Modelling premature cardiac aging with induced pluripotent stem cells from a hutchinson-gilford Progeria Syndrome patient.

Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic disorder that causes accelerated aging and a high risk of cardiovascular complications. However, the underlying mechanisms of cardiac complications of this syndrome are not fully understood. This study modeled HGPS using cardiomyocytes (CM) derived from induced pluripotent stem cells (iPSC) derived from a patient with HGPS and characterized the biophysical, morphological, and molecular changes found in these CM compared to CM derived from a healthy donor. Electrophysiological recordings suggest that the HGPS-CM was functional and had normal electrophysiological properties. Electron tomography showed nuclear morphology alteration, and the 3D reconstruction of electron tomography images suggests structural abnormalities in HGPS-CM mitochondria, however, there was no difference in mitochondrial content as measured by Mitotracker. Immunofluorescence indicates nuclear morphological alteration and confirms the presence of Troponin T. Telomere length was measured using qRT-PCR, and no difference was found in the CM from HGPS when compared to the control. Proteomic analysis was carried out in a high-resolution system using Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). The proteomics data show distinct group separations and protein expression differences between HGPS and control-CM, highlighting changes in ribosomal, TCA cycle, and amino acid biosynthesis, among other modifications. Our findings show that iPSC-derived cardiomyocytes from a Progeria Syndrome patient have significant changes in mitochondrial morphology and protein expression, implying novel mechanisms underlying premature cardiac aging.

Overexpression of mir-135b and mir-210 in mesenchymal stromal cells for the enrichment of extracellular vesicles with angiogenic factors.

Extracellular vesicles (EVs) are known as molecular carriers involved in cell communication and the regulation of (patho)physiological processes. miRNAs and growth factors are the main contents of EVs which make them a good candidate for the treatment of diseases caused by ischemia, but the low production of EVs by a cell producer and a significant variation of the molecular contents in EVs according to the cell source are the main limitations of their widespread use. Here, we show how to improve the therapeutic properties of mesenchymal stromal cell (MSC)-derived EVs (MSC-EVs) by modifying MSCs to enrich these EVs with specific angiomiRs (miR-135b or miR-210) using lentiviral vectors carrying miR-135b or miR-210. MSCs were obtained from the mouse bone marrow and transduced with a corresponding lentivector to overexpress miR-135b or miR-210. The EVs were then isolated by ultracentrifugation and characterized using a flow cytometer and a nanoparticle tracking analyzer. The levels of 20 genes in the MSCs and 12 microRNAs in both MSCs and EVs were assessed by RT‒qPCR. The proangiogenic activity of EVs was subsequently assessed in human umbilical vein endothelial cells (HUVECs). The results confirmed the overexpression of the respective microRNA in modified MSCs. Moreover, miR-135b overexpression upregulated miR-210-5p and follistatin, whereas the overexpression of miR-210 downregulated miR-221 and upregulated miR-296. The tube formation assay showed that EVs from MSCs overexpressing miR-210-5p (EVmiR210) significantly promoted tubular structure formation in HUVECs. A significant increase in angiogenic proteins (PGF, endothelin 1, and artemin) and genes (VEGF, activin A, and IGFBP1) in HUVECs treated with VEmiR210 justifies the better tubular structure formation of these cells compared with that of EVmiR135b-treated HUVECs, which showed upregulated expression of only artemin. Collectively, our results show that the EV cargo can be modified by lentiviral vectors to enrich specific miRNAs to achieve a specific angiogenic potential.

The monoterpene 1,8-cineole prevents cerebral edema in a murine model of severe malaria.

1,8-Cineole is a naturally occurring compound found in essential oils of different plants and has well-known anti-inflammatory and antimicrobial activities. In the present work, we aimed to investigate its potential antimalarial effect, using the following experimental models: (1) the erythrocytic cycle of Plasmodium falciparum; (2) an adhesion assay using brain microvascular endothelial cells; and (3) an experimental cerebral malaria animal model induced by Plasmodium berghei ANKA infection in susceptible mice. Using the erythrocytic cycle of Plasmodium falciparum, we characterized the schizonticidal effect of 1,8-cineole. This compound decreased parasitemia in a dose-dependent manner with a half maximal inhibitory concentration of 1045.53 ± 63.30 µM. The inhibitory effect of 972 µM 1,8-cineole was irreversible and independent of parasitemia. Moreover, 1,8-cineole reduced the progression of intracellular development of the parasite over 2 cycles, inducing important morphological changes. Ultrastructure analysis revealed a massive loss of integrity of endomembranes and hemozoin crystals in infected erythrocytes treated with 1,8-cineole. The monoterpene reduced the adhesion index of infected erythrocytes to brain microvascular endothelial cells by 60%. Using the experimental cerebral malaria model, treatment of infected mice for 6 consecutive days with 100 mg/kg/day 1,8-cineole reduced cerebral edema with a 50% reduction in parasitemia. Our data suggest a potential antimalarial effect of 1,8-cineole with an impact on the parasite erythrocytic cycle and severe disease.

