Use of new recombinant proteins for ovarian stimulation in ruminants

Currently, gonadotropin products (follicle stimulating hormone, FSH, and luteinizing hormone, LH) used in animal reproduction are produced by extraction and purification from abattoir-derived pituitary glands. This method, relying on animal-derived materials, carries the potential risk of hormone contamination and pathogen transmission. Additionally, chorionic gonadotropins are extracted from the blood of pregnant mares (equine chorionic gonadotropin; eCG) or the urine of pregnant women (human chorionic gonadotropin; hCG). However, recent advancements have introduced recombinant gonadotropins for assisted animal reproduction therapies. The traditional use of FSH for superovulation has limitations, including labor requirements and variability in superovulation response, affecting the success of in vivo (SOV) and in vitro (OPU/IVEP) embryo production. FSH treatment for superstimulation before OPU can promote the growth of a homogenous follicular population and the recovery of competent oocytes suitable for IVEP procedures. At present, a single injection of a preparation of long-acting bovine recombinant FSH (rFSH) produced similar superovulation responses resulting in the production of good-quality in vivo and in vitro embryos. Furthermore, the treatment with eCG at FTAI protocol has demonstrated its efficacy in promoting follicular growth, ovulation, and P/AI, mainly in heifers and anestrous cows. Currently, treatment with recombinant glycoproteins with eCG-like activity (r-eCG) have shown promising results in increasing follicular growth, ovulation, and P/AI in cows submitted to P4/E2 -based protocols. Bovine somatotropin (bST) is a naturally occurring hormone found in cows. Recombinant bovine somatotropin (rbST), produced through genetic engineering techniques, has shown potential in enhancing reproductive outcomes in ruminants. Treatment with rbST has been found to improve P/IA, increase donor embryo production, and enhance P/ET in recipients. The use of recombinant hormones allows to produce non-animal-derived products, offering several advantages in assisted reproductive technologies for ruminants. This advancement opens up new possibilities for improving reproductive efficiency and success rates in the field of animal reproduction.(AU)

Hesperozygis ringens essential oil as an anesthetic for Colossoma macropomum during biometric handling / Óleo essencial de Hesperozygis ringens como anestésico para Colossoma macropomum durante manipulação biométrica

This study evaluated the effectiveness of the essential oil of Hesperozygisringens (EOHR) for anesthesia of Colossomamacropomum by documenting hematological and blood biochemical responses after biometric handling. In Experiment 1, juveniles (14.12 ± 3.53 g) were exposed to different concentrations of EOHR: 0 (control), 75, 150, 300 and 450 µL L-1 (n=10 fish for each concentration), to determine times for induction and recovery from anesthesia, as well as its effects on ventilatory frequency (VF). Based on these results, Experiment 2 evaluated the effects of 0 (control), 75 (with induction and recovery times outside that recommended for fish anesthesia) and 150 µL L-1 EOHR (within recommended times) on hematological and biochemical variables of juveniles (20.52 ± 3.47 g) after anesthesia and after 24 h of recovery (n = 6 fish for each concentration and collection time). Survival was 100%. Induction time showed a quadratic effect of EOHR concentration. Recovery time did not differ among EOHR concentrations. Concentrations between 150 and 450 µL L-1 EOHR caused rapid induction (< 3 min) and recovery (< 5 min). EOHR concentration affected VF. The concentration of 150 µL L-1 EOHR had little influence on hematological and biochemical parameters of C. macropomum of 20 g.

Este estudo avaliou a eficiência do óleo essencial de Hesperozygis ringens (EOHR) para anestesia de Colossoma macropomum, documentando as respostas hematológicas e bioquímicas do sangue após o manuseio biométrico. No experimento 1, juvenis (14,12 ± 3,53 g) foram expostos a diferentes concentrações de EOHR: 0 (controle), 75, 150, 300 e 450 µL L-1 (n = 10 peixes para cada concentração), para determinar os tempos de indução e recuperação da anestesia, bem como seus efeitos na frequência ventilatória (VF). Com base nesses resultados, o experimento 2 avaliou os efeitos de 0 (controle), 75 (com tempos de indução e recuperação fora do recomendado para anestesia de peixes) e 150 µL L-1 EOHR (dentro dos tempos recomendados) sobre variáveis hematológicas e bioquímicas de juvenis (20,52 ± 3,47 g) após a anestesia e após 24h de recuperação (n = 6 peixes para cada concentração e tempo de coleta). A sobrevivência foi de 100%. O tempo de indução mostrou efeito quadrático da concentração de EOHR. O tempo de recuperação não diferiu entre as concentrações de EOHR. Concentrações entre 150 e 450 µL L-1 EOHR causaram rápida indução (< 3 min) e recuperação (< 5 min). As concentrações de EOHR afetaram a VF. A concentração de 150 µL L-1 de EOHR teve pouca influência nos parâmetros hematológicos e bioquímicos de C. macropomum de 20 g.

