42 results on '"Ribeiro LFC"'
Search Results
2. Maternal periodontitis potentiates monosodium glutamate-obesity damage on Wistar offspring's fast-glycolytic muscle.
- Author
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Costa LNC, de Paula TP, Zazula MF, Naliwaiko K, Nassar CA, Bertolini GRF, Torrejais MM, Ribeiro LFC, and Costa RM
- Subjects
- Animals, Female, Rats, Pregnancy, Male, Prenatal Exposure Delayed Effects, Tumor Necrosis Factor-alpha metabolism, Cytokines metabolism, Sodium Glutamate adverse effects, Rats, Wistar, Muscle, Skeletal pathology, Obesity complications, Obesity metabolism, Periodontitis pathology, Periodontitis metabolism, Periodontitis complications
- Abstract
Objective: To evaluate the effects of magnifying the damage caused by obesity induced by monosodium glutamate, using a model of maternal periodontitis, on the structure of the anterior tibialis muscle of the offspring., Materials and Methods: Twenty-four female Wistar rats were divided into four experimental groups: control (n = 6), obese (n = 6), control with periodontitis (n = 6) and obese with periodontitis (n = 6). At 78 days of life, the rats were mated with males without any experimental intervention. The offspring of these rats (n = 1/L), at 120 days of life, were weighed and measured, then euthanized. Plasma was collected for analysis of cytokines IL-6, IL-10, IL-17 and TNF-α. Adipose tissues were collected and weighed, and the anterior tibial muscle was designated for histomorphological analyses (n = 6/group)., Results: Monosodium glutamate offspring showed significant muscle changes, such as a reduction in the size of fibres and neuromuscular junctions, and an increase in the nucleus and capillaries. However, all these changes were more expressed in monosodium glutamate-obese with periodontitis offspring., Conclusion: This leads us to suggest a magnifying effect promoted by periodontitis to the damage already well described by monosodium glutamate-obesity, determined by low-intensity inflammation, causing greater muscle damage., (© 2024 Wiley Periodicals LLC.)
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- 2024
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3. Remobilization with whole-body vibration improves functionality, histomorphometric parameters, and AQP1 expression in the soleus muscle of Wistar rats.
- Author
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Wutzke MLS, Zazula MF, Peretti AL, Marin E, Gomes JC, de Carvalho AR, Beu CCL, Costa RM, Ribeiro LFC, and Bertolini GRF
- Subjects
- Animals, Rats, Male, Immobilization methods, Rats, Wistar, Vibration, Aquaporin 1 metabolism, Muscle, Skeletal metabolism, Muscle, Skeletal physiology
- Abstract
Background: Whole-body vibration (WBV) is used to enhance physical performance in sports and rehabilitation. The present study analyzed the effects of remobilization with WBV on the soleus muscle of Wistar rats., Methods: Twenty-eight animals were separated into four experimental groups (n = 7): CON (control); IM (immobilized); FR (immobilization and free remobilization); and WBV (immobilization and remobilization with WBV). The immobilization of the pelvic limb was carried out according to the standard protocol using a plaster cast for 15 days. For remobilization with WBV, a Frequency of 60 Hz was applied for 10 min, five days a week, for two weeks. After the remobilization period, the animals were euthanized, and the right soleus muscle was dissected followed by processing for histomorphometric analysis and immunolocalization of Aquaporin 1 (AQP1)., Results: We observed a reduced larger diameter in IM compared to CON, with restored values in WBV. For the estimation of connective tissue, a significant increase was observed in the immobilized groups, while a reduction was noted in the remobilized groups. AQP1 expression decreased significantly in IM and increased in WBV., Conclusion: Immobilization caused morphofunctional damage to the soleus muscle, and remobilization with WBV is efficient and offers advantages over free remobilization.
- Published
- 2024
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4. Articular mobilization promotes improvement in functional and inflammatory parameters in a gouty arthritis model.
- Author
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Hoss IM, Pradal LA, Leal TSDS, Bertolini GRF, Costa RM, and Ribeiro LFC
- Subjects
- Humans, Rats, Animals, Male, Uric Acid, Rats, Wistar, Inflammation, Pain, Edema, Arthritis, Gouty chemically induced, Arthritis, Gouty therapy, Arthritis, Gouty metabolism
- Abstract
Objective: Gouty arthritis is characterized by painful inflammation due to the deposition of monosodium urate crystals in joint tissues. Despite available treatments, many patients experience ineffective management and adverse effects. This study evaluated a manual therapy protocol involving passive joint mobilization at the peak of inflammation in a gouty arthritis model using functional and inflammatory parameters., Methods: Twenty male Wistar rats, 12 weeks old, were divided into two groups (n=10 each): Gouty Arthritis and Control Groups, which were further subdivided into treated and untreated groups (n=5 each). The Gouty Arthritis Group received intraarticular knee injection of 50µL of monosodium urate crystals, while the Control Group received 50µL of phosphate buffered saline. The treatment involved a 9-minutes session of grade III joint mobilization (according to Maitland). Nociception, grip strength, and edema were evaluated before induction (EV0), 7 hours after assessment (EV1), immediately after treatment (EV2), and 1 hour after treatment (EV3). The animals were euthanized, and synovial fluid was collected to analyze leukocyte migration., Results: The model mimicked the signs of the Gouty Arthritis Group, with a decrease in the threshold of nociception and strength and an increase in edema and leukocyte count. The mobilization protocol significantly increased the nociceptive threshold and grip strength and reduced edema; however, it did not reverse the increase in leukocyte count., Conclusion: Our results suggest that mobilization promotes analgesia and may modulate the inflammatory process owing to reduced edema and subtle attenuation of cell migration, which contributes to strength gain.
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- 2023
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5. Resistance exercise and low-level laser therapy improves grip strength and morphological aspects in the ankle joint of Wistar rats with experimental arthritis.
- Author
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Retameiro ACB, Neves M, Tavares ALF, Boaro CT, Rodriguez DFS, Leal TSDS, Costa RM, Bertolini GRF, and Ribeiro LFC
- Subjects
- Humans, Rats, Male, Animals, Rats, Wistar, Ankle Joint, Hand Strength, Low-Level Light Therapy, Resistance Training, Arthritis, Experimental radiotherapy
- Abstract
Rheumatoid arthritis (RA) is an inflammatory disease mainly affecting synovial joints. Photobiomodulation through low-level laser therapy (LLLT) and resistance exercise may improve the inflammatory process. Therefore, we analyzed the effects of resistance exercise, LLLT, and the combination of both treatments on hind paw grip strength and ankle joint histomorphometric aspects of Wistar rats subjected to experimental RA. A total of 64 male Wistar rats were divided into eight groups: control, control LLLT, control exercise, control LLLT and exercise, arthritis, arthritis LLLT, arthritis exercise, and arthritis LLLT and exercise groups. The experimental RA was induced by a complete Freund's adjuvant injection into the knee joint cavity. Climbing exercises and LLLT (660 nm; 5 J/cm
2 per point) were performed as the treatment. In addition, muscle strength was evaluated using the grip strength test, and morphometric evaluations were performed on the ankle joint. Generalized mixed models and multivariate analysis of variance tests were used for statistical analysis. Statistical significance was set at a p-value of .05. Arthritis LLLT, exercise, and LLLT and exercise had positive effects on grip strength between the groups (F[7.56] = 5.8, p < .004) and within the groups (F[4.3] = 9.9, p < .002) throughout the evaluations. Morphometry revealed degenerative lesions in the ankle joint as subintima with angiogenesis, inflammatory cells, flocculated articular cartilage, chondrocyte disorganization, and pannus in the arthritis group (p < .001). The treated groups exhibited morphological characteristics similar to those of the control group. LLLT and resistance exercise restored muscle strength and morphological aspects of the ankle joint in rats with experimentally induced RA., (© 2022 American Association for Anatomy.)- Published
- 2023
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6. An Early and Sustained Inflammatory State Induces Muscle Changes and Establishes Obesogenic Characteristics in Wistar Rats Exposed to the MSG-Induced Obesity Model.
- Author
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Zazula MF, Saraiva DF, Theodoro JL, Maciel M, Sepulveda EVDS, Zanardini de Andrade B, Boaretto ML, Maciel JIHN, Bronczek GA, Soares GM, Sagae Schneider SC, Bertolini GRF, Torrejais MM, Ribeiro LFC, Fernandes LC, and Naliwaiko K
- Subjects
- Rats, Animals, Rats, Wistar, Muscles metabolism, Fibrosis, Sodium Glutamate adverse effects, Obesity metabolism
- Abstract
The model of obesity induced by monosodium glutamate cytotoxicity on the hypothalamic nuclei is widely used in the literature. However, MSG promotes persistent muscle changes and there is a significant lack of studies that seek to elucidate the mechanisms by which damage refractory to reversal is established. This study aimed to investigate the early and chronic effects of MSG induction of obesity upon systemic and muscular parameters of Wistar rats. The animals were exposed to MSG subcutaneously (4 mg·g
-1 b.w.) or saline (1.25 mg·g-1 b.w.) daily from PND01 to PND05 ( n = 24). Afterwards, in PND15, 12 animals were euthanized to determine the plasma and inflammatory profile and to assess muscle damage. In PND142, the remaining animals were euthanized, and samples for histological and biochemical analyses were obtained. Our results suggest that early exposure to MSG reduced growth, increased adiposity, and inducted hyperinsulinemia and a pro-inflammatory scenario. In adulthood, the following were observed: peripheral insulin resistance, increased fibrosis, oxidative distress, and a reduction in muscle mass, oxidative capacity, and neuromuscular junctions, increased fibrosis, and oxidative distress. Thus, we can conclude that the condition found in adult life and the difficulty restoring in the muscle profile is related to the metabolic damage established early on.- Published
- 2023
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7. Experimental model of periodontitis and rheumatoid arthritis potentiates the deleterious effects on functional capacity, leukocyte migration, synovial and periodontal tissues in Wistar rats.