Early Effects of Extracellular Vesicles Secreted by Adipose Tissue Mesenchymal Cells in Renal Ischemia Followed by Reperfusion: Mechanisms Rely on a Decrease in Mitochondrial Anion Superoxide Production.

Acute kidney injury (AKI) caused by ischemia followed by reperfusion (I/R) is characterized by intense anion superoxide (O2•-) production and oxidative damage. We investigated whether extracellular vesicles secreted by adipose tissue mesenchymal cells (EVs) administered during reperfusion can suppress the exacerbated mitochondrial O2•- formation after I/R. We used Wistar rats subjected to bilateral renal arterial clamping (30 min) followed by 24 h of reperfusion. The animals received EVs (I/R + EVs group) or saline (I/R group) in the kidney subcapsular space. The third group consisted of false-operated rats (SHAM). Mitochondria were isolated from proximal tubule cells and used immediately. Amplex Red™ was used to measure mitochondrial O2•- formation and MitoTracker™ Orange to evaluate inner mitochondrial membrane potential (Δψ). In vitro studies were carried out on human renal proximal tubular cells (HK-2) co-cultured or not with EVs under hypoxic conditions. Administration of EVs restored O2•- formation to SHAM levels in all mitochondrial functional conditions. The gene expression of catalase and superoxide dismutase-1 remained unmodified; transcription of heme oxygenase-1 (HO-1) was upregulated. The co-cultures of HK-2 cells with EVs revealed an intense decrease in apoptosis. We conclude that the mechanisms by which EVs favor long-term recovery of renal structures and functions after I/R rely on a decrease of mitochondrial O2•- formation with the aid of the upregulated antioxidant HO-1/Nuclear factor erythroid 2-related factor 2 system, thus opening new vistas for the treatment of AKI.

Three-dimensional architecture of Cyrilia lignieresi gametocyte-stage development inside red blood cells.

The Haemogregarinidae family (Apicomplexa: Adeleina) comprises hemoprotozoa that infect mammals, birds, amphibians, fish, and reptiles. Some morphological characteristics of the Cyrilia lignieresi have been described previously, but the parasite-erythrocyte relationship is still poorly understood. In order to understand the structural architecture of C. lignieresi-infected red blood cells, electron microscopy-based three-dimensional reconstruction was carried out using TEM as well as FIB-SEM tomography. Results showed that development of the macrogametocyte-stage inside the red blood cell is related to an increase in cleft-like structures in the host cell cytoplasm. Furthermore, other aspects related to parasite intraerythrocytic development were explored by 3D visualization techniques. We observed the invagination of a large extension of the Inner Membrane Complex (IMC) on the parasite body, which results from or induces a folding of the posterior end of the parasite. Small tubular structures were seen associated with areas related to IMC folding. Taken together, results provide new information on the remodeling of erythrocytes induced by the protozoan C. lignieresi.

Current Microscopy Strategies to Image Fungal Vesicles: From the Intracellular Trafficking and Secretion to the Inner Structure of Isolated Vesicles.

Extracellular vesicles (EVs) are nano-sized structures that play important roles in a variety of biological processes among members of the Eukaryota domain. They have been studied since the 1940s and a broader use of different microscopy techniques to image either isolated vesicles or vesicles within the intracellular milieu (trafficking) has been limited by their nanometric size, usually below the resolution limit of most standard light microscopes. The development of genetically encoded fluorescent proteins and fluorescent probes able to switch between "on" and "off" states, as well the improvement in computer-assisted microscopy, photon detector devices, illumination designs, and imaging strategies in the late Twentieth century, boosted the use of light microscopes to provide structural and functional information at the sub-diffraction resolution, taking advantage of a nondestructive analytical probe such light, and opening new possibilities in the study of life at the nanoscale. As well, traditional and novel electron microscopy techniques have been widely used in the characterization of subcellular compartments, either isolated or in situ, providing a comprehensive understanding of their functional role in many cellular processes. Here, we present basic aspects of some of these techniques that have already been applied and their potential application to the study of fungal vesicles.

Genetic parameters of morphometric measurements in Criollo horses.

