RESUMO
Aims This study aimed to identify the physiotherapy exercise rehabilitation services available to patients with cancer in Ireland and to identify barriers to the provision of services. Methods Physiotherapy department managers in specialised cancer centres, public and private hospitals and palliative care settings were surveyed to establish the availability of exercise rehabilitation services for patients with cancer. Results Of 40 managers contacted, 24 responded providing information about 26 services. Ten services employed a dedicated oncology physiotherapist. Exercise classes were offered to patients with cancer by five services, primarily within the palliative care setting. In the 17 hospitals which provided surgery, ten provided oncology specific post-operative exercise rehabilitation and one offered a prehabilitation programme. Limited human and physical resources and absence of established physiotherapy pathways were cited barriers to service provision. Conclusion Exercise rehabilitation is not an element of standard care for patients with cancer in Ireland.
Assuntos
Terapia por Exercício/estatística & dados numéricos , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Neoplasias/reabilitação , Cuidados Paliativos/estatística & dados numéricos , Serviço Hospitalar de Fisioterapia/estatística & dados numéricos , Modalidades de Fisioterapia/estatística & dados numéricos , Terapia por Exercício/métodos , Humanos , Entrevistas como Assunto , Irlanda/epidemiologia , Cuidados Paliativos/métodosRESUMO
The complexity of the human shoulder girdle enables the large mobility of the upper extremity, but also introduces instability of the glenohumeral (GH) joint. Shoulder movements are generated by coordinating large superficial and deeper stabilizing muscles spanning numerous degrees-of-freedom. How shoulder muscles are coordinated to stabilize the movement of the GH joint remains widely unknown. Musculoskeletal simulations are powerful tools to gain insights into the actions of individual muscles and particularly of those that are difficult to measure. In this study, we analyze how enforcement of GH joint stability in a musculoskeletal model affects the estimates of individual muscle activity during shoulder movements. To estimate both muscle activity and GH stability from recorded shoulder movements, we developed a Rapid Muscle Redundancy (RMR) solver to include constraints on joint reaction forces (JRFs) from a musculoskeletal model. The RMR solver yields muscle activations and joint forces by minimizing the weighted sum of squared-activations, while matching experimental motion. We implemented three new features: first, computed muscle forces include active and passive fiber contributions; second, muscle activation rates are enforced to be physiological, and third, JRFs are efficiently formulated as linear functions of activations. Muscle activity from the RMR solver without GH stability was not different from the computed muscle control (CMC) algorithm and electromyography of superficial muscles. The efficiency of the solver enabled us to test over 3600 trials sampled within the uncertainty of the experimental movements to test the differences in muscle activity with and without GH joint stability enforced. We found that enforcing GH stability significantly increases the estimated activity of the rotator cuff muscles but not of most superficial muscles. Therefore, a comparison of shoulder model muscle activity to EMG measurements of superficial muscles alone is insufficient to validate the activity of rotator cuff muscles estimated from musculoskeletal models.
Assuntos
Articulação do Ombro , Ombro , Humanos , Ombro/fisiologia , Articulação do Ombro/fisiologia , Fenômenos Biomecânicos , Músculo Esquelético/fisiologia , Eletromiografia , Amplitude de Movimento Articular/fisiologiaRESUMO
The apicomplexan cattle parasite Theileria parva is a major barrier to improving the livelihoods of smallholder farmers in Africa, killing over one million cattle on the continent each year. Although exotic breeds not native to Africa are highly susceptible to the disease, previous studies have illustrated that such breeds often show innate tolerance to infection by the parasite. The mechanisms underlying this tolerance remain largely unclear. To better understand the host response to T. parva infection we characterised the transcriptional response over 15 days in tolerant and susceptible cattle (n = 29) naturally exposed to the parasite. We identify key genes and pathways activated in response to infection as well as, importantly, several genes differentially expressed between the animals that ultimately survived or succumbed to infection. These include genes linked to key cell proliferation and infection pathways. Furthermore, we identify response expression quantitative trait loci containing genetic variants whose impact on the expression level of nearby genes changes in response to the infection. These therefore provide an indication of the genetic basis of differential host responses. Together these results provide a comprehensive analysis of the host transcriptional response to this under-studied pathogen, providing clues as to the mechanisms underlying natural tolerance to the disease.
