Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

Abstract The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.

Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.(AU)

Cutaneous myxoma in broiler chicken detected during slaughtering

Background: A cutaneous or superficial myxoma is a benign neoplasm of dermal or subcutaneous fibroblast origin. Although rare, it has been previously described in several species, including poultry. It presents as a single node or soft mass with a gelatinous cut surface. Histopathological analysis is essential for diagnosis and to differentiate it from other mesenchymal neoplasms and inflammatory or degenerative processes. Microscopically, it consists of dermal or subcutaneous lobules of plump, stellate, or spindle-shaped, bland-looking cells embedded in a basophilic myxoid matrix. This report describes the pathological findings in a rare case of cutaneous myxoma in a 42-day-old broiler flock. Cases: During ante mortem inspection of a 42-day-old broiler flock at a slaughterhouse under the authority of the Federal Inspection Service (southern Brazil), nodular lesions or encrusted areas with yellow and black areas were observed in the head skin of less than 1% of animals. These lesions, approximately 0.5 cm in diameter, were observed on the comb, in the periocular skin region, and close to the animals' nostrils. During the breeding period, no health or epidemiological events were observed. Fragments of the lesions in the comb and periocular skin were collected and fixed in buffered 10% formalin. The samples were sent to the laboratory, routinely processed, and stained with hematoxylin and eosin and Alcian blue. Microscopically, the lesions consisted of irregular multifocal proliferation of connective tissue showing spindle cells with poorly demarcated borders and scarce cytoplasm in a slightly basophilic myxoid aspect matrix. The adjacent epidermis is compressed due to neoplastic proliferation. No areas of epithelial hyperplasia or inclusion bodies were observed. According to the pathologic description and considering its descriptive epidemiology, our main clinical suspicion was cutaneous fowl pox, a pathology characterized by the appearance of nodules in regions devoid of feathers. However, the microscopic changes observed were compatible with those described for cutaneous myxomas. In addition, the extracellular matrix was positive for Alcian Blue staining, which is an indicator of myxoma. In the present case, the SIF did not report the same macroscopic lesions in other flocks of the same origin. Discussion: Connective tissue tumors, including myxomas, occur considerably less frequently under field conditions. In addition, these neoplasms are more frequent in mature birds and are not usually described in broilers, as observed in this report. The cutaneous myxoma described in broilers is usually a sporadic neoplasm that does not cause zootechnical losses, as observed in the case report. Its etiology is unknown and has been associated with various factors, such as local trauma and foreign bodies. Some fragments of plant material from the breeding environment were microscopically detected in the encrusted areas, which may indicate previous trauma or a foreign body. Myxoma has been associated with avian leukosis virus (ALV) subgroup A, but SIF did not report the same macroscopic lesions in other flocks of the same breeder hen's origin in the present case. Furthermore, sporadic connective tissue tumors associated with the virus occur in mature chickens but not in broilers. Myxoma lesions should be considered in the differential diagnosis of other connective tissue tumors and infectious agents that cause lesions in the skin and subcutaneous tissue.

Paclobutrazol in the in vitro conservation of cassava genotypes

In vitro germplasm conservation allows to extend the interval between subcultures without compromising the viability and genetic integrity of the plant, ensuring a backup of genotypes with high phytosanitary quality. Thus, this study aimed to verify the effect of four concentrations of Paclobutrazol® in inducing minimum growth in five Manihot esculenta accessions from the in vitro Active Germplasm Bank of Embrapa Cassava and Fruits. An experiment was installed using the Murashige and Skoog medium without addition and added with four concentrations of Paclobutrazol® (0.10, 0.20, 0.30, and 0.40 mg L-1), in five in vitro accessions of M. esculenta: BRS Jari (BGM 2041), Cigana (BGM 0264), BRS Poti Branca (BGM 2017), TME 14, and BRS Novo Horizonte. The statistical design was completely randomized in a 5 x 5 factorial scheme, with 15 repetitions. After 120 days of cultivation, the following variables were evaluated: plant height (cm), number of green leaves, number of senescent leaves, number of mini-cuttings, number of shoots, and fresh and dry mass of shoots and roots (mg). Paclobutrazol® caused a reduction in plant height and gain in root mass for all accessions, in addition to preserving the number of green leaves and decreasing leaf senescence for most genotypes. There was a strong dependence of the genotype in relation to the concentration of Paclobutrazol®. The concentration of 0.20 mg L-1 showed potential in the in vitro conservation of M. esculenta genotypes.(AU)

