Derivation of stem cell lines from domesticated animals has been of great interest as it benefits translational medicine, clinical applications to improve human and animal health and biotechnology. The main types of stem cells studied are Embryonic Stem Cells (ESCs), induced Pluripotent Stem Cells (iPSCs) and Mesenchymal Stem/Stromal Cells (MSCs). This thesis had two main aims: (I) The isolation of bovine MSCs from amniotic fluid (AF) at different trimesters of pregnancy and their characterization to study pluripotency markers expression. Stemness markers were studied also in MSCs isolated from equine AF, Wharton’s jelly (WJ) and umbilical cord blood (UCB) as continuation of the characterization of these cells previously performed by our research group; (II) The establishment and characterization of iPSCs lines in two attractive large animal models for biomedical and biotechnology research such as the bovine and the swine, and the differentiation into the myogenic lineage of porcine iPSCs. It was observed that foetal tissues in domestic animals such as the bovine and the horse represent a source of MSCs able to differentiate into the mesodermal lineage but they do not proliferate indefinitely and they lack the expression of many pluripotency markers, making them an interesting source of cells for regenerative medicine, but not the best candidate to elucidate pluripotency networks. The protocol used to induce pluripotency in bovine fibroblasts did not work, as well as the chemical induction of pluripotency in porcine fibroblasts, while the reprogramming protocol used for porcine iPSCs was successful and the line generated was amenable to being differentiated into the myogenic lineage, demonstrating that they could be addressed into a desired lineage by genetic modification and appropriated culture conditions. Only a few cell types have been differentiated from domestic animal iPSCs to date, so the development of a reliable directed-differentiation protocol represents a very important result.

La placenta è stata descritta come il "diario di vita intrauterina" e ha il potenziale per chiarire molti aspetti del periodo pre- e post-natale. La presente inchiesta mira a studiare e descrivere aspetti patologici macroscopici e istologici di placente canine a termine gravidanza, con le relative correlazioni clinico-patologiche tra i risultati placentari e outcome neonatale. Lo scopo di questo studio è stato applicare, per la prima volta nella specie canina, tecniche di analisi d'immagine (ImagJ) per andare a valutare alcuni aspetti istologici placentari, in particolare la microvascolarizzazione placentare. Lo studio ha anche lo scopo di verificare se queste modificazioni placentari possano influenzare lo stato di salute del cucciolo, nonché mettere in relazione parametri clinici del cucciolo, quali Apgar score, temperatura, glicemia e lattatemia a tempo zero e a due ore dalla nascita , tra di loro e con il tipo di parto svolto (spontaneo, cesareo d’urgenza e cesareo programmato). Sono rientrati nello studio dati clinici e materiale istologico placentare raccolto da 33 cagne e 144 cuccioli nel periodo gennaio 2013 - dicembre 2015. Ne è emerso che la placenta è un organo plastico e adattabile che risente “dell’effetto dimensione cucciolata” con aumento della densità capillare nelle placente più piccole ottenute in cucciolate più numerose. L’influenza delle lesioni istologiche lobulari sull’outcome del neonato sono da considerarsi irrilevanti, la stessa cosa non si può affermare per le lesioni multilobulari. Pare quanto riguarda i parametri clinici del cucciolo pare siano influenzate dal tipo di parto affrontato. Infatti la lattatemia e la temperatura alla nascita risultano essere più alte nel parto spontaneo, la glicemia alla nascita è più alta invece nel parto cesareo programmato. Al contrario, per quanto riguarda l’Apgar score e la sopravvivenza a 24 ore, pare che questi non siano influenzati dai diversi tipi di parto. / Placenta has been described as the "diary of intrauterine life" and it has the potential to clarify many aspects of pre- and post-natal period. The following investigation aims to study and describe the pathological macroscopic and histological aspects of canine placentas at the end of the pregnancy, with the related clinical and pathological correlations between placental findings and neonatal outcome. The purpose of this study was to apply, for the first time in the canine species, an image analysis technique (ImagJ) in order to assess some histological features of placenta, in particular the placental microvasculature. The study also aims to verify if these placental modifications can affect the puppy's health, as well as to correlate the clinical parameters of the newborn, such as Apgar score, temperature, blood glucose and lactate assessed at birth and two hours later, among themselves and with the type of delivery (spontaneous, emergency cesarean and planned caesarean). This study includes clinical data and histological material from placentas collected from 33 bitches and 144 puppies between January 2013 and December 2015. It emerged that placenta is a plastic and adaptable organ that suffers of the "effect of litter size" with an increase of the capillary density in smaller placentas obtained in bigger litters. The influence of lobular histological lesions on the outcome of the puppy should be considered irrelevant, the same cannot be said for multi-lobular ones. Moreover, it seems that the clinical parameters of the puppy are affected by the type of delivery. In fact, lactatemia and body temperature at birth appear to be higher in case of natural delivery, while glycaemia at birth is higher in case of planned caesarean section. On the contrary, it seems that the Apgar score and the survival at 24 hours are not influenced by the different types of birth.

