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AIMS: This study aims to demonstrate the potential of the lactic acid bacteria (LAB) Pediococcus pentosaceus LBM18 against the mycotoxin-producing Alternaria alternata TEF-1A and highlight its application as an effective grain silage inoculant to control mycotoxin contamination. METHODS AND RESULTS: The antifungal properties of Ped. pentosaceus lyophilized (PPL) were assessed by evaluating its effect on A. alternata TEF-1A grown in a corn silage-based medium, which included morphological changes by Scanning Electron Microscopy (SEM) observations, growth rate, conidia production assays, and inhibition of Tenuazonic acid (TeA) production by high-performance liquid chromatography (HPLC-MS/MS) analyses. Furthermore, TeA biosynthesis was monitored for changes at the molecular level by PKS gene expression. The growth and sporulation processes of A. alternata TEF-1A were affected by Ped. pentosaceus LBM18 in a concentration-dependent manner. Moreover, a significant inhibition of TeA production (74.3%) and the transcription level of the PKS gene (42.9%) was observed. CONCLUSIONS: Ped. pentosaceus is one of the promising LAB to be applied as an inoculant for corn silage preservation, aiming to inhibit mycotoxigenic fungi growth and their mycotoxin production. SIGNIFICANCE AND IMPACT OF THE STUDY: Ped. pentosaceus could be used as an inoculant to reduce fungal and mycotoxins contamination in grain silage production.
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Micotoxinas , Ácido Tenuazônico , Animais , Ácido Tenuazônico/análise , Pediococcus pentosaceus/metabolismo , Espectrometria de Massas em Tandem , Gado/metabolismo , Antifúngicos/farmacologia , Antifúngicos/metabolismo , Alternaria , Micotoxinas/metabolismo , Silagem/microbiologia , Zea mays/metabolismoRESUMO
The tongue is a fundamental organ in feeding, vocalization, and grooming. It is characterized by evolutionary adaptations reflected by diet, habitat, and function. Rodents are a very diverse mammalian order and the tongue's morphology varies in size, form, and presence of papillae. This work aimed to describe the morphological and ultrastructural aspects of the tongue of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831). Tongues of Spix's yellow-toothed cavies were analyzed with light microscopy, scanning, and transmission electron microscopy. The results showed that the tongue was divided into apex, body, and root. There were different types of papillae, such as vallate, foliate, laterally placed fungiform, fungiform, filiform, and robust filiform. The epithelium was organized into layers, including keratinized, granulous, spinous, and basal, below were lamina propria, and musculature, which evolved mucous and serous gland clusters. The tongue of Spix's yellow-toothed cavy was structurally and ultrastructurally similar to other rodents and had papillae with similar morphologies to other Caviidae species. However, the presence of robust filiform papillary lines and laterally placed fungiform papillae showed the main differences from other species. This was the first description of the tongue of Spix's yellow-toothed cavy.
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The mdx mouse is an experimental model of Duchenne muscular dystrophy, a genetic disorder characterized by progressive muscular degeneration which affects the oral cavity musculature, and promotes difficulty in swallowing. This study aimed to describe morphological, structural, and ultrastructural changes in the tongue mucosa and musculature of mdx mice. Forty six-month-old mice were divided into two groups: Control C57bl/10 (n = 20) and mdx C57bl/10mdx (n = 20). The tongue was dissected and analyzed with light microscopy, scanning electron microscopy, and transmission electron microscopy techniques. Our results showed conical and triangular filiform, fungiform, foliate, and vallate papillae, and their connective tissue cores. The epithelium layers identified were corneum, granulosum, spinosum, and basale. The mdx group had a thicker epithelium. Lamina propria was composed of reddish and greenish collagen. In mdx, collagen was present in the musculature of the tongue's body and in the muscular tissue between mucous and serous glands of the caudal region. Musculature was also characterized by a shorter length of sarcoplasmic invaginations, myocytolysis in mitochondrial groupings, and inflammatory focus. In conclusion, the tongue of 6-month-old mdx mice had morphology, structure, and ultrastructure revealed, showing higher wear of filiform papillae indirect reflex from the muscular degeneration process.