Experimental model of nephropathy associated with diabetes mellitus in mice

Purpose: Nontransmissible chronic diseases, such as diabetes mellitus (DM) and nephropathy, affect a significant portion of the population, often treated due to injuries that require healing and regeneration. To create an experimental model of associated comorbidities, for healing and regeneration studies, protocols for induction of nephropathy by ischemia and reperfusion (I/R) and induction of DM by injection of streptozotocin (STZ) were associated. Methods: Sixty-four mice (Mus musculus), female, adult, Swiss strain, weighing approximately 20 g, were divided into four groups: G1: control (n = 24), G2: nephropathy group (N) (n = 7), G3, DM (n = 9), and G4: N+DM (n = 24). Arteriovenous stenosis (I/R) of the left kidney was performed as the first protocol. The animals received a hyperlipidemic diet for 7 days after the injection of STZ (150 mg/kg, via i.p.) and an aqueous glucose solution (10%) for 24 h. The animals in the G3 and G4 groups were observed for 14 days before receiving the diet and STZ. The evolution of nephropathy was observed using a urine test strip and the DM, through the analysis of blood glucose with a reagent strip on a digital monitor. Results: The ischemic induction protocols of nephropathy and DM with STZ, associated, were sustainable, low-cost, and without deaths. There were alterations compatible with initial renal alterations, in the first 14 days, such as increased urinary density, pH alteration, presence of glucose, proteins and leukocytes, when compared to the control group. DM was confirmed by the presence of hyperglycemia 7 days after induction and its evolution after 14 days. The animals in the G4 group showed constant weight loss when compared to the other groups. It was possible to observe morphological alterations in the kidneys submitted to I/R, regarding coloration, during surgery and after the end of the observation period, in the volume and size of the left kidney, when compared to the contralateral kidney. Conclusion: It was possible to induce nephropathy and DM associated in the same animal, in a simple way, confirmed with rapid tests, without losses, providing a basis for future studies.

Leaf mineral concentration of peach after the use of resistance inducers / Concentração mineral da folha do pessegueiro após o uso de indutores de resistência

ABSTRACT: This research studied the effect of the inducers acibenzolar-S-Methyl, potassium phosphite, and potassium silicate on the leaf mineral concentration of peach. The experiment was carried out in Tezontepec, Puebla, in the years 2017 and 2018. The treatments consisted of the foliar application of the resistance inducers acibenzolar-S-Methyl, potassium phosphite, and potassium silicate. Inducers were applied at commercially recommended dose. The experimental design was in randomized blocks with four replicates. Foliar quantification of nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, copper and silicon was carried out. The use of resistance inducers increases the leaf concentrations of calcium. The interaction of resistance inducers and years of application modified the foliar concentration of magnesium, phosphorus and sulfur in peaches.

RESUMO: O objetivo desta pesquisa foi estudar o efeito dos indutores acibenzolar-S-Metil, fosfito de potássio e silicato de potássio sobre o teor de minerais foliares do pessegueiro. O experimento foi realizado em Tezontepec, Puebla, nos anos de 2017 e 2018. Os tratamentos consistiram na aplicação foliar dos indutores acibenzolar-S-Metil, fosfito de potássio e silicato de potássio. Os indutores foram aplicados na dose comercialmente recomendada. O delineamento experimental foi em blocos casualizados com quatro repetições. Foi realizada a quantificação foliar de nitrogênio, fósforo, potássio, cálcio, magnésio, enxofre, cobre e silício. A interação de indutores de resistência e anos de aplicação modificou a concentração foliar de magnésio, fósforo e enxofre nas folhas de pessegueiro.

Determination of the minimum alveolar concentration (MAC) of isoflurane and sevoflurane in Callithrix penicillatac

Background: The minimum alveolar concentration (MAC) is a measure of quantitative anesthetic potency and has become the standard index for the evaluation and comparison of volatile anesthetics, in addition to guiding dose administration. Black-tufted marmosets (Callithrix penicillata) are primates present in the clinical and surgical routine of veterinary hospitals, as well as experimental models, especially in neuroscience. Few studies have evaluated the potency of the main volatile anesthetics in this species. This study aimed to determine the MAC of isoflurane and sevoflurane in C. penicillata using the up-and-down method and to evaluate the effects of these drugs on the quality of anesthetic induction, maintenance, and recovery. Materials, Methods & Results: Twenty-four animals of undetermined age were used. All marmosets were healthy according to hematological and physical evaluation. The animals were randomly divided into 2 groups: ISOMAC and SEVOMAC. Each animal was induced to general anesthesia in an anesthetic box with oxygen (5 L/min) and sevoflurane at 7% in the SEVOMAC group or isoflurane at 5% in the ISOMAC. Upon reaching lateral decubitus, orotracheal intubation was performed. General anesthesia was maintained with isoflurane or sevoflurane diluted in oxygen (0.8 L/min) using a non-rebreathing delivery system under spontaneous ventilation. As defined in the pilot study, the first animal from ISOMAC started the maintenance of anesthesia with 2.6% isoflurane, while the first animal in SEVOMAC received 4% sevoflurane. After finishing the instrumentation to assess heart rate, respiratory rate, systolic blood pressure, pulse oximeter oxygen saturation, end-tidal carbon dioxide concentration, and rectal temperature, a 15-min wait to reach anesthetic equilibrium was allowed, and then an electrical noxious stimulation (50 mA and 50 Hz) was performed on the lateral aspect of the thigh (a faradic current of 3 consecutive single stimuli, followed by 2 continuous stimuli). The animals' responses to the electrical stimulus were observed. The presence of a positive response (gross movement of the limbs, head, or vocalization) or a negative response (absence of gross movements) determined the increase or reduction, respectively, of the inhalation anesthetic concentration by 10% in the subsequent marmoset. The quality of anesthetic induction and recovery from anesthesia was evaluated using a scale that measured the intensity of agitation, coughing, nausea, and vomiting. Physiological variables were recorded before (M0) and after (M1) applying the nociceptive stimulus. Isoflurane and sevoflurane MAC values in C. penicillata were 2.29 ± 0.10% and 3.93 ± 0.61 % respectively. Physiological parameters, quality of anesthetic induction and recovery did not differ significantly between groups. However, isoflurane caused irritation of the airway and ocular mucous membranes, more coughing episodes, and tearing at induction. There was no difference between groups for time to extubation and recovery time to regain sternal position. Discussion: Previous studies in primates found lower MAC values for both anesthetics, except for Lemur catta. Those findings may be explained by the use of different nociceptive stimuli and the MAC determination method employed, although no differences in MAC values have been described between bracketing or up-and-down methods in human primates and dogs. It is unlikely that the stimulus and technique alone are the determining factors for the high concentration of isoflurane and sevoflurane observed in the present study since MAC was high with both halogenates, indicating that dose extrapolation from other species can lead to the wrong anesthetic dosage.

