Tania Maria Ruffoni Ortiga
@ufrj.br
Professor
Universidade Federal do Rio de Janeiro
Scopus Publications
Scopus Publications
Daniela Pereira‐Carvalho, Alessandra Cristina Chagas Valim, Cherley Borba Vieira Andrade, Enrrico Bloise, Ariane Fontes Dias, Veronica Muller Oliveira Nascimento, Rakel Kelly Silva Alves, Ronan Christian dos Santos, Felipe Lopes Brum, Inácio Gomes Medeiros,et al.
Wiley
AbstractMaternal Zika virus (ZIKV) infection during pregnancy has been associated with severe intrauterine growth restriction (IUGR), placental damage, metabolism disturbances, and newborn neurological abnormalities. Here, we investigated the impact of maternal ZIKV infection on placental nutrient transporters and nutrient‐sensitive pathways. Immunocompetent (C57BL/6) mice were injected with Low (103 PFU‐ZIKVPE243) or High (5 × 107 PFU‐ZIKVPE243) ZIKV titers at gestational day (GD) 12.5, and tissue was collected at GD18.5 (term). Fetal–placental growth was impaired in male fetuses, which exhibited higher placental expression of the ZIKV infective marker, eukaryotic translation initiation factor 2 (eIF2α), but lower levels of phospho‐eIF2α. There were no differences in fetal–placental growth in female fetuses, which exhibited no significant alterations in placental ZIKV infective markers. Furthermore, ZIKV promoted increased expression of glucose transporter type 1 (Slc2a1/Glut1) and decreased levels of glucose‐6‐phosphate in female placentae, with no differences in amino acid transport potential. In contrast, ZIKV did not impact glucose transporters in male placentae but downregulated sodium‐coupled neutral amino acid 2 (Snat2) transporter expression. We also observed sex‐dependent differences in the hexosamine biosynthesis pathway (HBP) and O‐GlcNAcylation in ZIKV‐infected pregnancies, showing that ZIKV can disturb placental nutrient sensing. Our findings highlight molecular alterations in the placenta caused by maternal ZIKV infection, shedding light on nutrient transport, sensing, and availability. Our results also suggest that female and male placentae employ distinct coping mechanisms in response to ZIKV‐induced metabolic changes, providing insights into therapeutic approaches for congenital Zika syndrome.
Raissa Rilo Christoff, Débora Santos da Silva, Rafael Ferreira Lima, Ana Luiza Meneguci Moreira Franco, Luiza Mendonça Higa, Átila Duque Rossi, Carolina Batista, Cherley Borba Vieira de Andrade, Tania Maria Ortiga‐Carvalho, Lucas Ascari,et al.
Wiley
ABSTRACTZika virus (ZIKV) infection during pregnancy can lead to a set of congenital malformations known as Congenital ZIKV syndrome (CZS), whose main feature is microcephaly. The geographic distribution of CZS in Brazil during the 2015–2017 outbreak was asymmetrical, with a higher prevalence in the Northeast and Central‐West regions of the country, despite the ubiquitous distribution of the vector Aedes aegypti, indicating that environmental factors could influence ZIKV vertical transmission and/or severity. Here we investigate the involvement of the most used agrochemicals in Brazil with CZS. First, we exposed human neuroblastoma SK‐N‐AS cells to the 15 frequently used agrochemical molecules or derivative metabolites able to cross the blood–brain barrier. We found that a derived metabolite from a widely used herbicide in the Central‐West region, 2,4‐dichlorophenoxyacetic acid (2,4D), exacerbates ZIKV neurotoxic effects in vitro. We validate this observation by demonstrating vertical transmission leading to microcephaly in the offspring of immunocompetent C57BL/6J mice exposed to water contaminated with 0.025 mg/L of 2,4D. Newborn mice whose dams were exposed to 2,4D and infected with ZIKV presented a smaller brain area and cortical plate size compared to the control. Also, embryos from animals facing the co‐insult of ZIKV and 2,4D exposition presented higher Caspase 3 positive cells in the cortex, fewer CTIP2+ neurons and proliferative cells at the ventricular zone, and a higher viral load. This phenotype is followed by placental alterations, such as vessel congestion, and apoptosis in the labyrinth and decidua. We also observed a mild spatial correlation between CZS prevalence and 2,4D use in Brazil's North and Central‐West regions, with R2 = 0.4 and 0.46, respectively. Our results suggest that 2,4D exposition facilitates maternal vertical transmission of ZIKV, exacerbating CZS, possibly contributing to the high prevalence of this syndrome in Brazil's Central‐West region compared to other regions.