No description available.

Admission blood lactate concentration has been shown to be a useful indicator of disease severity in human medicine and numerous studies have associated hyperlactatemia with patients at high risk of death who should be treated aggressively regardless of the cause of the lactate generation. The degree and duration of hyperlactacidaemia also have been correlated with the subsequent development of organ failure. Similarly, in a small number of studies about equine colic, blood lactate concentration has been investigated as a useful prognostic variable . In neonatal foals blood lactate was studied first by Magdesian (2003) who described venous blood lactate concentration in 14 normal foals during the initial 48 hours post-partum. A preliminary study about lactate concentration in foals presenting to a neonatal intensive care unit reported that surviving foals had earlier lactate clearance. The measurement of blood lactate concentration is traditionally available with a wet chemistry laboratory method or with blood-gas analyzers, for clinicians working at university or large private hospital. But this methods may not be easily accessible to many practitioners in field conditions. Several relatively inexpensive, easy to use and rapid pocket size monitors to measure lactate concentration have been validated in human patients and athletes. None of these portable lactate analyzer have been evaluated in clinically normal neonatal foals or in foals referred to a neonatal intensive care unit. The aims of this study were to validate the Lactate Scout analyzer in neonatal foals, investigating the correlation between lactate concentration in whole blood measured with the portable monitor and measured in plasma with the reference laboratory analyzer. The effect of hematocrit (Hct) on the accuracy of Lactate Scout was also evaluated. Further, we determined the utility of venous lactate measurement in critically-ill foals, describing lactate values in the most frequent neonatal pathologies, evaluating serial blood lactate measurements during hospitalization and investigating its prognostic value. The study also describes normal range for lactate in healthy neonatal foals during the first 72 hours of life.

No description available.

Over the past few years, in veterinary medicine there has been an increased interest in understanding the biology of mesenchymal stem cells (MSCs). This interest comes from their potential clinical use especially in wound repair, tissue engineering and application in therapeutics fields, including regenerative surgery. MSCs can be isolated directly from bone marrow aspirates, adipose tissue, umbilical cord and various foetal tissues. In this study, mesenchymal stem cells were isolated from equine bone marrow, adipose tissue, cord blood, Wharton’s Jelly and, for the first time, amniotic fluid. All these cell lines underwent in vitro differentiation in chondrocytes, osteocytes and adipocytes. After molecular characterization, cells resulted positive for mesenchymal markers such as CD90, CD105, CD44 and negative for CD45, CD14, CD34 and CD73. Adipose tissue and bone marrow mesenchymal stem cells were successfully applied in the treatment of tendinitis in race horses. Furthermore, for the first time in the horse, skin wounds of septicemic foal, were treated applying amniotic stem cells. Finally, results never reported have been obtained in the present study, isolating mesenchymal stem cells from domestic cat foetal fluid and membranes. All cell lines underwent in vitro differentiation and expressed mesenchymal molecular markers.

