Transcriptomic profiling of trophoblast fusion using BeWo and JEG-3 cell lines.

In human placenta, alteration in trophoblast differentiation has a major impact on placental maintenance and integrity. However, little is known about the mechanisms that control cytotrophoblast fusion. The BeWo cell line is used to study placental function, since it forms syncytium and secretes hormones after treatment with cAMP or forskolin. In contrast, the JEG-3 cell line fails to undergo substantial fusion. Therefore, BeWo and JEG-3 cells were used to identify a set of genes responsible for trophoblast fusion. Cells were treated with forskolin for 48 h to induce fusion. RNA was extracted, hybridised to Affymetrix HuGene ST1.0 arrays and analysed using system biology. Trophoblast differentiation was evaluated by real-time PCR and immunocytochemistry analysis. Moreover, some of the identified genes were validated by real-time PCR and their functional capacity was demonstrated by western blot using phospho-specific antibodies and CRISPR/cas9 knockdown experiments. Our results identified a list of 32 altered genes in fused BeWo cells compared to JEG-3 cells after forskolin treatment. Among these genes, four were validated by RT-PCR, including salt-inducible kinase 1 (SIK1) gene which is specifically upregulated in BeWo cells upon fusion and activated after 2 min with forskolin. Moreover, silencing of SIK1 completely abolished the fusion. Finally, SIK1 was shown to be at the center of many biological and functional processes, suggesting that it might play a role in trophoblast differentiation. In conclusion, this study identified new target genes implicated in trophoblast fusion. More studies are required to investigate the role of these genes in some placental pathology.

Helicobacter pylori infection in children: should it be carefully assessed?

The prevalence of H. pylori infection, mainly acquired during childhood and may be persisting throughout life, has been found high in developing countries; this high prevalence is related to low socioeconomic status. The persistence of bacterium exposure is related to gastritis and other severe complications including peptic ulcer, lymphoma MALT and gastric cancer, which are rarely present in the pediatric age due to a lower inflammatory and immunological response. Virulence factors, host gastric mucosal factors, and the natural environment of patients are associated with the clinical outcome of H. pylori infection. The main bacterial virulence factors include adhesins (BabA, SabA), vacuolating cytotoxin VacA, and the products of the cag pathogenicity island (cag PAI). There are geographic differences between cagA, vacA status and H. pylori related diseases. The main criteria to evaluate H. pylori infection in children are gastrointestinal and extra gastrointestinal manifestations related to H. pylori infection, familial history of gastric cancer, peptic ulcer, lymphoma MALT, symptomatic children living in high prevalence regions, and immigrant or adopted children in developed countries. Early detection of H. pylori and its virulence factors, in addition to effective methods of eradication associated with prevention programs, may lead to the decrease of H. pylori incidence and gastritis, especially in endemic high-risk regions. The early assessment in children may prevent further severe complications in adulthood.

[Breast reconstruction for gigantomastia complicating pregnancy. A case report]. / Reconstruction mammaire après gigantomastie gestationnelle: à propos d'un cas exceptionnel.

The authors report a very rare case of gestational gigantomastia and the pregnancy could be carried out in term. The patient has been operated of bilateral mastectomy with immediate reconstruction. Surgery is the only radical treatment. Mastectomy with secondary reconstruction is reserved to cases where the intervention must be fast. Mastectomy with immediate reconstruction is the advised intervention if there is a later desire of pregnancy. Mammary reduction is more aesthetic, and allows only one intervention, but exposes to an important risk of recurrence and is not recommended if there is a later desire of pregnancy.

Involvement of renal cytochromes P450 and arachidonic acid metabolites in diabetic nephropathy.

Diabetic nephropathy (DN) is one of the most serious complications of type I and type II diabetes. DN is characterized by hyperfiltration, hypertrophy, extracellular matrix accumulation, and proteinuria. This advances into renal fibrosis and loss of renal function. Reactive oxygen species (ROS) and TGF-beta have been implicated in the pathogenesis of diabetic nephropathy. Early stages of diabetic nephropathy are also associated with alterations in renal sodium handling as well as hypertension; both are processes linked by involvement of the arachidonic acid (AA) metabolites, 20-hydroxyeicosatetraenoic acid (20-HETE, produced by cytochrome P450-4a, (CYP4A) and epoxyeicosatrienoic acids (EETs). Indeed, metabolism of AA is increased in a rat model of diabetes. In this study, we demonstrate that rats with streptozotocin-induced diabetes of 1 month duration develop renal hypertrophy and increased fibronectin and TGF-beta1 expression/cortical levels concomitant with an increase in CYP4A expression and 20 HETE production. These results were also paralleled by an increase in reactive oxygen species (ROS) production and NADPH oxidase activity. Treatment of diabetic rats with HET0016, selective inhibitor of CYP 4A, prevented all these changes. Our results suggest that diabetes-induced induction of CYP4A and 20-HETE production could be a major pathophysiological mechanism leading to activation of ROS through an NADPH dependent pathway and TGF-beta1 thus resulting in major renal pathology. Inhibitors of 20-HETE production could thus have an important therapeutic potential in the treatment of diabetic nephropathy.