Roberto de.A. Antunes, Maria do.C.B. Souza, Marcelo M. Souza, Gabriela P.S. Melo, Brenda M.L. Melo, Ana C.A. Mancebo, Flavia L. Conceição, and Tania M. Ortiga-Carvalho
Elsevier BV
Roberto A Antunes, Brenda M. L de Melo, Maria do Carmo Borges de Souza, Marcelo Marinho de Souza, Gabriela Palhano Sifuentes Melo, Thamires F. M. Jandre, Ana Cristina Allemand Mancebo, Flávia Lúcia Conceição, and Tânia Maria Ortiga-Carvalho
GN1 Sistemas e Publicacoes Ltd.
OBJECTIVE
Vitamin D (VD) is a fat-soluble steroid hormone, synthesized by the skin, most known for its role in bone mineral balance. Vitamin D receptors (VDR) are also found in the female reproductive system, but their role remains unclear. The objective of this study was to analyze the relationship between serum vitamin D levels and the number of oocytes retrieved after ovarian stimulation.
METHODS
This is a retrospective study involving 267 patients undergoing in vitro fertilization (IVF) carried out in the Fertipraxis clinic, a private practice facility. The patients were initially divided into two groups according to their VD levels. Group 1 included 152 patients with VD levels < 30 ng/mL and group 2 had 115 patients with VD levels > 30 ng/mL. They were further analyzed and separated considering their age, anthropometric data, ovarian reserve, amount of gonadotropin used, and follicles obtained until trigger day.
RESULTS
In our analysis, there were no difference in the number of follicles and oocytes retrieved, nor in the number of mature oocytes obtained from patients with both vitamin D deficiency and sufficiency.
CONCLUSIONS
The results of our study show no difference among number of follicles, oocytes retrieved and mature oocytes obtained after ovarian stimulation according to their vitamin D serum levels. Further higher-quality studies are needed to evaluate the possible roles of serum vitamin D levels in other stages of human fertilization process.
Diana Caroline da Silva Bastos, Maria Izabel Chiamolera, Renata Elen Silva, Maria Do Carmo Borges de Souza, Roberto Azevedo Antunes, Marcelo Marinho Souza, Ana Cristina Allemand Mancebo, Patrícia Cristina Fernandes Arêas, Fernando M. Reis, Edson Guimarães Lo Turco,et al.
Springer Science and Business Media LLC
AbstractHashimoto thyroiditis is an autoimmune disease characterized by hypothyroidism and a high level of anti-thyroid autoantibodies. It has shown to negatively impact female fertility; however, the mechanisms are unclear. Ovarian follicular fluid appears to be the key to understanding how Hashimoto thyroiditis affecst fertility. Thus, we aimed to evaluated the metabolic profile of follicular fluid and antithyroid autoantibody levels in the context of Hashimoto thyroiditis. We collected follicular fluid from 61 patients, namely 38 women with thyroid autoantibody positivity and 23 women as negative controls, undergoing in vitro fertilization treatment. Follicular fluid samples were analyzed using metabolomics, and thyroid autoantibodies were measured. Fifteen metabolites with higher concentrations in the follicular fluid samples from Hashimoto thyroiditis were identified, comprising five possible affected pathways: the glycerophospholipid, arachidonic acid, linoleic acid, alpha-linolenic acid, and sphingolipid metabolism pathways. These pathways are known to regulate ovarian functions. In addition, antithyroglobulin antibody concentrations in both serum and follicular fluid were more than tenfold higher in women with Hashimoto thyroiditis than in controls. Our data showed that the metabolic profile of follicular fluid is altered in women with Hashimoto thyroiditis, suggesting a potential mechanistic explanation for the association of this disease with female infertility.