The aim of this study was to estimate the genetic parameters for the morphometric measurements of withers height (WH), thoracic circumference (TC) and cannon bone circumference (CBC) of Criollo horses, stratified for maternal and paternal effects. Statistical genetic design of factor crossings was used to evaluate the offspring of full siblings and half-siblings. Fifty stallions were selected (n = 50) who had been crossed with six mares each (n = 6), to provide 300 parental pairings in which two offspring were born per mare (n = 600). WH in females and TC in males were highly influenced by additive genetic effects, while the other morphometric measures were found to be closely related to the general effects of the environment. WH had the greatest additive genetic influence on female offspring (47.06%), while TC presented the greatest additive genetic influence on male offspring (58.73%). When comparing the parental influence on the morphometric characteristics evaluated, TC and CBC had greater additive genetic influence from maternal effects, regardless of offspring sex. However, the WH in female offspring showed greater maternal additive genetic contribution, while in males this characteristic presented greater paternal additive genetic influence. For WH in male offspring, narrow sense heritability ( h r 2 : 0.26) was more influenced by paternal effects, while for WH in females ( h r 2 : 0.47), TC in females ( h r 2 : 0.08) and males ( h r 2 : 0.59), and CBC in females ( h r 2 : 0.07) and males ( h r 2 : 0.05) the maternal effects were the most important. Therefore, the phenotypic expression of WH in males and females and TC in males is influenced by the additive genetic effect of the genes. On the other hand, the TC in female progenies and CBC in progenies of both sexes are closely related to the effects of the environment. The results obtained in this study could be useful in the selection process of Criollo horses.

Eugenol disrupts Plasmodium falciparum intracellular development during the erythrocytic cycle and protects against cerebral malaria.

BACKGROUND: Malaria is a parasitic disease that compromises the human host. Currently, control of the Plasmodium falciparum burden is centered on artemisinin-based combination therapies. However, decreased sensitivity to artemisinin and derivatives has been reported, therefore it is important to identify new therapeutic strategies. METHOD: We used human erythrocytes infected with P. falciparum and experimental cerebral malaria (ECM) animal model to assess the potential antimalarial effect of eugenol, a component of clove bud essential oil. RESULTS: Plasmodium falciparum cultures treated with increasing concentrations of eugenol reduced parasitemia in a dose-dependent manner, with IC50 of 532.42 ± 29.55 µM. This effect seems to be irreversible and maintained even in the presence of high parasitemia. The prominent effect of eugenol was detected in the evolution from schizont to ring forms, inducing important morphological changes, indicating a disruption in the development of the erythrocytic cycle. Aberrant structural modification was observed by electron microscopy, showing the separation of the two nuclear membrane leaflets as well as other subcellular membranes, such as from the digestive vacuole. Importantly, in vivo studies using ECM revealed a reduction in blood parasitemia and cerebral edema when mice were treated for 6 consecutive days upon infection. CONCLUSIONS: These data suggest a potential effect of eugenol against Plasmodium sp. with an impact on cerebral malaria. GENERAL SIGNIFICANCE: Our results provide a rational basis for the use of eugenol in therapeutic strategies to the treatment of malaria.

Complete avulsion of the hoof capsule and subsequent testicular degeneration in a criollo stallion

Background: Complete avulsion of the hoof in horses, also known as exungulation, is not a commonly reported injury and usually leads to euthanasia due to the great amount of tissue loss, intense pain, secondary complications, expensive and lengthy treatment. It can involve deep structures and cause different complications leading to chronic lameness. In stallions affected by such injury, the reproductive tract and performance may also be affected. The aim of this study was to report a case of complete avulsion of the right front hoof in a Criollo stallion and subsequent bilateral testicular degeneration. Case: A 10-year-old Criollo stallion was referred to the Veterinary Clinical Hospital of the Federal University of Pelotas (HCV- UFPel) with a complete avulsion of the left front hoof. At admission, the stallion had clinical parameters compatible with intense pain and blood loss. Evaluation of the wound demonstrated that the distal end of the third phalanx (P3) was exposed but no fracture was detected on radiological evaluation. No other structure was apparently affected. Initially, anti-inflammatory (phenylbutazone) and opioid (morphine) was given for pain control and supportive fluid therapy was started to restore hydration. Antibiotic (Sulfamethoxazole with trimethoprim) was administered for 10 days. Continued therapy with phenylbutazone, pentoxifylline, omeprazole and supplementation with methionine, biotin and zinc was also given. Local treatment was carried out by cleaning the wound, applying an antimicrobial ointment and dressing it with a bandage. Wound management was adapted according to the evolution and healing process. The stallion was kept in stall rest during its hospitalization time. In the second month after the injury, accumulation of liquid in the scrotum was observed. Clinical and ultrasound evaluation lead to a presumptive diagnosis of testicular degeneration. The stallion was discharged after three months when the wound was almost healed and the hoof had started to grow. Six month later, a follow up by the referring vet showed that the hoof was almost completely grown and the x-ray assessment demonstrated a dorsal rotation and resorption of the distal end of the third phalanx. Discussion: The stallion of this report had a complete avulsion of the hoof capsule caused by trauma. Conservative treatment was established including wound cleaning and dressing to avoid contamination, control of pain and inflammation, antimicrobial care and supplementation to support hoof growth. Time period for wound healing and hoof growth was in agreement with other cases described previously. Bone sequestrum of the distal end of the third phalanx, and detachment of a fragment were observed in this case, followed by bone resorption. The stallion was closely monitored to prevent laminitis in the contralateral limb and no alterations were detected during the treatment period. Testicular degeneration was observed, probably caused as a consequence of hoof avulsion and due to a long period of stall rest. Degenerative alterations in testicles interfere with thermoregulation and spermatogenesis, affecting semen quality and reproductive performance. Rotation of the third phalanx was also observed six months later caused by the hoof loss. In conclusion, the patient of this report had a complete regrowth of the hoof capsule although a long intensive treatment was necessary to achieve this result. As a consequence, testicles degeneration may happen impairing its function as a stallion.(AU)

