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This work aims to study the major salivary gland morphology of peccaries during their growth. The glands were analyzed using macroscopic description, light microscopy, electron microscopy, histochemistry, and immunohistochemistry. Topographically, the salivary glands resemble other animals, including domestic animals and pigs. During growth, the parotid enlarges and mandibular gland loses weight. Histologically, the parotid has serous production, and sublingual has mucous production, resembles most species, however, mandibular gland produces mucous, unlike other animals, including pigs, which produce seromucous secretion. Histochemically, parotid produces more acidic mucins than pigs and it undergoes maturation during development; mandibular, and especially the sublingual gland, produce more acidic and basic mucopolysaccharides than pigs. The results found with transmission and scanning electron microscopy techniques corroborate the histological and histochemistry findings. The major salivary glands were positive to different lecithins (Com-A, BSA-I-B4, WGA and PNA), which were also more positive than in pigs and sheep. We conclude that collared peccaries have a salivary secretion that facilitates the digestion of carbohydrates, and biometric characteristics and positivity to lecithins that facilitate adaptation to foods with antinutritional factors.
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Cold atmospheric plasma (CAP) has been employed as a therapy against both acute and chronic skin lesions, contaminated or not, and has effects on angiogenesis and reepithelialization promoting healing. In this context, the present study aimed to evaluate the effects of a CAP jet associated with pharmacological treatment described by the 2015 AAHA/AAFP pain management guidelines and the 2022 WSAVA guidelines for the recognition, assessment, and treatment of pain, on the healing of chronic skin lesions caused by a pruritic reaction resulting from post-surgical neuropathic pain. To this end, a single CAP application was performed on a feline patient with a 6 months old recurrent contaminated cervical skin lesions along with administration of ketamine (10 µg/kg/min) following the prescription of prednisone (1 mg/kg, SID, 6 days), gabapentin (8 mg/kg, BID, 60 days) and amitriptyline (0.5 mg /kg, SID, 60 days). A single application of plasma associated with an NMDA antagonist, anti-inflammatory steroid, tricyclic antidepressant and gabapentinoid thus provided a significant improvement in the macroscopic appearance of the lesion within 10 days, and the owner reported the cessation of intense itching within the first four hours after treatment and a consequent improvement in the animal's quality of life. The medical treatment was finished almost a year since the writing of this paper, without clinical or reported recurrent signs of the condition. Therefore, we observed that single dose CAP application associated with ketamine, gabapentin, amitriptyline and prednisone leads to significant healing of chronically infected skin lesions resulting from post-surgical neuropathic pain.
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Analgésicos , Enfermedades de los Gatos , Ketamina , Neuralgia , Gases em Plasma , Animales , Gatos , Neuralgia/veterinaria , Neuralgia/tratamiento farmacológico , Neuralgia/etiología , Gases em Plasma/uso terapéutico , Gases em Plasma/farmacología , Enfermedades de los Gatos/tratamiento farmacológico , Ketamina/administración & dosificación , Ketamina/uso terapéutico , Analgésicos/uso terapéutico , Analgésicos/administración & dosificación , Dolor Postoperatorio/veterinaria , Dolor Postoperatorio/tratamiento farmacológico , Gabapentina/uso terapéutico , Gabapentina/administración & dosificación , Masculino , Amitriptilina/uso terapéutico , Amitriptilina/administración & dosificación , Prednisona/uso terapéutico , Prednisona/administración & dosificación , Terapia Combinada/veterinaria , FemeninoRESUMEN
While titanium dental implants have already been clinically established, ongoing research is continuously being conducted to advance the fields of osseointegration and bacterial resistance, seeking further improvements in these areas. In this study, we introduce an innovative method for treating titanium surfaces within tightly sealed packaging. Specifically, titanium discs, enclosed in surgical-grade packaging, underwent treatment using cold atmospheric plasma (CAP). The surfaces were thoroughly characterized in terms of wettability, crystalline structure, and chemical composition. Hemocompatibility analyses were conducted using blood diluted in sodium citrate (1:9) exposed to titanium discs for 30 min inside a CO2 incubator at 37 °C. Subsequently, various blood parameters were evaluated, including prothrombin time (PT), activated partial thromboplastin time (APTT), and platelet adhesion. Microbiological analyses were also performed using Pseudomonas aeruginosa (ATCC 27853) for 4 h at 37 °C. The treatment with CAP Jet resulted in a reduction in contact angle without causing any changes in the crystalline structure. No statistically significant differences were observed in the blood parameters. The plasma-treated samples exhibited lower PT and APTT values compared to those of the control group. The surfaces treated with CAP Jet showed increased platelet activation, platelet density, and thrombus formation when compared with the untreated samples. Moreover, the treated surfaces demonstrated lower bacterial colony formation compared with other surfaces.