Expression analysis of m6A-related genes in various tissues of Meishan pigs at different developmental stages

To characterize the N6-methyladenosine (m6A)-related gene expression profiles in various tissues of Meishan pigs at different stages, m6A modification-related genes (METTL3, METTL14, METTL16, WTAP, RBM15, and FTO) were detected from newborn to physical maturity of Meishan pigs at eight important developmental stages (1, 7, 14, 21, 28, 35, 134, and 158 days old). The expression of m6A-related genes was tissue-specific. Furthermore, the level of METTL3 messenger RNA (mRNA) was higher on day 35 than in other stages in most tissues, and the expression of METTL14 increased after day 35, and FTO exhibited a peak on day 14 in muscle, intestine, lymph nodes, thymus, and kidney. This study provided a reference for an in-depth study of the expression patterns of m6A modification-related genes in Meishan pigs.

Efficiency analysis of commercial polymeric membranes for bone regeneration in rat cranial defects

Purpose: To evaluate the in vivo efficiency of commercial polymeric membranes for guided bone regeneration. Methods: Rat calvarial critical size defects was treated with LuminaCoat (LC), Surgitime PTFE (SP), GenDerm (GD), Pratix (PR), Techgraft (TG) or control (C-) and histomorphometric analysis determined the percentage of new bone, connective tissue and biomaterial at 1 or 3 months. Statistical analysis used ANOVA with Tukey's post-test for means at same experimental time and the paired Student's t test between the two periods, considering p < 0.05. Results: New bone at 1 month was higher for SP, TG and C-, at 3 months there were no differences, and between 1 and 3 months PR had greater increase growthing. Connective tissue at 1 month was higher for C-, at 3 months for PR, TG and C-, and between 1 and 3 months C- had sharp decline. Biomaterial at 1 month was higher for LC, in 3 months for SP and TG, and between 1 and 3 months, LC, GD and TG had more decreasing mean. Conclusion: SP had greater osteopromotive capacity and limitation of connective ingrowth, but did not exhibit degradation. PR and TG had favorable osteopromotion, LC less connective tissue and GD more accelerated biodegradation.

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT Raman spectroscopy / Cultivo de células-tronco mesenquimais de rato sobre arcabouços de PHBV-PCL: análise do meio de cultura condicionado por espectroscopia de FT-Raman

Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37ºC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a [...].

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT-Raman spectroscopy

Abstract Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37oC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

Resumo As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a análise. Não houve variações nas leituras entre as amostras estudadas, concluindo-se que a FT-Raman não atendeu às expectativas nas condições estudadas.

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT-Raman spectroscopy / Cultivo de células-tronco mesenquimais de rato sobre arcabouços de PHBV-PCL: análise do meio de cultura condicionado por espectroscopia de FT-Raman

Abstract Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37oC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

Resumo As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a análise. Não houve variações nas leituras entre as amostras estudadas, concluindo-se que a FT-Raman não atendeu às expectativas nas condições estudadas.

Lesão facial grave em um gambá-de-orelha-branca (Didelphis albiventris) - tratamento com ozonioterapia / Severe facial wound in a white-eared opossum (Didelphis albiventris) - treatment with ozone therapy