- Author
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Schnaufer TC, Tavares AL, Hoss IM, Correia BR, Pradal LA, Piana EA, Wachter F, Stein T, Ayala TS, Menolli RA, Bertolini GRF, Meire Costa R, Ribeiro LFC, Nassar CA, and Nassar PO
- Subjects
- Animals, Male, Rats, Edema chemically induced, Leukocytes, Models, Theoretical, Rats, Wistar, Cell Movement, Interleukin-17, Arthritis, Rheumatoid complications, Periodontitis complications
- Abstract
Objective: This study aimed to evaluate whether ligature-induced periodontitis and rheumatoid arthritis (RA) potentiate the deleterious effects on functional capacity, periodontal and synovial tissues, leukocyte migration, and interleukin 17 (IL-17) levels, and to investigate the repercussions of single Freund's Complete Adjuvant (FCA) injection associated with periodontitis., Materials and Methods: Fifty-one male Wistar rats were randomised into six groups: control (CG, n = 8), RA (RAG, n = 9), periodontitis (PG, n = 9), periodontitis and RA (PRAG, n = 9), periodontitis and intradermal injection (PIDG, n = 9), and periodontitis and intra-articular injection (PIAG, n = 7). The animals underwent ligature placement and one or two injections with FCA to induce RA. Motor disability, nociceptive threshold, joint edema, and muscle strength were assessed, and the animals were euthanized on day 30. Synovial fluid, hemimandibles, and knee joints were collected., Results: PRAG showed no reduction of edema or improvement of muscle strength, whereas it showed most significant changes in leukocyte migration, morphological analyses of the synovial membrane (SM), and radiographic and histometric analyses of the jaw. The PIAG showed some alterations, though not permanent., Conclusion: Ligature-induced periodontitis and RA induced by two FCA injections accentuated the deleterious effects on functional capacity, leukocyte migration, synovial and periodontal tissues.
- Published
- 2023
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8. Physical Exercise and Low-Level Laser Therapy Systemic Effects on the Ankle Joint in an Experimental Rheumatoid Arthritis Model.
- Author
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Retameiro ACB, Neves M, Tavares ALF, Boaro CDT, Rodriguez DFS, Stein T, Costa RM, Bertolini GRF, and Ribeiro LFC
- Subjects
- Animals, Ankle Joint, Freund's Adjuvant, Male, Rats, Rats, Wistar, Arthritis, Rheumatoid therapy, Low-Level Light Therapy methods
- Abstract
Objective: The purpose of this study was to analyze the systemic effects of stair climbing exercise, low-level laser therapy (LLLT), and the association of both treatments on pelvic member functionality and ankle joint histomorphometric aspects of Wistar rats submitted to experimental rheumatoid arthritis (RA) protocol., Methods: Male Wistar rats were randomly distributed into the following 8 groups: control; control LLLT; control exercise; control LLLT and exercise; arthritis group; arthritis LLLT; arthritis exercise; and arthritis LLLT and exercise, with n = 8 for functional and n = 5 for histomorphometrical tests. The experimental RA was induced by complete Freund adjuvant injection in the knee joint cavity. Functionality was evaluated by proprioception and motor function using Sciatic Functional Index and maximum angle reached at an inclined plane. Histomorphometrical aspects were evaluated in the ankle joint after histological routine., Results: The arthritis LLLT and exercise group had positive effects in Sciatic Functional Index (F [3.96] = 11.3, P < .001) and in inclined plane (F [3.4] = 36.1, P < .001). The arthritis exercise group presented a greater number of chondrocytes in the tibia (Wald [1; 6605.6] = 25.2, P < .001) and talus (Wald [1; 15958.6] = 19.8, P = .006) in relation to the other groups. The arthritis group morphology showed significant degenerative lesions as subintima with angiogenesis, inflammatory cells, flocculated articular cartilage, chondrocytes disorganization and pannus. Even with the higher chondrocytes number, the arthritis exercise group had morphological characteristics more similar to the control group., Conclusion: Low-level laser therapy and exercise restored functionality, and exercise restored morphological aspects of tissues in experimentally induced RA in rats., (Copyright © 2022. Published by Elsevier Inc.)
- Published
- 2022
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9. Analysis of Wistar Rats Submitted to a Gout Model, Treated with Double Cryotherapy Protocol.
- Author
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Tavares ALF, Reginato A, Neves M, Pradal LA, Leal TSDS, Ribeiro LFC, Menolli RA, and Bertolini GRF
- Subjects
- Animals, Cryotherapy, Inflammation, Male, Rats, Rats, Wistar, Gout pathology, Gout therapy, Hypothermia, Induced
- Abstract
Gout arthritis commonly affects joint regions by deposition of crystals, promoting functional damage mainly during periods of exacerbation. Cryotherapy is a commonly used resource to contain inflammatory processes, however, its use during a gout crisis is not yet well understood. Therefore, the objective was to evaluate the parameters of Wistar rats submitted to an experimental gout model and treated with dual cryotherapy protocol. Twenty-one male Wistar rats were used, separated into three groups: control group (CG), lesion group (LG), and lesion + cryotherapy group (LCG). Gout model induction was through intra-articular injection, with urate crystal solution, in the right knee and cryoimmersion treatment was performed for 20 minutes at a temperature of 5° ± 2°C. Seven evaluations and two treatment moments were performed, and the following parameters were analyzed: joint edema, grip strength, joint disability, motor function, and leukocyte migration through synovial lavage. In the statistical analysis we used SPSS 20.0 with Generalized Linear Models, with least significant difference posttest, always with 5% significance level. The treatment reduced edema, promoted strength recovery, and was effective in reducing total leukocytes in the synovial fluid. No difference was observed between the injured groups for joint disability and motor function. Cryotherapy promoted edema reduction and increased pelvic limb grip strength in Wistar rats during the acute period.
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- 2022
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10. Land use as an effective factor on the occurrence of chromosomal diseases in Brazil.
- Author
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Cochak MR, Favalesso MM, Costa RM, Guimarães ATB, and Ribeiro LFC
- Abstract
Background: The occurrence of chromosomal diseases is a worldwide health problem. The use of agrochemicals, urbanization processes, and solar radiation can be predictive factors of the elevated risk of congenital malformations. In this sense, predicting the geographical potential of the distribution of chromosomal diseases has high relevance for public health., Objectives: This study aimed to describe chromosomal prevalence in Brazil regions, from 2005 to 2015, to model a potential distribution of chromosomal disease occurrence probability associated with land use., Methods: We used chromosomal prevalence to model a potential distribution of chromosomal diseases using machine learning algorithms. As the predictors of the models, we used the variables global forest canopy height, distance from the built-up area , and solar radiation . We characterized the predictive areas as potential occurrence of chromosomal diseases by land use and occupation., Results: Georeferenced data of 43,672 karyotypes detected 7,237 cases of chromosomal diseases and used 5,362 to build the models. The models generated were accurate (TSS>0.5)., Discussion: The areas with greater occurrence of chromosomal diseases present a significant association with pasture areas, crops and agroforestry systems, and urbanized areas. This research is the first Brazilian study with this approach that seems promising in predicting the potential distribution of chromosomal diseases. Therefore, it can be an excellent management tool in public health., Competing Interests: None., (IJMEG Copyright © 2021.)
- Published
- 2021
11. Bariatric surgery induces morphological changes in the extensor digitorum longus muscle in the offspring of obese rats.
- Author
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Kuhn C, Covatti C, Ribeiro LFC, Balbo SL, and Torrejais MM
- Subjects
- Animals, Connective Tissue pathology, Female, Muscle, Skeletal ultrastructure, Neuromuscular Junction pathology, Pregnancy, Rats, Wistar, Rats, Bariatric Surgery adverse effects, Muscle, Skeletal pathology, Obesity surgery
- Abstract
Introduction: The effects of the maternal nutritional environment on the growth and metabolism of the offspring, and its impacts on health in adult life are defined as metabolic programming. Thus, the objective of this study was to evaluate the effects of Roux-en-Y gastric bypass (RYGB) on the morphology of muscle fiber and neuromuscular junction (NMJ) of the offspring of rats submitted to RYGB., Methods: Three-week-old Wistar rats were separated into two groups: 1) CAF SHAM which received a cafeteria diet and was submitted to a sham operation and 2) CAF RYGB, which received a cafeteria diet and was submitted to RYGB. The first generation (F1) offspring (male) was named according to the treatment of mothers as CAF SHAM-F1 and CAF RYGB-F1 and received a standard diet after weaning. At 17 weeks, the animals were euthanized, and the extensor digitorum longus muscle (EDL) was collected and processed in light microscopy and transmission electron microscopy for morphological and morphometric analysis., Results: The CAF RYGB-F1 group showed a reduction in the weight of the EDL muscle and also a reduction in the area of type I, IIa and IIb fibers and a nucleus/fiber ratio. This same group also showed an increase in the capillary density and myofibrillar disorganization and in the Z-line, as well as a reduction in the area of the NMJs., Conclusion: The RYGB surgery in mothers produced morphological changes in the skeletal striated muscles of the offspring., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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12. Corrigendum to analysis of the phosphorylome of Trichoderma reesei cultivated on sugarcane bagasse suggests post-translational regulation of the secreted glycosyl hydrolase Cel7A.
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Pedersoli WR, Paula RG, Antoniêto ACC, Carraro CB, Taveira LC, Maués DB, Martins MP, Ribeiro LFC, Damasio ARL, Silva-Rocha R, Filho AR, and Silva RN
- Abstract
[This corrects the article DOI: 10.1016/j.btre.2021.e00652.]., (© 2021 The Authors.)
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- 2021
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13. Study of muscle fibers of the extensor digitorium longus and soleus muscles of C57BL/6 females exposed to glyphosate during pregnancy and lactation.
- Author
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Barbosa A, Oliveira MC, Kuhn-Fraga C, Ribeiro LFC, Balbo SL, and Torrejais MM
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- Animals, Female, Glycine analogs & derivatives, Lactation, Mice, Mice, Inbred C57BL, Pregnancy, Glyphosate, Muscle Fibers, Skeletal, Muscle, Skeletal
- Abstract
Objective: To evaluate the morphology and morphometry of the muscles extensor digitorium longus and soleus of C57BL/6 females, who were exposed to glyphosate during pregnancy and lactation., Methods: Twelve female mice from the C57BL/6 lineage were used. After detection of pregnancy, they were divided into a Control Group, which received only water, and a Glyphosate Group, which received water with 0.5% glyphosate during pregnancy and lactation. Both groups received ad libitum standard diet. After weaning, the females were euthanized and weighed; naso-anal length was measured, and fats were collected and weighed. The muscles extensor digitorium longus and soleus were collected, and their length and weight were measured. Then, the muscles were fixed in Methacarn to perform the histological study of muscle fibers., Results: Glyphosate Group presented lower weight gain during pregnancy and also lower final body weight and naso-anal length; however, the other body parameters evaluated did not present a significant difference in relation to the Control Group. Significant differences were also not observed in the analysis of muscle fibers and connective tissue., Conclusion: Exposure to 0.5% glyphosate during pregnancy and lactation resulted in lower weight gain during pregnancy, final weight, and naso-anal length. Despite not directly altering the morphology of muscle tissue, these results may indicate enough exposure to interfere with animal metabolism.
