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The stem cell-based research for reproductive biotechnology has been widely studied and shows promise for repairing defective tissue or degenerated cells to treat different diseases. The adipose tissue and amniotic membrane have awakened great interest in regenerative medicine and arises as a promising source of mesenchymal stem cells. Both types, adipose and amniotic derived mesenchymal stem cells (AMSCs) are multipotent cells with an enhanced ability to differentiate into multiple lineages.. We aimed to evaluate the effect of basal supplementation of exosomes in cell cultures with canine amniotic mesenchymal stem cells (MSCs). Mesenchymal stem cells derived from canine amniotic and adipose tissue were isolated and cultured performing cell passages until 80-90% confluence was reached. The growth curve was determined and peak cell growth was observed in the second passage. The cells were then characterized and differentiated into adipogenic, chondrogenic and osteogenic lineages. Extracellular vesicles from amnion were isolated using an ultracentrifugation protocol and characterized by nanosight analysis. To evaluate their ability to improve cellular viability in naturally inefficient passages, exosomes were co-cultures to the MSC cells. The results showed a 15-20% increase in the expansion rate of cultures supplemented with vesicles extracted in the first and second passages when compared to the control group. Statistical analysis using the Dunnett test (p ≤ 0.05) corroborated this result, showing a positive correlation between supplementation and expansion rate. These results indicate not only the importance of exosomes in the cell communication process but also the feasibility of the culture supplementation protocol for therapeutic purposes. The potential of the AMSCs for reproductive biotechnology is undoubted, however, their application to repair reproductive disorders and the involved mechanisms remain elusive. The strategies to enable the Adipose Stem Cells and AMSCs application in reproductive biotechnology and optimize their use for tissue regeneration open new venues using exosomes interactions.

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The course of ADHD from childhood up to young adulthood has been characterized in several studies. However, little is known about the course of symptoms into middle age and beyond. This study aims to evaluate predictors of ADHD trajectories in midlife based on three assessments. The follow-up sample comprised 323 adults with ADHD, evaluated at baseline and seven and thirteen years later, from the average ages of 34 up to 47 years old. ADHD status at reassessments was used to characterize trajectories. Demographics, ADHD features, comorbidities, and polygenic scores for ADHD and genetically correlated psychiatric disorders were evaluated to predict ADHD trajectories. Study retention rate was 67% at T2 (n = 216) and 62% at T3 (n = 199). Data from patients evaluated three times showed that 68.8% coursed stable, 25.5% unstable, and 5.7% remission trajectory of ADHD. Women, individuals with more severe syndromes, higher frequency of comorbidities at reassessments, and genetic liability to depression present a higher probability of a stable trajectory. Our findings shed light on midlife ADHD trajectories and their gender, genomic and clinical correlates.

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Extracellular vesicles (EVs) derived from stem cells (SCs) have regenerative potential and the possibility of being used in treating chronic diseases. EVs present lower risk of tumorigenicity and easily to isolation and storage. Therefore, this research aims to compare the morphological characteristics of the EVs (up to 150nm) derived from stem cells obtained from canine amniotic membranes in different passages during the in vitro culture. For this, cells from the amniotic membranes were isolated, cultured, and characterized. In order to answer our aim, the number of cells was normalized at each passage to generate conditioned media for EVs separation. The cells were differentiated into adipogenic, chondrogenic, and osteogenic tissue, to characterize these cells as mesenchymal stem cells (MSC). Moreover, flow cytometry analysis was performed and showed that the MSC were positive for CD90, CD105 and negative for CD34, CD45, mesenchymal and hematopoietic markers, respectively. For EVs analysis, MSC in different passages (P0-P2) were culture until 80% of confluence, then the medium was replaced by EVs depleted medium. After 48h, culture medium was collected and centrifuged to separate EVs, followed by nanoparticle tracking analysis. The EVs were also characterized by western blot and transmission electron microscopy (TEM). EVs were positive for Alix and negative for Cytochrome C as well as presented the traditional cup-shape by transmission electronic microscopy. Our results demonstrated that the concentration in the different passages was increased in P0 compared to P1 and P2 (p<0.05). No differences were found in EVs size (P0=132nm, P1=130nm and P2=120nm). Together, these results demonstrate that P0 of MSC is enriched of EVs when compared to later passages, suggesting that this passage would be the best to be applied in pre-clinical tests. Despite that, more studies are necessary to identify the EVs content and how the cells will respond to treatment with them.