In vitro cytotoxicity of curcuminoids against head and neck cancer HNO97 cell line

Abstract Oral squamous cell carcinoma (OSCC) is a malignant tumour of Head and Neck Cancer (HNC). The recent therapeutic approaches used to treat cancer have adverse side effects. The natural agents exhibiting anticancer activities are generally considered to have a robust therapeutic potential. Curcuminoids, one of the major active compounds of the turmeric herb, are used as a therapeutic agent for several diseases including cancer. In this study, the cytotoxicity of curcuminoids was investigated against OSCC cell line HNO97. Our data showed that curcuminoids significantly inhibits the proliferation of HNO97 in a time and dose-dependent manner (IC50=35 M). Cell cycle analysis demonstrated that curcuminoids increased the percentage of G2/M phase cell populations in the treated groups. Treating HNO97 cells with curcuminoids led to cell shrinking and increased detached cells, which are the typical appearance of apoptotic cells. Moreover, flow cytometry analysis revealed that curcuminoids significantly induced apoptosis in a time-dependent manner. Furthermore, as a response to curcuminoids treatment, comet tails were formed in cell nuclei due to the induction of DNA damage. Curcuminoids treatment reduced the colony formation capacity of HNO97 cells and induced morphological changes. Overall, these findings demonstrate that curcuminoids can in vitro inhibit HNC proliferation and metastasis and induce apoptosis.

Resumo O carcinoma de células escamosas oral (OSCC) é um tumor maligno do câncer de cabeça e pescoço (HNC). As recentes abordagens terapêuticas usadas para tratar o câncer têm efeitos colaterais adversos. Os agentes naturais que exibem atividades anticâncer são geralmente considerados como tendo um potencial terapêutico robusto. Curcuminoides, um dos principais compostos ativos da erva cúrcuma, são usados como agente terapêutico para várias doenças, incluindo câncer. Neste estudo, a citotoxicidade dos curcuminoides foi investigada contra a linha de células OSCC HNO97. Nossos dados mostraram que os curcuminoides inibem significativamente a proliferação de HNO97 de forma dependente do tempo e da dose (IC50 = 35 M). A análise do ciclo celular demonstrou que os curcuminoides aumentaram a porcentagem de populações de células da fase G2 / M nos grupos tratados. O tratamento das células HNO97 com curcuminoides levou ao encolhimento celular e ao aumento das células destacadas, que são a aparência típica das células apoptóticas. Além disso, a análise de citometria de fluxo revelou que os curcuminoides induziram significativamente a apoptose de uma maneira dependente do tempo. Além disso, em resposta ao tratamento com curcuminoides, caudas de cometa foram formadas nos núcleos das células devido à indução de danos ao DNA. O tratamento com curcuminoides reduziu a capacidade de formação de colônias das células HNO97 e induziu alterações morfológicas. No geral, esses achados demonstram que os curcuminoides podem inibir in vitro a proliferação e metástase de HNC e induzir apoptose.

Enhancement of Annexin V in response to combination of epigallocatechin gallate and quercetin as a potent arrest the cell cycle of colorectal cancer

Abstract Colorectal cancer (CRC) is one of the most common cancers leading to comorbidities and mortalities globally. The rational of current study was to evaluate the combined epigallocatechin gallate and quercetin as a potent antitumor agent as commentary agent for therapeutic protocol. The present study investigated the effect of epigallocatechin Gallate (EGCG) (150mg) and quercetin (200mg) at different proportions on proliferation and induction of apoptosis in human colon cancer cells (HCT-116). Cell growth, colonogenic, Annexin V in addition cell cycle were detected in response to phytomolecules. Data obtained showed that, the colony formation was inhibited significantly in CRC starting from the lowest concentration tested of 10 µg/mL resulting in no colonies as visualized by a phase-contrast microscope. Data showed a significant elevation in the annexin V at 100 µg/mL EGCG(25.85%) and 150 µg/mL quercetin (48.35%). Moreover, cell cycle analysis showed that this combination caused cell cycle arrest at the G1 phase at concentration of 100 µg/mL (72.7%) and 150 µg/mL (75.25%). The combined effect of epigallocatechin Gallate and quercetin exert antiproliferative activity against CRC, it is promising in alternative conventional chemotherapeutic agent.