- Published
- 2021
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14. Analysis of the phosphorylome of trichoderma reesei cultivated on sugarcane bagasse suggests post-translational regulation of the secreted glycosyl hydrolase Cel7A.
- Author
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Pedersoli WR, de Paula RG, Antoniêto ACC, Carraro CB, Taveira IC, Maués DB, Martins MP, Ribeiro LFC, Damasio ARL, Silva-Rocha R, Filho AR, and Silva RN
- Abstract
Trichoderma reesei is one of the major producers of holocellulases. It is known that in T. reesei , protein production patterns can change in a carbon source-dependent manner. Here, we performed a phosphorylome analysis of T. reesei grown in the presence of sugarcane bagasse and glucose as carbon source. In presence of sugarcane bagasse, a total of 114 phosphorylated proteins were identified. Phosphoserine and phosphothreonine corresponded to 89.6% of the phosphosites and 10.4% were related to phosphotyrosine. Among the identified proteins, 65% were singly phosphorylated, 19% were doubly phosphorylated, 12% were triply phosphorylated, and 4% displayed even higher phosphorylation. Seventy-five kinases were predicted to phosphorylate the sites identified in this work, and the most frequently predicted serine/threonine kinase was PKC1. Among phosphorylated proteins, four glycosyl hydrolases were predicted to be secreted. Interestingly, Cel7A activity, the most secreted protein, was reduced to approximately 60% after in vitro dephosphorylation, suggesting that phosphorylation might alter Cel7A structure, substrate affinity, and targeting of the substrate to its carbohydrate-binding domain. These results suggest a novel post-translational regulation of Cel7A., Competing Interests: All authors declare no financial and personal relationships with other people or organizations that could inappropriately influence the work., (© 2021 The Author(s).)
- Published
- 2021
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15. Low-Level Laser Effect on Peripheral Sciatic Regeneration Under the Systemic Inflammatory Condition of Periodontal Disease.
- Author
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Malanotte JA, Ribeiro LFC, Peretti AL, Kakihata CMM, Potulsky A, Guimarães ATB, Bertolini GRF, Nassar PO, and Nassar CA
- Abstract
Introduction: Periodontal disease (PD) is an inflammatory condition, which leads to tooth loss and promotes a systemic inflammatory state that can aggravate the nerve degeneration. As laser therapy may stimulate regeneration, this study aimed to evaluate the effect of the low-level laser (LLL) on peripheral nerve regeneration under the systemic inflammatory condition of PD. Methods: Thirry-two male rats were used, distributed in 4 groups: nerve injury (NIG); periodontal disease with nerve injury (PDNI); nerve injury and treatment (TNIG); periodontal disease with nerve injury and treatment (PDNIT). On the 7th day of the experiment, the animals had ligatures placed around the lower first molars. On the 22nd day, they underwent peripheral nerve damage, and on the 25th day, the LLL treatment was initiated, performed for two weeks. The sciatic functional index (SFI) was evaluated with subsequent euthanasia of all the animals on the 37th day of the experiment. The sciatic nerve was collected for morphological and oxidative stress analysis and the hemi jaws for radiographic analysis. Results: Regarding the SFI, there was no difference among the groups in the first evaluation (EV) pre-injury; as for theEV2, after injury, all the groups presented a decrease in these values, which remained in post-treatment. For the morphology of the PDNI, nerve tissue presented larger diameter fibers, whereas, for NIT and PDNIT, fibers had smaller diameters with endoneurial organization. When it comes to the antioxidant system, there was an increase in protein concentration, higher superoxide activity, and decreased glutathione transferase activity in the treated groups. Catalase and cholinesterase did not differ between the groups, and lipoperoxidation (LPO) increased in the PD groups. For the mandible radiographic analysis, it was possible to verify the induction of PD. Conclusion: As for the used parameters, the low-level laser was not effective in increasing the nociceptive threshold, but it contributed to the regeneration of nerve fibers, although the inflammation was still present in the site. However, the treatment was effective in protecting cells against oxidative damage due to increased SOD and increased protein, although the decrease in GST demonstrates the inhibition of this stage of the antioxidant system., (Copyright © 2020 J Lasers Med Sci.)
- Published
- 2020
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16. Morphological effects of whole-body vibration on remobilization of the tibialis anterior muscle of Wistar rats.
- Author
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Zazula MF, Wutzke MLS, da Costa JRG, Guimarães ATB, Costa RM, Bertolini GRF, and Ribeiro LFC
- Subjects
- Animals, Electromyography, Male, Muscular Atrophy etiology, Muscular Atrophy physiopathology, Rats, Rats, Wistar, Immobilization adverse effects, Muscle, Skeletal physiopathology, Muscular Atrophy therapy, Recovery of Function physiology, Vibration therapeutic use
- Abstract
Therapeutic immobilization is a common treatment for the locomotor system; however, it causes loss of muscle due to disuse, leading to protein degradation and generating atrophy of muscle cells, ultimately changing functionality. In this sense, it is important for remobilization to be initiated early and performed with appropriate therapeutic strategies that enable tissue and functional recovery. One method of remobilization is physical exercise, among which whole body vibration (WBV) has been highlighted and mainly applied in people with reduced mobility. However, there are gaps on the morphological effects WBV has on muscle tissue, so in this study we analyzed the histomorphometry of the tibialis anterior muscle (TA) of Wistar rats remobilized using WBV. For the experiment, 32 male Wistar rats were used and divided into four groups (n = 8/group). Groups consisted of: control (CG), immobilized (IG), immobilized and remobilized freely (FG), and immobilized and remobilized with WBV (WG). After the experimental period, the TA was collected and processed for analysis in light microscopy. When compared to the control group, significant morphological changes were observed, which characterize muscle atrophy and reduction of all histomorphometric parameters of the TA of the immobilized animals. Remobilized animals showed improvement in all parameters, and the WBV was not different from the free remobilization, except for the reduction of central nuclei, which can be related to acceleration of the process of tissue regeneration. Thus, we can conclude that the WBV can have an impact on the acceleration of the muscle regenerative process, and may be beneficial in people with reduced mobility., (© 2020 American Association for Anatomy.)
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- 2020
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17. Short-term effects of whole-body vibration on the soleus of ooforectomized rats: Histomorphometric analysis and oxidative stress in an animal model.
- Author
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Peretti AL, Kakihata CMM, Tavares ALF, de Oliveira CMT, Guimarães ATB, Costa RM, Ribeiro LFC, and Bertolini GRF
- Subjects
- Animals, Disease Models, Animal, Male, Muscular Atrophy metabolism, Muscular Atrophy pathology, Ovariectomy, Oxidative Stress genetics, Rats, Rats, Wistar, Muscle Fibers, Skeletal metabolism, Muscle Fibers, Skeletal pathology, Muscle, Skeletal metabolism, Muscle, Skeletal pathology, Oxidative Stress physiology, Vibration adverse effects
- Abstract
Purpose: The aim of this study was to analyze the effects of mechanical vibration on the histomorphometry and oxidative stress of oophorectomized rats., Method: Seventy-two Wistar rats were randomized to Pseudoophorectomy (P) and Oophorectomy (O) and subdivided into untreated animals and euthanized after four (P4 and O4) and eight (P8 and O8) weeks and animals treated during four (PT4 and OT4) and eight (PT8 and OT8) weeks. The treatment consisted of use of whole-body vibration for 10 min, three times a week. After euthanasia, the soleus muscle was collected. The general morphological analysis was performed in the right soleus muscle and then the cross-sectional area, the largest and the smallest diameter of the muscle fiber in 100 fibers per muscle, also the nuclei and capillary/fiber ratios, and percentage of connective tissue were measured. The left soleous was used for oxidative stress analysis., Results: PT4 presented higher values in cross-sectional area than P4 and PT8, while O8 was lower than O4, P8 and OT8; for the fiber diameters, the oophorectomized animals had lower values than the pseudo-oophorectomized animals and the treatments values higher than the ones that had no treatment. In oxidative stress, O8 and OT8 presented higher lipoperoxidation, without any alterations to the activities of superoxide dismutase, catalase and cholinesterase., Conclusion: Whole-body vibration induced muscle hypertrophy in the pseudo-oophorectomized rats after four weeks, as well as being able to reverse the changes caused by the surgery in eight weeks in that variable., (Copyright © 2020 Elsevier GmbH. All rights reserved.)
- Published
- 2020
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18. Low-Level Laser Therapy in Different Wavelengths on the Tibialis Anterior Muscle of Wistar Rats After Nerve Compression Injury.
- Author
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Neves M, Tavares ALF, Reginato A, Kakihata CMM, Bertolini GRF, and Ribeiro LFC
- Subjects
- Animals, Connective Tissue pathology, Rats, Rats, Wistar, Sciatic Nerve radiation effects, Sciatic Neuropathy physiopathology, Low-Level Light Therapy methods, Muscle, Skeletal radiation effects, Peripheral Nerve Injuries radiotherapy, Sciatic Neuropathy radiotherapy
- Abstract
Objective: Traumatic injuries are common and may promote disruption of neuromuscular communication, triggering phenomena that lead to nerve degeneration and affect muscle function. A laser accelerates tissue recovery; however, the parameters used are varied, making it difficult to compare studies. The purpose of this study was to evaluate the effect of low-level laser therapy, at 660- and 830-nm wavelengths, on the tibialis anterior muscle of Wistar rats after sciatic nerve compression., Methods: Twenty animals were separated into 4 groups: control, sciatic nerve injury, lesion + 660-nm laser, and lesion + 830-nm laser. In the lesion groups, the right sciatic nerve was surgically exposed and compressed with hemostatic forceps for 30 seconds. After the third postoperative day, the groups with laser therapy were submitted to treatment for 2 weeks totaling 10 applications, performed directly on the surgical scar of the nerve injury. Grip strength was analyzed before and after the nerve injury and during the treatment period. The tibialis anterior muscle was processed for light microscopy, area measurement, smaller diameter, number of fibers, nuclei, and connective tissue., Results: The animals submitted to the injury experienced muscular atrophy and morphological changes in the number of muscle fibers and nuclei. In the connective tissue morphometry, there was a decrease in the treated groups compared with the untreated groups., Conclusion: The laser treatment at different wavelengths showed no improvement in the tibialis anterior muscle of Wistar rats within the morphological and functional aspects evaluated., (Copyright © 2020. Published by Elsevier Inc.)