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The myotendinous junction (MTJ) is the interface between muscle and tendon, and it is the main area of force transmission of the locomotor apparatus. Dystrophic processes promote pathological injury which affects the skeletal muscle and can influence the morphology of the MTJ. This study aimed to investigate the adaptations in MTJ morphology of mdx mice in the tibialis anterior muscle. Male mice (n = 24) were divided into ControlC57bl/10 and mdxC57bl/10mdx (Duchenne muscular dystrophy experimental model). In the mdx group, centralized nuclei with a large area and greater deposition of type III collagen (fibrosis) were observed. Also, shorter sarcomeres and sarcoplasmatic projections of MTJ were observed. We concluded that the adaptations in mdx mice demonstrated extensive impairment in the MTJ region with reduced ultrastructures.
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The myotendinous junction (MTJ) is the muscle-tendon interface and constitutes an integrated mechanical unit to force transmission. Joint immobilization promotes muscle atrophy via disuse, while physical exercise can be used as an adaptative stimulus. In this study, we aimed to investigate the components of the MTJ and their adaptations and the associated elements triggered with aquatic training after joint immobilization. Forty-four male Wistar rats were divided into sedentary (SD), aquatic training (AT), immobilization (IM), and immobilization/aquatic training (IMAT) groups. The samples were processed to measure fiber area, nuclear fractal dimension, MTJ nuclear density, identification of telocytes, sarcomeres, and MTJ perimeter length. In the AT group, the maintenance of ultrastructure and elements in the MTJ region were observed; the IM group presented muscle atrophy effects with reduced MTJ perimeter; the IMAT group demonstrated that aquatic training after joint immobilization promotes benefits in the muscle fiber area and fractal dimension, in the MTJ region shows longer sarcomeres and MTJ perimeter. We identified the presence of telocytes in the MTJ region in all experimental groups. We concluded that aquatic training is an effective rehabilitation method after joint immobilization due to reduced muscle atrophy and regeneration effects on MTJ in rats.
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Adaptação Fisiológica , Imobilização , Articulações , Condicionamento Físico Animal , Esforço Físico , Tendões/fisiologia , Animais , Masculino , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/ultraestrutura , Ratos , Sarcômeros/ultraestrutura , Tendões/citologia , Tendões/ultraestruturaRESUMO
The palate is a fundamental region in food swallowing and presents different adaptations in species. This research aimed to describe structural and ultrastructural characteristics of the palatine epithelium and the connective tissue cores (CTCs) of ten red-rumped agoutis (Dasyprocta leporinaLinnaeus, 1758) using macroscopic, light microscopy, scanning electron microscopy, and transmission electron microscopy. We found nine palatine ridges in the diastema and hard palate, and a smooth surface in the soft palate. Stratified squamous keratinized epithelium with projections of lamina propria and soft palate had gland clusters. Epithelial removal revealed CTCs with a conical shape with high density in the hard palate and the sides of the soft palate. Near the CTCs were nerve fibers in the hard palate, and the soft palate had muscular tissue below the gland clusters. The structural and ultrastructural characteristics enable stability of the hard palate and fixation to the soft palate sides, while the soft palate center has greater mobility thus assisting in food swallowing. We concluded that structural characteristics are similar to other mammals, although the morphology of agouti's palate differs in the amount and disposition of palatine ridges, and the conical CTC's morphology.