Resumo O câncer colorretal (CCR) é um dos cânceres mais comuns, levando a comorbidades e mortalidade em todo o mundo. O racional do presente estudo foi avaliar a combinação de galato de epigalocatequina e quercetina como um agente antitumoral potente como agente de comentário para protocolo terapêutico. O presente estudo investigou o efeito de galato de epigalocatequina (EGCG) (150 mg) e quercetina (200 mg) em diferentes proporções na proliferação e indução de apoptose em células de câncer de cólon humano (HCT-116). O crescimento celular, colonogênico, anexina V, além do ciclo celular foram detectados em resposta a fitomoléculas. Os dados obtidos mostraram que a formação de colônias foi inibida significativamente no CRC a partir da concentração mais baixa testada de 10 µg/mL, resultando em nenhuma colônia conforme visualizado por um microscópio de contraste de fase. Os dados mostraram uma elevação significativa na anexina V a 100 µg/mL de EGCG (25,85%) e 150 µg/mL de quercetina (48,35%). Além disso, a análise do ciclo celular mostrou que essa combinação causou parada do ciclo celular na fase G1 na concentração de 100 µg/mL (72,7%) e 150 µg/mL (75,25%). O efeito combinado da epigalocatequina galato e quercetina exerce atividade antiproliferativa contra o CCR, é promissor como agente quimioterápico alternativo convencional.

Leaf mineral concentration of peach after the use of resistance inducers / Concentração mineral da folha do pessegueiro após o uso de indutores de resistência

This research studied the effect of the inducers acibenzolar-S-Methyl, potassium phosphite, and potassium silicate on the leaf mineral concentration of peach. The experiment was carried out in Tezontepec, Puebla, in the years 2017 and 2018. The treatments consisted of the foliar application of the resistance inducers acibenzolar-S-Methyl, potassium phosphite, and potassium silicate. Inducers were applied at commercially recommended dose. The experimental design was in randomized blocks with four replicates. Foliar quantification of nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, copper and silicon was carried out. The use of resistance inducers increases the leaf concentrations of calcium. The interaction of resistance inducers and years of application modified the foliar concentration of magnesium, phosphorus and sulfur in peaches.

O objetivo desta pesquisa foi estudar o efeito dos indutores acibenzolar-S-Metil, fosfito de potássio e silicato de potássio sobre o teor de minerais foliares do pessegueiro. O experimento foi realizado em Tezontepec, Puebla, nos anos de 2017 e 2018. Os tratamentos consistiram na aplicação foliar dos indutores acibenzolar-S-Metil, fosfito de potássio e silicato de potássio. Os indutores foram aplicados na dose comercialmente recomendada. O delineamento experimental foi em blocos casualizados com quatro repetições. Foi realizada a quantificação foliar de nitrogênio, fósforo, potássio, cálcio, magnésio, enxofre, cobre e silício. A interação de indutores de resistência e anos de aplicação modificou a concentração foliar de magnésio, fósforo e enxofre nas folhas de pessegueiro.

Glyceryl guaiacol ether for sedation of horses undergoing orchiectomy when standing / Éter gliceril-guaiacol para sedação de equinos submetidos à orquiectomia em pé

A study was designed to compare two sedation protocols to be used in horses undergoing orchiectomy when standing. In both protocols, the induction to the sedative state was performed with bolus detomidine at a dose of 10 µg/kg, intravenously (IV). In the first protocol (eight horses), the sedative state was maintained with 1% glyceryl guaiacol ether (GGE) in continuous infusion, at a dose of 1ml/kg/hour IV. In the second protocol (eight horses) this sedative effect was maintained with detomidine in continuous infusion at the same dose and induction route. Orchiectomy was performed on all animals. The two protocols allowed the surgeries to be performed when standing. However, horses kept under sedation by the GGE showed greater relaxation and a more intense degree of sedation. It should be noted that the use of GGE to maintain the sedative state in horses is unprecedented in the literature. The drug is used in anesthetic protocols in the species, but only in pre-anesthetic medication in general anesthesia. It was concluded that the two sedation protocols allowed the performance of orchiectomy with the horses when standing. However, the protocol in which 1% GGE was used showed more profound sedation, without adverse effects.

Foi realizado um estudo para comparar dois protocolos de sedação a serem utilizados em equinos submetidos à orquiectomia em estação. Em ambos os protocolos, a indução ao estado sedativo foi realizada com bolus de detomidina, na dose de 10µg/kg, por via intravenosa (IV). No primeiro protocolo (oito cavalos), o estado sedativo foi mantido com 1% de éter gliceril guaiacol (GGE) em infusão contínua, na dose de 1mL/kg/hora IV. No segundo protocolo (oito cavalos), esse efeito sedativo foi mantido com detomidina, em infusão contínua, na mesma dose e via de indução. Foi realizada orquiectomia em todos os animais. Os dois protocolos permitiram que as cirurgias fossem realizadas em estação No entanto, os cavalos mantidos sob sedação pelo GGE apresentaram maior relaxamento e grau de sedação mais intenso. Ressalta-se que o uso de GGE para manutenção do estado sedativo em equinos é inédito na literatura. O fármaco é utilizado em protocolos anestésicos na espécie, mas apenas na medicação pré-anestésica em anestesia geral. Concluiu-se que os dois protocolos de sedação permitiram a realização da orquiectomia com os cavalos em estação. No entanto, o protocolo em que foi utilizado GGE 1% apresentou sedação mais profunda, sem efeitos adversos.

