121 results on '"Motta MC"'
Search Results
2. Functional changes in Duchenne muscular dystrophy: a 12-month longitudinal cohort study
- Author
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MAZZONE E, VASCO G, SORMANI MP, TORRENTE Y, BERARDINELLI A, MESSINA S, D' AMICO A, DOGLIO L, CAVALLARO F, FROSINI S, BELLO L, BONFIGLIO S, ZUCCHINI E, DE SANCTIS R, SCUTIFERO M, BIANCO F, ROSSI F, MOTTA MC, SACCO A, DONATI MA, MONGINI T, PINI A, BATTINI R, PEGORARO E, PANE M, GASPERINI S, PREVITALI S, NAPOLITANO S, MARTINELLI D, BRUNO C, VITA G, COMI G, BERTINI E., POLITANO, Luisa, Mazzone, E, Vasco, G, Sormani, Mp, Torrente, Y, Berardinelli, A, Messina, S, D' AMICO, A, Doglio, L, Politano, Luisa, Cavallaro, F, Frosini, S, Bello, L, Bonfiglio, S, Zucchini, E, DE SANCTIS, R, Scutifero, M, Bianco, F, Rossi, F, Motta, Mc, Sacco, A, Donati, Ma, Mongini, T, Pini, A, Battini, R, Pegoraro, E, Pane, M, Gasperini, S, Previtali, S, Napolitano, S, Martinelli, D, Bruno, C, Vita, G, Comi, G, and Bertini, E.
- Abstract
OBJECTIVE: The aim of the study was to assess different outcome measures in a cohort of ambulant boys with Duchenne muscular dystrophy (DMD) over 12 months in order to establish the spectrum of possible changes in relation to age and steroid treatment. METHODS: The study is a longitudinal multicentric cohort study. A total of 106 ambulant patients with DMD were assessed using the 6-minute walk test (6MWT) and North Star Ambulatory Assessment (NSAA) at baseline and 12 months. Clinical data including age and steroid treatment were collected. RESULTS: During the 12 months of the study, we observed a mean decline of 25.8 meters in the 6MWT with a SD of 74.3 meters. On NSAA, the mean decline was 2.2 points with a SD of 3.7. Not all the boys with DMD in our cohort showed a decline over the 12 months, with young boys showing some improvement in their 6MWT and NSAA scores up to the age of 7. NSAA and the 6MWT had the highest correlation (r = 0.52, p < 0.001). CONCLUSIONS: This study provides longitudinal data of NSAA and 6MWT over a 12-month period. These data can be useful when designing a clinical trial.
- Published
- 2011
3. Psychological and practical difficulties among parents and healthy siblings of children with Duchenne vs Becker muscular dystrphy: an italian comparative study
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Magliano, L, D'Angelo, Mg, Vita, G, Pane, M, D'Amico, A, Balottin, U, Angelini, C, Battini, R, Politano, L, Patalano M: Sagliocchi, A, Civatio, F, Brighina, E, Vita, Gl, Messina, S, Sframeli, M, Lombardo, Me, Scalise, R, Colia, G, Catteruccia, M, Berardinelli, A, Motta, Mc, Gaiani, A, Semplicini, C, Bello, L, Astrea, G, Zaccaro, A, and Scutifero, M
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- 2014
4. The families of children with muscular dystrophies: burden, social network and professional support
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Politano, L, Scutifero, M, Zaccaro, A, Balottin, U, Berardinelli, A, Camia, M, Motta, Mc, Vita, Gianluca, Messina, Sonia, Sframeli, Maria, Vita, Giuseppe, Pane, M, Lombardo, Me, Scalise, R, D’Amico, A, Catteruccia, M, Colia, G, Angelici, C, Giani, A, Semplicioni, C, Battini, R, Astrea, G, Ricci, G, D’Angelo, Mg, Brighino, E, Civati, F, Catalano, M, Sagliocchi, A, and Magliano, L.
- Published
- 2013
5. Importance of SPP1 genotype as a covariate in clinical trials in Duchenne muscular dystrophy.
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Bello L, Piva L, Barp A, Taglia A, Picillo E, Vasco G, Pane M, Previtali SC, Torrente Y, Gazzerro E, Motta MC, Grieco GS, Napolitano S, Magri F, D'Amico A, Astrea G, Messina S, Sframeli M, Vita GL, and Boffi P
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- 2012
- Full Text
- View/download PDF
6. 'I have got something positive out of this situation': psychological benefits of caregiving in relatives of young people with muscular dystrophy
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Corrado Angelini, Roberta Battini, Luisa Politano, Michela Catteruccia, Giulia Colia, Gian Luca Vita, Guja Astrea, Claudio Semplicini, Adele D'Amico, Maria Chiara Motta, Maria Grazia D'Angelo, Erika Brighina, Maria Elena Lombardo, Antonella Zaccaro, Marianna Scutifero, Luca Bello, Lorenza Magliano, Roberta Scalise, Umberto Balottin, Giuseppe Vita, Alessandra Sagliocchi, Maria Sframeli, Giulia Ricci, Alessandra Gaiani, Marika Pane, Angela Berardinelli, Sonia Messina, Federica Civati, Melania Patalano, Magliano, Lorenza, Patalano, M, Sagliocchi, A, Scutifero, M, Zaccaro, A, D'Angelo, Mg, Civati, F, Brighina, E, Vita, G, Vita, Gl, Messina, S, Sframeli, M, Pane, M, Lombardo, Me, Scalise, R, D'Amico, A, Colia, G, Catteruccia, M, Balottin, U, Berardinelli, A, Motta, Mc, Angelini, C, Gaiani, A, Semplicini, C, Bello, L, Battini, R, Astrea, G, Ricci, G, and Politano, Luisa
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Male ,Activities of daily living ,psychological benefit ,Muscular Dystrophies ,Cost of Illness ,Surveys and Questionnaires ,Activities of Daily Living ,80 and over ,Young adult ,Child ,media_common ,Aged, 80 and over ,Social network ,Original Communication ,Middle Aged ,Professional support ,Test (assessment) ,Caregivers ,Italy ,Neurology ,Child, Preschool ,Caregiving ,Female ,medicine.symptom ,Clinical psychology ,Adult ,medicine.medical_specialty ,Weakness ,Adolescent ,media_common.quotation_subject ,Clinical Neurology ,MEDLINE ,Stress ,Young Adult ,Social support ,Perception ,medicine ,Humans ,Family ,Psychological benefits ,Preschool ,Psychiatry ,Muscular dystrophy ,Aged ,Analysis of Variance ,Social Support ,Stress, Psychological ,Neurology (clinical) ,business.industry ,Psychological ,business - Abstract
This paper focuses on the psychological benefits of caregiving in key relatives of patients with muscular dystrophies (MD), a group of rare diseases characterized by progressive weakness and restriction of the patient's functional abilities. We describe whether relatives perceived caregiving to be a positive experience and test whether relatives' perceptions vary in relation to their view of the patient as a valued person, the degree of involvement in care, and the level of support provided by social network and professionals. The study sample included 502 key relatives of patients aged 4-25 years, suffering from Duchenne, Becker, or limb-girdle MD, in treatment for at least 6 months to one of the eight participating centers, living with at least one relative aged 18-80 years. Of key relatives, 88 % stated that they had gotten something positive out of the situation, 96 % considered their patients to be sensitive, and 94 % viewed their patients as talented. Positive aspects of caregiving were more recognized by key relatives who were more convinced that the patient was sensitive and who perceived that they received higher level of professional help and psychological social support. These results suggest that most key relatives consider that their caregiving experience has had a positive impact on their lives, despite the practical difficulties of caring for patients with MD. Professionals should help relatives to identify the benefits of caregiving without denying its difficulties. Clinicians themselves should develop positive attitudes towards family involvement in the care of patients with long-term diseases.
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- 2013
7. Importance of SPP1 genotype as a covariate in clinical trials in Duchenne muscular dystrophy
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P Boffi, Corrado Angelini, Luisa Politano, Guja Astrea, Eugenio Mercuri, Adele D'Amico, Tiziana Mongini, Marika Pane, Alessandra Ferlini, Giuseppe Vita, Sara Napolitano, Mario Ermani, Yvan Torrente, Francesca Gualandi, Antonella Taglia, Gaetano S. Grieco, Gian Luca Vita, Gianni Sorarù, Eric P. Hoffman, Angela Berardinelli, Andrea Barp, Esther Picillo, Gessica Vasco, Claudio Bruno, Roberta Battini, Stefano C. Previtali, Enrico Bertini, Giacomo P. Comi, Francesca Magri, Sonia Messina, Luca Bello, Luisa Piva, Maria Chiara Motta, Maria Sframeli, Carlo Minetti, Elisabetta Gazzerro, Elena Pegoraro, Bello, L, Piva, L, Barp, A, Taglia, A, Picillo, E, Vasco, G, Pane, M, Previtali, Sc, Torrente, Y, Gazzerro, E, Motta, Mc, Grieco, G, Napolitano, S, Magri, F, D'Amico, A, Astrea, G, Messina, S, Sframeli, M, Vita, Gl, Boffi, P, Mongini, T, Ferlini, A, Gualandi, F, Soraru', G, Ermani, M, Vita, G, Battini, R, Bertini, E, Comi, Gp, Berardinelli, A, Minetti, C, Bruno, C, Mercuri, E, Politano, Luisa, Angelini, C, Hoffman, Ep, and Pegoraro, E.
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Adult ,medicine.medical_specialty ,Adolescent ,Duchenne muscular dystrophy ,Single-nucleotide polymorphism ,Polymorphism, Single Nucleotide ,Settore MED/39 - NEUROPSICHIATRIA INFANTILE ,Internal medicine ,Genotype ,medicine ,Humans ,Multicenter Studies as Topic ,Longitudinal Studies ,Muscular Dystrophy ,Polymorphism ,Child ,Preschool ,Retrospective Studies ,Clinical Trials as Topic ,business.industry ,Genetic Variation ,Retrospective cohort study ,Articles ,Single Nucleotide ,Duchenne ,medicine.disease ,Muscular Dystrophy, Duchenne ,Clinical trial ,Child, Preschool ,Ambulatory ,Cohort ,Physical therapy ,Osteopontin ,Neurology (clinical) ,business ,Student's t-test - Abstract
Objective: To test the effect of the single nucleotide polymorphism −66 T>G (rs28357094) in the osteopontin gene ( SPP1 ) on functional measures over 12 months in Duchenne muscular dystrophy (DMD). Methods: This study was conducted on a cohort of ambulatory patients with DMD from a network of Italian neuromuscular centers, evaluated longitudinally with the North Star Ambulatory Assessment (NSAA) and the 6-Minute Walk Test (6MWT) at study entry and after 12 months. Genotype at rs28357094 was determined after completion of the clinical evaluations. Patients were stratified in 2 groups according to a dominant model (TT homozygotes vs TG heterozygotes and GG homozygotes) and clinical data were retrospectively compared between groups. Results: Eighty patients were selected (age 4.1–19.3 years; mean 8.3 ± 2.7 SD). There were no differences in age or steroid treatment between the 2 subgroups. Paired t test showed a significant difference in both NSAA ( p = 0.013) and 6MWT ( p = 0.03) between baseline and follow-up after 12 months in patients with DMD carrying the G allele. The difference was not significant in the T subgroup. The analysis of covariance using age and baseline values as covariate and SPP1 genotype as fixed effect showed that these parameters are significantly correlated with the 12-month values. Conclusions: These data provide evidence of the role of SPP1 genotype as a disease modifier in DMD and support its relevance in the selection of homogeneous groups of patients for future clinical trials.
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- 2012
8. Functional changes in Duchenne muscular dystrophy: A 12-month longitudinal cohort study
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Luisa Politano, Silvia Frosini, MP Sormani, Tiziana Mongini, Enrico Bertini, Adele D'Amico, Angela Berardinelli, Sara Napolitano, Roberta Battini, Eugenio Mercuri, Giacomo P. Comi, Maria Chiara Motta, Maria Benedetta Donati, Stefano C. Previtali, Marika Pane, Serena Gasperini, Flaviana Bianco, Fabio Cavallaro, Luca Doglio, Gianluca Vita, Sonia Messina, Elena Pegoraro, Danilo Martinelli, Angela Sacco, Gessica Vasco, M Scutifero, Luca Bello, Yvan Torrente, Francesca Rossi, E. Zucchini, Cristina Bruno, Elena S. Mazzone, Antonella Pini, R. De Sanctis, S Bonfiglio, Mazzone, E, Vasco, G, Sormani, Mp, Torrente, Y, Berardinelli, A, Messina, S, D’Amico, A, Doglio, L, Politano, L, Cavallaro, F, Frosini, S, Bello, L, Bonfiglio, S, Zucchini, E, De Sanctis, R, Scutifero, M, Bianco, F, Rossi, F, Motta, Mc, Sacco, A, Donati, Ma, Mongini, T, Pini, A, Battini, R, Pegoraro, E, Pane, M, Gasperini, S, Previtali, S, Napolitano, S, Martinelli, D, Bruno, C, Vita, G, Comi, Giancarlo, Bertini, E, and Mercuri, E.