IFPA Meeting 2012 Workshop Report II: epigenetics and imprinting in the placenta, growth factors and villous trophoblast differentiation, role of the placenta in regulating fetal exposure to xenobiotics during pregnancy, infection and the placenta.

Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialized topics. At IFPA meeting 2012 there were twelve themed workshops, four of which are summarized in this report. These workshops related to various aspects of placental biology: 1) epigenetics and imprinting in the placenta; 2) growth factors and villous trophoblast differentiation; 3) role of the placenta in regulating fetal exposure to xenobiotics during pregnancy; 4) infection and the placenta.

[Resurfacing of a trochanteric pressure sore by a pedicled fasciocutaneous anterolateral thigh flap: a case report]. / Couverture d'une escarre trochantérienne par lambeau pédiculé fasciocutané antéro-latéral de cuisse.

The anterolateral thigh flap is usually used as a free flap for various kinds of reconstruction and resurfacing of distant areas. Cover of a deep trochanteric pressure sore is commonly made by muscular or musculocutaneous flaps such as tensor of fascia lata or vastus lateralis. We report the case of a trochanteric pressure sore covered by a fasciocutaneous pedicled anterolateral thigh flap after negative pressure therapy in a 58-year-old paraplegic patient. After 6 months, a good quality of coverage was obtained with minimal morbidity of donor site. The pedicled fasciocutaneous anterolateral flap appears as a reliable option for the treatment of trochanteric pressure sore.

[Breast repaired by silicone implant. Case report of necrobiosis lipoidica]. / Nécrobiose lipoïdique sur sein reconstruit par implant en silicone : à propos d'un cas.

The authors report a very rare case of necrobiosis lipoidica, histopathologically confirmed, which happened in the breast a few months following the insertion of a silicone implant (post-mastectomy reconstruction). This case raises two problems: the physiopathology (role of the silicone implant) and the treatment of this accident, resistant to current therapy.

Gender differences in presentation, management, and in-hospital outcomes for patients with AMI in a lower-middle income country: evidence from Egypt.

BACKGROUND: Many studies in high-income countries have investigated gender differences in the care and outcomes of patients hospitalized with acute myocardial infarction (AMI). However, little evidence exists on gender differences among patients with AMI in lower-middle-income countries, where the proportion deaths stemming from cardiovascular disease is projected to increase dramatically. This study examines gender differences in patients in the lower-middle-income country of Egypt to determine if female patients with AMI have a different presentation, management, or outcome compared with men. METHODS AND FINDINGS: Using registry data collected over 18 months from 5 Egyptian hospitals, we considered 1204 patients (253 females, 951 males) with a confirmed diagnosis of AMI. We examined gender differences in initial presentation, clinical management, and in-hospital outcomes using t-tests and χ(2) tests. Additionally, we explored gender differences in in-hospital death using multivariate logistic regression to adjust for age and other differences in initial presentation. We found that women were older than men, had higher BMI, and were more likely to have hypertension, diabetes mellitus, dyslipidemia, heart failure, and atrial fibrillation. Women were less likely to receive aspirin upon admission (p<0.01) or aspirin or statins at discharge (p = 0.001 and p<0.05, respectively), although the magnitude of these differences was small. While unadjusted in-hospital mortality was significantly higher for women (OR: 2.10; 95% CI: 1.54 to 2.87), this difference did not persist in the fully adjusted model (OR: 1.18; 95% CI: 0.55 to 2.55). CONCLUSIONS: We found that female patients had a different profile than men at the time of presentation. Clinical management of men and women with AMI was similar, though there are small but significant differences in some areas. These gender differences did not translate into differences in in-hospital outcome, but highlight differences in quality of care and represent important opportunities for improvement.

[Gigantomastia complicating pregnancy. A case report]. / Gigantomastie gestationnelle. À propos d'un cas exceptionnel.

The authors report a very rare case of gestational gigantomastia and the pregnancy could be carried out in term. The repetition being inescapable, the mastectomy is the advised intervention if there is a later desire of pregnancy, in other cases a post-partum mammoplasty can be discussed.