RESUMO

Extracellular vesicles (EVs) derived from stem cells (SCs) have regenerative potential and the possibility of being used in treating chronic diseases. EVs present lower risk of tumorigenicity and easily to isolation and storage. Therefore, this research aims to compare the morphological characteristics of the EVs (up to 150nm) derived from stem cells obtained from canine amniotic membranes in different passages during the in vitro culture. For this, cells from the amniotic membranes were isolated, cultured, and characterized. In order to answer our aim, the number of cells was normalized at each passage to generate conditioned media for EVs separation. The cells were differentiated into adipogenic, chondrogenic, and osteogenic tissue, to characterize these cells as mesenchymal stem cells (MSC). Moreover, flow cytometry analysis was performed and showed that the MSC were positive for CD90, CD105 and negative for CD34, CD45, mesenchymal and hematopoietic markers, respectively. For EVs analysis, MSC in different passages (P0-P2) were culture until 80% of confluence, then the medium was replaced by EVs depleted medium. After 48h, culture medium was collected and centrifuged to separate EVs, followed by nanoparticle tracking analysis. The EVs were also characterized by western blot and transmission electron microscopy (TEM). EVs were positive for Alix and negative for Cytochrome C as well as presented the traditional cup-shape by transmission electronic microscopy. Our results demonstrated that the concentration in the different passages was increased in P0 compared to P1 and P2 (p<0.05). No differences were found in EVs size (P0=132nm, P1=130nm and P2=120nm). Together, these results demonstrate that P0 of MSC is enriched of EVs when compared to later passages, suggesting that this passage would be the best to be applied in pre-clinical tests. Despite that, more studies are necessary to identify the EVs content and how the cells will respond to treatment with them.(AU)

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A série inglesa Black Mirror desnuda o humano a partir da sua complexa relação com a tecnologia de uma forma que impacta e provoca o telespectador. Algo inquietante sobre nós é revelado através do espelho da série, que convida a refletir sobre um uso destrutivo das novas ferramentas e suas possíveis consequências. O autor aceita o convite e examina a partir das principais temáticas da série, o que em nosso funcionamento psíquico pode levar a um uso mortífero da tecnologia. Na busca desse objetivo, este trabalho propõe um diálogo entre as narrativas de Black Mirror e conceitos psicanalíticos de Freud, Meltzer, Bion e Green.(AU)

The English series Black Mirror exposes the human from its complex relationship with technology in a way that impacts and stirs the viewer. Something uncanny about us is revealed through series mirror, which invites us to reflect on the destructive use of new tools and their possible consequences.The author accepts the invitation and examines it from the main themes of the series, which in our psychic functioning can lead to a deadly use of technology. In the pursuit of this objective, this work proposes a dialogue between Black Mirror narratives and psychoanalytic concepts of Freud, Meltzer, Bion and Green.(AU)

La serie inglesa Black Mirror despoja al ser humano en su compleja relación con la tecnología de una manera que impacta y provoca al espectador. Algo inquietante acerca de nosotros se revela a través del espejo de la serie, que nos invita a reflexionar sobre el uso destructivo de nuevas herramientas y sus posibles consecuencias. El autor acepta la invitación y la examina a partir de los principales temas de la serie, que en nuestro funcionamiento psíquico pueden derivar en un uso letal de la tecnología. En la búsqueda de este objetivo, este trabajo propone un diálogo entre las narrativas de Black Mirror y los conceptos psicoanalíticos de Freud, Meltzer, Bion y Green.(AU)

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In regenerative medicine stem cell biology has become one of the most interesting and more often studied subject. The amniotic membrane is the innermost layer of the fetal membranes and is considered a potential tool to treat many pathologies. It is used because it can be collected from discarded fetal material and is a rich source of stem cells with high proliferation and plasticity ratio capable of proliferating and differentiate in vitro. We propose to elucidate the characteristics and potencial clinical application of cells derived of amniotic membrane in veterinary medicine.