Extracellular Vesicles Derived from Induced Pluripotent Stem Cells Promote Renoprotection in Acute Kidney Injury Model.

Induced pluripotent stem cells (iPSC) have been the focus of several studies due to their wide range of application, including in cellular therapy. The use of iPSC in regenerative medicine is limited by their tumorigenic potential. Extracellular vesicles (EV) derived from stem cells have been shown to support renal recovery after injury. However, no investigation has explored the potential of iPSC-EV in the treatment of kidney diseases. To evaluate this potential, we submitted renal tubule cells to hypoxia-reoxygenation injury, and we analyzed cell death rate and changes in functional mitochondria mass. An in vivo model of ischemia-reperfusion injury was used to evaluate morphological and functional alterations. Gene array profile was applied to investigate the mechanism involved in iPSC-EV effects. In addition, EV derived from adipose mesenchymal cells (ASC-EV) were also used to compare the potential of iPSC-EV in support of tissue recovery. The results showed that iPSC-EV were capable of reducing cell death and inflammatory response with similar efficacy than ASC-EV. Moreover, iPSC-EV protected functional mitochondria and regulated several genes associated with oxidative stress. Taken together, these results show that iPSC can be an alternative source of EV in the treatment of different aspects of kidney disease.

Gestação gemelar em éguas (parte II): diagnóstico e intervenção / Twin pregnancies in mares, part II: diagnosis and management / Embarazos geme/ares en yeguas, parte II: diagnóstico y manejo

Gestações gemelares em equinos resultam em perdas embrionárias, aborto no terço final ou nascimento de potros debilitados. O objetivo desta revisão é discutir os métodos de diagnóstico e intervenção da gestação gemelar. O diagnóstico deve ser realizado na fase de migração embrionária, entre 12 e 15 dias. A ruptura de uma das vesículas é o método mais eficaz promovendo perdas similares as de uma gestação simples. Caso não seja possível intervir nesta fase, pode-se realizar o acompanhamento até cerca de 30 dias para observar a redução natural. Não ocorrendo a redução, existem técnicas de maior risco que podem ser realizadas, dentre elas a aspiração transvaginal, manipulação craniana do feto, deslocamento craniocervical ou injeção de intracardíaca-pulmonar.A eficiência da manipulação está relacionada a capacidade técnica do profissional e método utilizado, sempre buscando preservar a integridade da égua gestante.

Twin pregnancies in horses result in embryonic losses, abortion in the final third or birth of debilitated foals. The aim of this review is to discuss the diagnosis and intervention methods of twin pregnancy. The diagnosis should be made in the embryonic migration phase, between 12and 15 days. Rupture of one of the vesicles is the most effective method promoting losses similar to those of a single pregnancy. If it is not possible to manipulate at this stage, observations can be done for up to 30 days to observe the natural reduction. If there is no reduction, there are higher risk techniques that can be performed, including transvaginal aspiration, cranial manipulation of the fetus, craniocervical dislocation or intracardiac-pulmonary injection. The efficiency of manipulationis related to the professional's technical capacity and method, always seeking to preserve the integrity of the pregnant mare.

Los embarazos gemelares en yeguas resultan en pérdidas embrionarias, aborto involuntario en el tercio final o nacimiento de potros debilitados. EI objetivo de esta revisión es analizar el diagnóstico y los métodos de intervención dei embarazo gemelar. EI diagnóstico debe hacerse en Ia fase de migración embrionaria, entre 12 y 15 días. La ruptura de una de las vesículas es el método más efectivo que promueve pérdidas similares a las de un embarazo simple. Si no es posible intervenir en esta etapa, se puede hacer un seguimiento de hasta 30 días para observar la reducción natural. Si no hay reducción, se pueden realizar técnicas de mayor riesgo, que incluyen la aspiración transvaginal, la manipulación craneal del feto, la dislocación craneocervical o la inyección intracardiaco-pulmonar. La eficiencia del manejo está relacionada con la capacidad técnica y elmétodo dei profesional, siempre buscando preservar la integridad de la yegua prenada.