[Clitoridal reconstruction after female circumcision]. / Chirurgie réparatrice du clitoris après excision rituelle.

Ritual sexual mutilations affect 140million of women around the world with over three millions new cases per year. France is not spared with 55,000 mutilated women living on our territory. There is a simple, reliable and reproductible reconstructive surgical technique described by the French urologist Pierre Foldès. This technique is still unknown by patients and plastic surgeons. Through a clinical case, the authors discuss the principles, results and key points of this procedure.

Involvement of MAPK signalling in human villous trophoblast differentiation.

Human villous trophoblast differentiation is a complex and highly regulated process essential for the well-being of the pregnancy and fetal development. In this review, we present an overview of the role of MAPKs signalling in morphological and functional differentiation of villous trophoblast.

PP2 regulates human trophoblast cells differentiation by activating p38 and ERK1/2 and inhibiting FAK activation.

Throughout gestation, fetal growth and development depend, in part, on placental transfer of nutrients from the maternal circulation. This latter function depends on multinucleated, terminally differentiated syncytiotrophoblasts. In vitro, freshly isolated cytotrophoblast cells differentiate spontaneously into syncytiotrophoblast in the presence of fetal bovine serum (FBS). We have previously showed that trophoblast differentiation is regulated by ERK1/2 and p38. Moreover, we showed that PP2 [4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3, 4-d]pyrimidine], a Src family kinase (SFK) specific inhibitor, stimulates biochemical trophoblast cells differentiation while it inhibits cell adhesion and spreading without affecting cell fusion. Therefore, we examined the mechanisms by which PP2 modulates trophoblast cells differentiation. This study shows that PP2 stimulates ERK1/2 and p38 activation after 24h of treatments and up to 3 days while it inhibits focal adhesion kinase (FAK) phosphorylation at many sites including Tyr-397, 407, 576 and 577. Furthermore, we showed that transient activation of ERK1/2 by FBS is independent of SFK and that PP2 induces rapid activation of p38. Moreover, the kinase activity of SFK is negatively regulated by the phosphorylation of their carboxy (C)-terminal regulatory tyrosines by specific proteins called carboxyl-terminal Src kinase (Csk) and Csk homologous kinase (CHK). We showed the expression of Csk and CHK in human trophoblast cells. In summary, this study showed that PP2 stimulates the biochemical differentiation of trophoblast cells by stimulating p38 and ERK1/2 while it inhibits the morphological differentiation by inhibiting FAK activation.

Differential expression of membrane and soluble adenylyl cyclase isoforms in cytotrophoblast cells and syncytiotrophoblasts of human placenta.

Adenylyl cyclase (AC) activity is ubiquitous in mammalian cells, and various forms of this enzyme exist that widely differ with regard to tissue distribution, abundance, and modes of regulation. Human placenta is made, among others, of cytotrophoblast cells and syncytiotrophoblasts. This latter is a polynucleate structure that originates from the differentiation of proliferative mononucleated cytotrophoblast cells, the placental stem cell, into syncytiotrophoblasts. In vitro, this differentiation process is associated with a concomitant increase in cellular levels of cAMP and enhanced expression of genes representative of syncytiotrophoblasts endocrine activity. Thus, in this study we evaluated the differential distribution of AC isoforms in cytotrophoblast cells and syncytiotrophoblasts by reverse transcription-polymerase chain reaction (RT-PCR) using total RNA or purified mRNA. Our results demonstrate that all membrane and soluble AC mRNA isoforms are present in both cell types. Interestingly in syncytiotrophoblasts, AC4 and AC8 mRNA are highly expressed, while AC1, AC2 mRNA are less abundant when compared to cytotrophoblast cells. Additionally, the soluble AC is expressed in both trophoblast cells, but its expression is greatly reduced in differentiated cells, syncytiotrophoblasts. The presence of these AC proteins in both cells was confirmed by Western blotting. Taken together, these data help us to characterize the different AC isoforms in human cytotrophoblast cells and syncytiotrophoblasts, and demonstrate that the AC isoforms expression seems to be mainly modulated in groups 1 and 2. Moreover, the important decrease of the soluble AC isoform in syncytiotrophoblasts as compared to cytotrophoblast cells could suggest an important role of this AC in the extravillous trophoblast formation.

