Morphometric analysis of fetal development of Cavia porcellus (Linnaeus, 1758) by ultrasonography: pilot study

Measurements on the growth process and placental development of the embryo and fetuses of Cavia porcellus were carried out using ultrasonography. Embryo, fetus, and placenta were monitored from Day 15 after mating day to the end of gestation. Based on linear and quadratic regressions, the following morphometric analysis showed a good indicator of the gestational age: placental diameter, biparietal diameter, renal length, and crown rump. The embryonic cardiac beat was first detected at an average of 22.5 days. The placental diameter showed constant increase from beginning of gestation then remained to term and presented a quadratic correlation with gestational age (r2 = 0.89). Mean placental diameter at the end of pregnancy was 3.5 ± 0.23 cm. By Day 30, it was possible to measure biparietal diameter, which followed a linear pattern of increase up to the end of gestation (r2 = 0.95). Mean biparietal diameter in the end of pregnancy was 1.94 ± 0.03 cm. Kidneys were firstly observed on Day 35 as hyperechoic structures without the distinction of medullar and cortical layers, thus the regression model equation between kidney length and gestational age presents a quadratic relationship (r2 = 0.7). The crown rump presented a simple linear growth, starting from 15 days of gestation, displaying a high correlation with the gestational age (r2 = 0.9). The offspring were born after an average gestation of 61.3 days. In this study, we conclude that biparietal diameter, placental diameter, and crown rump are adequate predictive parameters of gestational age in guinea pigs because they present high correlation index.

Efeito do precondicionamento isquêmico remoto no transplante de intestino fetal em camundongos

Objetivo: avaliar os efeitos de precondicionamento isquêmico remoto (PCI-R) no modelo de transplante de intestino delgado fetal. Métodos: foram constituídos dois grupos: transplante isogênico (Iso, camundongos C57BL/6, n=24) e transplante alogênico (Alo, camundongos BALB/c, n=24). Em cada grupo, distribuíram-se os animais com e sem PCI-R, que foi realizado por oclusão da artéria femoral esquerda da fêmea prenhe durante 10 minutos, seguida por tempo igual de reperfusão. O imunossupressor utilizado foi Tacrolimo (Fk, 5 mg/kg/dia v.o.). Ao final obteve-se os seguintes subgrupos: Alo-Tx, Alo-Pci, Alo-Fk, Alo-Pci-Fk, Iso-Tx, Iso-Pci, Iso-Fk e Iso-Pci-Fk. O enxerto foi transplantado no espaço entre o músculo reto-abdominal e pré- peritoneal dos receptores a meio centímetro do apêndice xifóide, à esquerda da linha mediana. Após o sétimo dia de seguimento, o enxerto foi removido, fixado e embebido em parafina para avaliação histomorfológica (desenvolvimento e rejeição) e análise imunohistoquímica (anti-PCNA e anti-caspase-3 clivada). Os dados foram analisados usando ANOVA e testes complementares e foi considerado significante quando p <0.05. Resultados: A avaliação do desenvolvimento do enxerto no grupo de Iso mostrou que o PCI-R reduziu o desenvolvimento comparado com Iso-Tx (5,2±0,4 vs 9,0±0,8), o Fk e sua associação com PCI-R aumentaram o desenvolvimento do enxerto comparado com PCI-R (11,2±0,7 e 10,2±0,8, respectivamente). No grupo Alo, o Fk e/ou sua associação com PCI-R aumentaram o desenvolvimento comparado com Alo-Tx e Alo com PCI-R (6,0±0,8, 9,0±1,2, 0,0±0,0, 0,5±0,3, respectivamente). A expressão de PCNA foi maior no grupo ISO em animais tratados com Fk e PCI-R comparados a outros grupos (12,2±0,8 vs Tx: 8,8±0,9, PCI-R: 8,0±0,4 e Fk: 9,0±0,6). No grupo Alo, a expressão de PCNA não diferiu entre grupos. A rejeição do enxerto foi menor nos grupos tratados com PCI-R (-18%), Fk (- 68%) ou ambos (-61%) comparados com Alo-Tx. A expressão de caspase-3 clivada foi menor no grupo Iso em animais tratados com associação de PCI-R e Fk (6,2 ±0,9 vs Tx: 8,6±0,5; PCI-R: 5,8 ±0,9 e Fk: 6,0 ±0,3). Conclusão: O PCIR mostrou efeito benéfico sobre a lesão de isquemia e reperfusão do enxerto intestinal fetal nos transplantes isogênico e alogênico, aumentando o número de células caliciformes e a proliferação celular. No transplante alogênico, aumentou o desenvolvimento do enxerto, diminuiu o grau de rejeição aguda na ausência de imunossupressão, porém não apresentou efeito sinérgico com o imunossupressor. No transplante isogênico houve diminuição do grau de desenvolvimento do enxerto, porém foi efetivo na redução da apoptose.