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Male ,medicine.medical_specialty ,Pediatrics ,Adolescent ,Cross-sectional study ,Prednisolone ,Duchenne muscular dystrophy ,Statistics as Topic ,Anti-Inflammatory Agents ,Walking ,Severity of Illness Index ,Cohort Studies ,Settore MED/39 - NEUROPSICHIATRIA INFANTILE ,Pregnenediones ,Severity of illness ,medicine ,Humans ,Muscular Dystrophy ,Muscular dystrophy ,Child ,Preschool ,business.industry ,Reproducibility of Results ,drug therapy/physiopathology ,Duchenne ,medicine.disease ,Muscular Dystrophy, Duchenne ,Clinical trial ,Cross-Sectional Studies ,therapeutic use ,Child, Preschool ,physiology ,Ambulatory ,Cohort ,Disease Progression ,Physical therapy ,Female ,Neurology (clinical) ,business ,Adolescent, Anti-Inflammatory Agents ,therapeutic use, Child, Child ,Preschool, Cohort Studies, Cross-Sectional Studies, Disease Progression, Female, Humans, Male, Muscular Dystrophy ,drug therapy/physiopathology, Prednisolone ,therapeutic use, Pregnenediones ,therapeutic use, Reproducibility of Results, Severity of Illness Index, Statistics as Topic, Walking ,Cohort study - Abstract
Objective: The aim of the study was to assess different outcome measures in a cohort of ambulant boys with Duchenne muscular dystrophy (DMD) over 12 months in order to establish the spectrum of possible changes in relation to age and steroid treatment. Methods: The study is a longitudinal multicentric cohort study. A total of 106 ambulant patients with DMD were assessed using the 6-minute walk test (6MWT) and North Star Ambulatory Assessment (NSAA) at baseline and 12 months. Clinical data including age and steroid treatment were collected. Results: During the 12 months of the study, we observed a mean decline of 25.8 meters in the 6MWT with a SD of 74.3 meters. On NSAA, the mean decline was 2.2 points with a SD of 3.7. Not all the boys with DMD in our cohort showed a decline over the 12 months, with young boys showing some improvement in their 6MWT and NSAA scores up to the age of 7. NSAA and the 6MWT had the highest correlation ( r = 0.52, p Conclusions: This study provides longitudinal data of NSAA and 6MWT over a 12-month period. These data can be useful when designing a clinical trial.
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- 2011
9. North Star Ambulatory Assessment, 6-minute walk test and timed items in ambulant boys with Duchenne muscular dystrophy
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Anna Maria Bonetti, Elisabetta Zucchini, Giuseppe Vita, Elena Pegoraro, Filippo Cavallaro, Tiziana Mongini, Adelina Carlesi, Gessica Vasco, Luca Bello, Adele D'Amico, Sonia Messina, Luisa Politano, Marianna Scutifero, Francesca Rossi, Annalisa Sacco, Chiara De Waure, Elisabetta Pasquini, Diego Martinelli, Claudio Bruno, Silvia Frosini, Enrico Bertini, Eugenio Mercuri, Maria Alice Donati, Elena S. Mazzone, Flaviana Bianco, Marika Pane, Marion Main, Luca Doglio, Angela Berardinelli, Antonella Pini, Roberto De Sanctis, Roberta Battini, Maria Chiara Motta, Mazzone, E, Martinelli, D, Berardinelli, A, Messina, S, D'Amico, A, Vasco, G, Main, M, Doglio, L, Politano, Luisa, Cavallaro, F, Frosini, S, Bello, L, Carlesi, A, Bonetti, Am, Zucchini, E, DE SANCTIS, R, Scutifero, M, Bianco, F, Rossi, F, Motta, Mc, Sacco, A, Donati, Ma, Mongini, T, Pini, A, Battini, R, Pegoraro, E, Pane, M, Pasquini, E, Bruno, C, Vita, G, DE WAURE, C, Bertini, E, and Mercuri, E.
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Male ,medicine.medical_specialty ,Duchenne muscular dystrophy ,Walking ,Physical medicine and rehabilitation ,medicine ,Humans ,6-minute walk test ,Prospective Studies ,Prospective cohort study ,Child ,Muscle, Skeletal ,Gait ,Genetics (clinical) ,business.industry ,Patient specific ,medicine.disease ,Test (assessment) ,Muscular Dystrophy, Duchenne ,Neurology ,Pediatrics, Perinatology and Child Health ,Ambulatory ,Cohort ,Physical therapy ,Exercise Test ,Neurology (clinical) ,business - Abstract
The North Star Ambulatory Assessment is a functional scale specifically designed for ambulant boys affected by Duchenne muscular dystrophy (DMD). Recently the 6-minute walk test has also been used as an outcome measure in trials in DMD. The aim of our study was to assess a large cohort of ambulant boys affected by DMD using both North Star Assessment and 6-minute walk test. More specifically, we wished to establish the spectrum of findings for each measure and their correlation. This is a prospective multicentric study involving 10 centers. The cohort included 112 ambulant DMD boys of age ranging between 4.10 and 17 years (mean 8.18±2.3 DS). Ninety-one of the 112 were on steroids: 37/91 on intermittent and 54/91 on daily regimen. The scores on the North Star assessment ranged from 6/34 to 34/34. The distance on the 6-minute walk test ranged from 127 to 560.6 m. The time to walk 10 m was between 3 and 15 s. The time to rise from the floor ranged from 1 to 27.5 s. Some patients were unable to rise from the floor. As expected the results changed with age and were overall better in children treated with daily steroids. The North Star assessment had a moderate to good correlation with 6-minute walk test and with timed rising from floor but less with 10 m timed walk/run test. The 6-minute walk test in contrast had better correlation with 10 m timed walk/run test than with timed rising from floor. These findings suggest that a combination of these outcome measures can be effectively used in ambulant DMD boys and will provide information on different aspects of motor function, that may not be captured using a single measure.
- Published
- 2010
10. 24 Month Longitudinal Data in Ambulant Boys with Duchenne Muscular Dystrophy
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Claudio Bruno, Enrico Bertini, Silvia Frosini, Gianluca Vita, Flaviana Bianco, Luisa Politano, Sara Napolitano, Maria Pia Sormani, Adele D'Amico, Stefano C. Previtali, Marika Pane, Angela Berardinelli, Emanuela Viggiano, Francesca Rossi, Filippo Cavallaro, Sonia Messina, Elena Pegoraro, Tiziana Mongini, Yvan Torrente, Antonella Pini, Roberto De Sanctis, Gessica Vasco, Eugenio Mercuri, Elena S. Mazzone, Francesca Magri, Enrica Rolle, Roberta Scalise, Paola D'Ambrosio, Roberta Battini, Giacomo P. Comi, Luca Bello, Maria Chiara Motta, Maria Alice Donati, Luca Doglio, Serena Bonfiglio, Michele Sacchini, Mazzone, E, Pane, M, Sormani, Mp, Scalise, R, Berardinelli, A, Messina, S, Torrente, Y, D'Amico, A, Doglio, L, Viggiano, E, D'Ambrosio, P, Cavallaro, F, Frosini, S, Bello, L, Bonfiglio, S, De Sanctis, R, Rolle, E, Bianco, F, Magri, F, Rossi, F, Vasco, G, Vita, G, Motta, Mc, Donati, Ma, Sacchini, M, Mongini, T, Pini, A, Battini, R, Pegoraro, E, Previtali, S, Napolitano, S, Bruno, C, Politano, Luisa, Comi, Gp, Bertini, E, and Mercuri, E.
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Male ,Time Factors ,Outcome Assessment ,Non-Clinical Medicine ,Duchenne muscular dystrophy ,Anti-Inflammatory Agents ,Duchenne Muscular Dystrophy ,Walking ,Outcome assessment ,Cohort Studies ,0302 clinical medicine ,Pregnenediones ,Risk Factors ,Outcome Assessment, Health Care ,Medicine ,Muscular Dystrophy ,Longitudinal Studies ,Muscular dystrophy ,10. No inequality ,Child ,0303 health sciences ,Multidisciplinary ,Statistics ,3. Good health ,Child, Preschool ,Ambulatory ,outcome ,Risk assessment ,Research Article ,Test Evaluation ,medicine.medical_specialty ,Adolescent ,Longitudinal data ,Clinical Research Design ,Science ,Biostatistics ,Natural history of disease ,Risk Assessment ,03 medical and health sciences ,Settore MED/39 - NEUROPSICHIATRIA INFANTILE ,Diagnostic Medicine ,Humans ,6-minute walk test ,Clinical Trials ,Analysis of Variance ,Exercise Test ,Glucocorticoids ,Logistic Models ,Muscular Dystrophy, Duchenne ,Prednisone ,Preschool ,030304 developmental biology ,Clinical Genetics ,Health Care Policy ,business.industry ,Health Risk Analysis ,X-Linked ,Duchenne ,medicine.disease ,Health Care ,Physical therapy ,business ,030217 neurology & neurosurgery ,Mathematics - Abstract
ObjectivesThe aim of the study was i) to assess the spectrum of changes over 24 months in ambulant boys affected by Duchenne muscular dystrophy, ii) to establish the difference between the first and the second year results and iii) to identify possible early markers of loss of ambulation.MethodsOne hundred and thirteen patients (age range 4.1-17, mean 8.2) fulfilled the inclusion criteria, 67 of the 113 were on daily and 40 on intermittent steroids, while 6 were not on steroids. All were assessed using the 6 Minute Walk Test (6MWT), the North Star Ambulatory Assessment (NSAA) and timed test.ResultsOn the 6MWT there was an average overall decline of -22.7 (SD 81.0) in the first year and of -64.7 (SD 123.1) in the second year. On the NSAA the average overall decline was of -1.86 (SD 4.21) in the first year and of -2.98 (SD 5.19) in the second year. Fourteen children lost ambulation, one in the first year and the other 13 in the second year of the study. A distance of at least 330 meters on the 6MWT, or a NSAA score of 18 at baseline reduced significantly the risk of losing ambulation within 2 years.ConclusionsThese results can be of help at the time of using inclusion criteria for a study in ambulant patients in order to minimize the risk of patients who may lose ambulation within the time of the trial.
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- 2013
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11. Psychological and practical difficulties among parents and healthy siblings of children with Duchenne vs. Becker muscular dystrophy: An Italian comparative study
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Magliano, Lorenza, D Angelo, Maria Grazia, Vita, Giuseppe, Pane, Marika, D Amico, Adele, Balottin, Umberto, Angelini, Corrado, Battini, Roberta, Politano, Luisa, Patalano, Melania, Sagliocchi, Alessandra, Civati, Federica, Brighina, Erika, Vita, Gian Luca, Messina, Sonia, Sframeli, Maria, Lombardo, Maria Elena, Scalise, Roberta, Colia, Giulia, Catteruccia, Maria, Berardinelli, Angela, Motta, Maria Chiara, Gaiani, Alessandra, Semplicini, Claudio, Bello, Luca, Guja Astrea, Zaccaro, Antonella, Scutifero, Marianna, Magliano, Lorenza, D'Angelo, Mg, Vita, G, Pane, M, D'Amico, A, Balottin, U, Angelini, C, Battini, R, Politano, Luisa, Patalano, M, Sagliocchi, A, Civati, F, Brighina, E, Vita, Gl, Messina, S, Sframeli, M, Lombardo, Me, Scalise, R, Colia, G, Catteruccia, M, Berardinelli, A, Motta, Mc, Gaiani, A, Semplicini, C, Bello, L, Astrea, G, Zaccaro, A, and Scutifero, M.
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musculoskeletal diseases ,Duchenne muscular dystrophy ,Adult ,Parents ,muscular dystrophy ,congenital, hereditary, and neonatal diseases and abnormalities ,Adolescent ,healthy siblings ,burden ,Cost of Illness ,parents, healthy siblings ,Humans ,Becker muscular dystrophy ,social network ,Child ,Child, Preschool ,Family ,Italy ,Middle Aged ,Siblings ,Social Support ,Socioeconomic Factors ,Caregivers ,Family Health ,Muscular Dystrophy, Duchenne ,Preschool ,Original Articles ,Duchenne ,family burden - Abstract
This study explored the burden in parents and healthy siblings of 4-17 year-old patients with Duchenne (DMD) and Becker (BMD) muscular dystrophies, and whether the burden varied according to clinical aspects and social resources. Data on socio-demographic characteristics, patient's clinical history, parent and healthy children burden, and on parent's social resources were collected using self-reported questionnaires administered to 336 parents of patients with DMD (246) and BMD (90). Parents of patients with DMD reported higher burden than those of patients with BMD, especially concerning feeling of loss (84.3% DMD vs. 57.4% BMD), stigma (44.2% DMD vs. 5.5% BMD) and neglect of hobbies (69.0% DMD vs. 32.5% BMD). Despite the burden, 66% DMD and 62.4% BMD parents stated the caregiving experience had a positive impact on their lives. A minority of parents believed MD has a negative influence on the psychological well-being (31.0% DMD vs. 12.8% BMD), and social life of unaffected children (25.7% vs. 18.4%). In the DMD group, burden correlated with duration of illness and parent age, and burden was higher among parents with lower social contacts and support in emergencies. In DMD, difficulties among healthy children were reported as higher by parents who were older, had higher burden and lower social contacts. In both groups, burden increased in relation to patient disability. These findings underline that the psychological support to be provided to parents of patients with MD, should take into account clinical features of the disease.