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Obesity is characterized by excess adipose tissue and chronic inflammation and promotes extensive changes that can compromise skeletal muscles' structural and functional integrity. Obesity can seriously impact the force transmission region between the muscle and the tendon, the myotendinous junction (MTJ). The present study aimed to investigate the plasticity of muscle fibers and MTJ regions in high-fat diet-induced obesity in rat tibialis anterior (TA) and soleus (SO) muscles. Wistar rats were divided into control and obese groups (induced by a high-fat diet). The samples of TA and SO muscles were prepared for histochemical and ultrastructural analysis (sarcomeres and MTJ projection). In the muscle fiber, similar adaptations were observed between the muscles of the smaller fiber (types I and IIa) in the obesity results. The MTJ region demonstrated different adaptations between the analyzed muscles. The TAMTJ region has shorter ultrastructures, while in the SOMTJ region, the ultrastructures were larger. We conclude that obesity induced by a high-fat diet promotes similar adaptation in the muscle fibers; however, in the MTJ region, the sarcoplasmatic projections and adjacent sarcomere demonstrate different adaptations according to distinct muscle phenotypes.
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Shark skin is predominantly specialized for swimming and protection, with the dermal denticle being the main structure associated with these abilities. The dermal denticle is a mineral structure with a unique morphology for each species, which allows its use as a taxonomic tool. Few studies have investigated the microscopy aspects of skin and dermal denticles, considering the high diversity of sharks. Here, we investigated the three-dimensional morphoquantitative aspects and mineral composition of dermal denticles in different regions of the angular angel shark, Squatina guggenheim, using scanning electron microscopy and dispersive energy system. With the microscopy, we were able to observe that the dermal denticle morphology changes according to the area it is located. It was possible to describe the dermal denticles individually, from root to the crown, highlighting all of their individualities. Through the dispersive energy system, we showed the proportions of each mineral found in the denticle, by area, demonstrating the composition and the particularities of crown, body, and root, where whitlockite was described for the first time in elasmobranchs. In this way, the present study presented the specificities of the dermal denticles of S. guggenheim, as well sought to understand the different structure functions for the animal, thus assisting future research in animal morphology.
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Calcificações da Polpa Dentária , Tubarões , Animais , Microscopia Eletrônica de Varredura , Minerais , PeleRESUMO
The morphology of the oral cavity of mammals relates to diet, habitat, and function. The palate is an important region with adaptations for oral somatosensation and mechanical loads due to the pressure of the tongue with food. The research aimed to describe the structural and ultrastructural characteristics of the epithelium and the connective tissue cores of the guinea pig palate using macroscopic, light microscopy, scanning electron microscopy, and transmission electron microscopy analysis. The hard palate had conical and filiform papillae, and the soft palate had open salivary ducts. After the removal of the epithelium, the connective tissue cores revealed thin filaments and laminar projections in the hard palate, and opening ducts were evidenced in the soft palate. The palatine epithelium was keratinized and organized by layers, lamellated corpuscles were found in lamina propria of the hard palate. In contrast, the soft palate had glands clusters associated with nerve fibers, and in both regions were identified telocytes. We concluded that the hard palate presented conical and filiform papillae that differ from other mammals. Besides, it is a new description of the connective tissue cores morphology and the first record of the telocytes in this anatomical region for mammals.
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Palato , Telócitos , Animais , Tecido Conjuntivo , Epitélio , Cobaias , Microscopia Eletrônica de Varredura , LínguaRESUMO
The present study shows chronic adjustments in the myotendinous junction (MTJ) in response to different ladder-based resistance training (LRT) protocols. Thirty adult male Wistar rats were divided into groups: sedentary (S), calisthenics (LRT without additional load [C]), and resistance-trained (LRT with extra weight [R]). We demonstrated longer lengths of sarcoplasmatic invaginations in the trained groups; however, evaginations were seen mainly in group R. We showed a greater thickness of sarcoplasmatic invaginations in groups C and R, in addition to greater evaginations in R. We also observed thinner basal lamina in trained groups. The support collagen layer (SCL) adjacent to the MTJ and the diameters of the transverse fibrils were larger in R. We also discovered a niche of telocytes in the MTJ with electron micrographs of the plantar muscle and with immunostaining with CD34+ in the gastrocnemius muscle near the blood vessels and pericytes. We concluded that the continuous adjustments in the MTJ ultrastructure were the result of tissue plasticity induced by LRT, which is causally related to muscle hypertrophy and, consequently, to the remodeling of the contact interface. Also, we reveal the existence of a collagen layer adjacent to MTJ and discover a new micro anatomic location of telocytes.