Lo sviluppo e la funzionalità della placenta influenzano direttamente la crescita ed il benessere del feto all'interno dell'utero, quindi qualsiasi problema strutturale o funzionale della placenta influenzerà lo sviluppo del feto. Lo scopo di questa tesi è stato quello di approfondire diversi aspetti clinici e clinico-patologici dell’insufficienza placentare nella specie equina, con l’intento di individuare dei parametri che possano essere di ausilio per l’identificazione precoce del puledro a rischio e della necessità di interventi terapeutici. La valutazione della concentrazione di lattato nel sangue e nel liquido amniotico potrebbe essere un utile strumento diagnostico per la diagnosi di acidosi metabolica associata ad ipossia/ischemia nel puledro e per identificare la necessità di un intervento precoce alla nascita. La risposta all’ipossia sembra essere mediata dall’HIF-1 e dall’HSF-1 anche nel puledro neonato, e se questi dati venissero confermati su un numero maggiore di animali, i due marcatori proteici e la MDA potrebbero essere utilizzati per la diagnosi di PAS nel puledro. L’esame di tutta l’unità placentare riveste un ruolo di fondamentale importanza per l’acquisizione di informazioni riguardo all’ambiente di vita intrauterino del puledro, ed è quindi auspicabile nella pratica ostetrica routinaria una maggiore attenzione all’esame della placenta, soprattutto in caso di patologie materno-fetali. Tra i parametri biochimici valutati al momento della nascita, la creatininemia e la glicemia possono fornire informazioni sull’efficienza dello scambio placentare ed essere quindi utilizzati per individuare puledri a rischio. Infine, lo sviluppo di una macro per il software ImageJ porta alla luce uno strumento nuovo, semplice da usare ed economico, per la valutazione morfometrica dell’arborizzazione dei villi placentari; tuttavia la ricerca necessità ulteriori indagini su un numero maggiore di animali per valutare le differenze morfometriche tra placente normali e patologiche. / Fetal health and growth are directly influenced by the development and function of the placenta. For this reason, structural or functional problems of the placenta may affect fetal development. The aim of this thesis s to investigate clinicopathological features of placental insufficiency in horses, in order to detect the most useful parameters in identifying foals at risk and the need for therapeutic interventions. The evaluation of blood and amniotic fluid lactate concentrations could be a useful tool to diagnose metabolic acidosis associated with hypoxia/ischemia in foal, and to identify the need for early intervention at birth. The body’s reactions to hypoxia are mediated by HIF-1α and HSF-1 also in the newborn foal, and if these data were confirmed on a greater number of animals, HIF-1α and HSF-1 expression and MDA concentration could be used for clinical diagnosis of PAS. Examination of fetal membranes plays a key role to acquire information concerning the intrauterine environment, and a careful examination of the placenta is recommended in routine obstetrical practice, especially in case of maternal and/or fetal diseases. Among biochemical parameters evaluated at birth, blood creatinine and glucose levels can provide information about the efficiency of placental exchange and thus it could be used for identifying foals at risk. At the end, the development of a macro for the software ImageJ provides a new, simple to use and inexpensive tool for the morphometric evaluation of the chorionic villi, but additional studies on a larger number of animals are required to evaluate the morphometric differences between normal and pathological placentas.