Assuntos
Doenças dos Bovinos , Theileria parva , Theileriose , Bovinos , Animais , Theileria parva/genética , Theileriose/parasitologia , Doenças dos Bovinos/genética , Doenças dos Bovinos/parasitologia , Perfilação da Expressão Gênica , ÁfricaRESUMO
Structural variants (SV) have been linked to important bovine disease phenotypes, but due to the difficulty of their accurate detection with standard sequencing approaches, their role in shaping important traits across cattle breeds is largely unexplored. Optical mapping is an alternative approach for mapping SVs that has been shown to have higher sensitivity than DNA sequencing approaches. The aim of this project was to use optical mapping to develop a high-quality database of structural variation across cattle breeds from different geographical regions, to enable further study of SVs in cattle. To do this we generated 100X Bionano optical mapping data for 18 cattle of nine different ancestries, three continents and both cattle sub-species. In total we identified 13,457 SVs, of which 1,200 putatively overlap coding regions. This resource provides a high-quality set of optical mapping-based SV calls that can be used across studies, from validating DNA sequencing-based SV calls to prioritising candidate functional variants in genetic association studies and expanding our understanding of the role of SVs in cattle evolution.
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Bovinos , Genômica , Animais , Fases de Leitura Aberta , Fenótipo , Análise de Sequência de DNARESUMO
Despite only 8% of cattle being found in Europe, European breeds dominate current genetic resources. This adversely impacts cattle research in other important global cattle breeds, especially those from Africa for which genomic resources are particularly limited, despite their disproportionate importance to the continent's economies. To mitigate this issue, we have generated assemblies of African breeds, which have been integrated with genomic data for 294 diverse cattle into a graph genome that incorporates global cattle diversity. We illustrate how this more representative reference assembly contains an extra 116.1 Mb (4.2%) of sequence absent from the current Hereford sequence and consequently inaccessible to current studies. We further demonstrate how using this graph genome increases read mapping rates, reduces allelic biases and improves the agreement of structural variant calling with independent optical mapping data. Consequently, we present an improved, more representative, reference assembly that will improve global cattle research.
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Bovinos/genética , Variação Genética , Genoma , África , Alelos , Animais , Mapeamento Cromossômico , Europa (Continente) , Genômica , MasculinoRESUMO
OBJECTIVE: Robotic endoscopes have the potential to dramatically improve endoscopy procedures, however current attempts remain limited due to mobility and sensing challenges and have yet to offer the full capabilities of traditional tools. Endoscopic intervention (e.g., biopsy) for robotic systems remains an understudied problem and must be addressed prior to clinical adoption. This paper presents an autonomous intervention technique onboard a Robotic Endoscope Platform (REP) using endoscopy forceps, an auto-feeding mechanism, and positional feedback. METHODS: A workspace model is established for estimating tool position while a Structure from Motion (SfM) approach is used for target-polyp position estimation with the onboard camera and positional sensor. Utilizing this data, a visual system for controlling the REP position and forceps extension is developed and tested within multiple anatomical environments. RESULTS: The workspace model demonstrates accuracy of 5.5% while the target-polyp estimates are within 5 mm of absolute error. This successful experiment requires only 15 seconds once the polyp has been located, with a success rate of 43% using a 1 cm polyp, 67% for a 2 cm polyp, and 81% for a 3 cm polyp. CONCLUSION: Workspace modeling and visual sensing techniques allow for autonomous endoscopic intervention and demonstrate the potential for similar strategies to be used onboard mobile robotic endoscopic devices. SIGNIFICANCE: To the authors' knowledge this is the first attempt at automating the task of colonoscopy intervention onboard a mobile robot. While the REP is not sized for actual procedures, these techniques are translatable to devices suitable for in vivo application.