12. Morphological analysis and lipid composition of the stable fly Stomoxys calcitrans salivary glands reinforce important features of feeding behaviour.
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Florencio M, Oilveira Magalhães FR, Araujo Zuma A, Lima Oliveira Lugathe C, Rosa D, Riguete K, Machado Motta MC, Alves do Nascimento A, Reis Dos Santos Mallet J, Correa Atella G, and Fampa P
- Abstract
Stomoxys calcitrans L. (Diptera: Muscidae), the stable fly, is a hematophagous insect of great veterinary importance, because it is a mechanical vector of diverse pathogens in livestock. The saliva of blood-feeding insects presents important pharmacologically active molecules that impair blood clotting, promote vasodilation and modulate the host immune system response, crucial processes for successful feeding. These properties also enable pathogens' transmission. In the present work, we describe an efficient protocol to dissect S. calcitrans salivary glands, their morphological characteristics and lipid profile. The mean length of the tubular gland is 3.23 mm with a bulbous posterior end and a narrow anterior end. Histological analysis revealed a monolayer of large polygonal epithelial cells with voluminous nuclei and high lipid content in their cytoplasm. Ultrastructural analysis showed that the epithelium is rich in mitochondria, free ribosomes, Golgi complex cisternae, presenting a great extension of rough endoplasmic reticulum that contains an electron-dense material. Lipid analysis by thin-layer chromatography showed that neutral fatty acids and phosphatidylcholine are predominant in the fly salivary glands. Lysophosphatidylcholine, an important signalling biomolecule involved in different metabolic processes, including host's immunomodulation and pathogens proliferation and differentiation, is also present., (© 2024 Royal Entomological Society.)
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- 2024
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13. The antiproliferative effect of FGF2 in K-Ras-driven tumor cells involves modulation of rRNA and the nucleolus.
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de Luna Vitorino FN, Levy MJ, Mansano Wailemann RA, Lopes M, Silva ML, Sardiu ME, Garcia BA, Machado Motta MC, Oliveira CC, Armelin HA, Florens LA, Washburn MP, and Pinheiro Chagas da Cunha J
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- RNA, Ribosomal genetics, RNA, Ribosomal metabolism, Transcription, Genetic, DNA, Ribosomal genetics, Chromatin genetics, Chromatin metabolism, Fibroblast Growth Factor 2 genetics, Fibroblast Growth Factor 2 pharmacology, Fibroblast Growth Factor 2 metabolism, Cell Nucleolus metabolism
- Abstract
The nucleolus is sensitive to stress and can orchestrate a chain of cellular events in response to stress signals. Despite being a growth factor, FGF2 has antiproliferative and tumor-suppressive functions in some cellular contexts. In this work, we investigated how the antiproliferative effect of FGF2 modulates chromatin-, nucleolus- and rDNA-associated proteins. The chromatin and nucleolar proteome indicated that FGF2 stimulation modulates proteins related to transcription, rRNA expression and chromatin-remodeling proteins. The global transcriptional rate and nucleolus area increased along with nucleolar disorganization upon 24 h of FGF2 stimulation. FGF2 stimulation induced immature rRNA accumulation by increasing rRNA transcription. The rDNA-associated protein analysis reinforced that FGF2 stimulus interferes with transcription and rRNA processing. RNA Pol I inhibition partially reversed the growth arrest induced by FGF2, indicating that changes in rRNA expression might be crucial for triggering the antiproliferative effect. Taken together, we demonstrate that the antiproliferative FGF2 stimulus triggers significant transcriptional changes and modulates the main cell transcription site, the nucleolus., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)
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- 2023
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14. Antileishmanial activity of the essential oils of Myrcia ovata Cambess. and Eremanthus erythropappus (DC) McLeisch leads to parasite mitochondrial damage.
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Amorim Gomes G, Martins-Cardoso K, Dos Santos FR, Florencio M, Rosa D, Araujo Zuma A, Pinheiro Santiago GM, M Motta MC, Carvalho MG, and Fampa P
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- Animals, Mice, Mice, Inbred BALB C, Antiprotozoal Agents pharmacology, Asteraceae, Leishmania mexicana, Oils, Volatile pharmacology, Parasites
- Abstract
Leishmania amazonensis is a species causative of cutaneous and anergic diffuse cutaneous leishmaniasis, treatment-resistant form, in the New World. Plants essential oils exhibit great potential as microbicide agents. We described the composition of the essential oils of two plants native from Brazil, Myrcia ovata , with geranial and neral as major constituents, and Eremanthus erythropappus, with α-bisabolol. In vitro effects of these essential oils on L. amazonensis promastigotes growth and ultrastructure were analysed as well as their cytotoxicity to murine macrophages. Both oils were highly active with IC
50 /96 h of 8.69 and 9.53 µg/mL for M. ovata and E. erythropappus against promastigotes and caused ultrastructural alterations including mitochondrial enlargement. Cytotoxicity for murine macrophages varied with the oil concentrations. The IC50 low values of both M. ovata and E. erythropappus oils against L. amazonensis and their relative low cytotoxicity to mammal host cells support their potential use against cutaneous leishmaniasis.- Published
- 2021
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15. The effect of the biflavonoid 2″,3″-dihydroochnaflavone on Trypanosoma cruzi Y strain.
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Florencio M, Tomás Nery E, Rosa D, Auxiliadora Nascimento Ribeiro T, de Brito Braz Moraes J, Araujo Zuma A, da Silva Trindade JD, Dutra Barbosa da Rocha RF, Decote-Ricardo D, Pinto-da-Silva LH, M Motta MC, de Carvalho MG, and Fampa P
- Subjects
- Animals, Lymphocytes drug effects, Macrophages, Peritoneal drug effects, Mice, Mice, Inbred BALB C, Plant Extracts chemistry, Plant Extracts pharmacology, Toxicity Tests, Biflavonoids pharmacology, Ochnaceae chemistry, Trypanocidal Agents pharmacology, Trypanosoma cruzi drug effects
- Abstract
Trypanosoma cruzi is the causative agent of Chagas disease which affects 8 million people in Latin America. The parasite possesses high capacity to evade host immune system and the available drugs to treat Chagas disease present low efficacy combined to serious side effects to patients. Therefore, the identification of alternative therapeutics is essential. Brazilian flora exhibits an immense diversity of metabolites with great potential to be developed into new drugs. We investigated the action of 2″,3″-dihydroochnaflavone a biflavonoid extracted from Luxemburgia nobilis Eichler ex Engl. (Ochnaceae) against T. cruzi (Y strain). Our experiments showed that this compound is effective against parasite epimastigote forms, presenting IC
50 value of (2.5 ± 0.1) μM after 96 h of treatment. Ultrastructure alterations were also detected in treated epimastigotes especially mitochondrial enlargement at the kinetoplast region. At the concentration of 30 μM, the compound killed (61.6 ± 3.37)% of the parasite in its amastigote form. In addition, at the same concentration, the compound killed all trypamastigotes growing within murine macrophages after 7-9 days of infection. Nonetheless, the biflavonoid concentrations were harmless to murine enriched population of lymphocytes and peritoneal macrophages. These results indicate that 2″,3″- dihydroochnaflavone presents activity against T. cruzi., Competing Interests: Declaration of competing interest On behalf of all authors, the corresponding author states that there is no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)- Published
- 2020
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16. HTLV-1-infected thymic epithelial cells convey the virus to CD4 + T lymphocytes.
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Carvalho Barros LR, Linhares-Lacerda L, Moreira-Ramos K, Ribeiro-Alves M, Machado Motta MC, Bou-Habib DC, and Savino W
- Subjects
- Apoptosis genetics, CD4-Positive T-Lymphocytes virology, Cell Adhesion Molecules genetics, Cell Adhesion Molecules metabolism, Cell Line, Chemokines genetics, Chemokines metabolism, Coculture Techniques, Disease Progression, Epithelial Cells physiology, Gene Expression Regulation, HTLV-I Infections immunology, Humans, Leukemia-Lymphoma, Adult T-Cell, Paraparesis, Spastic, Paraparesis, Tropical Spastic, Receptors, Virus metabolism, Virus Internalization, CD4-Positive T-Lymphocytes physiology, Epithelial Cells virology, HTLV-I Infections transmission, Human T-lymphotropic virus 1 physiology, Thymus Gland pathology
- Abstract
The human T-lymphotropic virus type-1 (HTLV-1) is the causative agent of adult T cell leukemia/lymphoma (ATL) and HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). CD4
+ T cells are the main target of HTLV-1, but other cell types are known to be infected, including immature lymphocytes. Developing T cells undergo differentiation in the thymus, through migration and interaction with the thymic microenvironment, in particular with thymic epithelial cells (TEC) the major component of this three dimensional meshwork of non-lymphoid cells. Herein, we show that TEC express the receptors for HTLV-1 and can be infected by this virus through cell-cell contact and by cell-free virus suspensions. The expression of anti-apoptosis, chemokine and adhesion molecules genes are altered in HTLV-1-infected TEC, although gene expression of antigen presentation molecules remained unchanged. Furthermore, HTLV-1-infected TEC transmitted the virus to a CD4+ T cell line and to CD4+ T cells from healthy donors, during in vitro cellular co-cultures. Altogether, our data point to the possibility that the human thymic epithelial cells play a role in the establishment and progression of HTLV-1 infection, functioning as a reservoir and transmitting the virus to maturing CD4+ T lymphocytes, which in turn will cause disease in the periphery., (Copyright © 2017. Published by Elsevier GmbH.)- Published
- 2017
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17. Integrated care of muscular dystrophies in Italy. Part 2. Psychological treatments, social and welfare support, and financial costs.
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Magliano L, Scutifero M, Patalano M, Sagliocchi A, Zaccaro A, Civati F, Brighina E, Vita G, Messina S, Sframeli M, Lombardo ME, Scalise R, Colia G, Catteruccia M, Berardinelli A, Motta MC, Gaiani A, Semplicini C, Bello L, Astrea G, Ricci G, D'Angelo MG, Vita G, Pane M, D'Amico A, Balottin U, Angelini C, Battini R, and Politano L
- Subjects
- Activities of Daily Living, Adolescent, Adult, Child, Child, Preschool, Delivery of Health Care, Integrated, Fees and Charges statistics & numerical data, Female, Humans, Italy, Male, Middle Aged, Muscular Dystrophies rehabilitation, Patient Education as Topic, Young Adult, Caregivers psychology, Cost of Illness, Muscular Dystrophies economics, Muscular Dystrophies psychology, Psychosocial Support Systems, Social Welfare
- Abstract
This paper describes the psycho-social treatments received by 502 patients with MDs and their relatives, and the costs for care sustained by the families in the previous six month period. Data were collected by the MD-Care Schedule (MD-CS) and the Family Problems Questionnaire (FPQ). Psycho-educational interventions were provided to 72 patients (14.3%), and social/welfare support to 331 patients (65.9%). Social/welfare support was higher in patients with DMD or LGMD, in those showing more severe disability, and in patients who were in contact with centres located in Northern Italy. Psycho-educational interventions were received by 156 (31%) relatives, and social/welfare support by 55 (10.9%) and mainly provided by Family/Patients Associations (83.6%). Relatives with higher educational levels, who spent more daily hours in the assistance of patients with DMD, and in contact with centres in Central Italy more frequently benefited from psycho-educational interventions. In the previous year, costs for care were sustained by 314 (63.9%) relatives. Financial difficulties related to patient's condition, were higher in families of patients who needed more intensive rehabilitation and daily hours of caregiving, and in families who lived further away from the reference's centre. These results showed that psycho-social aspects of MDs care are only partially met in Italy, and that ad hoc supportive interventions for these patients and their families should be potentiated.
- Published
- 2017
18. Integrated care of muscular dystrophies in Italy. Part 1. Pharmacological treatment and rehabilitative interventions.