Efecto inmediato de la analgesia epidural sobre el estado fetal y el trabajo de parto

Programa de doctorado: Patología quirúrgica, reproducción humana, factores psicológicos y el proceso de enfermar.

Caratteristiche biologiche e potenziale applicativo delle cellule staminali derivate da membrane fetali

La ricerca sulle cellule staminali apre nuove prospettive per approcci di terapia cellulare. Molta attenzione è concentrata sulle cellule staminali isolate da membrane fetali, per la facilità di recupero del materiale di partenza, le limitate implicazioni etiche e le caratteristiche delle popolazioni di cellule staminali residenti. In particolare a livello dell’epitelio amniotico si concentra una popolazione di cellule (hAECs) con interessanti caratteristiche di staminalità, pluripotenza e immunomodulazione. Restano però una serie di limiti prima di arrivare ad un’applicazione clinica: l’uso di siero di origine animale nei terreni di coltura e le limitate conoscenze legate alla reazione immunitaria in vivo. La prima parte di questo lavoro è focalizzata sulle caratteristiche delle hAECs coltivate in un terreno privo di siero, in confronto a un terreno di coltura classico. Lo studio è concentrato sull’analisi delle caratteristiche biologiche, immunomodulatorie e differenziative delle hAECs. L’interesse verso le caratteristiche immunomodulatorie è legato alla possibilità che l’uso di un terreno serum free riduca il rischio di rigetto dopo trapianto in vivo. La maggior parte degli studi in vivo con cellule isolate da membrane fetali sono stati realizzati con cellule di derivazione umana in trapianti xenogenici, ma poco si sa circa la sopravvivenza di queste cellule in trapianti allogenici, come nel caso di trapianti di cellule di derivazione murina in modelli di topo. La seconda parte dello studio è focalizzata sulla caratterizzazione delle cellule derivate da membrane fetali di topo (mFMSC). Le caratteristiche biologiche, differenziative e immunomodulatorie in vitro e in vivo delle mFMSC sono state confrontate con i fibroblasti embrionali di topo. In particolare è stata analizzata la risposta immunitaria a trapianti di mFMSC nel sistema nervoso centrale (CNS) in modelli murini immunocompetenti.

Retrospective comparison of maternal vs. HPA-matched donor platelets for treatment of fetal alloimmune thrombocytopenia

In fetal alloimmune thrombocytopenia (FAIT), transplacental maternal antibodies cause destruction of fetal platelets. FAIT is similar to fetal Rhesus haemolytic disease, but half of the affected fetuses are born to primiparous women. In 10-20% of cases, prenatal and perinatal intracranial haemorrhages are reported. Different therapeutic approaches have been described, including maternally administered high-dose intravenous immunoglobulin (high dose IVIG) without or with steroids or intrauterine transfusion (IUT) of compatible platelets. For the latter, the use of plasma-free maternal and donor platelets has been described, but a comparison of these two sources of platelets has not been reported.

Human prostate fibroblasts induce growth and confer castration resistance and metastatic potential in LNCaP Cells

The tumor microenvironment is important for progressive and metastatic disease.

Wound stimulation by growth-arrested human keratinocytes and fibroblasts: HP802-247, a new-generation allogeneic tissue engineering product

HP802-247 is a new-generation, allogeneic tissue engineering product consisting of growth-arrested, human keratinocytes (K) and fibroblasts (F) delivered in a fibrin matrix by a spray device.