Everton Luis dos Santos Cardoso, Fábio Cahuê, Iordan Emanuel Ferreira Miranda, Marcelo de Lima Sant’Anna, Cherley Borba Vieira Andrade, Raiana Andrade Quintanilha Barbosa, Tania Maria Ortiga-Carvalho, Mário Vaisman, and Verônica Pinto Salerno
Elsevier BV
Victor Jesus Huaringa Payano, Lara Verônica de Araújo Lopes, Larissa Rodrigues Peixoto, Keila Alves da Silva, Tania Maria Ortiga-Carvalho, Alexandre Tafuri, Annamaria Ravara Vago, and Enrrico Bloise
MDPI AG
The activin–follistatin system regulates several cellular processes, including differentiation and tumorigenesis. We hypothesized that the immunostaining of βA-activin and follistatin varies in neoplastic cervical lesions. Cervical paraffin-embedded tissues from 162 patients sorted in control (n = 15), cervical intraepithelial neoplasia (CIN) grade 1 (n = 38), CIN2 (n = 37), CIN3 (n = 39), and squamous cell carcinoma (SCC; n = 33) groups were examined for βA-activin and follistatin immunostaining. Human papillomavirus (HPV) detection and genotyping were performed by PCR and immunohistochemistry. Sixteen samples were inconclusive for HPV detection. In total, 93% of the specimens exhibited HPV positivity, which increased with patient age. The most detected high-risk (HR)-HPV type was HPV16 (41.2%) followed by HPV18 (16%). The immunostaining of cytoplasmatic βA-activin and follistatin was higher than nuclear immunostaining in all cervical epithelium layers of the CIN1, CIN2, CIN3, and SCC groups. A significant decrease (p < 0.05) in the cytoplasmic and nuclear immunostaining of βA-activin was detected in all cervical epithelial layers from the control to the CIN1, CIN2, CIN3, and SCC groups. Only nuclear follistatin immunostaining exhibited a significant reduction (p < 0.05) in specific epithelial layers of cervical tissues from CIN1, CIN2, CIN3, and SCC compared to the control. Decreased immunostaining of cervical βA-activin and follistatin at specific stages of CIN progression suggests that the activin–follistatin system participates in the loss of the differentiation control of pre-neoplastic and neoplastic cervical specimens predominantly positive for HPV.
Victor Machado de Mello Andrade, Amanda Fernandes de Moura, Katlen da Costa Chaves, Camilla Pereira Dias da Rocha, Cherley Borba Vieira de Andrade, Isis Hara Trevenzoli, Tania Maria Ortiga-Carvalho, Luciane Cláudia Barcellos, Mário Vaisman, and Verônica Pinto Salerno
Elsevier BV
C.B.V. Andrade, L.V.A. Lopes, T.M. Ortiga-Carvalho, S.G. Matthews, and E. Bloise
Elsevier BV
Lilian de Freitas Aguiar, Gisele dos Santos Pessanha da Cunha, Karoll Andrea Alfonso Torres Cordido, Francisco Augusto Colucci Coelho, and Tânia Maria Ortiga Carvalho
GN1 Sistemas e Publicacoes Ltd.
V. R. S. Monteiro, C. B. V. Andrade, H. R. Gomes, M. W. Reginatto, G. E. Império, K. N. Fontes, D. A. Spiess, W. S. Rangel-Junior, V. M. O. Nascimento, C. O. S. Lima,et al.