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Politano L, Scutifero M, Patalano M, Sagliocchi A, Zaccaro A, Civati F, Brighina E, Vita G, Messina S, Sframeli M, Lombardo ME, Scalise R, Colia G, Catteruccia M, Berardinelli A, Motta MC, Gaiani A, Semplicini C, Bello L, Astrea G, Ricci G, D'Angelo MG, Vita G, Pane M, D'Amico A, Balottin U, Angelini C, Battini R, and Magliano L
- Subjects
- Adolescent, Age Factors, Bone Density Conservation Agents therapeutic use, Breathing Exercises, Cardiotonic Agents therapeutic use, Child, Combined Modality Therapy, Dietary Supplements, Disability Evaluation, Female, Health Care Surveys, Humans, Italy, Male, Prednisone therapeutic use, Pregnenediones therapeutic use, Respiration, Artificial, Time Factors, Glucocorticoids therapeutic use, Muscular Dystrophies drug therapy, Muscular Dystrophies rehabilitation, Physical Therapy Modalities
- Abstract
This paper describes the pharmacological therapies and rehabilitative interventions received by 502 patients with Muscular Dystrophies, evaluated in relation to patient's socio-demographic and clinical variables, and geographical areas. Data were collected by the MD-Socio-Demographic and Clinical Schedule (MD-SC-CS) and by the Family Problems Questionnaire (FPQ). The most part of the enrolled patients were in drug treatment. The number of the medications increased in relation to patient's age, disability degree and duration of illness and was higher among patients with Duchenne Muscular Dystrophy (DMD) compared with Becker (BMD) or Limb-Girdle Muscular Dystrophies (LGMD). Steroids (deflazacort or prednisone) were the drug most frequently used, followed by cardiologic and bone metabolism drugs. In general, patients using steroids were younger and had a shorter duration of illness; patients using cardiac drugs and dietary supplements were older and had a longer duration of illness. Rehabilitative interventions were provided to about 70% (351/502) of patients, mainly DMD. Of these, physiotherapy was the more frequent treatment (96.6%) and was prevalently performed in rehabilitative centres (about 70% of patients) and at home in only 30%. Hydrokinetic-therapy was practiced by 6.8% of patients. Respiratory rehabilitation was provided to 47.0% of patients (165/351) and assisted mechanical ventilaventilation to 13.1% (46). The amount of rehabilitative interventions increased in relation to the patient's age, level of disability and duration of illness. Compared to Central and Northern Italy, in Southern Italy there was a higher attention to cardiological impairment as shown by a higher number of patients receiving heart drugs. No statistically significant differences concerning the possibility to have access to rehabilitative interventions were noted among the three geographical areas. However, patient living in Southern Italy tend to receive rehabilitation more often at home.
- Published
- 2017
19. Reduction of Tubulin Expression in Angomonas deanei by RNAi Modifies the Ultrastructure of the Trypanosomatid Protozoan and Impairs Division of Its Endosymbiotic Bacterium.
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Catta-Preta CM, Dos Santos Pascoalino B, de Souza W, Mottram JC, Motta MC, and Schenkman S
- Subjects
- Bacteria genetics, Cell Division, Protozoan Proteins metabolism, RNA Interference, Trypanosomatina genetics, Trypanosomatina metabolism, Trypanosomatina ultrastructure, Tubulin genetics, Tubulin metabolism, Bacteria cytology, Protozoan Proteins genetics, Symbiosis, Trypanosomatina microbiology
- Abstract
In the last two decades, RNA interference pathways have been employed as a useful tool for reverse genetics in trypanosomatids. Angomonas deanei is a nonpathogenic trypanosomatid that maintains an obligatory endosymbiosis with a bacterium related to the Alcaligenaceae family. Studies of this symbiosis can help us to understand the origin of eukaryotic organelles. The recent elucidation of both the A. deanei and the bacterium symbiont genomes revealed that the host protozoan codes for the enzymes necessary for RNAi activity in trypanosomatids. Here, we tested the functionality of the RNAi machinery by transfecting cells with dsRNA to a reporter gene (green fluorescent protein), which had been previously expressed in the parasite and to α-tubulin, an endogenous gene. In both cases, protein expression was reduced by the presence of specific dsRNA, inducing, respectively, a decreased GFP fluorescence and the formation of enlarged cells with modified arrangement of subpellicular microtubules. Furthermore, symbiont division was impaired. These results indicate that the RNAi system is active in A. deanei and can be used to further explore gene function in symbiont-containing trypanosomatids and to clarify important aspects of symbiosis and cell evolution., (© 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.)
- Published
- 2016
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20. Symbiont modulates expression of specific gene categories in Angomonas deanei.
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Penha LL, Hoffmann L, Souza SS, Martins AC, Bottaro T, Prosdocimi F, Faffe DS, Motta MC, Ürményi TP, and Silva R
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- Animals, Bacteria growth & development, Gene Expression Profiling, Genes, Protozoan, Genome, Protozoan, Genomics, Humans, RNA, Protozoan isolation & purification, Trypanosomatina metabolism, Gene Expression Regulation physiology, Gene Ontology, RNA, Protozoan genetics, Symbiosis genetics, Transcriptome genetics, Trypanosomatina genetics
- Abstract
Trypanosomatids are parasites that cause disease in humans, animals, and plants. Most are non-pathogenic and some harbor a symbiotic bacterium. Endosymbiosis is part of the evolutionary process of vital cell functions such as respiration and photosynthesis. Angomonas deanei is an example of a symbiont-containing trypanosomatid. In this paper, we sought to investigate how symbionts influence host cells by characterising and comparing the transcriptomes of the symbiont-containing A. deanei (wild type) and the symbiont-free aposymbiotic strains. The comparison revealed that the presence of the symbiont modulates several differentially expressed genes. Empirical analysis of differential gene expression showed that 216 of the 7625 modulated genes were significantly changed. Finally, gene set enrichment analysis revealed that the largest categories of genes that downregulated in the absence of the symbiont were those involved in oxidation-reduction process, ATP hydrolysis coupled proton transport and glycolysis. In contrast, among the upregulated gene categories were those involved in proteolysis, microtubule-based movement, and cellular metabolic process. Our results provide valuable information for dissecting the mechanism of endosymbiosis in A. deanei.
- Published
- 2016
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21. Social representations of nurses on tuberculosis.
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Rodrigues IL, Motta MC, and Ferreira Mde A
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- Female, Humans, Male, Middle Aged, Attitude to Health, Nursing, Social Perception, Tuberculosis nursing
- Abstract
Objective: to describe the social representation of nurses on tuberculosis and identify the implications on nursing care., Method: qualitative research with the participation of 52 nurses from 23 Basic Health Units of Belém, Pará. A semi-structured interview was conducted with subsequent analysis of the thematic content according to the Theory of Social Representations., Results: the social representations of tuberculosis were organized into two categories: infection, evidencing the clinical-epidemiological aspects of the disease, and stigma and prejudice, representing the social aspect. Care is affected by fear - a fact that explains the distance adopted by some nurses when handling ill people., Conclusion: the social representations of nurses on tuberculosis remain grounded in fear, leading professionals to keep a certain distance from patients and generating stigma and prejudice, which may affect adherence to treatment.
- Published
- 2016
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22. Glycosomal bromodomain factor 1 from Trypanosoma cruzi enhances trypomastigote cell infection and intracellular amastigote growth.
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Ritagliati C, Villanova GV, Alonso VL, Zuma AA, Cribb P, Motta MC, and Serra EC
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- Animals, Chlorocebus aethiops, Intracellular Fluid parasitology, Microbodies parasitology, Vero Cells, Intracellular Fluid metabolism, Membrane Proteins biosynthesis, Microbodies metabolism, Neuraminidase biosynthesis, Protozoan Proteins biosynthesis, Trypanosoma cruzi metabolism
- Abstract
Acetylation is a ubiquitous protein modification present in prokaryotic and eukaryotic cells that participates in the regulation of many cellular processes. The bromodomain is the only domain known to bind acetylated lysine residues. In the last few years, many bromodomain inhibitors have been developed in order to treat diseases caused by aberrant acetylation of lysine residues and have been tested as anti-parasitic drugs. In the present paper, we report the first characterization of Trypanosoma cruzi bromodomain factor 1 (TcBDF1). TcBDF1 is expressed in all life cycle stages, but it is developmentally regulated. It localizes in the glycosomes directed by a PTS2 (peroxisome-targeting signal 2) sequence. The overexpression of wild-type TcBDF1 is detrimental for epimastigotes, but it enhances the infectivity rate of trypomastigotes and the replication of amastigotes. On the other hand, the overexpression of a mutated version of TcBDF1 has no effect on epimastigotes, but it does negatively affect trypomastigotes' infection and amastigotes' replication., (© 2016 Authors; published by Portland Press Limited.)
- Published
- 2016
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23. Correction: Sirt1 Regulates Insulin Secretion by Repressing UCP2 in Pancreatic β Cells.
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Bordone L, Motta MC, Picard F, Robinson A, Jhala US, Apfeld J, McDonagh T, Lemieux M, McBurney M, Szilvasi A, Easlon EJ, Lin SJ, and Guarente L
- Published
- 2015
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24. A lupane-triterpene isolated from Combretum leprosum Mart. fruit extracts that interferes with the intracellular development of Leishmania (L.) amazonensis in vitro.
- Author
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Teles CB, Moreira-Dill LS, Silva Ade A, Facundo VA, de Azevedo WF Jr, da Silva LH, Motta MC, Stábeli RG, and Silva-Jardim I
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- Animals, Cytoplasm parasitology, DNA Topoisomerases, Type I drug effects, Female, Fruit chemistry, In Vitro Techniques, Leishmaniasis drug therapy, Mice, Mice, Inbred BALB C, Phytotherapy, Plant Extracts chemistry, Triterpenes isolation & purification, Combretum chemistry, Leishmania mexicana drug effects, Leishmaniasis parasitology, Macrophages, Peritoneal parasitology, Plant Extracts pharmacology, Triterpenes pharmacology
- Abstract
Background: 3beta,6beta,16beta-trihydroxylup-20(29)-ene is a lupane triterpene isolated from Combretum leprosum fruit. The lupane group has been extensively used in studies on anticancer effects; however, its possible activity against protozoa parasites is yet poorly known. The high toxicity of the compounds currently used in leishmaniasis chemotherapy stimulates the investigation of new molecules and drug targets for antileishmanial therapy., Methods: The activity of 3beta,6beta,16beta-trihydroxylup-20(29)-ene was evaluated against Leishmania (L.) amazonensis by determining the cytotoxicity of the compound on murine peritoneal macrophages, as well as its effects on parasite survival inside host cells. To evaluate the effect of this compound on intracellular amastigotes, cultures of infected macrophages were treated for 24, 48 and 96 h and the percentage of infected macrophages and the number of intracellular parasites was scored using light microscopy., Results: Lupane showed significant activity against the intracellular amastigotes of L. (L.) amazonensis. The treatment with 109 μM for 96 h reduced in 80 % the survival index of parasites in BALB/c peritoneal macrophages. At this concentration, the triterpene caused no cytotoxic effects against mouse peritoneal macrophages. Ultrastructural analyses of L. (L.) amazonensis intracellular amastigotes showed that lupane induced some morphological changes in parasites, such as cytosolic vacuolization, lipid body formation and mitochondrial swelling. Bioinformatic analyses through molecular docking suggest that this lupane has high-affinity binding with DNA topoisomerase., Conclusion: Taken together, our results have showed that the lupane triterpene from C. leprosum interferes with L. (L.) amazonensis amastigote replication and survival inside vertebrate host cells and bioinformatics analyses strongly indicate that this molecule may be a potential inhibitor of topoisomerase IB. Moreover, this study opens major prospects for the development of novel chemotherapeutic agents with leishmanicidal activity.
- Published
- 2015
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25. Endosymbiosis in trypanosomatid protozoa: the bacterium division is controlled during the host cell cycle.
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Catta-Preta CM, Brum FL, da Silva CC, Zuma AA, Elias MC, de Souza W, Schenkman S, and Motta MC
- Abstract
Mutualism is defined as a beneficial relationship for the associated partners and usually assumes that the symbiont number is controlled. Some trypanosomatid protozoa co-evolve with a bacterial symbiont that divides in coordination with the host in a way that results in its equal distribution between daughter cells. The mechanism that controls this synchrony is largely unknown, and its comprehension might provide clues to understand how eukaryotic cells evolved when acquiring symbionts that later became organelles. Here, we approached this question by studying the effects of inhibitors that affect the host exclusively in two symbiont-bearing trypanosomatids, Strigomonas culicis and Angomonas deanei. We found that inhibiting host protein synthesis using cycloheximide or host DNA replication using aphidicolin did not affect the duplication of bacterial DNA. Although the bacteria had autonomy to duplicate their DNA when host protein synthesis was blocked by cycloheximide, they could not complete cytokinesis. Aphidicolin promoted the inhibition of the trypanosomatid cell cycle in the G1/S phase, leading to symbiont filamentation in S. culicis but not in A. deanei. Treatment with camptothecin blocked the host protozoa cell cycle in the G2 phase and induced the formation of filamentous symbionts in both species. Oryzalin, which affects host microtubule polymerization, blocked trypanosomatid mitosis and abrogated symbiont division. Our results indicate that host factors produced during the cell division cycle are essential for symbiont segregation and may control the bacterial cell number.
- Published
- 2015
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26. Biochemical and phylogenetic analyses of phosphatidylinositol production in Angomonas deanei, an endosymbiont-harboring trypanosomatid.