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Fibras Musculares Esqueléticas/fisiologia , Músculo Esquelético/fisiologia , Condicionamento Físico Animal/fisiologia , Treinamento Resistido/métodos , Retículo Sarcoplasmático/fisiologia , Telócitos/fisiologia , Adaptação Fisiológica/fisiologia , Junções Aderentes/fisiologia , Animais , Membrana Basal/fisiologia , Adesão Celular , Movimento Celular/fisiologia , Junções Célula-Matriz/fisiologia , Colágeno/metabolismo , Masculino , Ratos , Ratos Wistar , Comportamento SedentárioRESUMO
This study analyzed the architecture of Platelet-Rich Fibrin (PRF) clots and assessed their elemental composition in order to provide new insight into this biomaterial. Five surplus PRF clots (2,700 RPM, 12â¯min.) donated by patients (63.6⯱â¯12.3 years old) were prepared for use in dental clinical procedures. The internal three-dimensional morphology of the red zones and the thirds of the yellow zones of the clots were analyzed by Variable Pressure Scanning Electron Microscope (VPSEM) after sample preparation by two methods: 1. Fixation (2.5% gluataraldehyde); and 2. Fixation with subsequent partial removal of extracellular elements (8â¯N, HCl). Semi-quantitative elemental analysis was performed by energy-dispersive X-ray spectrometry (EDX). VPSEM analysis showed erythrocytes in both the red zone and the yellow zone, which consisted mainly of fibrin. Removal of extracellular elements enriched the morphology of both zones; the organization of the fibrin was observed to differ in the thirds of the yellow zone, with increasing density and organization to distal. The elements that compose organic substances (C-Carbon, N-Nitrogen, O-Oxygen, Na-Sodium and P-Phosphorus) and halogens (Cl-Chloride and S-Sulfur) were detected; the highest concentrations were of C, followed by O (pâ¯<â¯0.05), in the proximal region of the fibrin. The results of the present study suggest organization of fibrin in the PRF clot, and also reveal the distribution of the elements present in the different regions of the clot. Improved understanding of these characteristics may favor the use of this biomaterial by increasing its efficiency and functionality.
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Coagulação Sanguínea , Elementos Químicos , Fibrina Rica em Plaquetas/química , Espaço Extracelular/química , Feminino , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Espectrometria por Raios XRESUMO
Pediococcus pentosaceus LBM 18 has shown potential as producer of an antibacterial and antifungal bacteriocin-like inhibitory substance (BLIS). BLIS inhibited the growth of spoilage bacteria belonging to Lactobacillus, Enterococcus and Listeria genera with higher activity than Nisaplin used as control. It gave rise to inhibition halos with diameters from 9.70 to 20.00 mm, with Lactobacillus sakei being the most sensitive strain (13.50-20.00 mm). It also effectively suppressed the growth of fungi isolated from corn grain silage for up to 25 days and impaired morphology of colonies by likely affecting fungal membranes. These results point out that P. pentosaceus BLIS may be used as a new promising alternative to conventional antibacterial and antifungal substances, with potential applications in agriculture and food industry as a natural bio-controlling agent. Moreover, cytotoxicity and cell death induction tests demonstrated cytotoxicity and toxicity of BLIS to human colon adenocarcinoma Caco-2cells but not to peripheral blood mononuclear cells, with suggests possible applications of BLIS also in medical-pharmaceutical applications.