Springer Science and Business Media LLC
AbstractLimited information is available about the effect of mid-pregnancy viral infections on the placental expression of efflux transporters and offspring behavior. We hypothesized that maternal exposure to polyinosinic-polycytidylic acid [poly(I:C)], a synthetic double-stranded RNA viral mimic, would impair placental cell turnover, the expression of selected ABC transporters and adult offspring behavior. C57BL/6 mice were administered poly(I:C) (10 mg/Kg;ip) or vehicle at gestational day (GD) 13.5 (mid-pregnancy). Dams were euthanized for blood collection 4 h after injection, fetal and placental collection at GD18.5 or allowed to deliver spontaneously at term. At GD 13.5, poly(I:C) induced an acute pro-inflammatory response characterized by an increase in maternal plasma levels of IL-6, CXCL-1 and CCL-2/MCP-1. At GD 18.5, poly(I:C) decreased cell proliferation/death in the labyrinthine and increased cell death in the junctional zones, characterizing a disruption of placental cell turnover. Abca1 and Abcg1 immunolabelling was decreased in the labyrinthine zone, whereas Abca1, Abcg1 and breast cancer resistance transporter (Bcrp) expression increased in the junctional zone. Moreover, adult offspring showed motor and cognitive impairments in the Rotarod and T-water maze tests. These results indicate that viral infection during mid-pregnancy may disrupt relevant placental efflux transporters, as well as placental cell turnover and offspring behavior in adult life.
Adriana Carvalho Natal de Moraes, Fernanda Oliveira Caires, Guinever Eustaquio Imperio, Rafael Henrique Nóbrega, Tania Maria Ortiga-Carvalho, and Valéria Freitas de Magalhães
Springer Science and Business Media LLC
Daniela Pereira Carvalho, Ariane Fontes Dias, Amanda Nancy Sferruzzi-Perri, and Tania Maria Ortiga-Carvalho
Oxford University Press (OUP)
Abstract Thyroid hormones (THs) are required for the growth and development of the fetus, stimulating anabolism, and oxygen consumption from the early stages of pregnancy to the period of fetal differentiation close to delivery. Maternal changes in the hypothalamic–pituitary–thyroid axis are also well known. In contrast, several open questions remain regarding the relationships between the placenta and the maternal and fetal TH systems. The exact mechanism by which the placenta participates in regulating the TH concentration in the fetus and mother and the role of TH in the placenta are still poorly studied. In this review, we aim to summarize the available data in the area and highlight significant gaps in our understanding of the ontogeny and cell-specific localization of TH transporters, TH receptors, and TH metabolic enzymes in the placenta in both human and rodent models. Significant deficiencies also exist in the knowledge of the contribution of genomic and nongenomic effects of TH on the placenta and finally, how the placenta reacts during pregnancy when the mother has thyroid disease. By addressing these key knowledge gaps, improved pregnancy outcomes and management of women with thyroid alterations may be possible.
Thamires Siqueira de Oliveira, Marilia Kimie Shimabukuro, Victoria Regina Siqueira Monteiro, Cherley Borba Vieira Andrade, Anita Boelen, Simone Magagnin Wajner, Ana Luiza Maia, Tania Maria Ortiga-Carvalho, and Flavia Fonseca Bloise
MDPI AG
Thyroid hormone (TH) signaling controls muscle progenitor cells differentiation. However, inflammation can alter muscle TH signaling by modulating the expression of TH transporters (Slc16a2), receptors (Thra1), and deiodinase enzymes (Dio2 and Dio3). Thus, a proinflammatory environment could affect myogenesis. The role of a low-grade inflammatory milieu in TH signaling during myogenesis needs further investigation. Herein, we aimed to study the impact of the bacterial lipopolysaccharide (LPS)-induced inflammatory stimulus on the TH signaling during myogenesis. C2C12 myoblasts differentiation was induced without (CTR) or with 10 ng/mL LPS presence. The myoblasts under LPS stimulus release the proinflammatory cytokines (IL-6 and IL-1β) and chemokines (CCL2 and CXCL-1). LPS decreases Myod1 expression by 28% during the initial myogenesis, thus reducing the myogenic stimulus. At the same time, LPS reduced the expression of Dio2 by 41% but doubled the D2 enzymatic activity. The late differentiation was not affected by inflammatory milieu, which only increased the Slc16a2 gene expression by 38%. LPS altered the intracellular metabolism of TH and reduced the initial myogenic stimulus. However, it did not affect late differentiation. Increased intracellular TH activation may be the compensatory pathway involved in the recovery of myogenic differentiation under a low-grade inflammatory milieu.