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Gases em Plasma , Titanio , Propiedades de Superficie , Titanio/farmacología , Titanio/química , Gases em Plasma/farmacología , Gases em Plasma/química , Humectabilidad , PlaquetasRESUMEN
This study aimed to investigate sexual dimorphism in stillborn hawksbill sea turtles (Eretmochelys imbricata) through gonadal morphological characterizations. Macroscopic, light microscopy, and transmission electron analyses were performed for 30 gonad-mesonephros complexes. Female gonads were spindle-shaped and present a translucent whitish appearance with a grainy texture. Male gonads were approximately ovoid with a smooth opaque white surface. A primary sexual difference concerns different marrow structures, with females presenting organized cellularity featuring oocytes, lacunae, and blood vessels, while males presented a distinct organizational medulla pattern marked by testicular cords extending throughout the gonad length. Ultrastructurally, female's stroma presented interstitial cells and an abundant cytoplasm rich in electrodense droplets and large oval germline cells, with a conspicuous and noncentral nucleus. Males, on the other hand, presented testicular cord cells containing small amounts of heterochromatin and approximately triangular apical and basal cytoplasms with an evident nucleolus characteristic of support cells. Additionally, there were cells with a large spherical nucleus compared with the cell size and a relatively scarce cytoplasm, identified as gonocytes. These findings indicate that macroscopic, microscopic, and ultrastructural evaluations are effective and reliable techniques for the sexual identification of stillborn E. imbricata hatchlings.
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Tortugas , Animales , Masculino , Femenino , Gónadas , Ovario , Oocitos , Folículo OváricoRESUMEN
BACKGROUND: Hystricomorpha rodents display a similar placentation model to humans. The present study was carried out considering the scarcity of information concerning the placental development in agouti. OBJECTIVE: Describe the microscopy of the placenta, subplacenta and yolk sac of agoutis in early pregnancy and report on the inversion of the yolk sac. METHODS: Fifteen females between the 14th-32nd day of gestation were used following euthanasia. Gestational buttons were collected, fixed, processed, stained to optical microscopy or immunohistochemistry. RESULTS: Chorioallantoic placenta (CP) ranged from conical to a half-sphere, as follows: from the 14th to 17th day, the CP displays an inverted "V" shape, predominantly formed by cytotrophoblasts; from 20 to 22 days, formed almost entirely by cytotrophoblasts; at 28 days, a half sphere, with distinct lobes and interlobular area, numerous maternal gaps delimited by syncytiotrophoblasts and trophoblast giant cells; at 32 days, globose and undergoing the maturation process. Subplacenta, located between decidua and CP, initially presents septa consisting of simple columnar epithelium and after 17 days, comprising stratified epithelium. Visceral yolk sac (VYS) is attached to two CP projections between 14 and 17 days, formed by a simple cubic epithelium and inverted. Between 20 and 22 days, the epithelium displays apical villous projections with cytoplasmic vacuoles and a vascularized mesoderm. After the 24th day, the VYS near the placenta is pleated, very vascularized and villous, with decreased villi sizes further away from the placenta. CONCLUSION: The agouti CP displays similar characteristics to other hystricomorpha, including placenta lobulation, a subplacenta and an inverted vitelline placenta.