Background: Ozone therapy (OT) is a type of treatment that uses ozone gas, which has high oxidizing power. OT has been utilized for various diseases and to accelerate the wound healing process and reduce inflammation. Additionally, OT is a low-cost treatment recommended for patients that can be easily restrained. There are several methods of ozone administration, including topical use through bagging, cupping, or rectal insufflation, in addition to ozonated autohemotherapy. Based on the benefits of OT and the scarcity of reports describing its effects on white-eared opossums (Didelphis albiventris), we report on the use of ozone in the treatment of a severe facial wound in a white-eared opossum. Case: An adult female white-eared opossum weighing 1.6 kg was rescued from a fire. The patient was pregnant, dehydrated, hypothermic and had an extensive burn wound in the facial region. Due to the stress of captivity and its aggressive behavior, the animal needed to be chemically restrained with dexmedetomidine and butorphanol. Following sedation, the cephalic vein was catheterized and intravenous fluid therapy with ringer lactate was started to treat dehydration. Next, enrofloxacin, meloxicam, and tramadol hydrochloride were administered. Body rewarming was performed with thermal bags. The microbial load of the wound was reduced with chlorhexidine degerming solution followed by washing with saline solution. A thin layer of silver sulfadiazine cream was applied to the wound area. Over the following days the patient was treated with OT using the cupping technique combined with ozonated sunflower oil. After the first applications there was an improvement in the appearance of the wound, which initially had a purulent aspect. After 20 days of treatment the wound showed good granulation and there were no signs of infection, and at 45 days of treatment it was completely healed. The patient was then released with her baby. Discussion: Ozone therapy was included in the patient's treatment plan to enhance the tissue repair process in order to reintroduce the animal to the wild in the shortest possible time. The strategy was successful in improving healing and reducing tissue inflammation and infection. Ozone is known to cause the inactivation of microorganisms as a result of the oxidation of phospholipids and lipoproteins present in the cell membrane of bacteria and to interrupt viral replication. Ozone helps in the perfusion of injured tissues, contributing to the entry of humoral components, which control inflammation. Moreover, ozone contributes to increase supply of oxygen and nutrients to tissues and to stimulate increased production of fibrinogen and formation of granulation tissue. Ozone increases the enzymatic antioxidant response, while also playing a role in immunoregulation. Ozone can be used in several ways and via various methods with potential routes including intravenous, urethral, intramuscular, subcutaneous, or intra-articular, in addition to rectal and intramammary insufflation. Immunomodulatory, anti-inflammatory, analgesic, and antiseptic effects can be achieved with different doses and concentrations, which enables its application in different types of pathologies. Because of the aggressive behavior of the animal, in the current case we opted for the cupping technique combined with ozonated oil, both techniques which can be administered easily with only physical restraint of the patient. Cupping allows for the provision of a higher concentration of ozone gas locally whereas the ozonated oil maintains the effect of the gas for slightly longer, enhancing its effect and promoting tissue repair. The present report showed that OT is an effective alternative for the treatment of burn wounds in opossums, without causing adverse effects to the patient or her baby.

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT Raman spectroscopy / Cultivo de células-tronco mesenquimais de rato sobre arcabouços de PHBV-PCL: análise do meio de cultura condicionado por espectroscopia de FT-Raman

Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37ºC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.(AU)

As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a [...].(AU)

Protocol for in vitro rooting of Pyrus comunnis rootstocks / Protocolo de enraizamento in vitro de porta-enxertos de Pyrus comunnis

Effective protocols for in vitro rooting for woody fruit trees are still a challenge for in vitro seedling production, especially when there is a need to insert new cultivars or rootstocks. These protocols are essential to accelerate studies in plant breeding programs and for seedling distribution. This study evaluated the use of 6-Benzylaminopurine (IBA) in in vitro rooting of Pyruscomunnis rootstocks, clones 'OHxF87' and Pyrodwarf. Explant exposure times (0, 24, 48, 72, and 96 hours) to 20 mg L-1 IBA were tested for in vitro rooting. The exposure to IBA resulted in rooting rates above 80%, surpassing some results reported in the literature. The 24-hour treatment provided 81,81% survival, leading to an average growth of five roots with 19 mm length, for 'OHxF87' rootstock. The same exposure time resulted in the highest survival rate (75%) and the highest mean root number, seven roots per plant with 10 mm length, for 'PDW' rootstock. Root formation did not occur in the absence of synthetic auxin. Therefore, it can be concluded that a 24-hour exposure at 20 mg L-1 IBA was sufficient to promote in vitro rooting in 'OHxF87' and Pyrodwarf rootstocks'.

Protocolos eficazes de enraizamento in vitro de frutíferas lenhosas ainda são um desafio para produção de mudas in vitro, especialmente quando há necessidade de inserção de novas cultivares ou porta-enxerto. Esses protocolos são essenciais para acelerar estudos nos programas de melhoramento genético e também para distribuição posterior das mudas. Nesse sentido, o objetivo deste estudo foi avaliar a utilização da 6-Benzilaminopurina no enraizamento in vitro de porta-enxerto Pirus comunnis, clones 'OHxF87' e Pyrodwarf. Para o enraizamento, foi testado o tempo de exposição dos explantes ao AIB. Para tanto, foram utilizados 20 mg L-1 do fitohormônio nas horas 0, 24, 48, 72 e 96 horas. A exposição ao AIB resultou em taxas de enraizamento acima de 80%, superando alguns resultados encontrados na literatura. Para o porta-enxerto 'OHxF87', o tratamento de 24 horas proporcionou 81,81% de sobrevivência, promovendo em média cinco raízes com comprimento de 19 mm. O mesmo tratamento para o porta-enxerto 'PDW' resultou na maior taxa de sobrevivência (75%), bem como no maior número médio de raízes, sete raízes por planta, com comprimento de 10 mm. Na ausência de auxina sintética, a formação de raízes não ocorreu. Assim sendo, podemos concluir que o tempo de exposição de 24 horas a 20 mg L-1 de IBA foi suficiente para promover o enraizamento de porta-enxertos Pyrus communis de 'OHxF87' e 'PDW'.