Thamires Siqueira Oliveira, Anderson Teixeira Santos, Cherley Borba Vieira Andrade, Johnatas Dutra Silva, Natália Blanco, Nazareth de Novaes Rocha, Juliana Woyames, Pedro Leme Silva, Patricia Rieken Macedo Rocco, Wagner Seixas da-Silva,et al.
Frontiers Media SA
Graphical AbstractSeptic diaphragm has impaired morphology and increased thickness that seems to be associated, at least in part, with decreased mitochondrial function related to reducing in Pgc1α expression, ATP production, mitochondrial number, and quality in the CLP mice compared with the control group.
Enrrico Bloise, Jair R. S. Braga, Cherley B. V. Andrade, Guinever E. Imperio, Lilian M. Martinelli, Roberto A. Antunes, Karina R. Silva, Cristiana B. Nunes, Luigi Cobellis, Flavia F. Bloise,et al.
MDPI AG
Assisted reproductive technologies (ART) may increase risk for abnormal placental development, preterm delivery and low birthweight. We investigated placental morphology, transporter expression and paired maternal/umbilical fasting blood nutrient levels in human term pregnancies conceived naturally (n = 10) or by intracytoplasmic sperm injection (ICSI; n = 11). Maternal and umbilical vein blood from singleton term (>37 weeks) C-section pregnancies were assessed for levels of free amino acids, glucose, free fatty acids (FFA), cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL), very low-density lipoprotein (VLDL) and triglycerides. We quantified placental expression of GLUT1 (glucose), SNAT2 (amino acids), P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) (drug) transporters, and placental morphology and pathology. Following ICSI, placental SNAT2 protein expression was downregulated and umbilical cord blood levels of citrulline were increased, while FFA levels were decreased at term (p < 0.05). Placental proliferation and apoptotic rates were increased in ICSI placentae (p < 0.05). No changes in maternal blood nutrient levels, placental GLUT1, P-gp and BCRP expression, or placental histopathology were observed. In term pregnancies, ICSI impairs placental SNAT2 transporter expression and cell turnover, and alters umbilical vein levels of specific nutrients without changing placental morphology. These may represent mechanisms through which ICSI impacts pregnancy outcomes and programs disease risk trajectories in offspring across the life course.
Mila W. Reginatto, Klaus Novaes Fontes, Victoria R. S. Monteiro, Natalia L. Silva, Cherley Borba Vieira Andrade, Hanailly Ribeiro Gomes, Guinever E. Imperio, Flavia Fonseca Bloise, George Eduardo Gabriel Kluck, Georgia Correa Atella,et al.
Frontiers Media SA
Infection alters the expression of transporters that mediate the placental exchange of xenobiotics, lipids and cytokines. We hypothesized that lipopolysaccharide (LPS) modifies the expression of placental transport systems and lipid homeostasis. LPS (150 μg/kg; i.p.) treatments were administered for 4 h or 24 h, animals were euthanized at gestational days (GD) 15.5 or 18.5, and maternal blood, fetuses and placentae were collected. Increased rates of fetal demise were observed at GD15.5 following LPS treatment, whereas at GD18.5, high rates of early labour occurred and were associated with distinct proinflammatory responses. Lipopolysaccharide did not alter ATP-binding cassette (ABC) transporter mRNA expression but decreased fatty acid binding protein associated with plasma membrane (Fabppm) at GD15.5 (LPS-4 h) and increased fatty acid translocase (Fat/Cd36) mRNA at GD18.5 (LPS-4 h). At the protein level, breast cancer-related protein (Bcrp) and ABC sub-family G member 1 (Abcg1) levels were decreased in the placental labyrinth zone (Lz) at GD15.5, whereas P-glycoprotein (P-gp) and Bcrp Lz-immunostaining was decreased at GD18.5. In the placental junctional zone (Jz), P-gp, Bcrp and Abcg1 levels were higher at GD18.5. Specific maternal plasma and placental changes in triacylglycerol, free fatty acid, cholesterol, cholesterol ester and monoacylglycerol levels were detected in a gestational age-dependent manner. In conclusion, LPS-increased risk of fetal death and early labour were associated with altered placental ABC and lipid transporter expression and deranged maternal plasma and placental lipid homeostasis. These changes may potentially modify fetal xenobiotic exposure and placental lipid exchange in cases of bacterial infection.