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The Ocelot (Leopardus pardalis) is the largest species of this genus, despite having broad distribution in the Americas; it is included in the main list of endangered species. Their conservation is widely studied, but there is a lack of studies about their morphology. In order to contribute to the knowledge of its reproductive system, five male and female ocelots were examined macro- and microscopically by histological techniques. Macroscopic analysis of the male reproductive system revealed presence of prostate and bulbourethral gland located caudally to the urinary bladder and a penis with small spicules. Microscopically, the testes were encased by the tunica albuginea and divided it into lobules with 5-10 tubules per lobe. In females, macroscopic analysis demonstrated two ovaries position dorsally in the sublumbar region and caudal to the kidneys. The bicornuate uterus is composed by uterine horns (12 to 14 cm in length), which travels from the ovaries in a caudal direction to form a small uterine body (4 cm in length). The ovary analysis revealed, in longitudinal section, medullary region composed of loose connective tissue, a stroma rich in blood vessels, and an external parenchymal region surrounded by a tunica albuginea. The results of the study confirmed the similarity between ocelot's reproductive system as domestic cat's ones and showing for the first time the complete morphological tool to highlight these organs and tissue in this male and female endangered wild felid specie. The present study open venue for other researchers to consider morphological and preservationist features and aimed to help at long-term conservation of wild felines.

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Objective: This study evaluated the hypothesis that methylphenidate immediate release (MPH-IR) treatment would improve Default Mode Network (DMN) within-connectivity. Method: Resting-state functional connectivity of the main nodes of DMN was evaluated in a highly homogeneous sample of 18 drug-naive male adult participants with ADHD. Results: Comparing resting-state functional connectivity functional magnetic resonance imaging (R-fMRI) scans before and after MPH treatment focusing exclusively on within-DMN connectivity, we evidenced the strengthening of functional connectivity between two nodes of the DMN: posterior cingulate cortex (PCC) and left lateral parietal cortex (LLP). Conclusion: Our results contribute to the further understanding on how MPH affects functional connectivity within DMN of male adults with ADHD and corroborate the hypothesis of ADHD being a delayed neurodevelopmental disorder.

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The Ocelot (Leopardus pardalis) is the largest species of this genus, despite having broad distribution in the Americas; it is included in the main list of endangered species. Their conservation is widely studied, but there is a lack of studies about their morphology. In order to contribute to the knowledge of its reproductive system, five male and female ocelots were examined macro- and microscopically by histological techniques. Macroscopic analysis of the male reproductive system revealed presence of prostate and bulbourethral gland located caudally to the urinary bladder and a penis with small spicules. Microscopically, the testes were encased by the tunica albuginea and divided it into lobules with 5-10 tubules per lobe. In females, macroscopic analysis demonstrated two ovaries position dorsally in the sublumbar region and caudal to the kidneys. The bicornuate uterus is composed by uterine horns (12 to 14 cm in length), which travels from the ovaries in a caudal direction to form a small uterine body (4 cm in length). The ovary analysis revealed, in longitudinal section, medullary region composed of loose connective tissue, a stroma rich in blood vessels, and an external parenchymal region surrounded by a tunica albuginea. The results of the study confirmed the similarity between ocelot's reproductive system as domestic cat's ones and showing for the first time the complete morphological tool to highlight these organs and tissue in this male and female endangered wild felid specie. The present study open venue for other researchers to consider morphological and preservationist features and aimed to help at long-term conservation of wild felines.(AU)