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Procedimentos Cirúrgicos Robóticos , Robótica , Colonoscopia , Endoscópios , Instrumentos CirúrgicosRESUMO
Substantial numbers of both alpha/beta and gamma/delta T cells are present in human fetal liver, which suggests a role of the fetal liver in T cell development. The diversity of fetal liver T cell receptor (TCR) gamma and delta chain rearrangements was examined among both CD4+CD8- and CD4-CD8- gamma/delta T cell clones. In addition, TCR delta chain transcripts from three fetal livers were sequenced after polymerase chain reaction amplification of TCR delta chains with V delta 1 or V delta 2 rearrangements. Five of six fetal liver gamma/delta T cell clones had a V delta 2-D delta 3-J delta 3 gene rearrangement with limited junctional diversity; three of these clones had an unusual CD4+CD8- phenotype. V delta 2-D delta 3-J delta 3 gene rearrangements were also common among both in-frame and out-of-frame transcripts from three fetal livers, indicating that they are the result of an ordered rearrangement process. TCR gamma chain sequences of the fetal liver gamma/delta T cell clones revealed V gamma 1-J gamma 2.3, V gamma 2-J gamma 1.2, and V gamma 3-J gamma 1.1 rearrangements with minimal incorporation of template-independent N region nucleotides. TCR gamma chain rearrangements found in these fetal liver T cell clones were different from those that have been observed among early thymic gamma/delta T cell populations, while similar TCR delta chain rearrangements are found among gamma/delta T cells from both sites. These data demonstrate that the fetal liver harbors gamma/delta T cell populations distinct from those found in the fetal thymus, suggesting that the fetal liver is a site of gamma/delta T cell development in humans. These unusual T cell populations may serve a specific function in the fetal immune system.
Assuntos
Rearranjo Gênico da Cadeia delta dos Receptores de Antígenos dos Linfócitos T , Rearranjo Gênico da Cadeia gama dos Receptores de Antígenos dos Linfócitos T , Fígado/embriologia , Receptores de Antígenos de Linfócitos T gama-delta/genética , Subpopulações de Linfócitos T/imunologia , Sequência de Aminoácidos , Sequência de Bases , Linfócitos T CD4-Positivos/imunologia , Células Clonais , Humanos , Dados de Sequência Molecular , Oligodesoxirribonucleotídeos/químicaRESUMO
Abstract This article assesses the value of using telecommunications with Promatoras (paraprofessional outreach workers) and an expert medical team of registered nurses (RNs) and endocrinologists in an at-risk type 2 diabetic Hispanic population recruited for a telemedicine feasibility project from a free clinic. Nineteen patients agreed to enter the program and 16 completed the program in 3.5 years of study. A Promatoras is the primary educator and the point of communication to patient or medical personnel overseeing each patient's home glucose monitoring, medical records, and medications, regularly communicating by telephone and e-mail with patients and diabetes specialists. Between clinic visits, all routine care, including body weight, blood glucose, and blood pressure monitoring, was shared over the Internet, and each patient was interviewed by audio and camera. The endocrinologist was in his office, while the primary care physician, patient, and Promotora volunteers were at the free clinic. Four variables were considered in this longitudinal study: weight, systolic blood pressure, diastolic blood pressure, and HbA1c. Estimates of means, correlations, t-tests, and slopes of the repeated measures were obtained, and comparisons were made between first and last values. The most important sign of improvement in the patients' situation was the significant decrease in HbA1c to 7.2% from 9.6% (p = 0.001).