- Published
- 2020
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19. Physical exercise and low-level laser therapy on the nociception and leukocyte migration of Wistar rats submitted to a model of rheumatoid arthritis.
- Author
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Neves M, Retameiro ACB, Tavares ALF, Reginato A, Menolli RA, Leal TSDS, Ribeiro LFC, and Bertolini GRF
- Subjects
- Animals, Arthritis, Rheumatoid complications, Arthritis, Rheumatoid diagnostic imaging, Edema complications, Inflammation pathology, Male, Nociception, Positron-Emission Tomography, Rats, Wistar, Synovial Fluid metabolism, Arthritis, Rheumatoid pathology, Arthritis, Rheumatoid radiotherapy, Cell Movement, Leukocytes cytology, Low-Level Light Therapy methods, Physical Conditioning, Animal
- Abstract
Rheumatoid arthritis denotes hyperplasia and intense inflammatory process. Treatment involves exercise protocols and use of resources such as low-level laser therapy (LLLT) to modulate the inflammatory process and maintain physical capacity. The objective was to investigate whether treatment with LLLT and exercise modulates the inflammatory process and peripheral functionality. Sample is composed of 128 male rats, separated into three groups, control, treated and untreated, in the acute and chronic period of the disease with 64 animals in each group, divided into 8 subgroups with n = 8. The animals were immunized with injection at the base of the tail and 7 days after intra-articular injection with complete Freund adjuvant (CFA) for lesion groups, and saline solution for the controls. Joint disability was evaluated by PET (paw elevation time) and joint edema and treated with LLLT and/or resisted stair climbing exercise. Normality Shapiro-Wilk test, ANOVA mixed for the functional analyses, and ANOVA one-way for the variables of cellular differentiation, with Bonferroni post hoc, p = 5% were used. For the evaluations of joint disability and nociception, there was a significant difference between the evaluations, the groups, and the interaction groups-evaluations. The treated groups showed recovery of functionality; it is still verified that laser therapy increased the nociceptive threshold of the chronic inflammatory period, and the exercise reflected in significant functional improvement and modulation of the inflammatory process both in the acute and chronic periods. LLLT, resistance exercise, or a combination of treatments had a positive effect on the modulation of the inflammatory process, reducing the migration of leukocytes, in addition to helping the return of peripheral functionality by reducing joint disability in a model of rheumatoid arthritis induced by CFA in rats.
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- 2020
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20. Morphometric Effects of Whole-Body Vibration on the Bone in a Rat Model of Postmenopausal Osteoporosis.
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Kakihata CMM, Peretti AL, Tavares ALF, Wutzke MLS, Ribeiro LFC, Costa RM, and Bertolini GRF
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- Animals, Bone Density, Female, Femur physiology, Humans, Random Allocation, Rats, Rats, Wistar, Biomechanical Phenomena physiology, Bone and Bones physiology, Exercise physiology, Osteoporosis, Postmenopausal prevention & control, Vibration therapeutic use
- Abstract
Objective: The purpose of this study was to analyze the morphometric effects of mechanical vibration with a duration of 4 or 8 weeks on the femur of oophorectomized Wistar rats., Methods: Sixty-four female rats were submitted to oophorectomy or a sham operation, and each of those 2 groups were randomized into 4 groups: untreated and euthanized at week 12, untreated and euthanized at week 16, treated for 4 weeks and euthanized at week 12, and treated for 8 weeks and euthanized at week 16. The vibration treatment was performed for 10 min/d, with a frequency of 60 Hz, 3 d/wk. The rats were then euthanized and the right femur dissected. Subsequently, histomorphometric analysis was performed on the proximal epiphysis and diaphysis of the spongy and cortical bone, respectively., Results: As expected, the oophorectomy groups presented reduction of spongy and cortical bone tissue. Further, the vibration therapy of 4 and 8 weeks' duration in the oophorectomized groups led to increased bone mass, observed as an increased percentage of spongy tissue, and increased thickness and percentage of cortical tissue. However, the variables of femoral neck diameter, mean area of the shaft, and number of osteocytes were not altered by oophorectomy and vibration., Conclusion: The mechanical vibration was effective in increasing the bone mass of the femur of oophorectomized Wistar rats, observed by increasing the percentage of spongy bone and increasing the percentage and thickness of cortical bone., (Copyright © 2020. Published by Elsevier Inc.)
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- 2020
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21. The Aspergillus fumigatus Phosphoproteome Reveals Roles of High-Osmolarity Glycerol Mitogen-Activated Protein Kinases in Promoting Cell Wall Damage and Caspofungin Tolerance.
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Mattos EC, Silva LP, Valero C, de Castro PA, Dos Reis TF, Ribeiro LFC, Marten MR, Silva-Rocha R, Westmann C, da Silva CHTP, Taft CA, Al-Furaiji N, Bromley M, Mortensen UH, Benz JP, Brown NA, and Goldman GH
- Subjects
- Aspergillus fumigatus genetics, Cell Wall drug effects, Fungal Proteins genetics, Fungal Proteins metabolism, Gene Expression Regulation, Fungal, Glycerol metabolism, Mitogen-Activated Protein Kinases genetics, Osmolar Concentration, Osmotic Pressure, Phosphorylation, Proteome, Signal Transduction, Antifungal Agents pharmacology, Aspergillus fumigatus drug effects, Aspergillus fumigatus enzymology, Caspofungin pharmacology, Cell Wall pathology, Mitogen-Activated Protein Kinases metabolism
- Abstract
The filamentous fungus Aspergillus fumigatus can cause a distinct set of clinical disorders in humans. Invasive aspergillosis (IA) is the most common life-threatening fungal disease of immunocompromised humans. The mitogen-activated protein kinase (MAPK) signaling pathways are essential to the adaptation to the human host. Fungal cell survival is highly dependent on the organization, composition, and function of the cell wall. Here, an evaluation of the global A. fumigatus phosphoproteome under cell wall stress caused by the cell wall-damaging agent Congo red (CR) revealed 485 proteins potentially involved in the cell wall damage response. Comparative phosphoproteome analyses with the Δ sakA , Δ mpkC , and Δ sakA Δ mpkC mutant strains from the osmotic stress MAPK cascades identify their additional roles during the cell wall stress response. Our phosphoproteomics allowed the identification of novel kinases and transcription factors (TFs) involved in osmotic stress and in the cell wall integrity (CWI) pathway. Our global phosphoproteome network analysis showed an enrichment for protein kinases, RNA recognition motif domains, and the MAPK signaling pathway. In contrast to the wild-type strain, there is an overall decrease of differentially phosphorylated kinases and phosphatases in Δ sakA , Δ mpkC , and Δ sakA Δ mpkC mutants. We constructed phosphomutants for the phosphorylation sites of several proteins differentially phosphorylated in the wild-type and mutant strains. For all the phosphomutants, there is an increase in the sensitivity to cell wall-damaging agents and a reduction in the MpkA phosphorylation upon CR stress, suggesting these phosphosites could be important for the MpkA modulation and CWI pathway regulation. IMPORTANCE Aspergillus fumigatus is an opportunistic human pathogen causing allergic reactions or systemic infections, such as invasive pulmonary aspergillosis in immunocompromised patients. The mitogen-activated protein kinase (MAPK) signaling pathways are essential for fungal adaptation to the human host. Fungal cell survival, fungicide tolerance, and virulence are highly dependent on the organization, composition, and function of the cell wall. Upon cell wall stress, MAPKs phosphorylate multiple target proteins involved in the remodeling of the cell wall. Here, we investigate the global phosphoproteome of the Δ sakA and Δ mpkC A. fumigatus and high-osmolarity glycerol (HOG) pathway MAPK mutants upon cell wall damage. This showed the involvement of the HOG pathway and identified novel protein kinases and transcription factors, which were confirmed by fungal genetics to be involved in promoting tolerance of cell wall damage. Our results provide understanding of how fungal signal transduction networks modulate the cell wall. This may also lead to the discovery of new fungicide drug targets to impact fungal cell wall function, fungicide tolerance, and virulence., (Copyright © 2020 Mattos et al.)
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- 2020
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22. Intrathecally injected tramadol reduces articular incapacitation and edema in a rat model of lipopolysaccharide (LPS)-induced reactive arthritis.
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Lucena F, Callado de Oliveira DMM, Norões MM, Mujica EMM, Melleu FF, Benedet PO, Stein T, Ribeiro LFC, and Tonussi CR
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- Animals, Arthralgia etiology, Arthralgia pathology, Arthritis, Experimental chemically induced, Arthritis, Experimental physiopathology, Arthritis, Reactive chemically induced, Arthritis, Reactive physiopathology, Disease Models, Animal, Edema etiology, Edema pathology, Injections, Spinal, Male, Rats, Rats, Wistar, Analgesics, Opioid administration & dosage, Arthralgia prevention & control, Arthritis, Experimental complications, Arthritis, Reactive complications, Edema prevention & control, Lipopolysaccharides toxicity, Tramadol administration & dosage
- Abstract
Aims: Intrathecal injection of morphine presents analgesic and antiedematogenic effects in rats. However, it is unknown whether tramadol, which possess a mixed mechanism of action, can also produce analgesic and antiedematogenic effects similarly., Main Methods: Male Wistar rats received carrageenan and LPS in the right knee joint. Tramadol (10 μg) was injected intrathecally 20 min before articular LPS injection. Incapacitation and articular edema were measured 5 h after LPS stimulation. Synovial fluid was collected for leukocyte counting and western blot analysis. Whole joint and lumbar spinal cord were also collected for histology and immunohistochemistry, respectively. Intrathecal pretreatments groups were with the NKCC1 blocker bumetanide, TRPV
1 agonist resiniferatoxin, μ-opioid receptor antagonist CTOP and serotonergic neurotoxin 5,7-DHT, all previously to tramadol., Key Findings: Tramadol treatment caused the reduction of incapacitation and edema. It also reduced c-Fos protein expression in the spinal cord dorsal horn and slightly reduced TNF-α levels in synovial fluid, but neither reduced cell migration nor tissue damage. Bumetanide and resiniferatoxin prevented the analgesic and antiedematogenic effects of tramadol. CTOP prevented the analgesic and the antiedematogenic effects, but 5,7-DHT prevented only tramadol-induced analgesia., Significance: Spinal NKCC1 cotransporter and peptidergic peripheral afferents seem to be important for the analgesic and antiedematogenic effects of tramadol, as well as μ-opioid receptor. However, the monoamine uptake inhibition effect of tramadol seems to be important only to the analgesic effect., (Copyright © 2019 Elsevier Inc. All rights reserved.)- Published
- 2019
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23. Engineered microbial host selection for value-added bioproducts from lignocellulose.