Perfil bioquímico do líquido amniótico de éguas com placentite ascendente e sua relação com parâmetros séricos do potro neonato / Biochemical profile of amniotic fluid in mares with ascending placentitis and its relationship with serum biochemical parameters in neonatal foals

O objetivo do presente estudo foi identificar os parâmetros bioquímicos do líquido amniótico (LA) no momento do parto de éguas com placentite ascendente e relacionar com parâmetros bioquímicos séricos dos neonatos. Foram utilizadas 36 éguas gestantes da raça Puro Sangue Inglês, divididas em dois grupos: placentite (n=5) e sadias (n=31). A placentite foi definida de acordo com a avaliação clínica, exame ultrassonográfico e histopatologia das placentas. Essas éguas foram submetidas a terapia antimicrobiana, antiinflamatórios e altrenogest. A coleta do LA foi realizada na segunda fase do parto e as amostras sanguíneas dos neonatos imediatamente após o nascimento. Foi observada menor concentração de creatinina no LA de éguas com placentite. Potros provenientes de éguas com placentite foram menores em peso e altura ao nascimento, e apresentaram concentrações elevadas de ureia e reduzidas de PPT na avaliação sérica. Foi observado correlação positiva moderada entre creatinina e ureia no LA e correlação positiva fraca entre a creatinina do LA e sérica dos potros. Concluiu-se que o perfil bioquímico do LA e sérico dos neonatos oriundos de éguas com placentite ascendente diferem de gestações sadias. Sendo que esses parâmetros podem ser utilizados como indicadores de alterações placentárias em éguas.

The aim of this study was to identify the biochemical parameters of amniotic fluid (AF) at parturition in mares with ascending placentitis, and to relate with biochemical parameters in neonatal foals’ serum. Thirty-six pregnant Thoroughbred mares were divided in 2 groups: mares with placentitis (n=5) and healthy mares (n=31). Placentitis was identified based on clinical signs, ultrasound evaluation and histopathological features in the placenta. Mares with placentitis were treated with antibiotics, antiinflammatories and altrenogest. AF was collected during the second stage of labor and blood samples were taken immediately after foaling. Lower concentrations of creatinine were found in the AF of mares with placentitis. Foals from mares with placentitis showed smaller height and lower birthweight, also showed increase of urea and decrease of total protein levels in blood serum. A moderate positive correlation between urea and creatinine in AF was observed, and a weak positive correlation was found between creatinine in AF and the serum of foals. In conclusion, biochemical composition of AF and of blood serum of neonatal foals from mares with ascending placentitis are different of healthy mares. So, those analysis could be useful to identify placental changes in mares.

Perfil bioquímico do líquido amniótico de éguas com placentite ascendente e sua relação com parâmetros séricos do potro neonato / Biochemical profile of amniotic fluid in mares with ascending placentitis and its relationship with serum biochemical parameters in neonatal foals

O objetivo do presente estudo foi identificar os parâmetros bioquímicos do líquido amniótico (LA) no momento do parto de éguas com placentite ascendente e relacionar com parâmetros bioquímicos séricos dos neonatos. Foram utilizadas 36 éguas gestantes da raça Puro Sangue Inglês, divididas em dois grupos: placentite (n=5) e sadias (n=31). A placentite foi definida de acordo com a avaliação clínica, exame ultrassonográfico e histopatologia das placentas. Essas éguas foram submetidas a terapia antimicrobiana, antiinflamatórios e altrenogest. A coleta do LA foi realizada na segunda fase do parto e as amostras sanguíneas dos neonatos imediatamente após o nascimento. Foi observada menor concentração de creatinina no LA de éguas com placentite. Potros provenientes de éguas com placentite foram menores em peso e altura ao nascimento, e apresentaram concentrações elevadas de ureia e reduzidas de PPT na avaliação sérica. Foi observado correlação positiva moderada entre creatinina e ureia no LA e correlação positiva fraca entre a creatinina do LA e sérica dos potros. Concluiu-se que o perfil bioquímico do LA e sérico dos neonatos oriundos de éguas com placentite ascendente diferem de gestações sadias. Sendo que esses parâmetros podem ser utilizados como indicadores de alterações placentárias em éguas.(AU)

The aim of this study was to identify the biochemical parameters of amniotic fluid (AF) at parturition in mares with ascending placentitis, and to relate with biochemical parameters in neonatal foals serum. Thirty-six pregnant Thoroughbred mares were divided in 2 groups: mares with placentitis (n=5) and healthy mares (n=31). Placentitis was identified based on clinical signs, ultrasound evaluation and histopathological features in the placenta. Mares with placentitis were treated with antibiotics, antiinflammatories and altrenogest. AF was collected during the second stage of labor and blood samples were taken immediately after foaling. Lower concentrations of creatinine were found in the AF of mares with placentitis. Foals from mares with placentitis showed smaller height and lower birthweight, also showed increase of urea and decrease of total protein levels in blood serum. A moderate positive correlation between urea and creatinine in AF was observed, and a weak positive correlation was found between creatinine in AF and the serum of foals. In conclusion, biochemical composition of AF and of blood serum of neonatal foals from mares with ascending placentitis are different of healthy mares. So, those analysis could be useful to identify placental changes in mares.(AU)

Plasmodium Infection Induces Dyslipidemia and a Hepatic Lipogenic State in the Host through the Inhibition of the AMPK-ACC Pathway.