Assuntos
Pessoal Técnico de Saúde , Diabetes Mellitus Tipo 2/terapia , Serviços de Assistência Domiciliar , Equipe de Assistência ao Paciente , Qualidade da Assistência à Saúde , Telecomunicações , Adulto , Idoso , California , Gerenciamento Clínico , Feminino , Hispânico ou Latino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The extent to which the impact of regulatory genetic variants may depend on other factors, such as the expression levels of upstream transcription factors, remains poorly understood. Here we report a framework in which regulatory variants are first aggregated into sets, and using these as estimates of the total cis-genetic effects on a gene we model their non-additive interactions with the expression of other genes in the genome. Using 1220 lymphoblastoid cell lines across platforms and independent datasets we identify 74 genes where the impact of their regulatory variant-set is linked to the expression levels of networks of distal genes. We show that these networks are predominantly associated with tumourigenesis pathways, through which immortalised cells are able to rapidly proliferate. We consequently present an approach to define gene interaction networks underlying important cellular pathways such as cell immortalisation.
Assuntos
Epistasia Genética/genética , Redes Reguladoras de Genes/genética , Linfócitos , Linhagem Celular , Proliferação de Células , Genótipo , Haplótipos , Humanos , Linfócitos/metabolismo , Modelos GenéticosRESUMO
INTRODUCTION: To evaluate the efficacy of a self-management support program including a 6 week self-management course, individualised phone support and goal setting in osteoarthritis patients on a waiting list for arthroplasty surgery. METHOD: Randomised controlled trial of 152 public hospital outpatients awaiting hip or knee replacement surgery who were not classified as requiring urgent surgery. Participants were randomised to a self-management program or to usual care. The primary outcome was change in the Health Education Intervention Questionnaire (HeiQ) from randomisation to 6 month follow-up. Quality of life and depressive symptoms were also measured. Changes in pain and function were assessed using the Western Ontario and McMaster Universities (WOMAC) Arthritis Index. RESULTS: At 6 month follow-up, health-directed behaviour was significantly greater in the intervention [mean 4.29, 95% confidence interval (CI) 3.99-4.58] than the control (mean 3.81, 95% CI 3.52-4.09; P=0.017). There was also a significant effect on skill and technique acquisition for the intervention (mean 4.37, 95% CI 4.19-4.55) in comparison to control (mean 4.11, 95% CI 3.93-4.29; P=0.036). There was no significant effect of the intervention on the remaining HeiQ subscales, WOMAC pain or disability, quality of life or depressive symptoms. DISCUSSION: The arthritis self-management program improved health-directed behaviours, skill acquisition and stiffness in patients on a joint replacement waiting list, although the observed effects were of modest size (Cohen's d between 0.36 and 0.42). There was no significant effect on pain, function or quality of life in the short term. Self-management programs can assist in maintaining health behaviours (particularly walking) in this patient group. Further research is needed to assess their impact on quality of life and over longer periods.
Assuntos
Artroplastia de Substituição/estatística & dados numéricos , Transtorno Depressivo/psicologia , Osteoartrite do Quadril/psicologia , Osteoartrite do Joelho/psicologia , Qualidade de Vida/psicologia , Idoso , Feminino , Financiamento Governamental , Humanos , Masculino , Osteoartrite do Quadril/cirurgia , Osteoartrite do Joelho/cirurgia , Medição da Dor , Autocuidado/métodos , Apoio Social , Inquéritos e Questionários , Listas de EsperaRESUMO
After the initiation of bud formation, cells of the yeast Saccharomyces cerevisiae direct new growth to the developing bud. We show here that this vectorial growth is facilitated by activity of the MYO2 gene. The wild-type MYO2 gene encodes an essential form of myosin composed of an NH2-terminal domain typical of the globular, actin-binding domain of other myosins. This NH2-terminal domain is linked by what appears to be a short alpha-helical domain to a novel COOH-terminal region. At the restrictive temperature the myo2-66 mutation does not impair DNA, RNA, or protein biosynthetic activity, but produces unbudded, enlarged cells. This phenotype suggests a defect in localization of cell growth. Measurements of cell size demonstrated that the continued development of initiated buds, as well as bud initiation itself, is inhibited. Bulk secretion continues in mutant cells, although secretory vesicles accumulate. The MYO2 myosin thus may function as the molecular motor to transport secretory vesicles along actin cables to the site of bud development.