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de Paula RG, Antoniêto ACC, Ribeiro LFC, Srivastava N, O'Donovan A, Mishra PK, Gupta VK, and Silva RN
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- Biomass, Hydrolysis, Lignin, Biofuels
- Abstract
Lignocellulose is a rich and sustainable globally available carbon source and is considered a prominent alternative raw material for producing biofuels and valuable chemical compounds. Enzymatic hydrolysis is one of the crucial steps of lignocellulose degradation. Cellulolytic and hemicellulolytic enzyme mixes produced by different microorganisms including filamentous fungi, yeasts and bacteria, are used to degrade the biomass to liberate monosaccharides and other compounds for fermentation or conversion to value-added products. During biomass pretreatment and degradation, toxic compounds are produced, and undesirable carbon catabolic repression (CCR) can occur. In order to solve this problem, microbial metabolic pathways and transcription factors involved have been investigated along with the application of protein engineering to optimize the biorefinery platform. Engineered Microorganisms have been used to produce specific enzymes to breakdown biomass polymers and metabolize sugars to produce ethanol as well other biochemical compounds. Protein engineering strategies have been used for modifying lignocellulolytic enzymes to overcome enzymatic limitations and improving both their production and functionality. Furthermore, promoters and transcription factors, which are key proteins in this process, are modified to promote microbial gene expression that allows a maximum performance of the hydrolytic enzymes for lignocellulosic degradation. The present review will present a critical discussion and highlight the aspects of the use of microorganisms to convert lignocellulose into value-added bioproduct as well combat the bottlenecks to make the biorefinery platform from lignocellulose attractive to the market., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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24. Sericin as treatment of obesity: morphophysiological effects in obese mice fed with high-fat diet.
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Kunz RI, Capelassi AN, Alegre-Maller ACP, Bonfleur ML, Ribeiro LFC, Costa RM, and Natali MRM
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- Adipose Tissue pathology, Animals, Anti-Obesity Agents pharmacology, Body Weight drug effects, Cholesterol analysis, Eating drug effects, Fatty Liver pathology, Gastrointestinal Transit drug effects, Glucose Tolerance Test, Liver metabolism, Male, Mice, Inbred C57BL, Mice, Obese, Obesity etiology, Obesity physiopathology, Reproducibility of Results, Sericins pharmacology, Time Factors, Treatment Outcome, Triglycerides analysis, Weight Gain drug effects, Anti-Obesity Agents therapeutic use, Diet, High-Fat adverse effects, Obesity drug therapy, Sericins therapeutic use
- Abstract
Objective: To investigate the effects of sericin extracted from silkworm Bombyx mori cocoon on morphophysiological parameters in mice with obesity induced by high-fat diet., Methods: Male C57Bl6 mice aged 9 weeks were allocated to one of two groups - Control and Obese, and fed a standard or high-fat diet for 10 weeks, respectively. Mice were then further subdivided into four groups with seven mice each, as follows: Control, Control-Sericin, Obese, and Obese-Sericin. The standard or high fat diet was given for 4 more weeks; sericin (1,000mg/kg body weight) was given orally to mice in the Control-Sericin and Obese-Sericin Groups during this period. Weight gain, food intake, fecal weight, fecal lipid content, gut motility and glucose tolerance were monitored. At the end of experimental period, plasma was collected for biochemical analysis. Samples of white adipose tissue, liver and jejunum were collected and processed for light microscopy analysis; liver fragments were used for lipid content determination., Results: Obese mice experienced significantly greater weight gain and fat accumulation and had higher total cholesterol and glucose levels compared to controls. Retroperitoneal and periepididymal adipocyte hypertrophy, development of hepatic steatosis, increased cholesterol and triglyceride levels and morphometric changes in the jejunal wall were observed., Conclusion: Physiological changes induced by obesity were not fully reverted by sericin; however, sericin treatment restored jejunal morphometry and increased lipid excretion in feces in obese mice, suggesting potential anti-obesity effects.
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- 2019
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25. Effects of Mechanical Vibration in Neuromuscular Junctions and Fiber Type of the Soleus Muscle of Oophorectomized Wistar Rats.
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Peretti AL, Kakihata CMM, Wutzke MLS, Torrejais MM, Ribeiro LFC, and Bertolini GRF
- Abstract
Objective To evaluate the neuromuscular junctions (NMJs) and the type of muscle fibers of the soleus muscle of oophorectomized Wistar rats submitted to a mechanical vibration protocol. Methods A total of 36 randomized rats were used in the pseudo-oophorectomy without and with treatment and oophorectomy without and with treatment groups. The treatment was performed with a vibratory platform, frequency of 60 Hz and duration of 10 minutes, 3 times a week, for 4 weeks. At the end of the intervention period, the animals were euthanized and the soleus muscles were collected and processed for analysis of the NMJs and fiber type. The data were analyzed for normality by the Shapiro-Wilk test and analysis of the 3-way variance using the post-hoc Tukey test, when necessary, and a significance level of 5% was adopted. Results In the analysis of the NMJs, the oophorectomy group presented a smaller area than the pseudo-oophorectomy group, but the oophorectomy with treatment group was equal to the pseudo-oophorectomy with treatment group. For the larger diameter of the joints, the oophorectomy group was also different from the others; however, the oophorectomy and treatment animals were larger than those of the pseudo-oophorectomy and treatment group. There was no distinction of the types of fibers, with the muscle presenting fibers of the oxidative type. Conclusion Hormonal deprivation reduced the area and diameter of the NMJs, with reversion of this process in the groups that underwent vibratory platform treatment for 4 weeks, and both surgery and treatment did not influence the type of soleus muscle fiber, composed of oxidative fibers.
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- 2019
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26. Microbial Community Profiling in Intensive Care Units Expose Limitations in Current Sanitary Standards.
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Ribeiro LF, Lopes EM, Kishi LT, Ribeiro LFC, Menegueti MG, Gaspar GG, Silva-Rocha R, and Guazzaroni ME
- Abstract
Hospital-associated infections (HAIs) are a leading cause of morbidity and mortality in intensive care units (ICUs) and neonatal intensive care units (NICUs). Organisms causing these infections are often present on surfaces around the patient. Given that microbiota may vary across different ICUs, the HAI-related microbial signatures within these units remain underexplored. In this study, we use deep-sequencing analyses to explore and compare the structure of bacterial communities at inanimate surfaces of the ICU and NICU wards of The Medical School Clinics Hospital (Brazil). The data revealed that NICU presents higher biodiversity than ICU and surfaces closest to the patient showed a peculiar microbiota, distinguishing one unit from the other. Several facultative anaerobes or obligate anaerobes HAI-related genera were classified as biomarkers for the NICU, whereas Pseudomonas was the main biomarker for ICU. Correlation analyses revealed a distinct pattern of microbe-microbe interactions for each unit, including bacteria able to form multi-genera biofilms. Furthermore, we evaluated the effect of concurrent cleaning over the ICU bacterial community. The results showed that, although some bacterial populations decreased after cleaning, various HAI-related genera were quite stable following sanitization, suggesting being well-adapted to the ICU environment. Overall, these results enabled identification of discrete ICU and NICU reservoirs of potentially pathogenic bacteria and provided evidence for the presence of a set of biomarkers genera that distinguish these units. Moreover, the study exposed the inconsistencies of the routine cleaning to minimize HAI-related genera contamination.
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- 2019
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27. Extracellular vesicles carry cellulases in the industrial fungus Trichoderma reesei .
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de Paula RG, Antoniêto ACC, Nogueira KMV, Ribeiro LFC, Rocha MC, Malavazi I, Almeida F, and Silva RN
- Abstract
Background: Trichoderma reesei is the most important industrial producer of lignocellulolytic enzymes. These enzymes play an important role in biomass degradation leading to novel applications of this fungus in the biotechnology industry, specifically biofuel production. The secretory pathway of fungi is responsible for transporting proteins addressed to different cellular locations involving some cellular endomembrane systems. Although protein secretion is an extremely efficient process in T. reesei , the mechanisms underlying protein secretion have remained largely uncharacterized in this organism., Results: Here, we report for the first time the isolation and characterization of T. reesei extracellular vesicles (EVs). Using proteomic analysis under cellulose culture condition, we have confidently identified 188 vesicular proteins belonging to different functional categories. Also, we characterized EVs production using transmission electron microscopy in combination with light scattering analysis. Biochemical assays revealed that T. reesei extracellular vesicles have an enrichment of filter paper (FPase) and β-glucosidase activities in purified vesicles from 24, 72 and 96, and 72 and 96 h, respectively. Furthermore, our results showed that there is a slight enrichment of small RNAs inside the vesicles after 96 h and 120 h, and presence of hsp proteins inside the vesicles purified from T. reesei grown in the presence of cellulose., Conclusions: This work points to important insights into a better understanding of the cellular mechanisms underlying the regulation of cellulolytic enzyme secretion in this fungus., Competing Interests: Competing interestsThe authors declare that they have no competing interests.
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- 2019
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28. Comprehensive Analysis of Aspergillus nidulans PKA Phosphorylome Identifies a Novel Mode of CreA Regulation.