La Sindrome da Asfissia Perinatale (PAS) è una delle più comuni patologie che colpiscono il puledro neonato nelle prime 72 h di vita. È una patologia difficile da diagnosticare in quanto non esistono parametri o segni clinici specifici, la sintomatologia è molto variabile in base alla durata e all’intensità dell’insulto ipossico ischemico e al tipo di organo maggiormente colpito. Lo scopo di questo studio è la ricerca e la valutazione di alcuni parametri biochimico-clinici e di alcuni biomarkers per la diagnosi precoce e il corretto trattamento dei puledri affetti da PAS. Nei puledri neonati che presentano questa patologia è stata riscontrata un’ipermagnesiemia al momento del ricovero associata a prognosi infausta, probabilmente causata da un grave danno cellulare con rilascio in circolo del magnesio intracellulare. La PAS potrebbe essere un’ulteriore causa di Euthyroid Sick Syndrome, in quanto abbiamo riscontrato una diminuzione delle concentrazioni di T3 e T4 nei puledri malati rispetto ai sani della stessa età, come avviene in altre malattie sistemiche. Lo studio del profilo proteomico ha permesso di separare le più importanti frazioni proteiche del liquido amniotico di cavalla, mettendo in evidenza similitudini e differenze qualitative e quantitative nei ferogrammi dei puledri sani e di quelli affetti da PAS ed una maggiore variabilità è stata riscontrata nei profili dei liquidi amniotici dei puledri malati. Il glutatione è risultato poco espresso nel puledro neonato, i puledri sani presentano concentrazioni più basse sia rispetto ai malati della stessa età sia agli adulti ma con una tendenza all’aumento nelle prime 24 ore di vita per i sani ed un calo nei malati. La somministrazione della terapia antiradicalica non influisce sulle concentrazioni di glutatione totale ed i puledri deceduti presentano concentrazioni più alte. / Perinatal Asphyxia Syndrome (PAS) that occurs during the perinatal period is one of the most common diseases affecting foals within the first 72 h of life. The diagnosis of PAS is very difficult and relies on an accurate history, identification of neurological signs, and exclusion of other cause of neurological deficits. No specific clinical-pathological and laboratory findings have been highly suggestive of neonatal asphyxia, many of the noted clinical signs can occur with other clinical conditions and depending on the duration and the intensity of hypoxic insult and the tissue injury. The aim of this study was to discover Perinatal Asphyxia Syndrome biomarkers in neonatal foals having an early identification and interventions of the foals at highest risk developing this syndrome. Foals affected by PAS present hypermagnesaemia at admission compared to healthy foals, may be the results of serious tissue damage with cell death and release of intracellular Mg. Higher concentrations is associated to poor outcome. Data obtained in this trial suggest that PAS may cause lower T3 and T4 concentrations in affected foals than in age-matched healthy foals, as reported for other systemic illnesses. PAS could be a cause of Euthyroid Sick Syndrome. The differences revealed between electrophoretic patterns of healthy and sick foals amniotic fluids have showed a qualitative and quantitative variability in the proteomic profile. We found a high proteomic profiling variability also among sick foals. The acute period of hypoxia-ischemia is followed by a period of reperfusion, the second tissue injury occurs in this phase. The cell damage is caused by post-ischemic release of oxygen radicals. Glutathione (endogenous antiradical) concentration (tGSH) is lower in healthy foals compared to sick foals and adults. The antiradical therapy didn’t change the tGSH concentration and no-surviving foals had higher concentration.

Made available in DSpace on 2014-06-11T19:29:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-27Bitstream added on 2014-06-13T18:58:52Z : No. of bitstreams: 1 coutinho_ln_me_jabo.pdf: 485979 bytes, checksum: 7ced96cedd272d7e86ada92ea92c5c39 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Realizou-se o exame ultrassonográfico de macacos-da-noite para avaliar os volumes uterino e ovariano e analisar a interação entre diferentes faixas etárias e número de partos. Foram realizados exames ultrassonográficos da região pélvica, a fim de comparar as dimensões uterinas com o peso, idade (infantil, juvenil, subadulta, adulta jovem e adulta) e número de partos (nulípara, primípara e multípara) e comparar as dimensões ovarianas com o peso e a idade. O volume uterino (VU) foi diretamente proporcional ao número de partos, fator mais importante no crescimento uterino das fêmeas adultas (p < 0,05). O peso e a idade demonstraram uma correlação positiva com o VU (r = 0.5354, r = 0.6489, p < 0.01), respectivamente. O volume dos ovários cresceu proporcionalmente a idade das fêmeas (p < 0,05). A puberdade foi o período de maior crescimento tanto do útero como do ovário / We aimed to evaluate the uterine and ovarian volumes of owl monkeys in different age groups with different numbers of live births and to analyze the interaction between both. We performed pelvic ultrasound exams to compare the uterine measurements with weight, age (infant, juvenile, subadult, young adults, and adults) and the number of live births (nulliparous, primiparous, and multiparous) and to compare the ovarian measurements with weight and age. The uterine volume (UV) was directly proportional to the number of parturitions, which was the most important factor in the uterine growth of adult females (P < 0.05). The body weight and age of the animals showed a high positive correlation with UV (r = 0.5354, r = 0.6489, P < 0.01), respectively. The volume of the ovaries grew in proportion to the age of the females (P < 0.05). Puberty was the period of greatest uterine and ovarian growth