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Dasyproctidae , Placentación , Embarazo , Femenino , Animales , Humanos , Placenta , Roedores , Saco VitelinoRESUMEN
The greater rhea, Rhea americana, is a wild ratite of high scientific importance and significant and zootechnical value, especially considering the current development state of Brazilian poultry production, where research aimed at increasing the productivity of these animals has become extremely relevant. Studies concerning fetal attachments and embryonic development are paramount, as they can provide essential information concerning reproductive and nutritional animal management. However, a lack of information on greater rhea fetal morphology is noted. Therefore, the aim of the present study was to establish a standard model for fetal attachments in this species. Greater rhea eggs were incubated from 0 to 36 days, and macroscopic and microscopic embryonic attachment characterizations were performed. Histologically, all embryonic annexes exhibit germ layers, namely the ectoderm (outer layer), mesoderm (middle layer) and endoderm (inner layer). The findings indicate that greater rhea development patterns are similar to other birds.
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Cardiogenesis is similar in all vertebrates, but differences in the valvuloseptal morphogenesis among non-crocodilian reptiles, birds, and mammals are noted. The origin of mesenchymal structures such as valves that regulate the passage of blood and the formation of partial septa that prevent the complete mixing of oxygen-rich and low-oxygen blood present in adult chelonians are essential in the evolutionary understanding of complete septation, endothermy and malformations, even in mammals. In this context, this study analyzed the heart morphogenesis of Podocnemis unifilis (Testudines: Podocnemididae) from the 4th to the 60th day of incubation. We identified the tubular heart stage, folding of the cardiac tube and expansion of the atrial and ventricular compartments followed by atrial septation by the septum primum, ventricle septation by partial septa, outflow tract septation and the formation of bicuspid valves with cartilage differentiation at the base. The formation of the first atrial septum with the mesenchymal cap is noted during the development of the atrial septum, joining the atrioventricular cushion on the 17th day and completely dividing the atria. Small secondary perforations appeared in the mid-cranial part, observed up to the 45th day. Partial ventricle septation into the pulmonary, venous, and arterial subcompartments takes place by trabeculae carneae thickening and grouping on the 15th day. The outflow tract forms the aorticopulmonary and interaortic septa on the 16th day and the bicuspid valves, on the 20th day. Therefore, after the first 20 days, the heart exhibits a general anatomical conformation similar to that of adult turtles.
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Fibrilación Atrial , Humanos , Desarrollo EmbrionarioRESUMEN
Morphological studies of the oropharyngeal cavity of chelonians have become an interesting tool in the understanding of evolutionary processes associated with feeding habits in aquatic animals and the transition from aquatic to terrestrial forms. In this context, the aim of the present study was to describe the oropharyngeal cavity floor morphology of hawksbill sea turtle (Eretmochelys imbricata) hatchlings. Ten dead hatchlings of undefined sex were obtained from nests hatched on the coast of the state of Rio Grande do Norte, Brazil. The heads of each specimen were fixed, dissected, and analyzed at the macroscopic and microscopic levels. The oropharyngeal cavity floor of the hawksbill sea turtle hatchlings is formed by the tongue, pharynx, floor muscles, and hyolingual skeleton, delimited in the rostral and lateral directions by a keratinized beak, called the rhamphotheca, and in the caudal region at the limit between the pharynx and the esophagus. The tongue muscles and the muscles that support the floor of the oral cavity comprise the following: m. hypoglossohyoideus, m. hypoglossoglossus, m. hyoglossus, m. genioglossus, m. constrictor laryngis, m. geniohyoideus pars lateralis, and m. intermandibularis. The oropharyngeal cavity floor mucosa is formed by keratinized stratified squamous epithelium and the lamina propria is formed by loose connective tissue. The floor mucosa is devoid of taste buds. We believe that the basic oropharyngeal cavity floor characteristics in hawksbill sea turtle hatchlings may comprise indications that these animals are plesiomorphic and that semiaquatic and terrestrial turtles may have undergone adaptations to feed out of water.