Anestesia intravenosa parcial (AIVP) em uma lobo-guará (Chrysocyon brachyurus) neonata / Partial intravenous anesthesia (PIVA) in a neonate maned wolf (Chrysocyon brachyurus)

Background: The maned wolf (Chrysocyon brachyurus) is the largest canid in South America and the only representative of its genus. The scientific literature presents studies referring to the management and anaesthesia of the species but is scarce in case of emergency procedures, and to date, there are no reports of anaesthesia for emergency procedures in a neonatal maned wolf. Thus, this study aimed to report xenotransfusion and emergency anaesthesia for thoracic limb amputation in an approximately 8-days-old maned wolf pup. Case: A maned wolf pup, approximately 8-day-old, with a history of an accident with an agricultural machine, was referred to the Veterinary Hospital of the Federal University of Paraná - Palotina Sector (HVP - UFPR) with a grade III open fracture of left radius and ulna. At the physical evaluation, bullous rales were observed in the right caudal lobe on pulmonary auscultation, hypoglycemia and severe dehydration, the latter being determined by the occurrence of enophthalmos, increased capillary refill time and skin turgor (3 s). The hemogram revealed regenerative hypochromic macrocytic anaemia, thrombocytopenia and leukopenia, and the chest radiograph showed alveolar pattern opacification, associated with the presence of air bronchograms in the caudal lobes, more evident on the right side, suggestive of pulmonary contusion. Considering the laboratory alterations and the need for amputation of the thoracic limb, xenotransfusion was chosen before the anaesthetic procedure. The animal was pre-medicated with methadone 0.2 mg/kg and anaesthetic induction was performed with propofol titrated to effect, requiring 10 mg/kg. This was followed by endotracheal intubation and anaesthetic maintenance using the partial intravenous technique with an infusion of 5 µg/kg/h of remifentanil and Isoflurane vaporised through a non-rebreathing system in oxygen at 0.6. Anaesthetic monitoring included heart rate and electrocardiogram tracing, respiratory rate, oxyhemoglobin saturation, end-tidal partial pressure of carbon dioxide, systemic blood pressure by the oscillometric method, and oesophageal body temperature. Although the blood tests after the transfusion showed an increase in hematocrit, changes suggestive of a delayed hemolytic reaction because of the transfusion were also noticed. Discussion: Preanesthetic stabilisation is critical, as anaesthesia of unstable patients may result in a higher risk of anaesthetic complications. Critically ill patients may present systemic imbalances that can trigger pharmacokinetic and pharmacodynamic changes of anaesthetics and analgesics. Neonates have several marked physiological differences, since some organs are still immature, have a high body volume of water, and present a reduced amount of total fat and concentrations of circulating proteins, making it necessary to adjust protocols and doses used for these patients. Blood transfusion between the same species is always the best option, though xenotransfusion becomes an option when there is no homologous donor available. However, it can present a great risk to life, as there is a lack of studies regarding the blood typing of the species and blood compatibility tests. We concluded that there was an increase in hematocrit after 24 h of xenotransfusion and, even with signs of delayed hemolytic reaction observed in the blood test, the patient did not show specific clinical signs of transfusion reaction. The pup was sensitive to methadone but required a high dose of propofol for anaesthetic induction. The use of methadone as a pre-anaesthetic agent and the infusion of remifentanil provided adequate analgesia based on the parameters evaluated.

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.

Minimal and normal-flow general anesthesia in patients undergoing surgery in prone position: impact on hemodynamics and regional cerebral oxygenation

Purpose: In this study, the aim to assess the combined effects of prone-positioning (PP) and minimal-flow (MF) general anesthesia on regional cerebral oxygenation (RCO) and systemic hemodynamics. Methods: This is a randomized prospective study aiming to evaluate changes in cerebral oxygenation and hemodynamic parameters in MF systemic anesthesia in patients undergoing surgery in PP. Patients were randomized to MF or normal-flow (NF) anesthesia. In the operating room, pulse rate, mean arterial pressure (MAP), peripheral hemoglobin oxygen saturation (spO2), and right- and left-side RCO (assessed by nearinfrared spectroscopy, NIRS) were measured perioperatively. Results: Overall, 46 patients were included (24 in the MF group and 22 in the NF group). The amount of anesthetic gas consumption was significantly lower in the low-flow (LF) group. In both groups, the mean pulse rate showed a decrease after PP. Before induction, RCO was significantly higher both at the right- and left-sides in the LF group compared to the NF group. This difference continued throughout the operation on the left-side and disappeared 10 min after intubation on the right-side. On the left side, mean RCO decreased after PP in both groups. Conclusion: MF anesthesia in PP did not reduce cerebral oxygenation compared to NF and was safe in terms of systemic hemodynamics and cerebral oxygenation.

Origin, evolution and strategies for the genetic improvement of physalis / Origem, evolução e estratégias para o melhoramento genético de fisális

ABSTRACT: Physalisperuviana L. (physalis) has significant economic potential by virtue of the unique flavor of its fruit. However, the productivity of Brazilian plantations is low because of the limited number of varieties or cultivars available. The main obstacle in the selection of superior genotypes is the lack of information about genetic variability within- and between- populations and limited genetic basis that has likely resulted from evolutionary, domestication and selection processes of the natural or artificial populations. Physalis currently cultivated in Brazil is tetraploid, and such polyploidy may have led to the reproductive isolation of the species, preventing the occurrence of intraspecific hybridization. Moreover, cultivated populations derive from a common gene pool and have undergone a long process of domestication and selection carried out empirically by farmers. In Colombia and other Andean countries there are wild populations that exhibit genetic diversity which; although, fundamental for the conservation of the species, have low potential for the development of genotypes with superior agronomic traits. In order to create and expand the genetic variability of physalis, breeders have employed various strategies including induction of mutation, chromosome duplication, and interspecific and intraspecific hybridization. Furthermore, the production of double haploid lines from in vitro anther cultures has shown good results in the selection of hybrids. The mutant genotypes and/or hybrids obtained using these methods in association with those of wide genomic selection can generate cultivars with superior agronomic traits.