Made available in DSpace on 2014-06-11T19:29:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2005Bitstream added on 2014-06-13T18:58:57Z : No. of bitstreams: 1 carvalho_lm_me_botfmvz.pdf: 178812 bytes, checksum: 797042795544766c3d38f3b7f7288cad (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O desvio ou divergência folicular é um momento importante da seleção folicular, onde pela primeira vez, nota-se uma diferença significativa na taxa de crescimento entre os dois maiores folículos ovarianos. A seleção folicular tem sido estudada intensamente em raças européias (Bos taurus taurus), porém pouco estudada em raças zebuínas (Bos taurus indicus). Objetivou-se com o presente trabalho determinar as características morfológicas do desvio associadas a primera onda folicular do ciclo estral de vacas da raça Nelore. A partir do dia da ovulação, os três maiores folículos (F1, F2 e F3) tiveram seu desenvolvimento acompanhado por ultra-sonografia transretal, realizada a cada 12 h, até o sexto dia após a ovulação. Em cada animal (n= 11), o momento do desvio foi determinado graficamente usando inspeção visual do perfil de crescimento dos F1, F2 e as vezes F3 (desvio observado) e matematicamente, usando análise de regressão linear segmentada para comparar as diferenças de diâmetro entre F1 e F2, e as vezes F3 (desvio calculado). Os resultados da determinação do momento do desvio usando os métodos matemático e observado, não foram significativamente diferentes. Utilizando-se a média dos resultados dos dois métodos, o momento do desvio ocorreu 2,4 dias após a ovulação, quando o F1 atingiu 6,1 mm. Concluiu-se que, o diâmetro do futuro folículo dominante no momento do desvio folicular foi de aproximadamente 6,1 mm em vacas da raça Nelore, cerca de 2,4 dias após a ovulação. / Follicle diameter deviation is defined as the beginning of the differential change in growth rates between the largest and next largest follicles subsequent to wave emergence and is considered a key component of follicle selection. Follicle selection has been extensively studied in European breeds of cattle (Bos taurus) but has not been critically studied in Zebu breeds (Bos indicus). The objective of the present study was to determine the morphological characteristics of deviation associated with the first post-ovulatory wave (Wave 1) of the estrous cycle in nonlactating cows. Beginning on the day of ovulation (day 0), the three largest follicles (F1-F3, respectively) were individually tracked every 12 h for 6 d using transrectal ultrasonography. In individual animals (n=11), deviation was determined graphically using visual inspection of the diameter profiles of F1, F2 and sometimes F3 (observed deviation) and mathematically using segmented regression analysis of the diameter differences between F1 and F2 or sometimes F3 (calculated deviation). The results of determining the beginning of deviation in cows using the observed and calculated methods were not significantly different. Averaged over both methods, diameter deviation occurred 2.4 d after ovulation when F1 reached 6.1 mm in cows. It is concluded that the future dominant follicle has 6.1 mm at the time of deviation in Nelore cows, 2.4 days after ovulation.