Assuntos

RESUMO

The Ocelot (Leopardus pardalis) is the largest species of this genus, despite having broad distribution in the Americas; it is included in the main list of endangered species. Their conservation is widely studied, but there is a lack of studies about their morphology. In order to contribute to the knowledge of its reproductive system, five male and female ocelots were examined macro- and microscopically by histological techniques. Macroscopic analysis of the male reproductive system revealed presence of prostate and bulbourethral gland located caudally to the urinary bladder and a penis with small spicules. Microscopically, the testes were encased by the tunica albuginea and divided it into lobules with 5-10 tubules per lobe. In females, macroscopic analysis demonstrated two ovaries position dorsally in the sublumbar region and caudal to the kidneys. The bicornuate uterus is composed by uterine horns (12 to 14 cm in length), which travels from the ovaries in a caudal direction to form a small uterine body (4 cm in length). The ovary analysis revealed, in longitudinal section, medullary region composed of loose connective tissue, a stroma rich in blood vessels, and an external parenchymal region surrounded by a tunica albuginea. The results of the study confirmed the similarity between ocelot's reproductive system as domestic cat's ones and showing for the first time the complete morphological tool to highlight these organs and tissue in this male and female endangered wild felid specie. The present study open venue for other researchers to consider morphological and preservationist features and aimed to help at long-term conservation of wild felines.

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RESUMO

The immune system is mainly responsible for protecting the organism against agents that may interfere in its homeostasis. Thus, understand how this system develops and operates is very important, for create new therapies to assist this system in its operation, such as its failure. In domestic dogs, few studies show how actually occurs the development, maturation and functioning of the immune system. Therefore, this study demonstrates the development and possible activation of it on dog fetus from late gestational period by in situ and microscopic analyzes.

RESUMO

The immune system is mainly responsible for protecting the organism against agents that may interfere in its homeostasis. Thus, understand how this system develops and operates is very important, for create new therapies to assist this system in its operation, such as its failure. In domestic dogs, few studies show how actually occurs the development, maturation and functioning of the immune system. Therefore, this study demonstrates the development and possible activation of it on dog fetus from late gestational period by in situ and microscopic analyzes.(AU)

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RESUMO

In regenerative medicine stem cell biology has become one of the most interesting and more often studied subject. The amniotic membrane is the innermost layer of the fetal membranes and is considered a potential tool to treat many pathologies. It is used because it can be collected from discarded fetal material and is a rich source of stem cells with high proliferation and plasticity ratio capable of proliferating and differentiate in vitro. We propose to elucidate the characteristics and potencial clinical application of cells derived of amniotic membrane in veterinary medicine.(AU)

Assuntos

RESUMO

In regenerative medicine stem cell biology has become one of the most interesting and more often studied subject. The amniotic membrane is the innermost layer of the fetal membranes and is considered a potential tool to treat many pathologies. It is used because it can be collected from discarded fetal material and is a rich source of stem cells with high proliferation and plasticity ratio capable of proliferating and differentiate in vitro. We propose to elucidate the characteristics and potencial clinical application of cells derived of amniotic membrane in veterinary medicine.

Assuntos

RESUMO

The immune system is mainly responsible for protecting the organism against agents that may interfere in its homeostasis. Thus, understand how this system develops and operates is very important, for create new therapies to assist this system in its operation, such as its failure. In domestic dogs, few studies show how actually occurs the development, maturation and functioning of the immune system. Therefore, this study demonstrates the development and possible activation of it on dog fetus from late gestational period by in situ and microscopic analyzes.

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RESUMO

Traumas são a segunda maior causa de morte em gatos, perdendo apenas para envenenamentos. No presente trabalho, 15 tutores de gatos paraplégicos responderam um questionário online. A causa de maior prevalência da paraplegia foi maus tratos (40%). A média de idade foi de 1 ano e 7 meses. Em média, os animais fazem/fizeram tratamento durante 11 meses, sendo que 26,7% só utilizou medicação anti-inflamatória e analgésica, enquanto dentre os demais, 27,3% foram submetidos à cirurgia, 72,7% fizeram acupuntura, 45,5% fisioterapia, 9% terapia celular e 9% outros. Entretanto, 33,3% dos tutores não observaram melhora. As doenças concomitantes mais predominantes foram constipação e retenção urinária (53,5%), e cistites recorrentes (40%), seguidas de escaras (33,3%), incontinência fecal e urinária (26,7%), dermatite de contato na região da fralda (6,7%) e necrose de cauda (6,7%). 40% dos tutores cogitaram ou foram instruídos a realizar a eutanásia, sendo que 16,7% optaram pelo procedimento. Apesar da paraplegia, 93,3% dos tutores consideram que seus animais apresentam boa qualidade de vida.