Malaria is a major parasitic disease of humans and is a health public problem that affects more than 100 countries. In 2017, it caused nearly half a million deaths out of 219 million infections. Malaria is caused by the protozoan parasites of the genus Plasmodium and is transmitted by female mosquitoes of the genus Anopheles. Once in the bloodstream, Plasmodium merozoites invade erythrocytes and proliferate until the cells lyses and release new parasites that invade other erythrocytes. Remarkably, they can manipulate the vertebrate host's lipid metabolism pathways, since they cannot synthesize lipid classes that are essential for their development and replication. In this study, we show that mice infected with Plasmodium chabaudi present a completely different plasma profile from control mice, with marked hyperproteinemia, hypertriglyceridemia, hypoglycemia, and hypocholesterolemia. In addition, white adipose and hepatic tissue and analyses from infected animals revealed the accumulation of triacylglycerol in both tissues and free fatty acids and free cholesterol in the liver. Hepatic mRNA and protein expression of key enzymes and transcription factors involved in lipid metabolism were also altered by P. chabaudi infection, leading to a lipogenic state. The enzyme 5' AMP-activated protein kinase (AMPK), a master regulator of cell energetic metabolism, was also modulated by the parasite, which reduced AMPK phosphorylation levels upon infection. Pretreatment with metformin for 21 days followed by infection with P. chabaudi was effective in preventing infection of mice and also lowered the hepatic accumulation of lipids while activating AMPK. Together, these results provide new and important information on the specific molecular mechanisms induced by the malaria parasite to regulate hepatic lipid metabolism in order to facilitate its development, proliferation, and lifespan in its vertebrate host.

Immunolocalization of Leptin and its Receptor (ObR-b) in Equine Placenta at Term and Plasma Level Measurement in the Late Gestation.

Placental tissues from humans, rodents, and farm animal contain leptin and its receptor. Expression of leptin has already been described in horses, although there is no description about immunolocalization of leptin and its receptor. The aim of the present study was to investigate the presence and distribution of leptin and ObR-b in the equine placenta at term by immunofluorescence staining, and the changes on plasma leptin concentrations during late gestation. The present study involved eight Criollo-type mares carrying healthy pregnancies. Blood samples were collected during the third trimester of gestation, at foaling, and at 24 hours after foaling. Leptin concentrations were analyzed via radioimmunoassay. Plasma leptin concentrations did not change from the 8th to the 10th month of gestation and displayed a discrete decrease 24 hours after parturition (P = .07). The expression of leptin and ObR-b was observed in the cytoplasm of pseudostratified epithelial cells in the areolar region and in the epithelium of microcotyledons. Also, leptin receptor was allocated in the apical surface of the cells. The presence of leptin and its receptor (ObR-b) in the placenta of mares at term supports an endocrine and autocrine/paracrine action of leptin within this organ.

Adipose-Derived Mesenchymal Stromal Cells Under Hypoxia: Changes in Extracellular Vesicles Secretion and Improvement of Renal Recovery after Ischemic Injury.

BACKGROUND/AIMS: The therapeutic potential of extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) in kidney injury has been largely reported. However, new approaches are necessary to optimize the efficacy in the treatment of renal diseases. MSCs physiologically are under a low O2 partial pressure (pO2), and culturing adipose-derived MSCs (ADMSCs) in hypoxia alters their secretory paracrine properties. The aim of this study was to evaluate whether hypoxia preconditioning of ADMSCs alters the properties of secreted EVs to improve renal recovery after ischemia-reperfusion injury (IRI). METHODS: The supernatants of ADMSCs cultivated under 21% pO2 (control) or 1% pO2 (hypoxia) were ultracentrifuged for EVs isolation that were posteriorly characterized by flow cytometry and electron microscopy. The uptake and effects of these EVs were analyzed by using in vitro and in vivo models. HK-2 renal tubule cell line was submitted do ATP depletion injury model. Proteomic analyses of these cells treated with EVs after injury were performed by nano-UPLC tandem nano-ESI-HDMSE method. For in vivo analyses, male Wistar rats were submitted to 45 min bilateral ischemia, followed by renal intracapsular administration of ADMSC-EVs within a 72 h reperfusion period. Histological, immunohistochemical and qRT-PCR analysis of these kidneys were performed to evaluate cell death, inflammation and oxidative stress. Kidney function was evaluated by measuring the blood levels of creatinine and urea. RESULTS: The results demonstrate that hypoxia increases the ADMSCs capacity to secrete EVs that trigger different energy supply, antiapoptotic, immunomodulatory, angiogenic and anti-oxidative stress responses in renal tissue compared with EVs secreted in normoxia. Proteomic analyses of renal tubule cells treated with EVs from ADMSCs in normoxia and hypoxia give a specific signature of modulated proteins for each type of EVs, indicating regulation of distinct biological processes. CONCLUSION: In summary, hypoxia potentially offers an interesting strategy to enhance the properties of EVs in the treatment of acute kidney disease.