RESUMO: Physalis peruviana L. (fisális) apresenta grande potencial econômico devido ao sabor diferenciado de seus frutos. A produtividade dos pomares brasileiros é baixa em função do número limitado de variedades ou cultivares disponíveis. O principal entrave na seleção de genótipos superiores é a falta de informação sobre a variabilidade genética dentro e entre as populações de fisális e, possivelmente, a base genética limitada das mesmas, que pode ser explicada pelos processos evolutivos, de domesticação e de seleção das populações naturais ou artificiais. A fisális cultivada atualmente no Brasil é tetraploide e tal poliploidia pode ter levado ao isolamento reprodutivo da espécie, o qual impede a ocorrência de hibridação intraespecífica. As populações cultivadas provêm de um pool genético comum e, além disso, sofreram um longo processo de domesticação e seleção realizadas empiricamente pelos agricultores. Porém, na Colômbia e em outros países andinos existem populações silvestres que exibem diversidade genética que, embora sejam fundamentais para a conservação da espécie, apresentam baixo potencial para o desenvolvimento de genótipos com características agronômicas superiores. A fim de criar e ampliar a variabilidade genética de fisális, os melhoristas tem empregado várias estratégias incluindo a indução de mutação, a duplicação cromossômica e a hibridação inter e intraespecífica. Além disso, a produção de linhagens duplo-haploides a partir da cultura de anterasin vitro vem demonstrando bons resultados para a seleção de híbridos. Os genótipos mutantes e/ou híbridos obtidos através dos métodos citados em associação com os de seleção genômica ampla podem gerar cultivares com características agronômicas superiores.

Influence on clinical biochemistry values of black-tufted marmosets (Callithrix penicillata) anesthetized with isoflurane or sevoflurane / Influência sobre valores de bioquímica clínica de saguis-de-tufo-preto (Callithrix penicillata) anestesiados com isoflurano ou sevoflurano

The objective of this study was to evaluate the clinical biochemistry behavior of Black-Tufted Marmosets (Callithrix penicillatta) submitted to blood collection without sedation and after general anesthesia with anesthetics isoflurane or sevoflurane. Blood collections were performed on (M1) day before anesthesia by physical restraint, and (M2) after anesthesia. There were four groups: Isoflurane (GI) and Sevoflurane (GS) using an anesthetic box. GIM: isoflurane induction with mask for a shorter period. Control group (GP): physical restraint in both moments. Plasma was separated and frozen to measure clinic biochemistry values. Urea was higher at M2 in groups GI and GP. AST was higher in M2 in GI, GS, and GP and only GI showed an increase in CK in M2. Glucose was higher in M1 in the GI, GS, and GP groups and fructosamine was higher in M2 in the GI. Stress caused by physical restraint can cause biochemical changes and these must be considered when interpreting the exams. Both the inhalational anesthetic isoflurane and sevoflurane did not cause clinically significant changes in clinical biochemistry results.

O objetivo desse estudo foi avaliar o comportamento da bioquímica clínica de saguis-do-tufo-preto (Callithrix penicillatta) submetidos à coleta sanguínea sem sedação e após anestesia geral, com os anestésicos isoflurano ou sevoflurano. As coletas de sangue foram realizadas: (M1) dia antes da anestesia por contenção física e (M2) após anestesia. Foram definidos quatro grupos: isoflurano (GI) e sevoflurano (GS), utilizando caixa anestésica; GIM: indução com isoflurano com máscara por um período menor; grupo controle (GCF): contenção física em ambos os momentos. O plasma com EDTA foi separado e congelado para realizar dosagem da bioquímica clínica. A ureia foi maior no M2 nos grupos GI e GCF. A AST foi maior no M2 nos GI, GS e GCF e somente o GI apresentou aumento de CK no M2. A glicose foi maior no M1 nos grupos GI, GS e GCF, e a frutosamina foi maior no M2 no GI. O estresse causado pela contenção física pode causar alterações bioquímicas e essas devem ser levadas em consideração na interpretação dos exames. Tanto o anestésico inalatório isoflurano quanto o sevoflurano não causaram alterações clinicamente significantes nos resultados da bioquímica clínica.

Protocolo anestésico em guaxinim submetido à colocefalectomia / Anesthesic protocol in a raccoon submitted to colocephalectomy

Em guaxinins, comumente se observa a presença de osteoartrites acometendo a articulação coxofemoral. Em carnívoros domésticos, o tratamento pode ser conservativo ou cirúrgico e, dentre as técnicas cirúrgicas, cita-se a colocefalectomia. Por ser um procedimento que provoca dor moderada a severa, um protocolo de anestesia balanceada se faz necessário, podendo associar a técnicas de bloqueio locorregional, como a epidural. Protocolos descritos para a espécie são limitados, porém comumente empregados para contenções químicas, avaliações físicas e exames de imagem. Portanto, o objetivo deste trabalho é relatar a o protocolo anestésico realizado em guaxinim (Procyon cancrivorus) submetido à colocefalectomia. Foram utilizados dexmedetomidina (5µg/kg), cetamina (5mg/kg) e diazepam (0,3mg/kg) por via intramuscular como medicação pré-anestésica, indução com propofol e manutenção por anestesia inalatória com isoflurano. Para execução da anestesia epidural, realizou-se uma abordagem na região lombossacral, sendo administrados bupivacaína (1mg/kg) e morfina (0,1mg/kg). Não houve intercorrências e os parâmetros fisiológicos mantiveram-se estáveis. Descartou-se a necessidade de resgate analgésico e a recuperação anestésica foi rápida. Dessa forma, conclui-se que o manejo anestésico proposto promoveu estabilidade hemodinâmica e recuperação satisfatórias para a espécie.