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Anti-Infecciosos/farmacologia , Produtos Biológicos/farmacologia , Pediococcus pentosaceus/química , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Infecciosos/química , Antifúngicos/química , Antifúngicos/farmacologia , Bacteriocinas/química , Bacteriocinas/farmacologia , Produtos Biológicos/química , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Testes de Sensibilidade Microbiana , Análise EspectralRESUMO
The myofibrillary types establish to the skeletal muscle functional and adaptive properties that influence the sarcomeric arrangement during muscle contraction and may have repercussions on an important related force transmission region of the locomotor apparatus, the myotendinous junction (MTJ). This study aimed to describe changes in myofibrillary type and sarcomeric lengths in the belly muscle and MTJ of the soleus and plantaris muscles associated with training protocols in vertical ladder. Thirty adults male Wistar rats were divided into three groups (n = 10): Control (CTR), No-load Training (NLT), and Load Training (LT). Morphoquantitative analysis of different fibers types and sarcomere lengths were performed in distinct regions of plantaris and soleus muscles. In the plantaris muscle with both trainings, there was an increase in the cross-sectional area (CSA) in Type I and II fibers (p < .0001) while sarcomeric lengths revealed greater lengths in the proximal and distal sarcomeres of NLT, although in the LT we found greater lengths in the belly and MTJ sarcomeres. The soleus muscle showed an increase in CSA muscle fiber only in the NLT (p < .0001) and revealed alterations in belly and MTJ sarcomere lengths with training. We concluded that plantaris muscle has an adaptive effect directly associated with training load, with hypertrophy in both trainings and sarcomere length inverse from belly and MTJ, in LT associated with increased force generation and transmission at the MTJ, although soleus muscle has a lower adaptive response to training stimuli with variation in the belly and distal sarcomere of the MTJ.
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Sarcômeros , Tendões , Animais , Masculino , Fibras Musculares Esqueléticas , Músculo Esquelético , Ratos , Ratos WistarRESUMO
Psychological stress and occlusal alteration are important etiologic factors for temporomandibular/masticatory muscular disorders. In particular, the exact physiologic mechanism underlying the relation by occlusal alteration and temporomandibular disorders remains unclear. Our purpose was to test the hypothesis that benzodiazepine therapy is able to prevent metabolic and vascular changes in the medial pterygoid muscle of rats under chronic stress after 14 days of unilateral exodontia. Adult Wistar rats were submitted to unpredictable chronic mild stress (10 days) and/or unilateral exodontia and their plasma and medial pterygoid muscles were removed for analysis. A pre-treatment with diazepam was used to verify its effect on stress. The parameters evaluated included anxiety behavior, plasma levels of corticosterone, metabolic activity by succinate dehydrogenase, capillary density by laminin staining and ultrastructural findings by transmission electron microscopy. Occlusal instability induced anxiety-like behavior on elevated plus-maze test and diazepam administration blocked the appearance of this behavior. Unilateral exodontia promoted in the contralateral muscle an increase of oxidative fibers and capillaries and modification of sarcoplasmic reticulum. Chronic stress caused increased glycolytic metabolism, reduced capillary density and morphological changes in mitochondria on both sides. Association of both factors induced a glycolytic pattern in muscle and hemodynamic changes. Pharmacological manipulation with diazepam inhibited the changes in the medial pterygoid muscle after stress. Our results reveal a preventive benzodiazepine treatment for stress and occlusal instability conditions affecting masticatory muscle disorders. In addition, provide insights into the mechanisms by which chronic stress and exodontia might be involved in the pathophysiology of masticatory muscular dysfunctions.