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de Azevedo-Martins AC, Alves JM, de Mello FG, Vasconcelos AT, de Souza W, Einicker-Lamas M, and Motta MC
- Subjects
- Bacteria isolation & purification, Gene Expression Regulation physiology, Phylogeny, Symbiosis, Trypanosomatina genetics, Phosphatidylinositols metabolism, Trypanosomatina metabolism
- Abstract
Background: The endosymbiosis in trypanosomatids is characterized by co-evolution between one bacterium and its host protozoan in a mutualistic relationship, thus constituting an excellent model to study organelle origin in the eukaryotic cell. In this association, an intense metabolic exchange is observed between both partners: the host provides energetic molecules and a stable environment to a reduced wall symbiont, while the bacterium is able to interfere in host metabolism by enhancing phospholipid production and completing essential biosynthesis pathways, such as amino acids and hemin production. The bacterium envelope presents a reduced cell wall which is mainly composed of cardiolipin and phosphatidylcholine, being the latter only common in intracellular prokaryotes. Phosphatidylinositol (PI) is also present in the symbiont and host cell membranes. This phospholipid is usually related to cellular signaling and to anchor surface molecules, which represents important events for cellular interactions., Methods: In order to investigate the production of PI and its derivatives in symbiont bearing trypanosomatids, aposymbiotic and wild type strains of Angomonas deanei, as well as isolated symbionts, were incubated with [(3)H]myo-inositol and the incorporation of this tracer was analyzed into inositol-containing molecules, mainly phosphoinositides and lipoproteins. Gene searches and their phylogenies were also performed in order to investigate the PI synthesis in symbiontbearing trypanosomatids., Results: Our results showed that the bacterium did not incorporate the tracer and that both strains produced similar quantities of PI and its derivatives, indicating that the symbiont does not influence the production of these metabolites. Gene searches related to PI synthesis revealed that the trypanosomatid genome contains an inositol transporter, PI synthase and the myo-inositol synthase. Thus, the host is able to produce PI either from exogenous myo-inositol (inositol transporter) or from myo-inositol synthesized de novo. Phylogenetic analysis using other organisms as references indicated that, in trypanosomatids, the genes involved in PI synthesis have a monophyletic origin. In accordance with experimental data, sequences for myo-inositol transport or for myo-inositol and PI biosynthesis were not found in the symbiont., Conclusions: Altogether, our results indicate that the bacterium depends on the host to obtain PI.
- Published
- 2015
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27. Identification and ultrastructural characterization of the Wolbachia symbiont in Litomosoides chagasfilhoi.
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Chagas-Moutinho VA, Silva R, de Souza W, and Motta MC
- Subjects
- Animals, Female, Filarioidea physiology, Male, Microscopy, Electron, Transmission, Phylogeny, Subcutaneous Tissue microbiology, Wolbachia genetics, Wolbachia ultrastructure, Filarioidea microbiology, Symbiosis, Wolbachia isolation & purification, Wolbachia physiology
- Abstract
Background: Filarial nematodes are arthropod-transmitted parasites of vertebrates that affect more than 150 million people around the world and remain a major public health problem throughout tropical and subtropical regions. Despite the importance of these nematodes, the current treatment strategies are not efficient in eliminating the parasite. The main strategy of control is based on chemotherapy with diethylcarbamazine, albendazole and ivermectin. In the 1970s, it was found that some filarids possess endosymbiotic bacteria that are important for the development, survival and infectivity of the nematodes. These bacteria belong to the genus Wolbachia, which is a widespread and abundant intracellular symbiont in worms. Knowledge about the structure of the bacteria and their relationship with their nematode hosts may allow new perspectives for the control of filarial nematodes., Methods: In this study, we used transmission electron microscopy combined with three-dimensional approaches to observe the structure of the endosymbiont of the filarial nematode Litomosoides chagasfilhoi, an experimental model for the study of lymphatic filariasis. In addition, the bacterium was classified based on PCR analyses., Results: The bacterium was mainly found in the hypodermis and in the female reproductive system in close association with host cell structures, such as the nucleus and endoplasmic reticulum. Our ultrastructural data also showed that the symbiont envelope is composed of two membrane units and is enclosed in a cytoplasmic vacuole, the symbiosome. Molecular data revealed that the bacterium of L. chagasfilhoi shares 100% identity with the Wolbachia endosymbiont of Litomosoides galizai., Conclusions: Here we described ultrastructural aspects of the relationship of the Wolbachia with the filarial nematode Litomosoides chagasfilhoi and the findings lead us to consider this relationship as a mutualistic symbiosis.
- Published
- 2015
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28. Unveiling the effects of berenil, a DNA-binding drug, on Trypanosoma cruzi: implications for kDNA ultrastructure and replication.
- Author
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Zuma AA, Cavalcanti DP, Zogovich M, Machado AC, Mendes IC, Thiry M, Galina A, de Souza W, Machado CR, and Motta MC
- Subjects
- Cell Proliferation drug effects, Cell Survival drug effects, Chagas Disease parasitology, Diminazene pharmacology, Membrane Potential, Mitochondrial drug effects, Oxygen metabolism, Reactive Oxygen Species metabolism, Trypanosoma cruzi genetics, Trypanosoma cruzi growth & development, Trypanosoma cruzi ultrastructure, Chagas Disease drug therapy, DNA Replication drug effects, Diminazene analogs & derivatives, Trypanocidal Agents pharmacology, Trypanosoma cruzi drug effects
- Abstract
Trypanosoma cruzi, the etiological agent of Chagas disease, exhibits a single mitochondrion with an enlarged portion termed kinetoplast. This unique structure harbors the mitochondrial DNA (kDNA), composed of interlocked molecules: minicircles and maxicircles. kDNA is a hallmark of kinetoplastids and for this reason constitutes a valuable target in chemotherapeutic and cell biology studies. In the present work, we analyzed the effects of berenil, a minor-groove-binding agent that acts preferentially at the kDNA, thereby affecting cell proliferation, ultrastructure, and mitochondrial activity of T. cruzi epimastigote form. Our results showed that berenil promoted a reduction on parasite growth when high concentrations were used; however, cell viability was not affected. This compound caused significant changes in kDNA arrangement, including the appearance of membrane profiles in the network and electron-lucent areas in the kinetoplast matrix, but nuclear ultrastructure was not modified. The use of the TdT technique, which specifically labels DNA, conjugated to atomic force microscopy analysis indicates that berenil prevents the minicircle decatenation of the network, thus impairing DNA replication and culminating in the appearance of dyskinetoplastic cells. Alterations in the kinetoplast network may be associated with kDNA lesions, as suggested by the quantitative PCR (qPCR) technique. Furthermore, parasites treated with berenil presented higher levels of reactive oxygen species and a slight decrease in the mitochondrial membrane potential and oxygen consumption. Taken together, our results reveal that this DNA-binding drug mainly affects kDNA topology and replication, reinforcing the idea that the kinetoplast represents a potential target for chemotherapy against trypanosomatids.
- Published
- 2015
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29. Mitochondrial respiration and genomic analysis provide insight into the influence of the symbiotic bacterium on host trypanosomatid oxygen consumption.
- Author
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Azevedo-Martins AC, Machado AC, Klein CC, Ciapina L, Gonzaga L, Vasconcelos AT, Sagot MF, DE Souza W, Einicker-Lamas M, Galina A, and Motta MC
- Subjects
- Bacteria metabolism, Biological Evolution, Electron Transport genetics, Electron Transport physiology, Gene Expression Regulation, Trypanosomatina genetics, Bacteria classification, Mitochondria metabolism, Oxygen Consumption physiology, Symbiosis physiology, Trypanosomatina microbiology, Trypanosomatina physiology
- Abstract
Certain trypanosomatids co-evolve with an endosymbiotic bacterium in a mutualistic relationship that is characterized by intense metabolic exchanges. Symbionts were able to respire for up to 4 h after isolation from Angomonas deanei. FCCP (carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone) similarly increased respiration in wild-type and aposymbiotic protozoa, though a higher maximal O2 consumption capacity was observed in the symbiont-containing cells. Rotenone, a complex I inhibitor, did not affect A. deanei respiration, whereas TTFA (thenoyltrifluoroacetone), a complex II activity inhibitor, completely blocked respiration in both strains. Antimycin A and cyanide, inhibitors of complexes III and IV, respectively, abolished O2 consumption, but the aposymbiotic protozoa were more sensitive to both compounds. Oligomycin did not affect cell respiration, whereas carboxyatractyloside (CAT), an inhibitor of the ADP-ATP translocator, slightly reduced O2 consumption. In the A. deanei genome, sequences encoding most proteins of the respiratory chain are present. The symbiont genome lost part of the electron transport system (ETS), but complex I, a cytochrome d oxidase, and FoF1-ATP synthase remain. In conclusion, this work suggests that the symbiont influences the mitochondrial respiration of the host protozoan.
- Published
- 2015
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30. Therapeutic itineraries and explanations for tuberculosis: an indigenous perspective.
- Author
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Nogueira LM, Teixeira E, Basta PC, and Motta MC
- Subjects
- Adult, Aged, Aged, 80 and over, Brazil, Female, Humans, Interviews as Topic, Male, Middle Aged, Tuberculosis diagnosis, Tuberculosis therapy, Young Adult, Culture, Health Knowledge, Attitudes, Practice, Indians, South American, Medicine, Traditional, Tuberculosis ethnology
- Abstract
Objective: To analyze explanations for tuberculosis and therapeutic itineraries of Brazilian indigenous people., Methods: Case study with a qualitative-descriptive approach. We conducted semi-structured interviews with 11 Munduruku indigenous, including direct observation of treatment for tuberculosis in the municipality of Jacareacanga, south-western region of the state of Para, Brazil. To identify explanations for tuberculosis and therapeutic itineraries, we performed thematic content analysis., Results: Traditional medicine was the first therapeutic option chosen by the indigenous. However, biomedicine was also employed, which indicates a circulation between different therapeutic contexts and health concepts among the Munduruku. The explanations provided ranged from recognition of the signs and symptoms specific to tuberculosis to the attribution of the disease to a spirit that leaves the body and wanders in the woods, returning ill into the body. Unlike the biomedical model, which links tuberculosis transmission strictly to interpersonal contact, in closed spaces without natural lighting and ventilation (preferably domestic environments), the Munduruku associate the disease to an indirect contact between people socially distant (enemies or adversaries) in public and open places., Conclusions: The explanations made by the indigenous are unique and deserve the attention of those who are responsible for developing health public policies, as well as of the teams who work on the villages. To guarantee an efficient control of tuberculosis in these regions, it is necessary that the developed actions integrate biomedicine knowledge and the traditional medicine of the indigenous people, in addition to respecting and welcoming local culture manifestations.
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- 2015
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31. Psychological and practical difficulties among parents and healthy siblings of children with Duchenne vs. Becker muscular dystrophy: an Italian comparative study.
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Magliano L, D'Angelo MG, Vita G, Pane M, D'Amico A, Balottin U, Angelini C, Battini R, Politano L, Patalano M, Sagliocchi A, Civati F, Brighina E, Vita GL, Messina S, Sframeli M, Lombardo ME, Scalise R, Colia G, Catteruccia M, Berardinelli A, Motta MC, Gaiani A, Semplicini C, Bello L, Astrea G, Zaccaro A, and Scutifero M
- Subjects
- Adolescent, Adult, Child, Child, Preschool, Cost of Illness, Family, Humans, Italy, Middle Aged, Social Support, Socioeconomic Factors, Caregivers psychology, Caregivers statistics & numerical data, Family Health, Muscular Dystrophy, Duchenne physiopathology, Muscular Dystrophy, Duchenne psychology, Parents psychology, Siblings psychology
- Abstract
This study explored the burden in parents and healthy siblings of 4-17 year-old patients with Duchenne (DMD) and Becker (BMD) muscular dystrophies, and whether the burden varied according to clinical aspects and social resources. Data on socio-demographic characteristics, patient's clinical history, parent and healthy children burden, and on parent's social resources were collected using self-reported questionnaires administered to 336 parents of patients with DMD (246) and BMD (90). Parents of patients with DMD reported higher burden than those of patients with BMD, especially concerning feeling of loss (84.3% DMD vs. 57.4% BMD), stigma (44.2% DMD vs. 5.5% BMD) and neglect of hobbies (69.0% DMD vs. 32.5% BMD). Despite the burden, 66% DMD and 62.4% BMD parents stated the caregiving experience had a positive impact on their lives. A minority of parents believed MD has a negative influence on the psychological well-being (31.0% DMD vs. 12.8% BMD), and social life of unaffected children (25.7% vs. 18.4%). In the DMD group, burden correlated with duration of illness and parent age, and burden was higher among parents with lower social contacts and support in emergencies. In DMD, difficulties among healthy children were reported as higher by parents who were older, had higher burden and lower social contacts. In both groups, burden increased in relation to patient disability. These findings underline that the psychological support to be provided to parents of patients with MD, should take into account clinical features of the disease.