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Tortugas , Animales , Tortugas/anatomía & histología , Adaptación Fisiológica , Aclimatación , Membrana Mucosa , EpitelioRESUMEN
Duchenne Muscular Dystrophy (DMD) reproductive alterations and the influence of antioxidant treatments may aid in understanding morphometry testicular quantification. In this context, the aim of the present study was to characterize the intertubular compartment (ITC) morphometry of animal testes in mdx mice supplemented with ascorbic acid (AA). Sixteen mice were used, namely the C57BL/10 (non-dystrophic) and C57BL/10Mdx (dystrophic) lineages, distributed into the following groups: Control (C60), Dystrophic (D60), Control supplemented with AA (CS60), Dystrophic supplemented with AA (DS60). A total of 200 mg/kg of AA were administered to mice for 30 days. Subsequently, the testicles were collected, weighed, and fragmented. The obtained fragments were fixed in Karnovsky's solution (pH 7.2) and embedded in historesin for morphometric and transmission electron microscopy assessments. Leydig cells were hypertrophic in the D60 group, but was reverted by AA supplementation in the DS60 group. The DS60 group also exhibited increased intertubular volume compared to the CS60 group. The ultrastructural images identified multilamellar bodies in dystrophic animals (lipid storage) and telocyte cells (transport substances) in both control and dystrophic animals. Morphometric alterations were, therefore, noted in the intertubular compartment due to Duchenne muscular dystrophy (DMD), with AA administration capable of altering Leydig cells in this condition.
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Morphological studies concerning the digestive system can further information on animal diets, thus aiding in the understanding of feeding behavior. Given the scarcity of information on sea turtle digestive system morphology, the aim of the present study was to describe the digestive tube (DT) morphology of Eretmochelys imbricata hatchlings to further understand the diet of these individuals in the wild. DT samples from 10 stillborn turtles (undefined sex) were analyzed at the macro and microscopic levels. The esophagus, stomach, small intestine (SI), and large intestine (LI) are described. Histologically, the DT is formed by four tunics, the mucosa, submucosa, muscular, and adventitia or serosa. The esophagus is lined by keratinized stratified squamous epithelium, while the remainder of the DT is lined by a simple columnar epithelium. The esophagus mucosa is marked by conical, pointed papillae. The stomach comprises three regions, the cardiac, fundic, and pyloric and is covered by neutral mucous granular cells. The intestinal mucosa presents absorptive cells with microvilli, neutral and acidic goblet cells, and mucosa-associated lymphoid tissue. The SI is significantly longer than the LI (p value = 0.006841). These morphological findings are strong indications of adaptations to a carnivorous diet in this hawksbill turtle age group.
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Sex steroid hormones are critical in gonadal differentiation in turtles. The gonads are not the only organs responsible for producing these hormones during this phase. Mesonephros play an important role in steroidogenesis. The present study aimed to investigate the presence of steroidogenic cells in mesonephros of Podocnemis expansa during gonadal differentiation and to evaluate their morphology and ultrastructure. Ten embryos of P. expansa were collected from 5 nests on day 36 of incubation, during spawning period on an artificial beach. Embryos were extracted from eggs by slicing the shell and euthanized. They were dissected under a stereoscopic microscope to collect the gonad-mesonephro complex, in which were fixed and subsequently processed for light microscopy, immunohistochemistry and transmission electron microscopy analysis. During histological analysis was observed mesonephros has typical morphological structure. Immunohistochemistry showed immunoreaction to aromatase in cells of intertubular space. Confirming these findings, it was possible to observe a type of intertubular cell in several regions of mesonephro, being more predominant in region close to blood vessels, distal and proximal tubules. In ultrastructural analysis these cells were characterized by having a clear, large, and rounded nucleus with evident nucleolus and cytoplasm rich in electron-dense droplets. This study demonstrated for the first time the presence of cells with morphological, immunohistochemical and ultrastructural characteristics similar to steroid-producing cells in P. expansa mesonephrons, suggesting that this organ may contribute to gonadal differentiation in this species.
