification, plays critical roles in plant growth, development, and stress responses. However, its functional role in tomato fruit ripening has not been extensively investigated. Here, key protein glycosylation-related genes involved in tomato fruit ripening were identified by genome-wide screen and subsequently functional characterization. First, a dataset comprising 242 glycosylation-related proteins was established based on Gene Ontology annotations in tomato, combined with sequence homology to protein glycosylation-related proteins from Arabidopsis thaliana and Homo sapiens. Then, Subsequently, 28 genes encoding highly expressed glycosylation-related proteins (RPKM > 30) at the breaker (BR) stage were selected for functional screening, and subsequently 6 genes were identified as regulators of fruit ripening by method of virus-induced gene silencing (VIGS). Among them, Solyc03g098600 (STT3B), Solyc01g109410 (OST48), Solyc04g082670 (RPN1), and Solyc08g076460 (DAD1) functioned as positive regulators of tomato fruit ripening, whereas Solyc04g005340 (UAM2) and Solyc08g075340 (XEG113), acted as negative regulators. The expression of these genes responded dynamically to multiple ripening-related cues, including temperature, light, ethylene, and transcription factors. Furthermore, silencing of these genes individually affected the expression of genes involved in fruit ripening, including ethylene biosynthesis genes (ACS2, ACS4, ACO1, and ACO3), ripening-associated transcription factors (RIN, NOR, NOR-LIKE1, FUL1, and FUL2), and the key gene (PSY1) of lycopene biosynthesis pathway. Collectively, these findings demonstrate that protein glycosylation plays an important role in tomato fruit ripening by modulating ethylene signaling, ripening-associated transcriptional regulation, and lycopene biosynthesis.
Fruit ripening↗
Stepwise Humanization of the Yeast TRAPP Core Enables Functional Analysis of TRAPP Variants.
The Transport Protein Particle (TRAPP) complex is a highly conserved multi-subunit tethering complex that plays a critical role in membrane trafficking. Mutations in TRAPP complex subunits have been implicated in a growing spectrum of rare genetic disorders, yet the molecular mechanisms underlying variant pathogenicity often remain unclear. Here, we developed a humanized yeast platform to enable systematic functional characterization of TRAPP complex variants of uncertain significance. Using a stepwise gene replacement strategy in Saccharomyces cerevisiae, we constructed a strain in which five yeast TRAPP core subunits were replaced with their human orthologues. The integration of human subunits was validated through quantitative RT-PCR and Western blotting. Growth assays revealed that partial humanization of the core complex recapitulates key functional aspects of TRAPP assembly and enables the functional investigation of variants of uncertain significance in vivo. Structural modeling and clash analysis provided insights into the impact of specific mutations on complex stability and subunit interactions. TRAPPC3 has not yet been definitively associated with human disease. Introduction of TRAPPC3 variants of uncertain clinical significance into the humanized strain resulted in pronounced growth defects and predicted structural clashes. This work demonstrates the power of humanized yeast as a model for elucidating potential genotype-phenotype relationships in TRAPPopathy disorders and provides a versatile platform to support variant interpretation, mechanistic studies, and potential therapeutic screening.
Saccharomyces cerevisiae↗
Methylation patterns associated with TTV load in geriatric hospitalized patients: an exploratory functional analysis.
Torque Teno Virus (TTV) is a widespread commensal virus within the human virome, characterized by a high prevalence in human population and an unclear pathogenic role. Over the past three decades, TTV has garnered increasing attention due to its ability to establish lifelong chronic viremia, which intriguingly fluctuates among individuals in relation to immune competence status, with a typical peak after an organ transplantation, followed by a plateau and a slow decrease. The regulatory mechanisms underlying TTV infection remain elusive, and factors influencing its interactions with the immune system have yet to be identified. To explore this complex interplay, we analyzed DNA methylation patterns associated with TTV load in older adult hospitalized patients (mean age: 83.15 ± 7.49) from the PROMOTERA cohort. In this study, we present for the first time the identification of differentially methylated probes (DMPs) correlated to TTV load in our cohort. The statistically significant DMPs were located in genes involved in immune regulation and lipid metabolism. To further characterize these findings, we performed an exploratory enrichment analysis by applying several p-value thresholds, which yielded multiple gene lists derived from the sets of significant probes. Genes associated with this epigenetic signature were found to enrich functional pathways related to immune activation, leukocyte differentiation, and cytokine production, while additional significantly enriched gene sets were involved in cell-cell adhesion and cell migration processes. Since our analysis followed an exploratory approach, these results should be interpreted as hypothesis-generating and warrant further investigation.
Humans↗
Letter: Split-renal-function analysis without ureteral catheterization in renovascular hypertension.
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Glomerular Filtration Rate↗