Traumas are the second largest cause of death in cats, behind only for poisonings. In the present study, 15 owners of paraplegic cats answered an online questionnaire. The greatest prevalence of paraplegia was animal cruelty (40%). The mean age was 1 year and 7 months. On average, the animals treated for 11 months, with 26.7% only using anti-inflammatory and analgesic medication, while among the others, 27.3% underwent surgery, 72.7% acupuncture, 45.5 % physiotherapy, 9% cell therapy and 9% others. However, 33.3% of the owners did not notice improvement. The most frequent concomitant diseases were constipation and urinary retention (53.5%), and recurrent cystitis (40%), followed by pressure sores (33.3%), fecal and urinary incontinence (26.7%), diaper dermatitis (6.7%) and tail necrosis (6.7%). 40% of the owners considered or were instructed to perform euthanasia, and 16.7% opted for the procedure. Despite paraplegia, 93.3% of the owners considered that their animals had a good quality of life.

Traumatismos son la segunda causa de muerte en gatos, perdiendo sólo para envenenamientos. En el presente trabajo, 15 tutores de gatos parapléjicos respondieron un cuestionario en línea. La causa de la mayor prevalencia de la paraplejia fue maltrato (40%). EI promedio de edad fue de 1 ano y 7 meses. En promedio, los animales hacen/han hecho un tratamiento durante 11 meses, siendo que el 26,7% sólo utilizó medicación antiinflamatoria y analgésica, mientras que los demás, el 27,3% fueron sometidos a la cirugía, el 72,7% hicieron acupuntura, 45,5 % fisioterapia, 9% terapia celular y 9% otros. Sin embargo, el 33,3% de los pacientes no observaron mejoría. Las enfermedades concomitantes más predominantes fueron constipación y retención de orina (53,5%), y cistitis recurrentes (40%), seguidas de escaras (33,3%), incontinencia fecal y urinaria (26,7%), dermatitis de contacto región del panal (6,7%) y necrosis de cola (6,7%). EI 40% de los tutores pensó o fue instruido para realizar una eutanasia, y el 16,7% optó por el procedimiento. A pesar de la paraplejia, el 93,3% de los derechos de autor en sus animales presentan buena calidad de vida.

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RESUMO

Traumas são a segunda maior causa de morte em gatos, perdendo apenas para envenenamentos. No presente trabalho, 15 tutores de gatos paraplégicos responderam um questionário online. A causa de maior prevalência da paraplegia foi maus tratos (40%). A média de idade foi de 1 ano e 7 meses. Em média, os animais fazem/fizeram tratamento durante 11 meses, sendo que 26,7% só utilizou medicação anti-inflamatória e analgésica, enquanto dentre os demais, 27,3% foram submetidos à cirurgia, 72,7% fizeram acupuntura, 45,5% fisioterapia, 9% terapia celular e 9% outros. Entretanto, 33,3% dos tutores não observaram melhora. As doenças concomitantes mais predominantes foram constipação e retenção urinária (53,5%), e cistites recorrentes (40%), seguidas de escaras (33,3%), incontinência fecal e urinária (26,7%), dermatite de contato na região da fralda (6,7%) e necrose de cauda (6,7%). 40% dos tutores cogitaram ou foram instruídos a realizar a eutanásia, sendo que 16,7% optaram pelo procedimento. Apesar da paraplegia, 93,3% dos tutores consideram que seus animais apresentam boa qualidade de vida.(AU)

Traumas are the second largest cause of death in cats, behind only for poisonings. In the present study, 15 owners of paraplegic cats answered an online questionnaire. The greatest prevalence of paraplegia was animal cruelty (40%). The mean age was 1 year and 7 months. On average, the animals treated for 11 months, with 26.7% only using anti-inflammatory and analgesic medication, while among the others, 27.3% underwent surgery, 72.7% acupuncture, 45.5 % physiotherapy, 9% cell therapy and 9% others. However, 33.3% of the owners did not notice improvement. The most frequent concomitant diseases were constipation and urinary retention (53.5%), and recurrent cystitis (40%), followed by pressure sores (33.3%), fecal and urinary incontinence (26.7%), diaper dermatitis (6.7%) and tail necrosis (6.7%). 40% of the owners considered or were instructed to perform euthanasia, and 16.7% opted for the procedure. Despite paraplegia, 93.3% of the owners considered that their animals had a good quality of life.(AU)