- Published
- 2014
32. Identification of a novel nucleocytoplasmic shuttling RNA helicase of trypanosomes.
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Inoue AH, Serpeloni M, Hiraiwa PM, Yamada-Ogatta SF, Muniz JR, Motta MC, Vidal NM, Goldenberg S, and Avila AR
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- Amino Acid Sequence, Axenic Culture, Catalytic Domain, Cell Nucleus enzymology, Conserved Sequence, Cytoplasm enzymology, Models, Molecular, Molecular Sequence Data, Nuclear Pore enzymology, Protein Transport, Protozoan Proteins chemistry, Protozoan Proteins genetics, RNA Helicases chemistry, RNA Helicases genetics, RNA Transport, RNA, Messenger metabolism, Ribonucleoproteins chemistry, Ribonucleoproteins genetics, Ribonucleoproteins metabolism, Protozoan Proteins metabolism, RNA Helicases metabolism, Trypanosoma cruzi enzymology
- Abstract
Gene expression in trypanosomes is controlled mostly by post-transcriptional pathways. Little is known about the components of mRNA nucleocytoplasmic export routes in these parasites. Comparative genomics has shown that the mRNA transport pathway is the least conserved pathway among eukaryotes. Nonetheless, we identified a RNA helicase (Hel45) that is conserved across eukaryotes and similar to shuttling proteins involved in mRNA export. We used in silico analysis to predict the structure of Trypanosoma cruzi Hel45, including the N-terminal domain and the C-terminal domain, and our findings suggest that this RNA helicase can form complexes with mRNA. Hel45 was present in both nucleus and cytoplasm. Electron microscopy showed that Hel45 is clustered close to the cytoplasmic side of nuclear pore complexes, and is also present in the nucleus where it is associated with peripheral compact chromatin. Deletion of a predicted Nuclear Export Signal motif led to the accumulation of Hel45ΔNES in the nucleus, indicating that Hel45 shuttles between the nucleus and the cytoplasm. This transport was dependent on active transcription but did not depend on the exportin Crm1. Knockdown of Mex67 in T. brucei caused the nuclear accumulation of the T. brucei ortholog of Hel45. Indeed, Hel45 is present in mRNA ribonucleoprotein complexes that are not associated with polysomes. It is still necessary to confirm the precise function of Hel45. However, this RNA helicase is associated with mRNA metabolism and its nucleocytoplasmic shuttling is dependent on an mRNA export route involving Mex67 receptor.
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- 2014
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33. Effects of camptothecin derivatives and topoisomerase dual inhibitors on Trypanosoma cruzi growth and ultrastructure.
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Lacombe OK, Zuma AA, da Silva CC, de Souza W, and Motta MC
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- Humans, Treatment Outcome, Trypanosoma cruzi drug effects, Trypanosoma cruzi enzymology, Camptothecin analogs & derivatives, Camptothecin pharmacology, Topoisomerase I Inhibitors pharmacology, Topoisomerase II Inhibitors pharmacology, Trypanosoma cruzi growth & development, Trypanosoma cruzi ultrastructure
- Abstract
Background: Trypanosoma cruzi is the etiological agent of Chagas' disease that is an endemic disease in Latin America and affects about 8 million people. This parasite belongs to the Trypanosomatidae family which contains a single mitochondrion with an enlarged region, named kinetoplast that harbors the mitochondrial DNA (kDNA). The kinetoplast and the nucleus present a great variety of essential enzymes involved in DNA replication and topology, including DNA topoisomerases. Such enzymes are considered to be promising molecular targets for cancer treatment and for antiparasitic chemotherapy. In this work, the proliferation and ultrastructure of T. cruzi epimastigotes were evaluated after treatment with eukaryotic topoisomerase I inhibitors, such as topotecan and irinotecan, as well as with dual inhibitors (compounds that block eukaryotic topoisomerase I and topoisomerase II activities), such as baicalein, luteolin and evodiamine. Previous studies have shown that such inhibitors were able to block the growth of tumor cells, however most of them have never been tested on trypanosomatids., Results: Considering the effects of topoisomerase I inhibitors, our results showed that topotecan decreased cell proliferation and caused unpacking of nuclear heterochromatin, however none of these alterations were observed after treatment with irinotecan. The dual inhibitors baicalein and evodiamine decreased cell growth; however the nuclear and kinetoplast ultrastructures were not affected., Conclusions: Taken together, our data showed that camptothecin is more efficient than its derivatives in decreasing T. cruzi proliferation. Furthermore, we conclude that drugs pertaining to a certain class of topoisomerase inhibitors may present different efficiencies as chemotherapeutical agents.
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- 2014
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34. Trypanosoma cruzi bromodomain factor 3 binds acetylated α-tubulin and concentrates in the flagellum during metacyclogenesis.
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Alonso VL, Villanova GV, Ritagliati C, Machado Motta MC, Cribb P, and Serra EC
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- Acetylation, Cytoplasm metabolism, Flagella ultrastructure, Microtubules metabolism, Protein Binding, Protein Structure, Tertiary, Protein Transport, Protozoan Proteins chemistry, Protozoan Proteins genetics, Transcription Factors chemistry, Transcription Factors genetics, Trypanosoma cruzi genetics, Trypanosoma cruzi growth & development, Flagella metabolism, Life Cycle Stages, Protein Processing, Post-Translational, Protozoan Proteins metabolism, Transcription Factors metabolism, Trypanosoma cruzi metabolism, Tubulin metabolism
- Abstract
Bromodomains are highly conserved acetyl-lysine binding domains found mainly in proteins associated with chromatin and nuclear acetyltransferases. The Trypanosoma cruzi genome encodes at least four bromodomain factors (TcBDFs). We describe here bromodomain factor 3 (TcBDF3), a bromodomain-containing protein localized in the cytoplasm. TcBDF3 cytolocalization was determined, using purified antibodies, by Western blot and immunofluorescence analyses in all life cycle stages of T. cruzi. In epimastigotes and amastigotes, it was detected in the cytoplasm, the flagellum, and the flagellar pocket, and in trypomastigotes only in the flagellum. Subcellular localization of TcBDF3 was also determined by digitonin extraction, ultrastructural immunocytochemistry, and expression of TcBDF3 fused to cyan fluorescent protein (CFP). Tubulin can acquire different posttranslational modifications, which modulate microtubule functions. Acetylated α-tubulin has been found in the axonemes of flagella and cilia, as well as in the subpellicular microtubules of trypanosomatids. TcBDF3 and acetylated α-tubulin partially colocalized in isolated cytoskeletons and flagella from T. cruzi epimastigotes and trypomastigotes. Interaction between the two proteins was confirmed by coimmunoprecipitation and far-Western blot assays with synthetic acetylated α-tubulin peptides and recombinant TcBDF3., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)
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- 2014
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35. Unveiling benznidazole's mechanism of action through overexpression of DNA repair proteins in Trypanosoma cruzi.
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Rajão MA, Furtado C, Alves CL, Passos-Silva DG, de Moura MB, Schamber-Reis BL, Kunrath-Lima M, Zuma AA, Vieira-da-Rocha JP, Garcia JB, Mendes IC, Pena SD, Macedo AM, Franco GR, de Souza-Pinto NC, de Medeiros MH, Cruz AK, Motta MC, Teixeira SM, and Machado CR
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- Animals, Cell Survival, Chagas Disease drug therapy, Chagas Disease genetics, Chagas Disease parasitology, DNA Glycosylases genetics, DNA Repair drug effects, DNA, Protozoan drug effects, Guanine analogs & derivatives, Guanine metabolism, Real-Time Polymerase Chain Reaction, Trypanosoma cruzi genetics, DNA Repair Enzymes genetics, Drug Resistance genetics, Nitroimidazoles pharmacology, Protozoan Proteins genetics, Trypanocidal Agents pharmacology, Trypanosoma cruzi drug effects
- Abstract
Benznidazole (BZ) is the most commonly used drug for the treatment of Chagas disease. Although BZ is known to induce the formation of free radicals and electrophilic metabolites within the parasite Trypanosoma cruzi, its precise mechanisms of action are still elusive. Here, we analyzed the survival of T. cruzi exposed to BZ using genetically modified parasites overexpressing different DNA repair proteins. Our results indicate that BZ induces oxidation mainly in the nucleotide pool, as heterologous expression of the nucleotide pyrophosphohydrolase MutT (but not overexpression of the glycosylase TcOgg1) increased drug resistance in the parasite. In addition, electron microscopy indicated that BZ catalyzes the formation of double-stranded breaks in the parasite, as its genomic DNA undergoes extensive heterochromatin unpacking following exposure to the drug. Furthermore, the overexpression of proteins involved in the recombination-mediated DNA repair increased resistance to BZ, reinforcing the idea that the drug causes double-stranded breaks. Our results also show that the overexpression of mitochondrial DNA repair proteins increase parasite survival upon BZ exposure, indicating that the drug induces lesions in the mitochondrial DNA as well. These findings suggest that BZ preferentially oxidizes the nucleotide pool, and the extensive incorporation of oxidized nucleotides during DNA replication leads to potentially lethal double-stranded DNA breaks in T. cruzi DNA., (Copyright © 2013 Wiley Periodicals, Inc.)
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- 2014
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36. Structural characterization of the cell division cycle in Strigomonas culicis, an endosymbiont-bearing trypanosomatid.
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Brum FL, Catta-Preta CM, de Souza W, Schenkman S, Elias MC, and Motta MC
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- Bacteria, Cell Cycle physiology, Cell Division physiology, DNA, Protozoan analysis, DNA, Protozoan chemistry, Microscopy, Fluorescence, Organelles chemistry, Organelles microbiology, Trypanosomatina chemistry, Trypanosomatina cytology, Symbiosis physiology, Trypanosomatina microbiology, Trypanosomatina physiology
- Abstract
Strigomonas culicis (previously referred to as Blastocrithidia culicis) is a monoxenic trypanosomatid harboring a symbiotic bacterium, which maintains an obligatory relationship with the host protozoan. Investigations of the cell cycle in symbiont harboring trypanosomatids suggest that the bacterium divides in coordination with other host cell structures, particularly the nucleus. In this study we used light and electron microscopy followed by three-dimensional reconstruction to characterize the symbiont division during the cell cycle of S. culicis. We observed that during this process, the symbiotic bacterium presents different forms and is found at different positions in relationship to the host cell structures. At the G1/S phase of the protozoan cell cycle, the endosymbiont exhibits a constricted form that appears to elongate, resulting in the bacterium division, which occurs before kinetoplast and nucleus segregation. During cytokinesis, the symbionts are positioned close to each nucleus to ensure that each daughter cell will inherit a single copy of the bacterium. These observations indicated that the association of the bacterium with the protozoan nucleus coordinates the cell cycle in both organisms.
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- 2014
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37. How Trypanosoma cruzi handles cell cycle arrest promoted by camptothecin, a topoisomerase I inhibitor.
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Zuma AA, Mendes IC, Reignault LC, Elias MC, de Souza W, Machado CR, and Motta MC
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- Apoptosis drug effects, DNA Damage drug effects, Trypanosoma cruzi cytology, Camptothecin pharmacology, Cell Cycle Checkpoints drug effects, Topoisomerase I Inhibitors pharmacology, Trypanosoma cruzi drug effects
- Abstract
The protozoan Trypanosoma cruzi is the etiological agent of Chagas disease, which affects approximately 8 million people in Latin America. This parasite contains a single nucleus and a kinetoplast, which harbors the mitochondrial DNA (kDNA). DNA topoisomerases act during replication, transcription and repair and modulate DNA topology by reverting supercoiling in the DNA double-strand. In this work, we evaluated the effects promoted by camptothecin, a topoisomerase I inhibitor that promotes protozoan proliferation impairment, cell cycle arrest, ultrastructure alterations and DNA lesions in epimastigotes of T. cruzi. The results showed that inhibition of cell proliferation was reversible only at the lowest drug concentration (1μM) used. The unpacking of nuclear heterochromatin and mitochondrion swelling were the main ultrastructural modifications observed. Inhibition of parasite proliferation also led to cell cycle arrest, which was most likely caused by nuclear DNA lesions. Following camptothecin treatment, some of the cells restored their DNA, whereas others entered early apoptosis but did not progress to late apoptosis, indicating that the protozoa stay alive in a "senescence-like" state. This programmed cell death may be associated with a decrease in mitochondrial membrane potential and an increase in the production of reactive oxygen species. Taken together, these results indicate that the inhibition of T. cruzi proliferation is related to events capable of affecting cell cycle, DNA organization and mitochondrial activity., (Copyright © 2014. Published by Elsevier B.V.)
- Published
- 2014
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38. "I have got something positive out of this situation": psychological benefits of caregiving in relatives of young people with muscular dystrophy.
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Magliano L, Patalano M, Sagliocchi A, Scutifero M, Zaccaro A, D'Angelo MG, Civati F, Brighina E, Vita G, Vita GL, Messina S, Sframeli M, Pane M, Lombardo ME, Scalise R, D'Amico A, Colia G, Catteruccia M, Balottin U, Berardinelli A, Motta MC, Angelini C, Gaiani A, Semplicini C, Bello L, Battini R, Astrea G, Ricci G, and Politano L
- Subjects
- Activities of Daily Living, Adolescent, Adult, Aged, Aged, 80 and over, Analysis of Variance, Child, Child, Preschool, Cost of Illness, Female, Humans, Italy, Male, Middle Aged, Social Support, Stress, Psychological, Surveys and Questionnaires, Young Adult, Caregivers psychology, Family psychology, Muscular Dystrophies nursing, Muscular Dystrophies psychology
- Abstract
This paper focuses on the psychological benefits of caregiving in key relatives of patients with muscular dystrophies (MD), a group of rare diseases characterized by progressive weakness and restriction of the patient's functional abilities. We describe whether relatives perceived caregiving to be a positive experience and test whether relatives' perceptions vary in relation to their view of the patient as a valued person, the degree of involvement in care, and the level of support provided by social network and professionals. The study sample included 502 key relatives of patients aged 4-25 years, suffering from Duchenne, Becker, or limb-girdle MD, in treatment for at least 6 months to one of the eight participating centers, living with at least one relative aged 18-80 years. Of key relatives, 88 % stated that they had gotten something positive out of the situation, 96 % considered their patients to be sensitive, and 94 % viewed their patients as talented. Positive aspects of caregiving were more recognized by key relatives who were more convinced that the patient was sensitive and who perceived that they received higher level of professional help and psychological social support. These results suggest that most key relatives consider that their caregiving experience has had a positive impact on their lives, despite the practical difficulties of caring for patients with MD. Professionals should help relatives to identify the benefits of caregiving without denying its difficulties. Clinicians themselves should develop positive attitudes towards family involvement in the care of patients with long-term diseases.