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Ribeiro LFC, Chelius C, Boppidi KR, Naik NS, Hossain S, Ramsey JJJ, Kumar J, Ribeiro LF, Ostermeier M, Tran B, Ah Goo Y, de Assis LJ, Ulas M, Bayram O, Goldman GH, Lincoln S, Srivastava R, Harris SD, and Marten MR
- Subjects
- Aspergillus nidulans chemistry, Fungal Proteins genetics, Gene Deletion, Gene Expression Profiling, Proteome analysis, Repressor Proteins genetics, Aspergillus nidulans genetics, Aspergillus nidulans metabolism, Cyclic AMP-Dependent Protein Kinases metabolism, Fungal Proteins metabolism, Gene Expression Regulation, Bacterial, Phosphoproteins analysis, Protein Processing, Post-Translational, Repressor Proteins metabolism
- Abstract
In filamentous fungi, an important kinase responsible for adaptation to changes in available nutrients is cyclic AMP (cAMP)-dependent protein kinase (protein kinase A [PKA]). This kinase has been well characterized at a molecular level, but its systemic action and direct/indirect targets are generally not well understood in filamentous fungi. In this work, we used a pkaA deletion strain (Δ pkaA ) to identify Aspergillus nidulans proteins for which phosphorylation is dependent (either directly or indirectly) on PKA. A combination of phosphoproteomic and transcriptomic analyses revealed both direct and indirect targets of PKA and provided a global perspective on its function. One of these targets was the transcription factor CreA, the main repressor responsible for carbon catabolite repression (CCR). In the Δ pkaA strain, we identified a previously unreported phosphosite in CreA, S319, which (based on motif analysis) appears to be a direct target of Stk22 kinase (AN5728). Upon replacement of CreA S319 with an alanine (i.e., phosphonull mutant), the dynamics of CreA import to the nucleus are affected. Collectively, this work provides a global overview of PKA function while also providing novel insight regarding significance of a specific PKA-mediated phosphorylation event. IMPORTANCE The cyclic AMP (cAMP)-dependent protein kinase A (PKA) signaling pathway is well conserved across eukaryotes, and previous work has shown that it plays an important role in regulating development, growth, and virulence in a number of fungi. PKA is activated in response to extracellular nutrients and acts to regulate metabolism and growth. While a number of components in the PKA pathway have been defined in filamentous fungi, current understanding does not provide a global perspective on PKA function. Thus, this work is significant in that it comprehensively identifies proteins and functional pathways regulated by PKA in a model filamentous fungus. This information enhances our understanding of PKA action and may provide information on how to manipulate it for specific purposes., (Copyright © 2019 Ribeiro et al.)
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- 2019
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29. Phosphoproteomic and transcriptomic analyses reveal multiple functions for Aspergillus nidulans MpkA independent of cell wall stress.
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Chelius CL, Ribeiro LFC, Huso W, Kumar J, Lincoln S, Tran B, Goo YA, Srivastava R, Harris SD, and Marten MR
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- Aspergillus nidulans enzymology, Aspergillus nidulans growth & development, Cell Cycle Proteins genetics, Cell Wall genetics, Cell Wall metabolism, Gene Expression Regulation, Fungal genetics, Iron metabolism, Sequence Deletion genetics, Signal Transduction genetics, Aspergillus nidulans genetics, Mitogen-Activated Protein Kinases genetics, Phosphoproteins genetics, Transcriptome genetics
- Abstract
The protein kinase MpkA plays a prominent role in the cell wall integrity signaling (CWIS) pathway, acting as the terminal MAPK activating expression of genes which encode cell wall biosynthetic enzymes and other repair functions. Numerous studies focus on MpkA function during cell wall perturbation. Here, we focus on the role MpkA plays outside of cell wall stress, during steady state growth. In an effort to seek other, as yet unknown, connections to this pathway, an mpkA deletion mutant (ΔmpkA) was subjected to phosphoproteomic and transcriptomic analysis. When compared to the control (isogenic parent of ΔmpkA), there is strong evidence suggesting MpkA is involved with maintaining cell wall strength, branching regulation, and the iron starvation pathway, among others. Particle-size analysis during shake flask growth revealed ΔmpkA mycelia were about 4 times smaller than the control strain and more than 90 cell wall related genes show significantly altered expression levels. The deletion mutant had a significantly higher branching rate than the control and phosphoproteomic results show putative branching-regulation proteins, such as CotA, LagA, and Cdc24, have a significantly different level of phosphorylation. When grown in iron limited conditions, ΔmpkA had no difference in growth rate or production of siderophores, whereas the control strain showed decreased growth rate and increased siderophore production. Transcriptomic data revealed over 25 iron related genes with altered transcript levels. Results suggest MpkA is involved with regulation of broad cellular functions in the absence of stress., (Copyright © 2019 Elsevier Inc. All rights reserved.)
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- 2019
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30. Sericin silk protein in peripheral nervous repair associated with the physical exercise of swimming in Wistar rats.
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Debastiani JC, Santana AJ, Ribeiro LFC, Brancalhão RMC, and Bertolini GRF
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- Animals, Disease Models, Animal, Pain Measurement, Pain Threshold drug effects, Pain Threshold physiology, Rats, Rats, Wistar, Sciatic Nerve pathology, Sciatic Neuropathy physiopathology, Sericins metabolism, Severity of Illness Index, Statistics, Nonparametric, Treatment Outcome, Physical Conditioning, Animal methods, Sciatic Neuropathy drug therapy, Sciatic Neuropathy rehabilitation, Sericins therapeutic use, Swimming
- Abstract
Objective: To verify the action of sericin associated to swim exercise with overload, on sciatic nerve repair in Wistar rats, after 22 days of nerve compression., Methods: Forty animals been composed of five groups: control, injury, injury-sericin, injury-swimming and injury-sericin-swimming. During the lesion procedure, sericin, in hydrolyzed form, applied directly to the injury in the injury-sericin and injury-sericin-swimming groups. Injury-swimming and injury-sericin-swimming groups underwent to 5 days per week for 3 weeks, with a 10% overload of the animal's body weight, and a weekly progressive evolution of swimming time, lasting 15, 20 and 25 min/day. Pre and throughout the treatment period the animals performed evaluation of sciatic functional index and pressure pain threshold with digital von Frey filament. Euthanasia was performed on the 22nd postoperative day, and two fragments of the nerve were collected and prepared for descriptive and quantitative analysis., Results: The sciatic functional index assessment showed significant differences in the motor function of the control until the 14th day. Regarding the allodynia, there was revealed a significant improvement in injury-swimming performance relative to injury, injury-sericin and injury-sericin-swimming, and the number of viable and non-viable nerve fibers smaller than 4 μm in diameter was significantly higher in the injury-sericin-swimming., Conclusion: swimming showed a better evolution of the nociceptive threshold and allodynia. Sericin treatment had exacerbated pro-inflammatory characteristics. On the other hand, the association of sericine and swimming showed a possible regulatory effect by resting swimming exercise, with a significant increase of fibers of smaller diameter.
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- 2019
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31. Converting a Periplasmic Binding Protein into a Synthetic Biosensing Switch through Domain Insertion.
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Ribeiro LF, Amarelle V, Ribeiro LFC, and Guazzaroni ME
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- Protein Domains, Biosensing Techniques methods, Periplasm chemistry, Periplasmic Binding Proteins chemistry, Protein Engineering methods
- Abstract
All biosensing platforms rest on two pillars: specific biochemical recognition of a particular analyte and transduction of that recognition into a readily detectable signal. Most existing biosensing technologies utilize proteins that passively bind to their analytes and therefore require wasteful washing steps, specialized reagents, and expensive instruments for detection. To overcome these limitations, protein engineering strategies have been applied to develop new classes of protein-based sensor/actuators, known as protein switches, responding to small molecules. Protein switches change their active state (output) in response to a binding event or physical signal (input) and therefore show a tremendous potential to work as a biosensor. Synthetic protein switches can be created by the fusion between two genes, one coding for a sensor protein (input domain) and the other coding for an actuator protein (output domain) by domain insertion. The binding of a signal molecule to the engineered protein will switch the protein function from an "off" to an "on" state (or vice versa) as desired. The molecular switch could, for example, sense the presence of a metabolite, pollutant, or a biomarker and trigger a cellular response. The potential sensing and response capabilities are enormous; however, the recognition repertoire of natural switches is limited. Thereby, bioengineers have been struggling to expand the toolkit of molecular switches recognition repertoire utilizing periplasmic binding proteins (PBPs) as protein-sensing components. PBPs are a superfamily of bacterial proteins that provide interesting features to engineer biosensors, for instance, immense ligand-binding diversity and high affinity, and undergo large conformational changes in response to ligand binding. The development of these protein switches has yielded insights into the design of protein-based biosensors, particularly in the area of allosteric domain fusions. Here, recent protein engineering approaches for expanding the versatility of protein switches are reviewed, with an emphasis on studies that used PBPs to generate novel switches through protein domain insertion.
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- 2019
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32. New Genomic Approaches to Enhance Biomass Degradation by the Industrial Fungus Trichoderma reesei .
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de Paula RG, Antoniêto ACC, Ribeiro LFC, Carraro CB, Nogueira KMV, Lopes DCB, Silva AC, Zerbini MT, Pedersoli WR, Costa MDN, and Silva RN
- Abstract
The filamentous fungi Trichoderma reesei is one of the most well-studied cellulolytic microorganisms. It is the most important fungus for the industrial production of enzymes to biomass deconstruction being widely used in the biotechnology industry, mainly in the production of biofuels. Here, we performed an analytic review of the holocellulolytic system presented by T. reesei as well as the transcriptional and signaling mechanisms involved with holocellulase expression in this fungus. We also discuss new perspectives about control of secretion and cellulase expression based on RNA-seq and functional characterization data of T. reesei growth in different carbon sources, which comprise glucose, cellulose, sophorose, and sugarcane bagasse.
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- 2018
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33. Analysis of the muscle tissue of Wistar rats submitted to the sciatic nerve compression model and cryotherapy.