Traumatismos son la segunda causa de muerte en gatos, perdiendo sólo para envenenamientos. En el presente trabajo, 15 tutores de gatos parapléjicos respondieron un cuestionario en línea. La causa de la mayor prevalencia de la paraplejia fue maltrato (40%). EI promedio de edad fue de 1 ano y 7 meses. En promedio, los animales hacen/han hecho un tratamiento durante 11 meses, siendo que el 26,7% sólo utilizó medicación antiinflamatoria y analgésica, mientras que los demás, el 27,3% fueron sometidos a la cirugía, el 72,7% hicieron acupuntura, 45,5 % fisioterapia, 9% terapia celular y 9% otros. Sin embargo, el 33,3% de los pacientes no observaron mejoría. Las enfermedades concomitantes más predominantes fueron constipación y retención de orina (53,5%), y cistitis recurrentes (40%), seguidas de escaras (33,3%), incontinencia fecal y urinaria (26,7%), dermatitis de contacto región del panal (6,7%) y necrosis de cola (6,7%). EI 40% de los tutores pensó o fue instruido para realizar una eutanasia, y el 16,7% optó por el procedimiento. A pesar de la paraplejia, el 93,3% de los derechos de autor en sus animales presentan buena calidad de vida.(AU)

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Duchenne Muscular Dystrophy (DMD) is the most common X-linked muscular disease affecting humans. The Golden Retriever Muscular Dystrophy model (GRMD) is considerthe most suitable for several studies. This assay aims to quantify lymphocyte subpopulations CD4, CD5, and CD8, and standardize, the serum electrophoretic profile, to understand their contribution to the pathologic process in normal Golden Retriever dogs (GR group) and dystrophic´s (GRMD group), through the umbilical cord blood, in dogs aged from 2 to 3 months (GR II and GRMD II), and in dogs over 1 year of age (GR III and GRMD III). No significant differences were observed between the CD8+ lymphocyte subpopulations of the groups studied. The CD4+ and CD5+ lymphocyte subpopulations were significantly higher in the GRMD III group compared to the GR III group. Twenty-two different proteins in the gel were identified. The serum concentrations of the proteins belonging to the GR I and GRMD I groups were significantly lower than those of the other groups. We show that expression of acute phase proteins are worst during the aging of the dogs. We hope to expand knowledge to better understand the GRMD model and the translational data.

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Attention-Deficit/Hyperactivity Disorder (ADHD) is a common and highly heritable neuropsychiatric disorder. Despite the high heritability, the unraveling of specific genetic factors related to ADHD is hampered by its considerable genetic complexity. Recent evidence suggests that gene-gene interactions can explain part of this complexity. We examined the impact of strongly supported interaction effects between the LPHN3 gene and the NTAD gene cluster (NCAM1-TTC12-ANKK1-DRD2) in a 7-year follow-up of a clinical sample of adults with ADHD, addressing associations with susceptibility, symptomatology and stability of diagnosis. The sample comprises 548 adults with ADHD and 643 controls. Entropy-based analysis indicated a potential interaction between the LPHN3-rs6551665 and TTC12-rs2303380 SNPs influencing ADHD symptom counts. Further analyses revealed significant interaction effects on ADHD total symptoms (p=0.002), and with hyperactivity/impulsivity symptom counts (p=0.005). In the group composed by predominantly hyperactive/impulsive and combined presentation, the presence of LPHN3-rs6551665 G allele was related to increased ADHD risk only in individuals carrying the TTC12-rs2303380 AA genotype (p=0.026). Also, the same allelic constellation is involved in maintenance of ADHD in a predominantly hyperactive/impulsive or combined presentation after a 7-year follow-up (p=0.008). These observations reinforce and replicate previous evidence suggesting that an interaction effect between the LPHN3 gene and the NTAD cluster may have a role in the genetic substrate associated to ADHD also in adults. Moreover, it is possible that the interactions between LPHN3 and NTAD are specific factors contributing to the development of an ADHD phenotype with increased hyperactivity/impulsivity that is maintained throughout adulthood.

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