Monitoramento da gestação em éguas – o que fazer na rotina de campo / Monitoring pregnancy in mares – What to do in the field routine

As complicações gestacionais em éguas, em especial no terço final da gestação, representam um grande problema para a indústria equina, podendo acarretar em consequências maternas, fetais e em implicações no próprio potro neonato. A garantia de uma gestação adequada abrange as trocas metabólicas entre o potro e a égua, e a manutenção do adequado ambiente intra-uterino para o desenvolvimento fetal. Assim, objetivo do presente é apresentar métodos de monitoramento de éguas gestantes, com ênfase nas gestações de risco, buscando o nascimento de um potro saudável. Os procedimentos iniciam pela avaliação do histórico e exame regular das éguas. Essa conduta consiste no exame clínico obstétrico, ultrassonografia transretal e transabdominal. Além da avaliação de alguns biomarcadores séricos, ainda que controversos na literatura, como a dosagem de progesterona, estrógenos (estrógenos totais e/ou 17β- estradiol), amiloide A sérica e alfa-feto proteína quando disponíveis.

Complications in pregnant mares, especially in late gestation, remains a major cause of economic loss for horse breeders. Those could lead maternal, fetal and neonatal implications. Healthy intrauterine environment and on placental efficiency are essential for fetal development. Therefore, the aim of this review is described methods of monitoring pregnant mares to seek a healthy neonatal foal, particularly in a high-risk pregnancy. The first step is the collection of medical histories and systematic clinical examination of the mares. This regular examination realizes clinical obstetric evaluation, transrectal and transabdominal ultrasound exam. In addition to peripheral blood markers assessment, although sometimes controversial, as progesterone, estrogens (total estrogen and/or 17βestradiol), serum amyloide A and alpha-fetoprotein when available.

Gestação gemelar em éguas (parte 1): Desenvolvimento e Fatores de Risco / Twin pregnancies in mares, part I: Development and risk factors / Embarazos gemelares en yeguas, parte l: desarrollo y factores de riesgo

Gestações gemelares em equinos são responsáveis por abortos no terço final de gestação ou nascimento de potros debilitados. O objetivo desta revisão é discutir os fatores de risco para o de gêmeos, sendo mais frequentes em éguas Puro Sangue Inglês, de Polo Argentino e Brasileiro e de Hipismo quando comparadas a Árabes e Pôneis. Éguas solteiras terão maior incidência do múltiplas ovulações e gestações gemelares e éguas mais velhas possuem maiores taxas de múltiplas ovulações, mas não de gêmeos. O uso de indutores de ovulação pode elevar as taxas de ovulações duplas e à medida que a estação reprodutiva progride, aumenta a incidência de gestações gemelares. O conhecimento desses fatores de risco é importante para a realização do diagnóstico e intervenção adequada para evitar prejuízos causados por essas gestações.

Twin pregnancies in horses are responsible for abortion in the late gestation or birth of debilitated foals. The aim of this review is to discuss the risk factors for the development of twin pregnancies. The multiple ovulations are responsible for the occurrence of twins, being more frequent in Thoroughbred, Polo Argentino and Brazilian Sport Horse when compared to Arabs and Ponies. Single mares will have a higher incidence of multiple ovulations and twin pregnancies and older mares have higher rates of multiple ovulations but not twins. The use of ovulation inducers may increase double ovulation rates and as the reproductive season progresses, the incidence of twin pregnancies increases. Knowledge of these risk factors is important for proper diagnosis and intervention to avoid loses caused by this pregnancies.

Los embarazos gemelares en yeguas son pueden causar abortos en la gestación tardía o el nacimiento de potros debilitados. El objetivo de esta revisión es analizar los factores de riesgo para el desarrollo de embarazos gemelares. Las ovulaciones múltiples son responsables por la ocurrencia de gemelos, siendo más frecuentes en yeguas Pura Sangre Inglés, de Polo Argentino y Brasileiro de Hipismo en comparación con las Árabes y Ponis. Las yeguas solteras tendrán una mayor incidencia de ovulaciones múltiples y embarazos gemeiares, y las yeguas mayores tienen tasas más altas de ovulaciones múltiples pero no gemelos. El uso de inductores de la ovulación puede aumentar las tasas de ovulación doble y a medida que avanza la temporada reproductiva aumenta la incidencia de embarazos gemelares. El conocimiento de estos factores de riesgo es importante para un diagnóstico e intervención adecuados para eviar daños causados por estos embarazos.