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The determination of arterial vascular distribution patterns can contribute to more detailed knowledge on arterial systems. In this context, the aim of the present study was to describe the collateral aortic branches in the collared peccary, aiming to define a standard model for this species, providing information through comparative anatomy for analysis of some evolutionary aspects of the order Artiodactyla. Ten young male animals were employed. The vascular system was washed with a saline solution and then perfused with Neoprene 450 latex coloured with red or yellow pigment, followed by collateral aorta branch dissections and analyses. The contrasted digital radiography technique was applied to two animals, which were initially perfused with a barium sulphate solution (1 g/ml) and Neoprene 450 latex at a 1:3 ratio, and subsequently, the digital radiographic examination was performed. The aortic arch of the collared peccary emitted the brachiocephalic trunk and the left subclavian artery, while eight symmetric pairs of dorsal intercostal arteries originated from the thoracic aorta. The abdominal aorta, in turn, exhibited the celiac, cranial mesenteric, renal, caudal mesenteric, testicular, external iliac, internal iliac and sacral arteries as the collateral branches in all studied animals. Therefore, no relevant variations were observed regarding the arrangement of the arterial distribution of the aorta, thus suggesting a static standard model in the collared peccary.
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Artiodáctilos , Látex , Animales , Aorta Abdominal , Evolución Biológica , Masculino , NeoprenoRESUMEN
Sex steroid hormones are critical in gonadal differentiation in turtles. The gonads are not the only organs responsible for producing these hormones during this phase. Mesonephros play an important role in steroidogenesis. The present study aimed to investigate the presence of steroidogenic cells in mesonephros of Podocnemis expansa during gonadal differentiation and to evaluate their morphology and ultrastructure. Ten embryos of P. expansa were collected from 5 nests on day 36 of incubation, during spawning period on an artificial beach. Embryos were extracted from eggs by slicing the shell and euthanized. They were dissected under a stereoscopic microscope to collect the gonad-mesonephro complex, in which were fixed and subsequently processed for light microscopy, immunohistochemistry and transmission electron microscopy analysis. During histological analysis was observed mesonephros has typical morphological structure. Immunohistochemistry showed immunoreaction to aromatase in cells of intertubular space. Confirming these findings, it was possible to observe a type of intertubular cell in several regions of mesonephro, being more predominant in region close to blood vessels, distal and proximal tubules. In ultrastructural analysis these cells were characterized by having a clear, large, and rounded nucleus with evident nucleolus and cytoplasm rich in electron-dense droplets. This study demonstrated for the first time the presence of cells with morphological, immunohistochemical and ultrastructural characteristics similar to steroid-producing cells in P. expansa mesonephrons, suggesting that this organ may contribute to gonadal differentiation in this species.(AU)
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Animales , Tortugas/fisiología , Procesos de Determinación del Sexo/fisiología , Factor Esteroidogénico 1/análisis , Aromatasa/inmunologíaRESUMEN
Duchenne Muscular Dystrophy (DMD) reproductive alterations and the influence of antioxidant treatments may aid in understanding morphometry testicular quantification. In this context, the aim of the present study was to characterize the intertubular compartment (ITC) morphometry of animal testes in mdx mice supplemented with ascorbic acid (AA). Sixteen mice were used, namely the C57BL/10 (non-dystrophic) and C57BL/10Mdx (dystrophic) lineages, distributed into the following groups: Control (C60), Dystrophic (D60), Control supplemented with AA (CS60), Dystrophic supplemented with AA (DS60). A total of 200 mg/kg of AA were administered to mice