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Karvat J, Kakihata CMM, Vieira L, Silva JLDC, Ribeiro LFC, Brancalhão RMC, and Bertolini GRF
- Subjects
- Animals, Disease Models, Animal, Hypertrophy physiopathology, Male, Muscle Weakness physiopathology, Nerve Compression Syndromes physiopathology, Random Allocation, Rats, Wistar, Reference Values, Reproducibility of Results, Sciatic Nerve physiopathology, Sciatic Nerve surgery, Sciatic Neuropathy physiopathology, Time Factors, Treatment Outcome, Cryotherapy methods, Nerve Compression Syndromes pathology, Nerve Compression Syndromes therapy, Sciatic Nerve pathology, Sciatic Neuropathy pathology, Sciatic Neuropathy therapy
- Abstract
Objective: To evaluate the effects of right sciatic nerve compression and cryotherapy on muscle tissue., Methods: We used 42 male Wistar rats, subdivided in the following Groups Control, Injury 3, Injury 8 and Injury 15 submitted to nerve compression and euthanized in the 3rd, 8th and 15th day after surgery. The Cryotherapy Injury 3 was entailed treatment with cryotherapy by immersion of the animal in recipient for 20 minutes during 1 day, then animals were euthanized at the 3rd day after surgery, and the Cryotherapy Injury 8 and the Cryotherapy Injury 15 was treated for 6 days, and euthanized at the 8th and 15th day after surgery. Functional evaluation was performed by the grasping strength of the right pelvic limb. The right tibialis anterior muscles were evaluated for mass, smaller diameter and cross-sectional area. In the Cryotherapy Injury 8 and the Cryotherapy Injury 15 groups, the hydroxyproline was dosed in the right soles., Results: In the compression there was a significant difference in the Injury Groups compared with the Control Group (p<0.05). In the smaller diameter, the compression in Control Group was higher than Injury 8 (p=0.0094), Injury 15 (p=0.002) and Cryotherapy Injury 15 (p<0.001) groups. The comparison between groups with euthanasia in the same post-operative period, a significant difference (p=0.0363) was seen in day 8th after surgery, and this result in Cryotherapy Injury Group was greater than Injury Group. In the fiber area, Control Group was also higher than the Injury 8 (p=0.0018), the Injury 15 (p<0.001) and the Cryotherapy Injury 15 (p<0.001). In hydroxyproline, no significant difference was seen between groups., Conclusion: Nerve damage resulted in decreased muscle strength and trophism, the cryotherapy delayed hypotrophy, but this effect did not persist after cessation of treatment.
- Published
- 2018
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34. Protein Engineering Strategies to Expand CRISPR-Cas9 Applications.
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Ribeiro LF, Ribeiro LFC, Barreto MQ, and Ward RJ
- Abstract
The development of precise and modulated methods for customized manipulation of DNA is an important objective for the study and engineering of biological processes and is essential for the optimization of gene therapy, metabolic flux, and synthetic gene networks. The clustered regularly interspaced short palindromic repeat- (CRISPR-) associated protein 9 is an RNA-guided site-specific DNA-binding complex that can be reprogrammed to specifically interact with a desired DNA sequence target. CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes. Although Cas9 has been successfully used as a precise tool in all these applications, some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence, (ii) aberrant off-target activity, (iii) the large size of Cas9 is problematic for CRISPR delivery, and (iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing. These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications. Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation. Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
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- 2018
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35. Altered secretion patterns and cell wall organization caused by loss of PodB function in the filamentous fungus Aspergillus nidulans.
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Boppidi KR, Ribeiro LFC, Iambamrung S, Nelson SM, Wang Y, Momany M, Richardson EA, Lincoln S, Srivastava R, Harris SD, and Marten MR
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- Aspergillus nidulans genetics, Aspergillus nidulans growth & development, Cell Wall ultrastructure, Fungal Proteins genetics, Gene Expression Regulation, Fungal, Genotype, Hyphae ultrastructure, Mutation genetics, Phenotype, Proteomics, Temperature, Unfolded Protein Response, Up-Regulation, Aspergillus nidulans cytology, Aspergillus nidulans metabolism, Cell Wall metabolism, Fungal Proteins metabolism
- Abstract
Filamentous fungi are widely used in the production of a variety of industrially relevant enzymes and proteins as they have the unique ability to secrete tremendous amounts of proteins. However, the secretory pathways in filamentous fungi are not completely understood. Here, we investigated the role of a mutation in the POlarity Defective (podB) gene on growth, protein secretion, and cell wall organization in Aspergillus nidulans using a temperature sensitive (Ts) mutant. At restrictive temperature, the mutation resulted in lack of biomass accumulation, but led to a significant increase in specific protein productivity. Proteomic analysis of the secretome showed that the relative abundance of 584 (out of 747 identified) proteins was altered due to the mutation. Of these, 517 were secreted at higher levels. Other phenotypic differences observed in the mutant include up-regulation of unfolded protein response (UPR), deformation of Golgi apparatus and uneven cell wall thickness. Furthermore, proteomic analysis of cell wall components in the mutant revealed the presence of intracellular proteins in higher abundance accompanied by lower levels of most cell wall proteins. Taken together, results from this study suggest the importance of PodB as a target when engineering fungal strains for enhanced secretion of valuable biomolecules.
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- 2018
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36. Neurotrophin expression and histomorphometric evaluation in Wistar rats subjected to neural mobilization after compression of the median nerve.
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Marcioli MAR, Silva JLDC, Ribeiro LFC, Brancalhão RMC, and Bertolini GRF
- Abstract
Objective: To evaluate the neurotrophin mRNA expression and axon count in the median nerve of Wistar rats submitted to neural mobilization (NM) after nerve compression., Methods: Eighteen animals were randomly divided into G1 (nerve compression only), G2 (NM for 1 min), and G3 (NM for 3 min). For NM, the animals were anesthetized and the right scapula received the mobilization, adapted as indicated for humans, on alternate days, from the third to the 13th postoperative (PO) day, totaling six days of therapy. On the 14th PO day, animals were anesthetized and euthanized. Fragments of the median nerve, distal to the compression procedure, were removed for histomorphometric analysis and expression of neurotrophins, nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) by RT-PCR., Results: Histomorphometric analysis revealed differences in the number of axons in the injured side, which was significantly lower in the injured limb nerve compared to the control limb, whereas the RT-PCR analysis showed no significant differences in the expression of NGF or BDNF., Conclusion: NM treatment did not affect median nerve regeneration, which maintained normal recovery rates.
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- 2018
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37. Sericin and swimming on histomorphometric parameters of denervated plantar muscle in Wistar rats.
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Santana AJ, Debastiani JC, Buratti P, Peretti AL, Kunz RI, Brancalhão RMC, Ribeiro LFC, Torrejais MM, and Bertolini GRF
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- Animals, Disease Models, Animal, Lower Extremity injuries, Lower Extremity pathology, Muscle, Skeletal injuries, Muscle, Skeletal pathology, Nerve Crush, Random Allocation, Rats, Rats, Wistar, Lower Extremity innervation, Muscle, Skeletal innervation, Physical Conditioning, Animal physiology, Sericins pharmacology, Swimming physiology
- Abstract
Objective To analyze the combined effects of the silk protein sericin and swimming exercise on histomorphometry of the plantar muscle in Wistar rats. Methods Forty adult rats were randomly allocated into 5 groups comprising 8 animals each, as follows: Control, Injury, Sericin, Swim, and Swim plus Sericin. Three days after crushing of the sciatic nerve the rats in the Swim and Swim plus Sericin Groups were submitted to swimming exercise for 21 days. Rats were then euthanized and the plantar muscle harvested and processed. Results Cross-sectional area, peripheral nuclei and muscle fiber counts, nucleus/fiber ratio and smallest muscle fiber width did not differ significantly between groups. Morphological analysis revealed hypertrophic fibers in the Swim Group and evident muscle damage in the Swim plus Sericin and Injury Groups. The percentage of intramuscular collagen was apparently maintained in the Swim Group compared to remaining groups. Conclusion Combined treatment with sericin and swimming exercise did not improve muscle properties. However, physical exercise alone was effective in maintaining intramuscular connective tissue and preventing progression of deleterious effects of peripheral nerve injury.
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- 2018
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38. Experimental Periodontitis in the Potentialization of the Effects of Immobilism in the Skeletal Striated Muscle.
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Leite MA, de Mattia TM, Kakihata CMM, Bortolini BM, de Carli Rodrigues PH, Bertolini GRF, Brancalhão RMC, Ribeiro LFC, Nassar CA, and Nassar PO
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- Animals, Muscle Strength, Muscle, Skeletal blood supply, Muscular Atrophy pathology, Nociceptive Pain, Rats, Wistar, Immobilization adverse effects, Muscle, Skeletal pathology, Periodontitis pathology
- Abstract
This study aims to evaluate if ligature-induced periodontitis can potentiates the deleterious effects of immobilization in the skeletal striated muscle, contributing to the development of muscle atrophy due to disuse. Forty Wistar rats were divided into four groups: (1) Control Group (CG), (2) Periodontal Disease (PDG), (3) Immobilized (IG), and (4) Immobilized with Periodontal Disease (IPDG). Periodontal disease was induced for 30 days, with ligature method, and the immobilization was performed with cast bandage for 15 days. Prior to euthanasia, nociceptive threshold and muscular grasping force were evaluated. Afterwards, the soleus muscle was dissected and processed for sarcomere counting and morphological/morphometric analysis. For data analysis, was used the one-way ANOVA and post-test Tukey (p < 0.05). The IG and IPDG presented lower muscle weight, lower muscular grip strength, and less number of sarcomeres compared to CG. The PDG showed reduction of muscle strength and nociceptive threshold after 15 days of periodontal disease and increased connective tissue compared to CG. The IPDG presented lower muscle length and nociceptive threshold. The IG presented reduction in cross-sectional area and smaller diameter, increase in the number of nuclei and a nucleus/fiber ratio, decrease in the number of capillaries and capillary/fiber ratio, with increase in connective tissue. The IPDG had increased nucleus/fiber ratio, decreased capillaries, and increased connective tissue when compared to the IG. The IPDG presented greater muscle tissue degeneration and increased inflammatory cells compared to the other groups. Ligature-induced periodontitis potentiated the deleterious effects of immobilization of the skeletal striated muscle.
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- 2017
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39. Therapeutic ultrasound after sciatic nerve compression of Wistar rats.