Biobanking and use of gonadal tissues - a promising strategy for conserving wildlife from the Caatinga biome

Biological Resource Banks (BRB) or Genetic Resource Banks (GRB) are critical tools for the conservation of animal biodiversity. According to the International Union for Conservation of Nature, more than 38,500 species are threatened with extinction, out of a total of 138,300 surveyed species. These banks are repositories of biological samples and data recovered and preserved for the long term by zoos, universities, research centers and other conservation organizations. In recent years, BRB have increasingly included ovarian and testicular tissues as additional options to rescue and propagate wild species, especially those at risk of extinction. After in vitro culture or grafting, gonadal tissues are potential sources of matured gametes that can be used for Assisted Reproduction Technologies while informing about gametogenesis or mechanisms involved in infertility. It therefore is crucial to properly recover, cryopreserve, and culture these tissues using species-specific protocols. Developing BRBs is currently one of the strategies to preserve species from the Caatinga biome - an exclusively Brazilian biome with a rich wild fauna that suffers from anthropogenic activities. Among wild species from this biome, studies have been primarily conducted in collared peccaries, agoutis, cavies, and armadillos to preserve their ovarian and testicular tissues. Additionally, domestic species such as the domestic cat and donkeys have been proposed as models for wild species that are phylogenetically close. This review addresses the main technical aspects involved in obtaining BRB derived from gonadal tissues in some wild species of the Caatinga biome. It reports recent advances and perspectives to use these biological materials for wildlife conservation.(AU)

Activated charcoal in the control of oxidation and in vitro growth of Dioscorea spp. nodal segments

In vitrooxidationisaproblem forsome herbaceous and woody species and can causedarkening of tissues and consequently death of explantsand plants Therefore, this study aimed to assess the effect of activated charcoal on in vitro yamcultivation, aiming at reducing or eliminating explant oxidation and optimizing the growth of the genotypes Dioscorea alata var.purpurea(Roxb.) A. Pouchet and Dioscorea rotundataPoir. Nodal segments of approximately 1 cm, extracted from plants previously grown in vitro, were introduced into test tubes containing 10 mL of 2GGC culture medium, plus 30 g L-1sucrose, solidified with 2.2 g L-1Phytagel®and pH adjusted to 5.8 before autoclaving, containing activated charcoal doses of 0, 1, 2, 3 and 4 g L-1. Plants were maintained for 90 days in a growth room, with temperature of 27 ± 1ºC, photon flux density of 30 µmol m-2s-1and photoperiod of 16 hours, after which their development variables were evaluated. Activated charcoal, at the concentration of 4 g L-1considerably promoted the best development of plants, and the species D. alatavar.purpureashowed higher means for all variables studied.(AU)

Development and characterization of poultry collagen-based hybrid hydrogels for bone regeneration

Purpose: Poultry by-products can contribute as an innovative natural source for the development of composites based on polymers and minerals aiming at bone regeneration. The objective of this study was the physicochemical and biological characterization of collagen-based hydrogels crosslinked with ultraviolet (UV)-riboflavin. Methods: Pure hydrogels of 100% collagen (G1) or hybrid hydrogels, 90% collagen:10% apatite (G2), 90% collagen:10% nanokeratin (G3), and 90% collagen:5% apatite:5% nanokeratin (G4) were characterized by scanning electron microscope, Fourier-transform infrared spectroscopy, differential scanning calorimetry, swelling degree and quali-quantitative histological analysis. Ectopic implantation in subcutaneous tissue in mice at one, three and nine weeks allowed to assess the inflammation (neutrophils, lymphocytes, macrophages, and giant cells) and repair (neovascularization, and connective tissue) to determine biocompatibility and the integrity of biomaterials to score their biodegradability. Histomorphometry on critical size defects in rat calvaria at one and three months evaluated the percentage of bone, connective tissue, and biomaterials in all groups. Results: The hydrogels presented porous microstructure, water absorption and physicochemical characteristics compatible with their polymeric and/or mineral composition. All materials exhibited biocompatibility, biodegradability, and low osteoconductivity. G2 showed greater density of new bone and biomaterial than the G1, G3 and G4. Conclusions: The collagen-apatite group formulation suggests potential for development as osteopromoting membrane.