Cherley Borba Vieira Andrade, Victoria Regina de Siqueira Monteiro, Sharton Vinicius Antunes Coelho, Hanailly Ribeiro Gomes, Ronny Paiva Campos Sousa, Veronica Muller de Oliveira Nascimento, Flavia Fonseca Bloise, Stephen Giles Matthews, Enrrico Bloise, Luciana Barros Arruda,et al.
Frontiers Media SA
Congenital Zika virus (ZIKV) infection can induce fetal brain abnormalities. Here, we investigated whether maternal ZIKV infection affects placental physiology and metabolic transport potential and impacts the fetal outcome, regardless of viral presence in the fetus at term. Low (103 PFU-ZIKVPE243; low ZIKV) and high (5x107 PFU-ZIKVPE243; high ZIKV) virus titers were injected into immunocompetent (ICompetent C57BL/6) and immunocompromised (ICompromised A129) mice at gestational day (GD) 12.5 for tissue collection at GD18.5 (term). High ZIKV elicited fetal death rates of 66% and 100%, whereas low ZIKV induced fetal death rates of 0% and 60% in C57BL/6 and A129 dams, respectively. All surviving fetuses exhibited intrauterine growth restriction (IUGR) and decreased placental efficiency. High-ZIKV infection in C57BL/6 and A129 mice resulted in virus detection in maternal spleens and placenta, but only A129 fetuses presented virus RNA in the brain. Nevertheless, pregnancies in both strains produced fetuses with decreased head sizes (p&lt;0.05). Low-ZIKV-A129 dams had higher IL-6 and CXCL1 levels (p&lt;0.05), and their placentas showed increased CCL-2 and CXCL-1 contents (p&lt;0.05). In contrast, low-ZIKV-C57BL/6 dams had an elevated CCL2 serum level and increased type I and II IFN expression in the placenta. Notably, less abundant microvilli and mitochondrial degeneration were evidenced in the placental labyrinth zone (Lz) of ICompromised and high-ZIKV-ICompetent mice but not in low-ZIKV-C57BL/6 mice. In addition, decreased placental expression of the drug transporters P-glycoprotein (P-gp) and breast cancer resistance protein (Bcrp) and the lipid transporter Abca1 was detected in all ZIKV-infected groups, but Bcrp and Abca1 were only reduced in ICompromised and high-ZIKV ICompetent mice. Our data indicate that gestational ZIKV infection triggers specific proinflammatory responses and affects placental turnover and transporter expression in a manner dependent on virus concentration and maternal immune status. Placental damage may impair proper fetal-maternal exchange function and fetal growth/survival, likely contributing to congenital Zika syndrome.
L.M. Martinelli, M.W. Reginatto, K.N. Fontes, C.B.V. Andrade, V.R.S. Monteiro, H.R. Gomes, F.R.C.L. Almeida, F.F. Bloise, S.G. Matthews, T.M. Ortiga-Carvalho,et al.
Elsevier BV
Lilian M. Martinelli, Klaus N. Fontes, Mila W. Reginatto, Cherley B. V. Andrade, Victoria R. S. Monteiro, Hanailly R. Gomes, Joao L. Silva‐Filho, Ana A. S. Pinheiro, Annamaria R. Vago, Fernanda R. C. L. Almeida,et al.