- Published
- 2014
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39. Biosynthesis of vitamins and cofactors in bacterium-harbouring trypanosomatids depends on the symbiotic association as revealed by genomic analyses.
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Klein CC, Alves JM, Serrano MG, Buck GA, Vasconcelos AT, Sagot MF, Teixeira MM, Camargo EP, and Motta MC
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- Betaproteobacteria metabolism, Biological Factors genetics, Biological Factors metabolism, Genome, Protozoan genetics, Genomics methods, Phylogeny, Trypanosoma metabolism, Vitamins genetics, Vitamins metabolism, Betaproteobacteria genetics, Biological Factors biosynthesis, Biosynthetic Pathways genetics, Symbiosis genetics, Trypanosoma genetics, Trypanosoma microbiology, Vitamins biosynthesis
- Abstract
Some non-pathogenic trypanosomatids maintain a mutualistic relationship with a betaproteobacterium of the Alcaligenaceae family. Intensive nutritional exchanges have been reported between the two partners, indicating that these protozoa are excellent biological models to study metabolic co-evolution. We previously sequenced and herein investigate the entire genomes of five trypanosomatids which harbor a symbiotic bacterium (SHTs for Symbiont-Haboring Trypanosomatids) and the respective bacteria (TPEs for Trypanosomatid Proteobacterial Endosymbiont), as well as two trypanosomatids without symbionts (RTs for Regular Trypanosomatids), for the presence of genes of the classical pathways for vitamin biosynthesis. Our data show that genes for the biosynthetic pathways of thiamine, biotin, and nicotinic acid are absent from all trypanosomatid genomes. This is in agreement with the absolute growth requirement for these vitamins in all protozoa of the family. Also absent from the genomes of RTs are the genes for the synthesis of pantothenic acid, folic acid, riboflavin, and vitamin B6. This is also in agreement with the available data showing that RTs are auxotrophic for these essential vitamins. On the other hand, SHTs are autotrophic for such vitamins. Indeed, all the genes of the corresponding biosynthetic pathways were identified, most of them in the symbiont genomes, while a few genes, mostly of eukaryotic origin, were found in the host genomes. The only exceptions to the latter are: the gene coding for the enzyme ketopantoate reductase (EC:1.1.1.169) which is related instead to the Firmicutes bacteria; and two other genes, one involved in the salvage pathway of pantothenic acid and the other in the synthesis of ubiquinone, that are related to Gammaproteobacteria. Their presence in trypanosomatids may result from lateral gene transfer. Taken together, our results reinforce the idea that the low nutritional requirement of SHTs is associated with the presence of the symbiotic bacterium, which contains most genes for vitamin production.
- Published
- 2013
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40. Endosymbiosis in trypanosomatids: the genomic cooperation between bacterium and host in the synthesis of essential amino acids is heavily influenced by multiple horizontal gene transfers.
- Author
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Alves JM, Klein CC, da Silva FM, Costa-Martins AG, Serrano MG, Buck GA, Vasconcelos AT, Sagot MF, Teixeira MM, Motta MC, and Camargo EP
- Subjects
- Betaproteobacteria physiology, Biological Evolution, Genome, Bacterial, Phylogeny, Trypanosomatina classification, Trypanosomatina metabolism, Amino Acids, Essential biosynthesis, Betaproteobacteria genetics, Gene Transfer, Horizontal, Symbiosis, Trypanosomatina genetics, Trypanosomatina microbiology
- Abstract
Background: Trypanosomatids of the genera Angomonas and Strigomonas live in a mutualistic association characterized by extensive metabolic cooperation with obligate endosymbiotic Betaproteobacteria. However, the role played by the symbiont has been more guessed by indirect means than evidenced. Symbiont-harboring trypanosomatids, in contrast to their counterparts lacking symbionts, exhibit lower nutritional requirements and are autotrophic for essential amino acids. To evidence the symbiont's contributions to this autotrophy, entire genomes of symbionts and trypanosomatids with and without symbionts were sequenced here., Results: Analyses of the essential amino acid pathways revealed that most biosynthetic routes are in the symbiont genome. By contrast, the host trypanosomatid genome contains fewer genes, about half of which originated from different bacterial groups, perhaps only one of which (ornithine cyclodeaminase, EC:4.3.1.12) derived from the symbiont. Nutritional, enzymatic, and genomic data were jointly analyzed to construct an integrated view of essential amino acid metabolism in symbiont-harboring trypanosomatids. This comprehensive analysis showed perfect concordance among all these data, and revealed that the symbiont contains genes for enzymes that complete essential biosynthetic routes for the host amino acid production, thus explaining the low requirement for these elements in symbiont-harboring trypanosomatids. Phylogenetic analyses show that the cooperation between symbionts and their hosts is complemented by multiple horizontal gene transfers, from bacterial lineages to trypanosomatids, that occurred several times in the course of their evolution. Transfers occur preferentially in parts of the pathways that are missing from other eukaryotes., Conclusion: We have herein uncovered the genetic and evolutionary bases of essential amino acid biosynthesis in several trypanosomatids with and without endosymbionts, explaining and complementing decades of experimental results. We uncovered the remarkable plasticity in essential amino acid biosynthesis pathway evolution in these protozoans, demonstrating heavy influence of horizontal gene transfer events, from Bacteria to trypanosomatid nuclei, in the evolution of these pathways.
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- 2013
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41. Acriflavine treatment promotes dyskinetoplasty in Trypanosoma cruzi as revealed by ultrastructural analysis.
- Author
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Manchester T, Cavalcanti DP, Zogovich M, DE Souza W, and Motta MC
- Subjects
- Cell Proliferation drug effects, DNA Replication drug effects, DNA, Kinetoplast genetics, Histocytochemistry, Microscopy, Atomic Force, Microscopy, Electron, Transmission, Mitochondria drug effects, Trypanosoma cruzi drug effects, Trypanosoma cruzi metabolism, Acriflavine pharmacology, DNA, Kinetoplast drug effects, Mitochondria ultrastructure, Trypanosoma cruzi ultrastructure
- Abstract
Trypanosomatid mitochondrial DNA is structured as a giant network of thousands of interlocked DNA molecules enclosed within the kinetoplast. The structure and replication mechanism of kinetoplast DNA (kDNA) is unique, thereby making it an excellent chemotherapeutic target. Alteration in the structural organization of kDNA can give rise to dyskinetoplastic (Dk) strains. In Dk cells, the kDNA is dispersed in clumps throughout the mitochondrial matrix and not organized into a network. In this work, Trypanosoma cruzi epimastigotes were treated with acriflavine, a DNA intercalating drug, which promoted a decrease in cell proliferation and induced the appearance of Dk protozoa. In treated cells, the kinetoplast lost its normal disc-shaped structure because the fibrillar arrangement was reduced to a compact, amorphous mass within the mitochondrion. Moreover, basic proteins associated with kDNA were redistributed throughout the Dk protozoal kinetoplast. We sought to understand how the disruption of the kDNA leads to the emergence of the Dk phenotype with atomic force microscopy (AFM) analysis of isolated networks. Our results demonstrate that the detachment of minicircles from the kDNA disk promotes the disassembly of the network, thereby generating Dk cells. Our data strongly suggest that acriflavine inhibits T. cruzi multiplication by interfering with kDNA replication.
- Published
- 2013
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42. The presence of a symbiotic bacterium in Strigomonas culicis is related to differential ecto-phosphatase activity and influences the mosquito-protozoa interaction.
- Author
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Catta-Preta CM, Nascimento MT, Garcia MC, Saraiva EM, Motta MC, and Meyer-Fernandes JR
- Subjects
- Animals, Female, Gastrointestinal Tract parasitology, Salivary Glands parasitology, Trypanosomatina enzymology, Aedes parasitology, Bacteria growth & development, Bacterial Physiological Phenomena, Phosphoric Monoester Hydrolases metabolism, Symbiosis, Trypanosomatina microbiology, Trypanosomatina physiology
- Abstract
Strigomonas culicis is a monoxenous trypanosomatid that co-evolves with a symbiotic bacterium in a mutualistic relationship that is characterized by intense metabolic exchanges between both partners. S. culicis infects and colonizes the Aedes aegypti mosquito midgut, reaches its hemocoel and then invades the salivary glands. An artificial aposymbiotic strain is unable to colonize insects, reinforcing the idea that the bacterium influences the protozoan surface composition and cell interaction. Here, we report the characterization of the hydrolytic activity of ecto-phosphatases evaluated in symbiont-bearing and aposymbiotic strains of S. culicis by incubating the protozoa with p-nitrophenyl phosphate (pNPP) at different pH levels, in the presence of phosphatase inhibitors, and with several divalent metals. The symbiont-bearing and aposymbiotic cells differ in their ecto-phosphatase enzymes, based on their activities and specificities. Furthermore, the ability of the protozoan to bind to the mosquito midgut and salivary glands was impaired by ecto-phosphatase inhibition. Taken together, our data suggest that the symbiont influences the host protozoan ecto-phosphatase activity and indicate a possible role of this enzyme during mosquito tissue colonization by S. culicis., (Copyright © 2013 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2013
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43. 24 month longitudinal data in ambulant boys with Duchenne muscular dystrophy.
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Mazzone ES, Pane M, Sormani MP, Scalise R, Berardinelli A, Messina S, Torrente Y, D'Amico A, Doglio L, Viggiano E, D'Ambrosio P, Cavallaro F, Frosini S, Bello L, Bonfiglio S, De Sanctis R, Rolle E, Bianco F, Magri F, Rossi F, Vasco G, Vita G, Motta MC, Donati MA, Sacchini M, Mongini T, Pini A, Battini R, Pegoraro E, Previtali S, Napolitano S, Bruno C, Politano L, Comi GP, Bertini E, and Mercuri E
- Subjects
- Adolescent, Analysis of Variance, Anti-Inflammatory Agents therapeutic use, Child, Child, Preschool, Glucocorticoids therapeutic use, Humans, Logistic Models, Longitudinal Studies, Male, Muscular Dystrophy, Duchenne drug therapy, Outcome Assessment, Health Care, Prednisone therapeutic use, Pregnenediones therapeutic use, Risk Assessment, Risk Factors, Time Factors, Exercise Test methods, Muscular Dystrophy, Duchenne physiopathology, Walking physiology
- Abstract
Objectives: The aim of the study was i) to assess the spectrum of changes over 24 months in ambulant boys affected by Duchenne muscular dystrophy, ii) to establish the difference between the first and the second year results and iii) to identify possible early markers of loss of ambulation., Methods: One hundred and thirteen patients (age range 4.1-17, mean 8.2) fulfilled the inclusion criteria, 67 of the 113 were on daily and 40 on intermittent steroids, while 6 were not on steroids. All were assessed using the 6 Minute Walk Test (6MWT), the North Star Ambulatory Assessment (NSAA) and timed test., Results: On the 6MWT there was an average overall decline of -22.7 (SD 81.0) in the first year and of -64.7 (SD 123.1) in the second year. On the NSAA the average overall decline was of -1.86 (SD 4.21) in the first year and of -2.98 (SD 5.19) in the second year. Fourteen children lost ambulation, one in the first year and the other 13 in the second year of the study. A distance of at least 330 meters on the 6MWT, or a NSAA score of 18 at baseline reduced significantly the risk of losing ambulation within 2 years., Conclusions: These results can be of help at the time of using inclusion criteria for a study in ambulant patients in order to minimize the risk of patients who may lose ambulation within the time of the trial.
- Published
- 2013
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44. Predicting the proteins of Angomonas deanei, Strigomonas culicis and their respective endosymbionts reveals new aspects of the trypanosomatidae family.