In raccoons, the presence of osteoarthritis affecting the hip joint is commonly observed. In domestic carnivores, the treatment can be conservative or surgical and, among the surgical techniques, colocephalectomy is mentioned. Since it is a procedure that causes moderate to severe pain, a balanced anesthesia protocol is necessary, which can be associated with locoregional block techniques such as the epidural. Protocols described for the species are limited, being commonly used for chemical restraints, physical assessments, and imaging exams. Therefore, this work aims to report the anesthetic protocol performed on a raccoon (Procyon cancrivorus) submitted to colocephalectomy. Dexmedetomidine (5µg/kg), ketamine (5mg/kg), and diazepam (0.3mg/kg) were used intramuscularly as pre-anesthetic medication, in addition to induction with propofol, and the maintenance by inhalational anesthesia with isoflurane. For epidural anesthesia, an approach was performed in the lumbosacral region, with the administration of bupivacaine (1mg/kg) and morphine (0.1mg/kg). There were no complications and the physiological parameters remained stable. The need for analgesic rescue was ruled out and the anesthetic recovery was fast. Thus, it is concluded that the proposed anesthetic management promoted hemodynamic stability and a satisfactory recovery for the species.

Recent advances in treatments for resynchronization of ovulation in small ruminants: a review

Hormonal methodologies to control small ruminants' estrous cycle are worldwide used and evolved, adjusting the application to the precise female physiological moments to enhance reproductive performance. The estrous cycle can be induced and/or synchronized, aiming for fixed-time artificial insemination, or based on estrus behavior signs for insemination, natural or guided mating. Successive protocols can be performed to resynchronize ovulation and increase reproductive outcomes in females that failed to conceive. These recently developed treatments aim to resynchronize the ovulation as earlier as non-pregnancy is detected. The present review aimed to summarize the recent advances and main findings regarding resynchronization protocols used in small ruminants. Lastly, we present future perspectives and new paths to be studied in the subject. The resynchronization treatment is still a growing field in small ruminant reproduction, nevertheless, some enhancements are found in the reproductive outcome, showing that such protocols can be successfully used in sheep and goat production.(AU)

In vitromorphogenesis of gypsophila: Response to exogenous phytohormones / Morfogênese de gypsophila in vitro: Resposta a fitormônios exógenos

The success of in vitro cultivation of plant species depends on factors associated with the induction and control of morphogenesis regarding the regeneration of shoots and roots in the organogenesis process, such as the culture medium composition. Thus, the objective of this study was to investigate if theconcentration of growth regulators in culture medium interferes with the in vitro morphogenesis of gypsophila. 'Golan' cultivar was subjected to three culture media (M), which constituted the study treatments: M1) Murashige and Skoog (MS) medium without the addition of growth regulators; M2) MS + 1 mg L-1of benzylaminopurine (BAP) + 0.05 mg L-1of naphthaleneacetic acid (NAA); M3) MS + 0.05 mg L-1BAP + 1 mg L-1NAA. After 45 days, the multiplication rate, plant height and root length were evaluated. The results showed that plantlets produced in M2 medium had a higher multiplication rate. Also, plantlets produced in M1 and M3 media showed higher shoot height and root length. It is concluded that there is a difference in the in vitromorphogenesis of gypsophila according to the concentration of growth regulators in the micropropagation medium.(AU)

O sucesso do cultivo in vitro de espécies vegetais depende de fatores associados à indução e ao controle da morfogênese quanto à regeneração de brotos e raízes no processo de organogênese, a exemplo da composição de meio de cultura. Assim, o objetivo do estudo foi investigar se a concentração de reguladores de crescimento em meio de cultura interfere na morfogênese in vitrode gipsofila. A cultivar 'Golan' foi submetida a três meios de cultura (M), que constituíram os tratamentos do estudo: M1) meio Murashige e Skoog (MS) sem adição de reguladores de crescimento; M2) MS + 1 mg.L-1de benzilaminopurina (BAP) + 0.05 mg.L-1de ácido naftalenoacético (ANA); M3) MS + 0.05 mg.L-1BAP + 1 mg.L-1de ANA. Após 45 dias avaliou-se a taxa de multiplicação, altura de plantas e comprimento de raízes. Os resultados mostraram que plântulas produzidas no meio M2 tiveram maior taxa de multiplicação. Ainda, plântulas produzidas nos meios M1 e M3 apresentaram maior altura de parte aérea e comprimento de raízes. Conclui-se que há diferença na morfogênese in vitrode gipsofila de acordo com a concentração de reguladores de crescimento em meio de cultura na micropropagação.(AU)

Ácido salicílico e cobertura de solo no cultivo de physalis / Salicylic acid and soil cover in the cultivation of physalis