Assuntos
Miosinas/genética , Saccharomyces cerevisiae/genética , Actinas/análise , Sequência de Aminoácidos , Sequência de Bases , Quitina/metabolismo , Grânulos Citoplasmáticos/metabolismo , Citoesqueleto/ultraestrutura , Genes Fúngicos , Glicosídeo Hidrolases/metabolismo , Fator de Acasalamento , Microscopia Eletrônica , Microtúbulos/ultraestrutura , Dados de Sequência Molecular , Mutação , Miosinas/metabolismo , Peptídeos/metabolismo , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/ultraestrutura , Temperatura , beta-FrutofuranosidaseRESUMO
The Cdc34 (Ubc3) ubiquitin-conjugating enzyme from Saccharomyces cerevisiae plays an essential role in the progression of cells from the G1 to S phase of the cell division cycle. Using a high-copy suppression strategy, we have identified a yeast gene (UBS1) whose elevated expression suppresses the conditional cell cycle defects associated with cdc34 mutations. The UBS1 gene encodes a 32.2-kDa protein of previously unknown function and is identical in sequence to a genomic open reading frame on chromosome II (GenBank accession number Z36034). Several lines of evidence described here indicate that Ubs1 functions as a general positive regulator of Cdc34 activity. First, overexpression of UBS1 suppresses not only the cell proliferation and morphological defects associated with cdc34 mutants but also the inability of cdc34 mutant cells to degrade the general amino acid biosynthesis transcriptional regulator, Gcn4. Second, deletion of the UBS1 gene profoundly accentuates the cell cycle defect when placed in combination with a cdc34 temperature-sensitive allele. Finally, a comparison of the Ubs1 and Cdc34 polypeptide sequences reveals two noncontiguous regions of similarity, which, when projected onto the three-dimensional structure of a ubiquitin-conjugating enzyme, define a single region situated on its surface. While cdc34 mutations corresponding to substitutions outside this region are suppressed by UBS1 overexpression, Ubs1 fails to suppress amino acid substitutions made within this region. Taken together with other findings, the allele specificity exhibited by UBS1 expression suggests that Ubs1 regulates Cdc34 by interaction or modification.
Assuntos
Ciclo Celular/genética , Proteínas de Ligação a DNA , Proteínas Fúngicas/genética , Genes Fúngicos , Genes Reguladores , Ligases/genética , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Complexos Ubiquitina-Proteína Ligase , Sequência de Aminoácidos , Ciclossomo-Complexo Promotor de Anáfase , Sequência de Bases , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Proteínas Quinases/metabolismo , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/crescimento & desenvolvimento , Homologia de Sequência de Aminoácidos , Supressão Genética , Enzimas de Conjugação de Ubiquitina , Ubiquitina-Proteína LigasesRESUMO
This work presents an analysis and comparison of the efficacy of two methods for pedicle screw placement during posterior spinal fusion surgery. A total of 100 screws (64 manual and 36 power driven), all placed utilizing a surgical navigation system, were analyzed and compared. Final screw placement was compared to initial surgical plans using the navigation system, and the final screw locations were analyzed on the basis of angular deviation from these planned trajectories as well as screw translation within a critical reference plane. The power driver was found to insignificantly decrease the resulting angular deviation of these pedicle screws with a mean deviation of 3.35 degrees compared to 3.44 degrees with the manual driver (p = 0.853). Conversely, the power driver was found to increase the translational distance in the critical region, with mean deviations of 2.45 mm for the power driver compared to 1.54 mm with the manual driver. The increase in translational deviation was significant (p = 0.002) indicating that there may be some loss in performance from the adoption of the power driver.