Anatomia e histologia do esôfago e vasos sanguíneos em Cães ­ Revisão de literatura / Anatomy and histology of the esophagus and blood vessels in dogs ­ Literature review

O sistema digestório é composto por órgãos de morfologia tubular, e glândulas associadas; tendo como principal função o processamento de alimentos e a absorção de nutrientes dos mesmos. Há um padrão estrutural comum entre órgãos tubulares deste sistema, o conhecimento desse padrão, facilita a compreensão das particularidades de cada órgão. Já os vasos sanguíneos são caracterizados e definidos por sua posição dentro do circuito vascular, pela estrutura individual, e mecanismos de controle da função vascular. Dessa forma, o objetivo do presente trabalho é aprofundar conhecimentos a respeito de anatomia e histologia do esôfago e vasos sanguíneos em cães.(AU)

The digestive system is composed of tubular morphology organs and associated glands; with the main function of food processing and nutrient absorption. There is a common structural pattern among tubular organs of this system, knowledge of this pattern facilitates the understanding of the particularities of each organ. Blood vessels are characterized and defined by their position within the vascular circuit, their individual structure, and the mechanisms that control vascular function. Thus, the objective of the present work is to deepen knowledge about the anatomy and histology of the esophagus and blood vessels.(AU)

Biobancos para a conservação da vida silvestre: desafios e perspectivas / Biobanks for wildlife conservation: challenges and prospects

Um repositório de amostras biológicas, ou biobanco, pode ser definido como uma coleção-base de amostras orgânicas de origem humana, animal, vegetal ou microbiana, destinados para a conservação da ampla variabilidade dos recursos genéticos, apresentando-se como uma alternativa para a manutenção da biodiversidade. Esses repositórios são instrumentos indispensáveis no proposito de salvaguardar e coordenar um conjunto de informações da fauna silvestre, dando origem a uma rede de dados para o desenvolvimento de estratégias para a conservação de espécimes com genótipos raros ou ameaçados. Portanto, esta revisão tem como objetivo ressaltar a importância e aplicabilidade dos biobancos para a conservação da vida silvestre, destacando o estado da arte, e enfatizando os aspectos técnicos, procedimentos necessários para sua implementação e quais os principais desafios e pespectivas para o aprimoramento e utilização desse recurso na biotecnologia reprodutiva.(AU)

A repository of biological samples or biobanks can be defined as a base collection of organic samples of human, animal, plant or microbial origin, intended for the conservation of the wide variability of genetic resources, presenting itself as an alternative for the maintenance of the biodiversity. These repositories are indispensable instruments for the purpose of safeguarding and coordinating a set of information on wildlife, giving rise to a data network for the development of strategies for the conservation of specimens with rare or endangered genotypes. Therefore, this review aims to highlight the importance and applicability of biobanks for the conservation of wildlife, highlighting the state of the art, and emphasizing the technical aspects, as well as the necessary procedures for its implementation and what are the main challenges and perspectives for the improvement and use of this resource in reproductive biotechnology.(AU)

Conventional vs. advanced echocardiographic parameters for predicting fluid responsiveness in healthy dogs

Background: Volume replacement is considered an essential component in the treatment of critically ill patients, but excess f luid administration can be as harmful as the lack of fluid therapy. Therefore, the volume replacement is only beneficial in patients deemed to be fluid responsive, that is, when the individual has an increase in their stroke volume after administration of a volume challenge. This study aim to assess whether conventional and advanced echocardiographic parameters for assessing left ventricular systolic function are able to predict fluid responsiveness in healthy spontaneously breathing dogs. The hypothesis was that some of these parameters would differ between responsive and non-responsive animals and could be used as a complementary measure for assessment of fluid responsiveness. Materials, Methods & Results: A prospective study was carried out in which 22 client-owned healthy dogs over 1 year of age that presented to the Veterinary Medical Teaching Hospital for elective neutering procedure were included. All dogs underwent conventional and advanced echocardiographic examination to obtain the left ventricular internal diameter in diastole normalized (LVIDDn), left ventricular internal diameter in systole normalized (LVIDSn), ejection fraction (EF) and shortening fraction (SF) according to the Teischholz methodology, global longitudinal strain (GLS) and tissue motion annular displacement (TMAD). There parameters were evaluated before and after administration of a volume challenge with 10 mL/kg lactate ringer intravenously for 20 min. Based on the variation in aortic velocity integral time, 31.82% of dogs were considered responsive and 68.18% were non-responsive to the volume challenge. For advanced echocardiography, TMAD indexed by body weight (mm/kg) > 0.89 (P = 0.004) and body surface area (mm/m2) >18.9 (P = 0.004) after volume challenge had the best area...