Antifibrotic effects of tocotrienols on human Tenon's fibroblasts

Antifibrotic effects of α- (40, 60, 80, 100, and 120 μM), γ- (10, 20, 30, and 40 μM) and δ-tocotrienol (10, 20, 30, and 40 μM) on hTf cultures were evaluated by performing proliferation, migration and collagen synthesis assays. Whereas for vitamin E the exposure time was set to 7 days to mimic subconjunctival application, cultures were exposed only 5 min to mitomycin C 100 μg/ml to mimic intraoperative administration. Cell morphology (phase contrast microscopy) as an assessment for cytotoxicity and cell density by measuring DNA content in a fluorometric assay to determine proliferation inhibition was performed on day 0, 4, and 7. Migration ability and collagen synthesis of fibroblasts were measured. Results All tested tocotrienol isoforms were able to significantly inhibit hTf proliferation in a dose-dependent manner (maximal inhibitory effect without relevant morphological changes at day 4 for α-tocotrienol 80 μM with 36.7% and at day 7 for α-tocotrienol 80 μM with 42.6% compared to control). Degenerative cell changes were observed in cultures with concentrations above 80 μM for α- and above 30 μM for γ- and δ-tocotrienol. The highest collagen synthesis inhibition has been found with 80 µM α-tocotrienol (62.4%) and no significant inhibition for mitomycin C (2.5%). Migration ability was significantly reduced in cultures exposed to 80 µM α- and 30 µM γ-tocotrienol (inhibition of 82.2% and 79.5%, respectively, compared to control) and also after mitomycin C treatment (60.0%). Complete growth inhibition without significant degenerative cell changes could only be achieved with mitomycin C. Conclusion In vitro, all tested tocotrienol isoforms were able to inhibit proliferation, migration and collagen synthesis of human Tenon’s fibroblasts and therefore may have the potential as an anti-scarring agent in filtrating glaucoma surger

Pericellular fibronectin is required for RhoA-dependent responses to cyclic strain in fibroblasts

To test the hypothesis that the pericellular fibronectin matrix is involved in mechanotransduction, we compared the response of normal and fibronectin-deficient mouse fibroblasts to cyclic substrate strain. Normal fibroblasts seeded on vitronectin in fibronectin-depleted medium deposited their own fibronectin matrix. In cultures exposed to cyclic strain, RhoA was activated, actin-stress fibers became more prominent, MAL/MKL1 shuttled to the nucleus, and mRNA encoding tenascin-C was induced. By contrast, these RhoA-dependent responses to cyclic strain were suppressed in fibronectin knockdown or knockout fibroblasts grown under identical conditions. On vitronectin substrate, fibronectin-deficient cells lacked fibrillar adhesions containing alpha5 integrin. However, when fibronectin-deficient fibroblasts were plated on exogenous fibronectin, their defects in adhesions and mechanotransduction were restored. Studies with fragments indicated that both the RGD-synergy site and the adjacent heparin-binding region of fibronectin were required for full activity in mechanotransduction, but not its ability to self-assemble. In contrast to RhoA-mediated responses, activation of Erk1/2 and PKB/Akt by cyclic strain was not affected in fibronectin-deficient cells. Our results indicate that pericellular fibronectin secreted by normal fibroblasts is a necessary component of the strain-sensing machinery. Supporting this hypothesis, induction of cellular tenascin-C by cyclic strain was suppressed by addition of exogenous tenascin-C, which interferes with fibronectin-mediated cell spreading.

Causes and mechanisms of intrauterine hypoxia and its impact on the fetal cardiovascular system: a review

Until today the role of oxygen in the development of the fetus remains controversially discussed. It is still believed that lack of oxygen in utero might be responsible for some of the known congenital cardiovascular malformations. Over the last two decades detailed research has given us new insights and a better understanding of embryogenesis and fetal growth. But most importantly it has repeatedly demonstrated that oxygen only plays a minor role in the early intrauterine development. After organogenesis has taken place hypoxia becomes more important during the second and third trimester of pregnancy when fetal growth occurs. This review will briefly adress causes and mechanisms leading to intrauterine hypoxia and their impact on the fetal cardiovascular system.

Techniques for assessing cardiac output and fetal cardiac function

Fetal echocardiography was initially used to diagnose structural heart disease, but recent interest has focused on functional assessment. Effects of extracardiac conditions on the cardiac function such as volume overload (in the recipient in twin-twin transfusion syndrome), a hyperdynamic circulation (arterio-venous malformation), cardiac compression (diaphragmatic hernia, lung tumours) and increased placental resistance (intrauterine growth restriction and placental insufficiency) can be studied by ultrasound and may guide decisions for intervention or delivery. A variety of functional tests can be used, but there is no single clinical standard. For some specific conditions, however, certain tests have shown diagnostic value.

Repeated intrauterine IgG infusions in fetal alloimmune thrombocytopenia do not increase foetal platelet counts

Foetal alloimmune thrombocytopenia (FNAIT) is often treated transplacentally with maternally administered i.v. immunoglobulins, but not all foetuses show a consistent platelet increase during such treatment.