RESUMO
Two tumors, human sarcoma #1 (HS #1) and human epidermoid carcinoma #3 (HEp #3), were cultured on the chorioallantoic membrane of chick embryos. Under experimental conditions, HS #1 does not metastasize, whereas HEp #3 metastasizes extensively to chick embryo lungs and other organs. The glycosphingolipid profiles of these tumors were studied and HEp #3 wad found to contain about 2.5-fold less lipid-bound sialic acid per 100 mg of total lipid extracted than did HS #1, due mainly to smaller levels of monosialoganglioside (3.7-fold) and disialoganglioside (3.8-fold) in HEp #3. The total amount of neutral glycosphingolipids was approximately the same in both tumors, but their profiles differed. Treatment of these tumors with 6,7,8,9-tetrahydro-1-mercapto-1,2,4-triazolo-[4,3-a]quinazolin-5-ol (2.5 mg/egg/tumor) completely inhibited the formation of metastases in HEp #3 and increased the total content of lipid-bound sialic acid in the tumor by 63% (hematoside, monosialoganglioside, and disialoganglioside by 71, 99, and 67%, respectively). No change was seen in the content of lipid-bound sialic acid in HS #1. Treatment of HEp #3 with a smaller dose of te quinazolinol derivative (1.25 mg/egg) caused an average of 88% inhibition of metastasis, with a 37% increase in lipid-bound sialic acid. Another compound, 2,5-diphenylthiazolo-[5,4-d]thiazole (500 microgram/egg), completely inhibited the formation of metastasis and caused a substantial increase in the amount of lipid-bound sialic acid (77%). The data showed the existence of a correlation between the level of gangliosides in HEp #3 and the ability of these tumors to metastasize.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Glicoesfingolipídeos/metabolismo , Metástase Neoplásica , Sarcoma/metabolismo , Animais , Carcinoma de Células Escamosas/patologia , Embrião de Galinha , Gangliosídeos/análise , Glicoesfingolipídeos/análise , Humanos , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Quinazolinas/farmacologia , Sarcoma/patologia , Tiazóis/farmacologiaRESUMO
INTRODUCTION: While autonomous surgical robotic systems exist primarily at the research level, recently these systems have made a strong push into clinical settings. The autonomous or semi-autonomous control of surgical robotic platforms may offer significant improvements to a diverse field of surgical procedures, allowing for high precision, intelligent manipulation of these systems and opening the door to advanced minimally invasive surgical procedures not currently possible. AREAS COVERED: This review highlights those experimental systems currently under development with a focus on in vivo modeling and control strategies designed specifically for the complex and dynamic surgical environment. Expert review: Novel methods for state estimation, system modeling and disturbance rejection, as applied to these devices, continues to improve the performance of these important surgical tools. Procedures such as Natural Orifice Transluminal Endoscopic Surgery and Laparo-Endoscopic Single Site surgery, as well as more conventional procedures such as Colonoscopy, serve to benefit tremendously from the development of these automated robotic systems, enabling surgeons to minimize tissue damage and shorten procedure times while avoiding the consequences of laparotomy.
Assuntos
Procedimentos Cirúrgicos Minimamente Invasivos/instrumentação , Movimento (Física) , Procedimentos Cirúrgicos Robóticos/instrumentação , Animais , Humanos , Miniaturização/instrumentaçãoRESUMO
T lymphocyte activation correlates with the transcriptional induction of a variety of genes that encode proteins that are believed to play a role in specific effector functions of the mature cells. Transcripts corresponding to members of the cytotoxic cell proteinase (CCP) family of genes accumulate with different kinetics depending upon the nature of the T cell stimulus. The profile of expression for each family member is unique. Sequences corresponding to the 5' and 3' flanking regions of each of the CCP genes were isolated and sequenced. A comparison of these sequences reveal regions of conservation that are consistent with the differential expression observed and indicate potential regulatory elements.