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Benzodiazepinas/administração & dosagem , Músculos da Mastigação/fisiopatologia , Estresse Psicológico/tratamento farmacológico , Transtornos da Articulação Temporomandibular/tratamento farmacológico , Animais , Benzodiazepinas/farmacologia , Estudos de Casos e Controles , Diazepam/efeitos adversos , Modelos Animais de Doenças , Masculino , Músculos da Mastigação/efeitos dos fármacos , Microscopia Eletrônica de Transmissão , Ratos , Ratos Wistar , Estresse Psicológico/induzido quimicamente , Transtornos da Articulação Temporomandibular/fisiopatologia , Extração Dentária , Resultado do TratamentoRESUMO
Plasmodium vivax is the most common species of human malaria parasite found outside Africa, with high endemicity in Asia, Central and South America, and Oceania. Although Plasmodium falciparum causes the majority of deaths, P. vivax can lead to severe malaria and result in significant morbidity and mortality. The development of a protective vaccine will be a major step toward malaria elimination. Recently, a formulation containing the three allelic variants of the P. vivax circumsporozoite protein (PvCSP-All epitopes) showed partial protection in mice after a challenge with the hybrid Plasmodium berghei (Pb) sporozoite, in which the PbCSP central repeats were replaced by the VK210 PvCSP repeats (Pb/Pv sporozoite). In the present study, the chimeric PvCSP allelic variants (VK210, VK247, and P. vivax-like) were fused with the mumps virus nucleocapsid protein in the absence (NLP-CSPR) or presence of the conserved C-terminal (CT) domain of PvCSP (NLP-CSPCT). To elicit stronger humoral and cellular responses, Pichia pastoris yeast was used to assemble them as nucleocapsid-like particles (NLPs). Mice were immunized with each recombinant protein adjuvanted with Poly (I:C) and presented a high frequency of antigen-specific antibody-secreting cells (ASCs) on days 5 and 30, respectively, in the spleen and bone marrow. Moreover, high IgG titers against all PvCSP variants were detected in the sera. Later, these immunized mice with NLP-CSPCT were challenged with Pb/Pv sporozoites. Sterile protection was observed in 30% of the challenged mice. Therefore, this vaccine formulation use has the potential to be a good candidate for the development of a universal vaccine against P. vivax malaria.
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The tongue is an important organ in species due to its feeding functions, and its structure is influenced by the habitat and diet. The Collared Peccary (Pecari tajacu, Linnaeus, 1758) is a terrestrial mammal that is distributed on the American continents and has an omnivorous diet. This study aimed to describe the morphological characteristics of the tongue, lingual papillae and the connective tissue cores (CTCs) of the Collared Peccary. Eight tongues were collected from the Wild Animals Multiplication Center. The samples (n = 6) were processed for three-dimensional analysis of their dorsal epithelium, and their CTCs by scanning electron microscopy and the other samples (n = 2) were used to observe ultrastructural characteristics by transmission electron microscopy. Filiform papillae were observed in the lingual apex and body with their conical CTC demonstrating ripples in their extent. Two types of fungiform papillae were observed, the first in the apex and body with a dome-shaped CTC and the second forming a dorsolateral line with a radial pattern CTC. The vallate papillae were bilateral in the caudal region with the CTCs, characterised by numerous projections, and foliate papilla were not observed. We found a new type of papilla anteriorly to the vallate papilla with an irregular groove and a CTC formed by triangular-shaped projections with a duct opening at the top. We concluded that the lingual papillae of the Collared Peccary resemble those papillae of other mammals, however, were revealed another form of their CTCs and a new lingual papillae morphological description.