for 30 days. Subsequently, the testicles were collected, weighed, and fragmented. The obtained fragments were fixed in Karnovsky's solution (pH 7.2) and embedded in historesin for morphometric and transmission electron microscopy assessments. Leydig cells were hypertrophic in the D60 group, but was reverted by AA supplementation in the DS60 group. The DS60 group also exhibited increased intertubular volume compared to the CS60 group. The ultrastructural images identified multilamellar bodies in dystrophic animals (lipid storage) and telocyte cells (transport substances) in both control and dystrophic animals. Morphometric alterations were, therefore, noted in the intertubular compartment due to Duchenne muscular dystrophy (DMD), with AA administration capable of altering Leydig cells in this condition.(AU)
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Animales , Masculino , Ratones , Túbulos Seminíferos/fisiopatología , Ratones Endogámicos mdx/fisiología , Células Intersticiales del Testículo/fisiología , Enfermedades Musculares/veterinariaRESUMEN
Glutamine is often used to treat metabolic changes associated with anorexia-cachexia syndrome in patients with malignant neoplasms. Walker 256 tumor is an excellent model for studying these changes associated with cancer in different organs, including injuries in testicular functions. However, the effects of supplementing glutamine on testicular morphometry in this model have not yet been investigated. Thus, the objective of this study was to evaluate the effect of L-glutamine supplementation on testicular morphometry in rats transplanted with Walker 256 tumor cells. Forty puberty Wistar rats were divided into four groups: control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The testicles were removed, weighed, fixed in Bouin, and included in paraffin for histomorphometric analysis. Walker 256 tumor caused quantitative changes in the tubular and intertubular compartments and tunica albuginea, with reductions in the percentages of lumen and tunica albuginea, number of Sertoli cells per gram of testis; number of Leydig cells; percentage of blood vessels and connective tissue in intertubule. However, glutamine supplementation prevented part of these changes caused by the tumor, presenting mainly a protective effect on the tunica albuginea and percentage of blood and lymph vessels in the intertubule. These results indicate the potential of L-glutamine was able to recover for testicular dysfunction associated with cancer.
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Células Intersticiales del Testículo , Telocitos/ultraestructura , Testículo/ultraestructura , Animales , Masculino , Ratones , Ratones Endogámicos mdx , Microscopía Electrónica de Transmisión/veterinaria , Telocitos/fisiología , Telopodos/fisiología , Telopodos/ultraestructura , Testículo/citologíaRESUMEN
Heart diseases in birds are frequent and generate significant production disorders. Morphometry is a valuable tool to provide fundamental information about heart conditions. Few studies have addressed morphological aspects of the heart of ratite birds, such as the Greater rhea. The present study aimed to analyse rhea heart morphometry, comparing young and adult subjects, in order to provide relevant information for the diagnosis of heart disease in this species. Hearts of young (n = 10) and adult (n = 10) female rheas were used in this research. Heart length and width and sternum length were measured using a caliper. Heart length and width and sternum length in adults were approximately three times greater than in young individuals. The left ventricular wall (LVW) was thicker than the right ventricular wall (RVW) at all ages, and the RVW was thicker in adults when compared to young subjects. The basal and middle RVW regions thicken with advancing age, and the thickness of the interventricular septum (ISW) occupies an intermediate position between the LVW and RVW. In general, an increase in rhea heart thickness and size relative to age is observed. The morphometric variations between young and adult rhea hearts observed in the present study may serve as a comparative subsidy for the diagnosis of cardiac abnormalities observed in this species.