O objetivo neste trabalho foi verificar a influência da aplicação de diferentes concentrações de ácido salicílico e distintas coberturas de solo no cultivo de physalis (Physalis peruviana L). O trabalho foi desenvolvido na área experimental da Universidade Federal da Fronteira Sul, Laranjeiras do Sul-PR. Foram utilizadas mudas de physalis produzidas em casa de vegetação. A campo foram submetidas a cinco concentrações de ácido salicílico (0, 0,001, 0,002, 0,003, 0,004 Mol L-¹), e três coberturas de solo (palhada de gramínea, eucalipto e testemunha sem palhada). Nas avaliações de pré-colheita, o delineamento experimental foi de blocos ao acaso, em esquema fatorial 3x5. Para as avaliações pós-colheita o delineamento experimental foi 3x5x2, sendo considerado também o mês de colheita. Não houve interação entre os fatores para as variáveis analisadas. Para altura e diâmetro, presença de insetos, incidência e severidade de doenças foliares, somente o fator concentrações de ácido salicílico influenciou. Na pós-colheita somente o mês de colheita interferiu nos fatores, sendo a colheita de setembro com maior número e massa dos frutos, mas também o maior ataque de insetos e doenças. Concluiu-se com este trabalho que a aplicação de ácido salicílico influenciou no desenvolvimento de plantas de physalis.(AU)

The objective of this work was to verify the influence of the application of different concentrations of salicylic acid and different soil coverings in the cultivation of physalis (Physalis peruvianaL). The work was developed in the experimentalarea of the Federal University of Fronteira Sul, Laranjeiras do Sul-PR. Physalis seedlings produced in a greenhouse were used. The field was subjected to five concentrations of salicylic acid (0, 0,001, 0,002, 0,003, 0,004 Mol L-¹), and three soil coverings (grass straw, eucalyptus and control without straw). In the pre-harvest evaluations, the experimental design was randomized blocks, in a 3x5 factorial scheme. For post-harvest evaluations, the experimental design was 3x5x2, and the month of harvest was also considered. There was no interaction between the factors for the analyzed variables. For height and diameter increase and presence of insects, incidence and severity of leaf diseases, only the factor of salicylic acid influences. In the post-harvestperiod, only the month of harvest interfered in the factors, with the September harvest having the largest number and mass of fruits, but also the greatest attack by insects and diseases. It was concluded with this work that the application of salicylic acid influenced the development of physalis plants.(AU)

Differential sprouting ability during micropropagation of Nidularium minutum Mez (Bromeliaceae)

Nidularium minutum is an ornamental bromeliad from the Brazilian Rainforest. The micropropagation of this species is essential for obtaining plants available for conservation programs or commercial use. Our study aimed to establish an efficient plant production method by in vitro sprouting. This bromeliad takes a long time to sprout in vitro, and 10% of the plants produce shoots in a culture medium without plant growth regulators (PGRs). When subcultured in a PGR-free medium, these individualized shoots can sprout like the mother plant. The Murashige and Skoog basal medium (MS) containing 1.0 mg L-1 of 6-benzylaminopurine (BAP) promoted the induction of adventitious shoots in greater than 90% of the plants after 240 days of culture with an average of more than eight shoots per plant. Approximately 100% of the in vitro-produced shoots survived after acclimatization, reaching the flowering stage. Therefore, our results showed that in vitro regeneration of N. minutum depends on the cultivation period and that plants with a higher sprouting capacity can be selected and used as micropropagation matrices, contributing to the production of this endangered bromeliad.(AU)

Complete bladder eversion in periparturient Nellore cow: case report / Eversão completa de bexiga no pré-parto em vaca Nelore: relato de caso

Partial or complete bladder eversion is a rare condition of poor prognosis in cows, commonly associated with intense tenesmus observed in the peripartum period. A 14-year-old obese Nellore cow at 280 days of gestation was referred with 24-hour bladder prolapse. The bladder was complete eversion through the vulvar vestibule showing a thick congested wall and small residual urine volume. After clinical examination, the cow received scopolamine butylbromide and intercoccygeal epidural anesthesia, and the externalized bladder segment was partially reduced, remaining only 10 cm externalized. The cow was maintained with an intravesical human gastric tube number 16 and constant monitoring. The eversion was fully reduced after 12 hours of local treatment, and as the cow presented subclinical ketosis, hypocalcemia and cystitis, antibiotic, glucose, calcium and propylene glycol therapy were performed. We opted for induction of parturition, and after 24 hours, a healthy 52kg calf was born and the placenta was delivered 16 hours after calving. The cow and calf were discharged on the sixth day of hospitalization, with no recurrences or secondary complications after treatment.

A eversão vesical parcial ou completa é uma condição rara de mau prognóstico em vacas, comumente associada a tenesmo intenso observado no período periparto. Uma vaca Nelore obesa de 14 anos de idade, com 280 dias de gestação, foi encaminhada com prolapso de bexiga de 24 horas. A bexiga apresentava eversão completa por meio do vestíbulo vulvar apresentando parede espessa e congestionada e pequeno volume residual de urina. Após exame clínico, a vaca recebeu butilbrometo de escopolamina e anestesia peridural intercoccígea, e o segmento vesical exteriorizado foi parcialmente reduzido, permanecendo apenas 10cm exteriorizado. A vaca foi mantida com sonda gástrica humana número 16 por via intravesical e monitorada constantemente. A eversão foi totalmente reduzida após 12 horas de tratamento local, e, como a vaca apresentava cetose subclínica, hipocalcemia e cistite, foi realizada antibioticoterapia, glicose, cálcio e propilenoglicol. Optou-se pela indução do parto. Após 24 horas, nasceu um bezerro saudável de 52kg e a placenta foi expelida 16 horas após o parto. A vaca e o bezerro receberam alta no sexto dia de internação, sem recidivas ou complicações secundárias após o tratamento.