Monitoramento da gestação em éguas o que fazer na rotina de campo / Monitoring pregnancy in mares What to do in the field routine

As complicações gestacionais em éguas, em especial no terço final da gestação, representam um grande problema para a indústria equina, podendo acarretar em consequências maternas, fetais e em implicações no próprio potro neonato. A garantia de uma gestação adequada abrange as trocas metabólicas entre o potro e a égua, e a manutenção do adequado ambiente intra-uterino para o desenvolvimento fetal. Assim, objetivo do presente é apresentar métodos de monitoramento de éguas gestantes, com ênfase nas gestações de risco, buscando o nascimento de um potro saudável. Os procedimentos iniciam pela avaliação do histórico e exame regular das éguas. Essa conduta consiste no exame clínico obstétrico, ultrassonografia transretal e transabdominal. Além da avaliação de alguns biomarcadores séricos, ainda que controversos na literatura, como a dosagem de progesterona, estrógenos (estrógenos totais e/ou 17β- estradiol), amiloide A sérica e alfa-feto proteína quando disponíveis.(AU)

Complications in pregnant mares, especially in late gestation, remains a major cause of economic loss for horse breeders. Those could lead maternal, fetal and neonatal implications. Healthy intrauterine environment and on placental efficiency are essential for fetal development. Therefore, the aim of this review is described methods of monitoring pregnant mares to seek a healthy neonatal foal, particularly in a high-risk pregnancy. The first step is the collection of medical histories and systematic clinical examination of the mares. This regular examination realizes clinical obstetric evaluation, transrectal and transabdominal ultrasound exam. In addition to peripheral blood markers assessment, although sometimes controversial, as progesterone, estrogens (total estrogen and/or 17βestradiol), serum amyloide A and alpha-fetoprotein when available.(AU)

Gestação gemelar em éguas (parte 1): Desenvolvimento e Fatores de Risco / Twin pregnancies in mares, part I: Development and risk factors / Embarazos gemelares en yeguas, parte l: desarrollo y factores de riesgo

Gestações gemelares em equinos são responsáveis por abortos no terço final de gestação ou nascimento de potros debilitados. O objetivo desta revisão é discutir os fatores de risco para o de gêmeos, sendo mais frequentes em éguas Puro Sangue Inglês, de Polo Argentino e Brasileiro e de Hipismo quando comparadas a Árabes e Pôneis. Éguas solteiras terão maior incidência do múltiplas ovulações e gestações gemelares e éguas mais velhas possuem maiores taxas de múltiplas ovulações, mas não de gêmeos. O uso de indutores de ovulação pode elevar as taxas de ovulações duplas e à medida que a estação reprodutiva progride, aumenta a incidência de gestações gemelares. O conhecimento desses fatores de risco é importante para a realização do diagnóstico e intervenção adequada para evitar prejuízos causados por essas gestações.(AU)

Twin pregnancies in horses are responsible for abortion in the late gestation or birth of debilitated foals. The aim of this review is to discuss the risk factors for the development of twin pregnancies. The multiple ovulations are responsible for the occurrence of twins, being more frequent in Thoroughbred, Polo Argentino and Brazilian Sport Horse when compared to Arabs and Ponies. Single mares will have a higher incidence of multiple ovulations and twin pregnancies and older mares have higher rates of multiple ovulations but not twins. The use of ovulation inducers may increase double ovulation rates and as the reproductive season progresses, the incidence of twin pregnancies increases. Knowledge of these risk factors is important for proper diagnosis and intervention to avoid loses caused by this pregnancies.(AU)

Los embarazos gemelares en yeguas son pueden causar abortos en la gestación tardía o el nacimiento de potros debilitados. El objetivo de esta revisión es analizar los factores de riesgo para el desarrollo de embarazos gemelares. Las ovulaciones múltiples son responsables por la ocurrencia de gemelos, siendo más frecuentes en yeguas Pura Sangre Inglés, de Polo Argentino y Brasileiro de Hipismo en comparación con las Árabes y Ponis. Las yeguas solteras tendrán una mayor incidencia de ovulaciones múltiples y embarazos gemeiares, y las yeguas mayores tienen tasas más altas de ovulaciones múltiples pero no gemelos. El uso de inductores de la ovulación puede aumentar las tasas de ovulación doble y a medida que avanza la temporada reproductiva aumenta la incidencia de embarazos gemelares. El conocimiento de estos factores de riesgo es importante para un diagnóstico e intervención adecuados para eviar daños causados por estos embarazos.(AU)