Assuntos
Serina Endopeptidases/genética , Linfócitos T Citotóxicos/enzimologia , Animais , Sequência de Bases , Northern Blotting , Linhagem Celular , DNA , Regulação da Expressão Gênica , Granzimas , Camundongos , Dados de Sequência Molecular , Homologia de Sequência do Ácido Nucleico , Linfócitos T Citotóxicos/citologia , Transcrição GênicaRESUMO
A family of serine proteinases is believed to be important in cell-mediated cytotoxicity. Presented here are the genomic sequences for three murine members of this cytotoxic cell proteinase (CCP) family: the CCP3, CCP4 and CCP5 genes. All three of these genes have introns inserted at the same codon sites and the same exon distribution of the active site residues. These characteristics are also shared with the CCP1 and CCP2 genes, the charter members of the CCP gene family. Phylogenetic analysis using intron and exon sequences suggests that all five genes arose by various duplication events. This analysis also indicates that the recently described HuCCPX and CCP2 genes originated from recombination events between genes of different lineages. A phylogenetic and Southern analysis of the recombinant HuCCPX gene suggests that the human genome contains an additional CCP gene that has yet to be described. Finally, evidence is presented suggesting that the cDNA clone originally describing the CCP5 gene was derived from an alternately spliced transcript.
Assuntos
Serina Endopeptidases/genética , Linfócitos T Citotóxicos/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Evolução Biológica , Northern Blotting , Clonagem Molecular , DNA/genética , Expressão Gênica , Genes , Granzimas , Camundongos , Dados de Sequência Molecular , Família Multigênica , Splicing de RNA , RNA Mensageiro/genética , Serina Endopeptidases/químicaRESUMO
A centrifugation procedure to enrich for enlarged cells has been used to isolate temperature-sensitive cdc mutants of the yeast Saccharomyces cerevisiae. Among these mutants are strains containing mutations that arrest proliferation at the regulatory step start. These new start mutations define two previously unidentified genes, CDC67 and CDC68, and reveal that a previously identified gene, DNA33 (here termed CDC65), can harbour start mutations. Each new start mutation permits significant biosynthetic activity after transfer of mutant cells to the non-permissive temperature. The cdc68-1 start mutation causes arrest of cell proliferation without inhibition of mating ability, while the cdc65-1 and cdc67-1 mutations inhibit zygote formation and successful conjugation. The identification of new start genes by a novel selection procedure suggests that the catalog of genes that influence start is large.
Assuntos
Genes Fúngicos , Saccharomyces cerevisiae/genética , Divisão Celular , Centrifugação , Clonagem Molecular , Conjugação Genética , DNA Fúngico/análise , Metanossulfonato de Etila/farmacologia , Proteínas Fúngicas/biossíntese , Teste de Complementação Genética , Genótipo , Mutação , Fenótipo , RNA Fúngico/biossíntese , Mapeamento por Restrição , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/metabolismo , TemperaturaRESUMO
The monoclonal antibodies, Ta1 and IOT15, define T cell activation cell surface markers and have been assigned to the CD26 leukocyte differentiation antigen cluster. Dipeptidyl peptidase IV (DPP IV, EC 3.4.14.5) is an exoaminopeptidase that, among leukocytes, is expressed almost exclusively on activated T cells. Comparative binding studies showed that the Ta1 mAb binds to DPP IV purified from human placenta as well as in extracts of the human YT lymphoid cell line and of CD3 stimulated normal human peripheral blood mononuclear cells. The mAb IOT15 did not bind to DPP IV from any source even upon repeated incubations. Western blot analysis of YT cell extracts revealed that Ta1 and IOT15 bound to distinctly different molecular weight molecules. Immunofluorescent cell surface capping experiments showed that capping of the IOT15 did not alter the surface distribution of the Ta1 fluorescence. The capping results combined with the DPP IV binding results indicate that IOT15 and Ta1 mAb's bind to different, apparently unassociated, molecules on the surface of T cells and that only Ta1 binds the T cell surface enzyme DPP IV.