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Artiodáctilos/anatomia & histologia , Língua/anatomia & histologia , Animais , Tecido Conjuntivo/anatomia & histologia , Tecido Conjuntivo/ultraestrutura , Imageamento Tridimensional/veterinária , Microscopia Eletrônica de Varredura/veterinária , Microscopia Eletrônica de Transmissão/veterinária , Língua/ultraestruturaRESUMO
BACKGROUND: The prolonged tendon-healing process, the high costs associated with treatment, the increase in the number of injuries over the past decades, and the lack of consensus on the optimal treatment of tendon injuries are a global problem. Restoring the normal tendon anatomy and decreasing the healing time are key factors for treatment advancement. HYPOTHESIS: Application of a purified protein from natural latex (PPNL) accelerates the healing process, increasing collagen synthesis and decreasing metalloproteinases. PPNL associated with a simpler suture technique should decrease the healing time. STUDY DESIGN: Controlled laboratory study. METHODS: Injury, surgery, and treatment with PPNL were conducted with male Sprague-Dawley rats. Two suture techniques were used: U-suture, a simpler and lesser traumatic technique, and Kessler-Tajima, to avoid strangulation of the microcirculation. Achilles tendons were completely sectioned, and 100 µL of 0.1% PPNL was applied on the tendon during surgery. Tendon morphology, distribution, and quantity of collagen types I and III, as well as expression of TIMP-1, TIMP-2, MMP-2, and MMP-9 and ultrastructural aspects of cells and collagen fibrils, were assessed after 2 and 4 weeks. RESULTS: PPNL treatment improved collagen type I synthesis and reduced MMP-2 expression. All groups showed a 6.8-times increase in tendon weight as compared with the control group after 2 weeks and a 5.2-times increase after 4 weeks. All groups showed an increase in diameter after 4 weeks, except for the ones treated with PPNL, which showed a slight reduction in diameter. The peak of concentration of collagen fibrils with a 80-nm diameter was 27.79% in the control group; all other experimental groups presented fibrils between 50 and 60 nm. However, the best results were observed with Kessler-Tajima suture associated with PPNL. CONCLUSION/CLINICAL RELEVANCE: There are no known medicines or substances capable of aiding the tendon healing process besides surgery. The discovery of a substance able to improve this process and decrease its duration represents an important advancement in orthopaedic medicine.
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Tendão do Calcâneo/lesões , Tendão do Calcâneo/cirurgia , Borracha/administração & dosagem , Suturas , Cicatrização/fisiologia , Animais , Colágeno Tipo I/metabolismo , Colágeno Tipo III/metabolismo , Hevea , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Modelos Animais , Ratos , Ratos Sprague-Dawley , Técnicas de Sutura , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-2/metabolismoRESUMO
The effects of low-level laser therapy (LLLT) and natural latex protein (F1, Hevea brasiliensis) were evaluated on crush-type injuries (15kg) to the sciatic nerve in the expressions of nerve growth factor (NGF) and vascular endothelium growth factor (VEGF) and ultrastructural morphology to associate with previous morphometric data using the same protocol of injury and treatment. Thirty-six male rats were allocated into six experimental groups (n = 6): 1-Control; 2-Exposed nerve; 3-Injured nerve; 4-LLLT (15J/cm2, 780nm, 30mW, Continuous Wave) treated injured nerve; 5-F1 (0,1mg) treated injured nerve; and 6-LLLT&F1 treated injured nerve. Four or eight weeks after, sciatic nerve samples were processed for analysis. NGF expression were higher (p<0.05) four weeks after in all injured groups in comparison to Control (Med:0.8; Q1:0; Q3:55.5%area). Among them, the Injured (Med:70.7; Q1:64.4; Q3:77.5%area) showed the highest expression, and F1 (Med:17.3; Q1:14.1; Q3:21.7%area) had the lowest. At week 8, NGF expressions decreased in the injured groups. VEGF was expressed in all groups; its higher expression was observed in the injured groups 4 weeks after (Injured. Med:29.5; F1. Med:17.7 and LLLT&F1. Med:19.4%area). At week 8, a general reduction of VEGF expression was noted, remaining higher in F1 (Med:35.1; Q1.30.6; Q3.39.6%area) and LLLT&F1 (Med:18.5; Q1:16; Q3:25%area). Ultrastructural morphology revealed improvements in the treated groups; 4 weeks after, the F1 group presented greater quantity and diameter of the nerve fibers uniformly distributed. Eight weeks after, the F1 and LLLT&F1 showed similar characteristics to the non-injured groups. In summary, these results and our previous studies indicated that F1 and LLLT may favorably influence the healing of nerve crush injury. Four weeks after nerve injury F1 group showed the best results suggesting recovery acceleration; at 8th week F1 and LLLT&F1 groups presented better features and higher vascularization that could be associated with VEGF maintenance.