Esofagítis cáustica en niños / Caustic esophageals in childrens

La ingestión de cáusticos es común en pediatría y afecta a menores de 5 años, siendo de gran importancia la detección y tratamiento adecuado de la lesión esofágica para prevenir y tratar las secuelas. Se presenta la experencia en ingestión cáustica, con análisis de la clínica, tratamiento y complicaciones observadas, en 92 casos de ingestión cáustica. De los cuales 57 eran varones y 34 hembras con edades entre 4 meses y 14 años. 91/92 pacientes consultaron en la primera semana post-ingesta de cáustica. Las sustancias cáusticas fueron: 43.4 por ciento, amoníaco 20,6 por ciento, soda cáustica 6,4 por ciento, formol en 4.2 por ciento, derivado de hidrocarburos 34.2 por ciento, acetona 2.2 por ciento, alcanfor 2.2 por ciento y 16.8 por ciento desconocidos. La clínica al ingreso fué: vómitos 34.9 por ciento, salivación 9.3 por ciento, respiratorio 8.1 por ciento, disfagia 6.9 por ciento, fiebre 2.3 por ciento. A todos los pacientes se les practicó al ingreso endoscopia digestiva superior, con hallazgos normales en 10 niños, esofagitis grado I en 44, grado II en 12, grado III en 26. La asociación de esofagitis y quemadura en orofaringe fué observada en 15 pacientes; 23 niños con esofagitis no presentaron lesiones en orofaringe. Los niños con esofagitis grado II y III se hospitalizaron y se realizaron dilataciones en forma precoz, 4 pacientes presentaron como complicación estenosis esofágica, lográndose en 3 de ellos un calibre adecuado con las dilataciones. Hubo baja incidencia de secuelas en nuestros pacientes 3/91 (3.2 por ciento), probablemente por la referencia precoz y la realización de la endoscopia y dilataciones en forma rutinaria

Uso del trien en el tratamiento de la enfermedad de Wilson / Use of trien in the treatment of wilson's disease

Enfermedad de Wilson es una afección autosómica, que se caracteriza por el acúmulo de cobre a nivel de los tejidos con afinidad especial por hígado y cerebro. con una incidencia baja de 1/200.000, en las poblaciones estudiadas, exceptuando el Japón con el 1 por ciento (isla Mikura). Estudios en Venezuela han demostrado un foco importante en el área central, que da un 20 por ciento de la población con afectación leve. La D-penicilamina primera droga utilizada en esta enfermedad pero que presenta el problema de los múltiples efectos secundarios, entre otros trombocitopenia, neutropenia y síndrome lupoide así como su alto costo, que dificultan el manejo del paciente. El trien nueva droga descrita desde 1968 aunque no disponible comercialmente, ha sido utilizada en 40 pacientes sin efectos secundarios. En Venezuela se ha utilizado en 5 pacientes con muy buenos resultados, a propósito presentamos los efectos en un paciente asintomático con cirrosis por enfermedad de Wilson, quien con dos años de tratamiento con 300 mg Tid., presentó normalización de las pruebas de funcionalismo hepático y mejoría leve de la imagen histopatológica y sin efectos secundarios. El costo del Trien es 50 veces más económico y sin efectos secundarios hasta ahora, de allí que su utilización es recomendable

Genetic stability of rotaviruses recovered from asymptomatic neonatal infections.

The sequence of the VP7 gene from 19 rotavirus strains recovered from asymptomatically infected newborn infants was determined by direct analysis of transcript RNAs synthesized from virus present in the stool. For five viruses the entire VP7 gene was sequenced, whereas in the remaining instances only a portion of the gene could be sequenced. In 19 specimens collected over a 4-year period, only five nucleotide substitutions were detected. None of them resulted in an amino acid substitution. Examination of a 306-nucleotide segment of gene 4 in 11 specimens yielded similar results. These results suggest that the mutation rate of rotaviruses in nature is lower than that of single-stranded RNA viruses such as poliovirus and influenza virus.

Reactogenicity and antigenicity of rhesus rotavirus vaccine (MMU-18006) in newborn infants in Venezuela.

The reactogenicity and antigenicity of the rhesus rotavirus vaccine, strain MMU18006, developed at the Laboratory of Infectious Diseases (National Institute of Allergy and Infectious Diseases, National Institutes of Health) were examined in a double blind, placebo-controlled study of 40 newborn infants in Caracas, Venezuela. The children were observed for the first few days after birth in the hospital nursery and by home visits for 10 days after vaccination to detect any adverse reactions. No reactions could be attributed to the vaccine. Serologic responses to the vaccine were evaluated in paired sera obtained at birth (cord blood) and 4 weeks after vaccination. Serologic responses to the vaccine were not observed by complement fixation, neutralization or a rhesus rotavirus VP7 epitope-specific competition assay. However, such responses were found in 9 of 14 tested infants by an immunoglobulin A-specific enzyme-linked immunosorbent assay. Seventeen of the 20 vaccinees also shed rhesus rotavirus vaccine in stool during the postvaccination period.