Pouteria ramiflora leaf extract on emulgel in wound healing activity in diabetic rats / Ação do extrato de folhas de Pouteria ramiflora em emulgel na atividade de cicatrização de feridas em ratos diabéticos

The genus Pouteria has been studied because it presents various activities, among which is its anti-inflammatory potential. The effects of Pouteria ramiflora Carbopol gel on the healing of skin wounds in diabetic rats were evaluated by microscopic imaging. Streptozotocin was administered intraperitoneally in animals that had fasted for 12 hours, a situation confirmed by the glycemic index (> 240 mg dL-¹). An excision on the back of the animals was performed and three groups were formed: Control (Gel), Ethanolic extract (Ext) and Gel + extract 2% (Ext+gel); the histopathological evaluation occurred on the 7th, 14th, 21st and 30th days after the post-operative period. The results of the phytochemical prospecting of P. ramiflora extract demonstrated the major presence of phenolic compounds and flavonoids; the assessment of the inflammatory infiltrate on the 7th day was higher on group Ext and Ext+gel when compared to group Control; on the 14th day control and Ext (p<0.05). The quantification of fibroblasts was higher on the 7th day among the three treatments, control and Ext (p<0.05), on the 21st day. Angiogenesis showeda higher number of vessels in Ext+gel group (p<0.05) on the 7th day; in Control, Ext and Ext+gel (p<0.05) on the 14th day; and Control and Ext (p<0.05)on the 21st day. The histopathological results showed that the formulation Ext+gel was efficient in tissue reparation and decrease in inflammatory cells on the diabetic's animals.

O gênero Pouteria apresenta várias aplicações terapêuticas e, dentre elas, grande potencial antiflamatório. Os efeitos do gel de Pouteria ramiflora sobre a cicatrização de feridas na pele de ratos diabéticos foram avaliados pela histomorfometria. A estreptozotocina foi administrada por via intraperitoneal em animais após jejum de 12 horas, a confirmação de indução da diabetes foi confirmada pelo índice glicêmico (> 240 mg dL-1). Foi realizada uma incisão no dorso do animal e foram criados 3 grupos de tratamento: controle (gel carbopol), extrato etanólico (Ext) e Gel + extrato etanólico à 2% (Ext+gel); a avaliação histopatológica foi realizada no 7º, 14º, 21º e 30º dias após o período pós operatório. Os resultados da prospecção fitoquímica dos extratos de P. ramiflora demonstraram majoritariamente a presença de compostos fenólicos e flavonóides; o infiltrado inflamatório avaliado no 7º dia foi maior para animais do grupo controle em relação aos grupos Ext (p<0.05) e Ext+gel 2% (p<0.05); no 14º dia o controle e Exp (p<0.05) apresentaram aumento significativo dos infiltrados inflamatórios. A presença de fibroblastos foi elevada no 7º dia em todos os tratamentos. O processo da angiogênese mostrou um maior número de vasos sanguíneos entre os grupos Ext e Ext+gel (p<0.05) no 7º dia; no 14º dia o grupo controle, Ext (p<0.05), Control e Ext+gel (p<0.05) apresentaram aumento de vascularização, e no 21º dia apenas os grupos controle e Ext (p<0.05). Os resultados histopatológicos mostraram que a formulação gel carbopol + extrato etanólico a 2% foi eficiente na reparação de tecidos e na diminuição de células inflamatórias nos animais diabéticos.

Phytotoxicity of plant extracts of Vismia japurensis cultivated in vivo and in vitro / Fitotoxicidade de extratos de plantas de Vismia japurensis cultivadas in vivo e in vitro

Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.

Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.