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Hevea/química , Terapia com Luz de Baixa Intensidade , Traumatismos dos Nervos Periféricos/terapia , Proteínas de Plantas/administração & dosagem , Nervo Isquiático/lesões , Animais , Lesões por Esmagamento/complicações , Modelos Animais de Doenças , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Látex/química , Masculino , Microscopia Eletrônica de Transmissão , Regeneração Nervosa/efeitos dos fármacos , Regeneração Nervosa/efeitos da radiação , Traumatismos dos Nervos Periféricos/etiologia , Proteínas de Plantas/isolamento & purificação , Ratos , Ratos Wistar , Recuperação de Função Fisiológica , Nervo Isquiático/fisiologia , Nervo Isquiático/ultraestrutura , Cicatrização/efeitos dos fármacos , Cicatrização/efeitos da radiaçãoRESUMO
The existence of neurogenesis in the adult brain is a widely recognized phenomenon, occurring in the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone of the dentate gyrus in several vertebrate species. Neural precursors originated in the SVZ migrate to the main olfactory bulb (MOB), originating the rostral migratory stream (RMS) in the process. To better understand the formation of the adult neurogenic niches in dogs, we investigated the cellular composition and morphological organization of these areas in 57 days-old dog fetuses. Using multiple immunohistochemical markers, we demonstrated that the SVZ in the canine fetus is remarkably similar to the adult SVZ, with glial GFAP-immunoreactive (-ir) cells, DCX-ir neuroblasts and SOX2-ir neuronal progenitors tangentially organized along the dorsal lateral ventricle. The fetal RMS has all the features of its adult counterpart and closely resembles the RMS of other mammalian species. The late-development canine MOB has most of the neurochemical features of the adult MOB, including an early-developed TH-ir population and maturing CALR-ir interneurons, but CALB-ir neurons in the granule cell layer will only appear in the post-partum period. Taken together, our results suggest that the canine fetal development of adult neurogenic niches closely resembles those of primates, and dogs may be suitable models of human adult neurogenesis. Anat Rec, 301:1570-1584, 2018. © 2018 Wiley Periodicals, Inc.
Assuntos
Movimento Celular/fisiologia , Ventrículos Laterais/embriologia , Células-Tronco Neurais/citologia , Neurogênese/fisiologia , Bulbo Olfatório/embriologia , Animais , Proliferação de Células/fisiologia , Cães , Ventrículos Laterais/citologia , Neurônios/citologia , Bulbo Olfatório/citologiaRESUMO
Diabetic peripheral neuropathy (DPN) is a nervous disorder caused by diabetes mellitus, affecting about 50% of patients in clinical medicine. Chronic pain is one of the major and most unpleasant symptoms developed by those patients, and conventional available treatments for the neuropathy, including the associated pain, are still unsatisfactory and benefit only a small number of patients. Photobiomodulation (PBM) has been gaining clinical acceptance once it is able to promote early nerve regeneration resulting in significant improvement in peripheral nerves disabilities. In this work, the effects of PBM (660 nm, 30 mW, 1.6 J/cm2 , 0.28 cm2 , 15 s in a continuous frequency) on treating DPN-induced pain and nerve damage were evaluated in an experimental model of diabetic-neuropathy induced by streptozotocin in mice. PBM-induced antinociception in neuropathic-pain mice was dependent on central opioids release. After 21 consecutive applications, PBM increased nerve growth factor levels and induced structural recovery increasing mitochondrial content and regulating Parkin in the sciatic nerve of DPN-mice. Taking together, these data provide new insights into the mechanisms involved in the effects of PBM-therapy emphasizing its therapeutic potential in the treatment of DPN.