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Motta MC, Martins AC, de Souza SS, Catta-Preta CM, Silva R, Klein CC, de Almeida LG, de Lima Cunha O, Ciapina LP, Brocchi M, Colabardini AC, de Araujo Lima B, Machado CR, de Almeida Soares CM, Probst CM, de Menezes CB, Thompson CE, Bartholomeu DC, Gradia DF, Pavoni DP, Grisard EC, Fantinatti-Garboggini F, Marchini FK, Rodrigues-Luiz GF, Wagner G, Goldman GH, Fietto JL, Elias MC, Goldman MH, Sagot MF, Pereira M, Stoco PH, de Mendonça-Neto RP, Teixeira SM, Maciel TE, de Oliveira Mendes TA, Ürményi TP, de Souza W, Schenkman S, and de Vasconcelos AT
- Subjects
- Bacteria metabolism, Base Composition, Base Sequence, Biological Evolution, Leishmania major genetics, Metabolic Networks and Pathways, Molecular Sequence Annotation, Molecular Sequence Data, Open Reading Frames, Protozoan Proteins metabolism, Sequence Alignment, Sequence Analysis, DNA, Trypanosomatina classification, Trypanosomatina metabolism, Trypanosomatina microbiology, Genes, Protozoan, Phylogeny, Protozoan Proteins genetics, Symbiosis genetics, Trypanosomatina genetics
- Abstract
Endosymbiont-bearing trypanosomatids have been considered excellent models for the study of cell evolution because the host protozoan co-evolves with an intracellular bacterium in a mutualistic relationship. Such protozoa inhabit a single invertebrate host during their entire life cycle and exhibit special characteristics that group them in a particular phylogenetic cluster of the Trypanosomatidae family, thus classified as monoxenics. In an effort to better understand such symbiotic association, we used DNA pyrosequencing and a reference-guided assembly to generate reads that predicted 16,960 and 12,162 open reading frames (ORFs) in two symbiont-bearing trypanosomatids, Angomonas deanei (previously named as Crithidia deanei) and Strigomonas culicis (first known as Blastocrithidia culicis), respectively. Identification of each ORF was based primarily on TriTrypDB using tblastn, and each ORF was confirmed by employing getorf from EMBOSS and Newbler 2.6 when necessary. The monoxenic organisms revealed conserved housekeeping functions when compared to other trypanosomatids, especially compared with Leishmania major. However, major differences were found in ORFs corresponding to the cytoskeleton, the kinetoplast, and the paraflagellar structure. The monoxenic organisms also contain a large number of genes for cytosolic calpain-like and surface gp63 metalloproteases and a reduced number of compartmentalized cysteine proteases in comparison to other TriTryp organisms, reflecting adaptations to the presence of the symbiont. The assembled bacterial endosymbiont sequences exhibit a high A+T content with a total of 787 and 769 ORFs for the Angomonas deanei and Strigomonas culicis endosymbionts, respectively, and indicate that these organisms hold a common ancestor related to the Alcaligenaceae family. Importantly, both symbionts contain enzymes that complement essential host cell biosynthetic pathways, such as those for amino acid, lipid and purine/pyrimidine metabolism. These findings increase our understanding of the intricate symbiotic relationship between the bacterium and the trypanosomatid host and provide clues to better understand eukaryotic cell evolution.
- Published
- 2013
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45. Effects of miltefosine on the proliferation, ultrastructure, and phospholipid composition of Angomonas deanei, a trypanosomatid protozoan that harbors a symbiotic bacterium.
- Author
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de Freitas-Junior PR, Catta-Preta CM, Andrade Ida S, Cavalcanti DP, de Souza W, Einicker-Lamas M, and Motta MC
- Subjects
- Bacteria drug effects, Bacteria growth & development, Cell Membrane drug effects, Cell Membrane metabolism, Cell Membrane ultrastructure, Cell Wall drug effects, Cell Wall metabolism, Choline-Phosphate Cytidylyltransferase metabolism, Crithidia metabolism, Crithidia ultrastructure, Microscopy, Electron, Transmission, Mitochondria drug effects, Mitochondria ultrastructure, Phosphatidylcholines biosynthesis, Phosphorus Isotopes metabolism, Phosphorylcholine pharmacology, Crithidia drug effects, Crithidia microbiology, Phosphorylcholine analogs & derivatives, Symbiosis
- Abstract
Some trypanosomatids, such as Angomonas deanei formerly named as Crithidia deanei, present an obligatory intracellular bacterium, which maintains a mutualistic relationship with the host. Phosphatidylcholine (PC) is the major phospholipid in eukaryotes and an essential component of cell membranes playing structural, biochemical, and physiological roles. However, in prokaryotes, PC is present only in those species closely associated with eukaryotes, either in symbiotic or pathogenic interactions. In trypanosomatids, the endosymbiont envelope is composed by a reduced cell wall and by two membrane units that lack sterols and present cardiolipin (CL) and PC as the major phospholipids. In this study, we tested the effects of miltefosine in A. deanei proliferation, as well as, on the ultrastrucuture and phospholipid composition considering that this drug inhibits the CTP-phosphocholine cytidyltransferase (CCT), a key enzyme in the PC biosynthesis. Besides the low effect of miltefosine in cellular proliferation, treated protozoa presented ultrastructural alterations such as plasma membrane shedding and blebbing, mitochondrial swelling, and convolutions of the endosymbiont envelope. The use of (32) Pi as a tracer revealed that the production of PC, CL, and phosphatidylethanolamine decreased while phosphatidylinositol production remained stable. Mitochondrion and symbiont fractions obtained from protozoa treated with miltefosine also presented a decrease in phospholipid production, reinforcing the idea that an intensive metabolic exchange occurs between the host trypanosomatid and structures of symbiotic origin., (© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)
- Published
- 2012
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46. DNA polymerase beta from Trypanosoma cruzi is involved in kinetoplast DNA replication and repair of oxidative lesions.
- Author
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Schamber-Reis BL, Nardelli S, Régis-Silva CG, Campos PC, Cerqueira PG, Lima SA, Franco GR, Macedo AM, Pena SD, Cazaux C, Hoffmann JS, Motta MC, Schenkman S, Teixeira SM, and Machado CR
- Subjects
- Microscopy, Fluorescence, Mitochondria chemistry, Mitochondria enzymology, Oxidative Stress, Trypanosoma cruzi chemistry, Trypanosoma cruzi genetics, DNA Polymerase beta metabolism, DNA Repair, DNA Replication, DNA, Kinetoplast metabolism, Trypanosoma cruzi enzymology
- Abstract
Specific DNA repair pathways from Trypanosoma cruzi are believed to protect genomic DNA and kinetoplast DNA (kDNA) from mutations. Particular pathways are supposed to operate in order to repair nucleotides oxidized by reactive oxygen species (ROS) during parasite infection, being 7,8-dihydro-8-oxoguanine (8oxoG) a frequent and highly mutagenic base alteration. If unrepaired, 8oxoG can lead to cytotoxic base transversions during DNA replication. In mammals, DNA polymerase beta (Polβ) is mainly involved in base excision repair (BER) of oxidative damage. However its biological role in T. cruzi is still unknown. We show, by immunofluorescence localization, that T. cruzi DNA polymerase beta (Tcpolβ) is restricted to the antipodal sites of kDNA in replicative epimastigote and amastigote developmental stages, being strictly localized to kDNA antipodal sites between G1/S and early G2 phase in replicative epimastigotes. Nevertheless, this polymerase was detected inside the mitochondrial matrix of trypomastigote forms, which are not able to replicate in culture. Parasites over expressing Tcpolβ showed reduced levels of 8oxoG in kDNA and an increased survival after treatment with hydrogen peroxide when compared to control cells. However, this resistance was lost after treating Tcpolβ overexpressors with methoxiamine, a potent BER inhibitor. Curiously, a presumed DNA repair focus containing Tcpolβ was identified in the vicinity of kDNA of cultured wild type epimastigotes after treatment with hydrogen peroxide. Taken together our data suggest participation of Tcpolβ during kDNA replication and repair of oxidative DNA damage induced by genotoxic stress in this organelle., (Copyright © 2012 Elsevier B.V. All rights reserved.)
- Published
- 2012
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47. Effect of topoisomerase inhibitors and DNA-binding drugs on the cell proliferation and ultrastructure of Trypanosoma cruzi.
- Author
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Zuma AA, Cavalcanti DP, Maia MC, de Souza W, and Motta MC
- Subjects
- Cell Proliferation drug effects, Humans, Trypanosoma cruzi growth & development, Trypanosoma cruzi isolation & purification, Trypanosoma cruzi ultrastructure, Trypanosomatina parasitology, Antiprotozoal Agents pharmacology, Intercalating Agents pharmacology, Topoisomerase Inhibitors pharmacology, Trypanosoma cruzi drug effects
- Abstract
Trypanosomatids present unusual organelles, such as the kinetoplast that contains the mitochondrial DNA arranged in catenated circles. The nucleus of these protozoa presents distinct domains during interphase as well as a closed mitosis. DNA topoisomerases modulate the topological state of DNA by regulating supercoiling of the double-stranded DNA during replication, transcription, recombination and repair. Because topoisomerases play essential roles in cellular processes, they constitute a potential target for antitumour and antimicrobial drugs. In this study, the effects of various topoisomerase inhibitors and DNA-binding drugs were tested on the cellular proliferation and ultrastructure of the Trypanosoma cruzi epimastigote form Blastocrithidia culicis was used as a comparative model, which has a more relaxed kinetoplast DNA (kDNA) organization. The results showed that the eukaryotic topoisomerase I inhibitors camptothecin and rebeccamycin were the most effective compounds in the arrest of T. cruzi proliferation. Of the eukaryotic topoisomerase II inhibitors, mitoxantrone, but not merbarone, was effective against cell proliferation. The prokaryotic topoisomerase II inhibitors norfloxacin and enoxacin targeted the kinetoplast specifically, thus promoting ultrastructural kDNA rearrangement in B. culicis. Of the DNA-binding drugs, berenil caused remarkable kDNA disorganization. With the exception of camptothecin, there have been no previous evaluations of the compounds tested here on trypanosomatid ultrastructure. In conclusion, inhibitors of the same class may have different effects on trypanosomatid proliferation and ultrastructure. The results obtained in this work may help to reveal the mechanism of action of different topoisomerase inhibitors in trypanosomatids., (Copyright © 2010 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.)
- Published
- 2011
- Full Text
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48. Genetic and biological characterization of a densovirus isolate that affects dengue virus infection.
- Author
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Mosimann AL, Bordignon J, Mazzarotto GC, Motta MC, Hoffmann F, and Santos CN
- Subjects
- Animals, Chlorocebus aethiops, Densovirus physiology, Mice, Mice, Inbred BALB C, Pest Control, Biological, Phylogeny, Dengue Virus growth & development, Densovirus genetics
- Abstract
Brevidensoviruses have an encapsidated, single-stranded DNA genome that predominantly has a negative polarity. In recent years, they have received particular attention due to their potential role in the biological control of pathogenic arboviruses and to their unnoticed presence in cell cultures as contaminants. In addition, brevidensoviruses may also be useful as viral vectors. This study describes the first genetic and biological characterization of a mosquito densovirus that was isolated in Brazil; moreover, we examined the phylogenetic relationship between this isolate and the other brevidensoviruses. We further demonstrate that this densovirus has the potential to be used to biologically control dengue virus (DENV) infection with in vitro co-infection experiments. The present study provides evidence that this densovirus isolate is a fast-spreading virus that affects cell growth and DENV infection.
- Published
- 2011
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49. Trypanosoma cruzi DNA replication includes the sequential recruitment of pre-replication and replication machineries close to nuclear periphery.
- Author
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Calderano SG, de Melo Godoy PD, Motta MC, Mortara RA, Schenkman S, and Elias MC
- Subjects
- Active Transport, Cell Nucleus, Cell Cycle Proteins metabolism, DNA, Protozoan metabolism, G1 Phase, Origin Recognition Complex metabolism, Proliferating Cell Nuclear Antigen metabolism, Protozoan Proteins metabolism, S Phase, Cell Nucleus metabolism, DNA Replication, DNA, Protozoan biosynthesis, Trypanosoma cruzi cytology, Trypanosoma cruzi metabolism
- Abstract
In eukaryotes, many nuclear processes are spatially compartmentalized. Previously, we have shown that in Trypanosoma cruzi, an early-divergent eukaryote, DNA replication occurs at the nuclear periphery where chromosomes remain constrained during the S phase of the cell cycle. We followed Orc1/Cdc6, a pre-replication machinery component and the proliferating cell nuclear antigen (PCNA), a component of replication machinery, during the cell cycle of this protozoon. We found that, at the G(1) stage, TcOrc1/Cdc6 and TcPCNA are dispersed throughout the nuclear space. During the G(1)/S transition, TcOrc1/Cdc6 migrates to a region close to nuclear periphery. At the onset of S phase, TcPCNA is loaded onto the DNA and remains constrained close to nuclear periphery. Finally, in G(2), mitosis and cytokinesis, TcOrc1/Cdc6 and TcPCNA are dispersed throughout the nuclear space. Based on these findings, we propose that DNA replication in T. cruzi is accomplished by the organization of functional machineries in a spatial-temporal manner.
- Published
- 2011
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50. Barriers related to screening examinations for prostate cancer.
- Author
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Paiva EP, Motta MC, and Griep RH
- Subjects
- Aged, Cross-Sectional Studies, Humans, Male, Middle Aged, Surveys and Questionnaires, Health Knowledge, Attitudes, Practice, Prostatic Neoplasms diagnosis
- Abstract
With the aim of describing barriers to screening for prostate cancer, a domicile survey was carried out covering 160 men of a Family Health Strategy (FHS) area. Slightly over half had undergone the examination. Regarding beliefs related to the disease, 95% of the men believed there is a cure if detected early, 29.4% mentioned the possibility of a normal life while ill, 56.3% believed it may be asymptomatic, 36.1% agreed/disagreed that the treatment is worse than the disease and 34.4% agreed that the examination affects masculinity and that if you are well it is not necessary to perform it. Regarding barriers, 15% reported that the physician had never requested it, 10.9% did not consider it important and 16.9% were afraid to take the examination. While not the sole determinant, the dissemination of adequate knowledge regarding the examination can constitute a key strategy for the formation of a positive attitude in relation to early detection.
- Published
- 2011
- Full Text
- View/download PDF
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