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Reiformes , Animales , Aves , Femenino , CorazónAsunto(s)
Animales , Masculino , Testículo/ultraestructura , Telocitos/ultraestructura , Células Intersticiales del Testículo , Testículo/citología , Ratones Endogámicos mdx , Microscopía Electrónica de Transmisión/veterinaria , Telopodos/fisiología , Telopodos/ultraestructura , Telocitos/fisiologíaRESUMEN
Abstract Glutamine is often used to treat metabolic changes associated with anorexia-cachexia syndrome in patients with malignant neoplasms. Walker 256 tumor is an excellent model for studying these changes associated with cancer in different organs, including injuries in testicular functions. However, the effects of supplementing glutamine on testicular morphometry in this model have not yet been investigated. Thus, the objective of this study was to evaluate the effect of L-glutamine supplementation on testicular morphometry in rats transplanted with Walker 256 tumor cells. Forty puberty Wistar rats were divided into four groups: control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The testicles were removed, weighed, fixed in Bouin, and included in paraffin for histomorphometric analysis. Walker 256 tumor caused quantitative changes in the tubular and intertubular compartments and tunica albuginea, with reductions in the percentages of lumen and tunica albuginea, number of Sertoli cells per gram of testis; number of Leydig cells; percentage of blood vessels and connective tissue in intertubule. However, glutamine supplementation prevented part of these changes caused by the tumor, presenting mainly a protective effect on the tunica albuginea and percentage of blood and lymph vessels in the intertubule. These results indicate the potential of L-glutamine was able to recover for testicular dysfunction associated with cancer.
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Esse estudo objetivou caracterizar o desenvolvimento e a diferenciação gonadal de Podocnemis unifilis em temperatura controlada de 32°C em incubadora artificial e com controle de umidade feito por um higrômetro interno. Foram realizadas análises macroscópicas, por microscopia de luz e por microscopia eletrônica de transmissão. O desenvolvimento gonadal foi agrupado em três etapas: células germinativas primordiais (CGP), estabelecimento da gônada indiferenciada e diferenciação gonadal. As CGP só foram visualizadas a partir do 7º dia de incubação, e se encontravam na região caudal do corpo do embrião, e entre os dias 9 e 11 de incubação foi possível observar a migração das CGP a partir do saco vitelínico em direção à região ventromedial do mesonefro. Com 20 dias de desenvolvimento, a gônada indiferenciada estava estabelecida, sendo identificadas duas regiões distintas: a região cortical externa, caracterizada pela presença de CGP, e a região medular interna, marcada por cordões sexuais primitivos. Nesse estudo, a diferenciação gonadal só ocorreu em testículo. A diferenciação teve início após 35 dias de incubação, com o início da organização dos cordões sexuais em túbulos seminíferos. As etapas de diferenciação gonadal foram semelhantes às de outras espécies de quelônios com determinação do sexo pela temperatura, no entanto, neste estudo, usando a temperatura pivotal descrita para a espécie, todos os indivíduos analisados diferenciaram-se em machos, sendo necessária, portanto, a realização de mais estudos com diferentes temperaturas.
This study aimed to characterize the development and gonadal differentiation of Podocnemis unifilis at a controlled temperature of 32° C in artificial incubator and with humidity control performed by an internal hygrometer. Macroscopic analyzes, light microscopy and transmission electron microscopy were performed. We grouped gonadal development into three stages: primordial germ cells (PGC), establishment of the undifferentiated gonad and gonadal differentiation. We only visualized the PGC from the 7th day of incubation, and in the caudal region of the embryo body; between days 9 and 11 of incubation we were able to observe the migration of the PGC from the viteline sac towards the ventromedial region of the mesonefro. At 20 days of development, the undifferentiated gonad was established and two distinct regions were identified: the external cortical region characterized by the presence of PGC, and the internal medullary region marked by primitive sexual cords. In this study, only gonadal differentiation occurred in the testicle. Gonadal differentiation began after 35 days of incubation with the beginning of the organization of sexual cords in the seminiferous tubules. The stages of gonadal differentiation were similar to those of other species of turtles with temperature determined sex, however in this study, using the pivotal temperature described for the species all the individuals analyzed were males, and it is there for necessary to carry out further studies using a higher incubation temperature.