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Bertolini GRF, Karvat J, Kakihata CMM, Ribeiro LFC, and Brancalhão RMC
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- Animals, Disease Models, Animal, Exercise Test, Follow-Up Studies, Hand Strength physiology, Male, Pain Measurement, Rats, Rats, Wistar, Sciatic Neuropathy physiopathology, Severity of Illness Index, Statistics, Nonparametric, Time Factors, Treatment Outcome, Sciatic Neuropathy therapy, Ultrasonography, Doppler, Pulsed methods
- Abstract
Objectives: The present study analyzed the effect of therapy with therapeutic ultrasound on the sciatic nerve after compression injury, comparing two similar doses of SATA., Methods: In total, 32 Wistar rats were used, divided into the following groups: CG - control; IG - compression injury of the sciatic nerve; IGCU - injury and continuous ultrasound; and IGPU - injury and 20% pulsed ultrasound. The treatment with ultrasound started on the 3rd postoperative day, with a frequency of 1 MHz, 0.4 W/cm² (SATA) for IGCU. IGPU received 2.0 W/cm
2 (SATP), with 20% of the active cycle, for 3 minutes. The treatment was performed on a daily basis, totaling 15 days of therapy. Evaluations were performed for functional, histological, and morphometric forms., Results: Both the Sciatic Functional Index and the withdrawal threshold and grip strength failed to show an advantage of using therapeutic ultrasound. For the morphometric evaluations of nerve fiber diameter and axons, myelin sheath thickness, and G quotient and nerve fiber estimates, IGPU values were estimated to be significantly lower. The morphological analysis revealed intense inflammatory response and neovascularization, as well as degeneration of axons and the myelin sheath, for the injury group and IGCU; however, IGPU showed greater tissue disorganization., Conclusion: There were no significant differences, showing functional or nocicepitive recovery of the treated groups, including with characteristics pointing to the pulsed group with worse results.- Published
- 2017
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40. Insights regarding fungal phosphoproteomic analysis.
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Ribeiro LFC, Chelius CL, Harris SD, and Marten MR
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- Biological Evolution, Catalytic Domain, Fungi pathogenicity, Phosphorylation, Phosphotransferases metabolism, Proteomics, Fungal Proteins metabolism, Fungi metabolism, Phosphoproteins metabolism
- Abstract
Protein phosphorylation is a major means of regulation for cellular processes, and is important in cell signaling, growth, and cell proliferation. To study phosphorylated proteins, high throughput phosphoproteomic technologies, such as reverse phase protein array, phospho-specific flow cytometry, and mass spectrometry (MS) based technologies, have been developed. Among them, mass spectrometry has become the primary tool employed for the identification of phosphoproteins and phosphosites in fungi, leading to an improved understanding of a number of signaling pathways. Using mass spectrometry techniques, researchers have discovered new kinase substrates, established connections between kinases and fungal pathogenicity, and studied the evolutionary lineage of kinases between different fungal species. Further, many specific phosphorylation sites recognized by individual kinases have been described. In this review, we will focus on recent discoveries made in yeast and filamentous fungi using phosphoproteomic analysis., (Copyright © 2017 Elsevier Inc. All rights reserved.)
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- 2017
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41. Action of vanillin (Vanilla planifolia) on the morphology of tibialis anterior and soleus muscles after nerve injury.
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Peretti AL, Antunes JS, Lovison K, Kunz RI, Castor LRG, Brancalhão RMC, Bertolini GRF, and Ribeiro LFC
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- Animals, Connective Tissue pathology, Humans, Male, Models, Animal, Muscle Fibers, Skeletal drug effects, Muscle, Skeletal pathology, Random Allocation, Rats, Wistar, Sciatic Neuropathy rehabilitation, Anti-Inflammatory Agents pharmacology, Benzaldehydes pharmacology, Connective Tissue drug effects, Muscle, Skeletal drug effects, Sciatic Neuropathy pathology
- Abstract
Objective: To evaluate the action of vanillin (Vanilla planifolia) on the morphology of tibialis anterior and soleus muscles after peripheral nerve injury., Methods: Wistar rats were divided into four groups, with seven animals each: Control Group, Vanillin Group, Injury Group, and Injury + Vanillin Group. The Injury Group and the Injury + Vanillin Group animals were submitted to nerve injury by compression of the sciatic nerve; the Vanillin Group and Injury + Vanillin Group, were treated daily with oral doses of vanillin (150mg/kg) from the 3rd to the 21st day after induction of nerve injury. At the end of the experiment, the tibialis anterior and soleus muscles were dissected and processed for light microscopy and submitted to morphological analysis., Results: The nerve compression promoted morphological changes, typical of denervation, and the treatment with vanillin was responsible for different responses in the studied muscles. For the tibialis anterior, there was an increase in the number of satellite cells, central nuclei and fiber atrophy, as well as fascicular disorganization. In the soleus, only increased vascularization was observed, with no exacerbation of the morphological alterations in the fibers., Conclusion: The treatment with vanillin promoted increase in intramuscular vascularization for the muscles studied, with pro-inflammatory potential for tibialis anterior, but not for soleus muscle., Objetivo: Avaliar a ação da vanilina (Vanilla planifolia) sobre a morfologia dos músculos tibial anterior e sóleo após lesão nervosa periférica., Métodos: Ratos Wistar foram divididos em quatro grupos, com sete animais cada, sendo Grupo Controle, Grupo Vanilina, Grupo Lesão e Grupo Lesão + Vanilina. Os animais dos Grupos Lesão e Grupo Lesão + Vanilina foram submetidos à lesão nervosa por meio da compressão do nervo isquiático, e os Grupos Vanilina e Grupo Lesão + Vanilina foram tratados diariamente com doses orais de vanilina (150mg/kg) do 3o ao 21o dia após a indução da lesão nervosa. Ao término do experimento, os músculos tibial anterior e sóleo foram dissecados e seguiram o processamento de rotina em microscopia de luz, para posterior análise morfológica., Resultados: A compressão nervosa promoveu alterações morfológicas características de denervação, sendo que o tratamento com vanilina foi responsável por respostas distintas nos músculos estudados. Para o tibial anterior, houve aumento do número de células satélites, núcleos centrais e atrofia das fibras, bem como desorganização fascicular. Já no sóleo, houve apenas aumento da vascularização, sem exacerbação das alterações morfológicas nas fibras., Conclusão: O tratamento com vanilina promoveu o aumento da vascularização intramuscular para os músculos estudados, com potencial pró-inflamatório para o tibial anterior, o que não ocorreu no músculo sóleo.
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- 2017
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42. Jumping in aquatic environment after sciatic nerve compression: nociceptive evaluation and morphological characteristics of the soleus muscle of Wistar rats.
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Malanotte JA, Kakihata CMM, Karvat J, Brancalhão RMC, Ribeiro LFC, and Bertolini GRF
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- Animals, Male, Physical Conditioning, Animal physiology, Random Allocation, Rats, Wistar, Reference Values, Reproducibility of Results, Time Factors, Treatment Outcome, Hydrotherapy methods, Muscle, Skeletal physiopathology, Nerve Compression Syndromes physiopathology, Nerve Compression Syndromes therapy, Nociception physiology, Sciatic Neuropathy physiopathology, Sciatic Neuropathy therapy
- Abstract
Objective: To evaluate the effect of jumping in aquatic environment on nociception and in the soleus muscle of trained and not trained Wistar rats, in the treatment of compressive neuropathy of the sciatic nerve., Methods: Twenty-five Wistar rats were distributed into five groups: Control, Lesion, Trained + Lesion, Lesion + Exercise, and Trained + Lesion + Exercise. The training was jumping exercise in water environment for 20 days prior to injury, and treatment after the injury. Nociception was evaluated in two occasions, before injury and seven after injury. On the last day of the experiment, the right soleus muscles were collected, processed and analyzed as to morphology and morphometry., Results: In the assessment of nociception in the injury site, the Control Group had higher average than the rest, and the Lesion Group was larger than the Trained + Lesion and Lesion + Exercise Groups. The Control Group showed higher nociceptive threshold in paw, compared to the others. In the morphometric analysis, in relation to Control Group, all the injured groups showed decreased muscle fiber area, and in the Lesion Group was lower than in the Lesion + Exercise Group and Trained + Lesion Group. Considering the diameter of the muscle fiber, the Control Group had a higher average than the Trained + Lesion Group and the Trained + Lesion + Exercise Group; and the Lesion Group showed an average lower than the Trained + Lesion and Lesion + Exercise Groups., Conclusion: Resistance exercise produced increased nociception. When performed prior or after nerve damage, it proved effective in avoiding hypotrophy. The combination of the two protocols led to decrease in diameter and area of the muscle fiber., Objetivo: Avaliar os efeitos do salto em meio aquático, na nocicepção e no músculo sóleo, em ratos Wistar treinados e não treinados, no tratamento de neuropatia compressiva do nervo isquiático., Métodos: Foram distribuídos em cinco grupos 25 ratos Wistar: Controle, Lesão, Treinado + Lesão, Lesão + Exercício e Treinado + Lesão + Exercício. O treino foi com exercício de salto em meio aquático durante 20 dias, prévio à lesão, e o tratamento ocorreu após a lesão. Foram realizadas avaliações da nocicepção, sendo uma pré-lesão e sete pós-lesão. No último dia de experimento, os músculos sóleos direitos foram coletados, processados e analisados por meio de morfologia e morfometria., Resultados: Na avaliação da nocicepção no local da lesão, o Grupo Controle apresentou média maior que os demais, e o Grupo Lesão foi maior que os Grupos Treinado + Lesão e Lesão + Exercício. O Grupo Controle apresentou limiar nociceptivo na pata maior com relação aos demais. Nas análises morfométricas, em relação ao Grupo Controle, todos os grupos lesionados apresentaram diminuição da área da fibra muscular; o Grupo Lesão apresentou-se menor que os Grupos Treinado + Lesão e Lesão + Exercício. No diâmetro da fibra muscular, o Grupo Controle apresentou média maior que os Grupos Treinado + Lesão e Treinado + Lesão + Exercício, e o Grupo Lesão apresentou média menor que os Grupos Treinado + Lesão e Lesão + Exercício., Conclusão: O exercício físico resistido produziu aumento da nocicepção. Quando realizado previamente ou após a lesão nervosa, mostrou-se eficaz em evitar a hipotrofia. A associação dos dois protocolos levou à diminuição do diâmetro e da área da fibra muscular.
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- 2017
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