Wiley
Malaria in pregnancy (MiP) induces intrauterine growth restriction (IUGR) and preterm labour (PTL). However, its effects on yolk sac morphology and function are largely unexplored. We hypothesized that MiP modifies yolk sac morphology and efflux transport potential by modulating ABC efflux transporters. C57BL/6 mice injected with Plasmodium berghei ANKA (5 × 105 infected erythrocytes) at gestational day (GD) 13.5 were subjected to yolk sac membrane harvesting at GD 18.5 for histology, qPCR and immunohistochemistry. MiP did not alter the volumetric proportion of the yolk sac's histological components. However, it increased levels of Abcb1a mRNA (encoding P‐glycoprotein) and macrophage migration inhibitory factor (Mif chemokine), while decreasing Abcg1 (P < 0.05); without altering Abca1, Abcb1b, Abcg2, Snat1, Snat2, interleukin (Il)‐1β and C‐C Motif chemokine ligand 2 (Ccl2). Transcripts of Il‐6, chemokine (C‐X‐C motif) ligand 1 (Cxcl1), Glut1 and Snat4 were not detectible. ABCA1, ABCG1, breast cancer resistance protein (BCRP) and P‐gp were primarily immunolocalized to the cell membranes and cytoplasm of endodermic epithelium but also in the mesothelium and in the endothelium of mesodermic blood vessels. Intensity of P‐gp labelling was stronger in both endodermic epithelium and mesothelium, whereas ABCA1 labelling increased in the endothelium of the mesodermic blood vessels. The presence of ABC transporters in the yolk sac wall suggests that this fetal membrane acts as an important protective gestational barrier. Changes in ABCA1 and P‐gp in MiP may alter the biodistribution of toxic substances, xenobiotics, nutrients and immunological factors within the fetal compartment and participate in the pathogenesis of malaria‐induced IUGR and PTL.
Marianna Ferreira Gonçalves, Karina Dutra Asensi, Anna Luiza Lima Nascimento, Julia Helena Oliveira de Barros, Rosana de Almeida Santos, Cherley Borba Vieira de Andrade, Tais Hanae Kasai-Brunswick, Marcel Frajblat, Tania M. Ortiga-Carvalho, and Regina Coeli dos Santos Goldenberg
Mary Ann Liebert Inc
There is a constant need for improving embryo culture conditions in assisted reproduction. One possibility is to use mesenchymal stem/stromal cells derived from menstrual blood (mbMSCs), with an endometrial origin. In this study, we sought to analyze the expansion of mouse embryos in a direct coculture model with mbMSCs. Our results showed that after five passages, mbMSCs presented a spindle-shaped morphology, with surface markers that were comparable with the normal mesenchymal cell phenotype. mbMSCs could differentiate into adipogenic and osteogenic lineages and secrete angiopoetin-2 and hepatocyte growth factor. The coculture experiments employed 103 two-cell-stage embryos that were randomly divided into two groups: control (n = 50), embryos cultured in GV-Blast medium, and cocultured mbMSCs (n = 53), embryos cocultured with GV-Blast and mbMSCs. Typically, two to three embryos were placed in a well with 200 μL of culture medium and observed until developmental day 5. After 5 days, the cocultured group had more embryos in the blastocyst stage (69.8%) when compared with the control group (30%) (p < 0.001). It was also found that nearly 57% of blastocysts in the cocultured group reached the hatching stage, while only 13% achieved this stage in the control group (p < 0.001). Analyses of cultured mbMSCs and growth media, in the presence or absence of an embryo, were also performed. Immunofluorescence detected similar levels of collagen I and III and fibronectin in both mbMSCs and cocultured mbMSCs, and similar amounts of growth factors, VEGF, PDGF-AA, and PDGF-BB, were also observed in the conditioned medium, regardless of embryo presence. The present study describes, for the first time, an easy, noninvasive, and autologous method that could potentially increase blastocyst growth rates during assisted reproductive procedures (i.e., in vitro fertilization). It is proposed that this mbMSC coculture strategy enriches the embryonic microenvironment and promotes embryo development. This technique may complement or replace existing assisted reproduction methods and is directly relevant to the field of personalized medicine. Impact statement The study demonstrates a novel and potentially personalized assisted reproduction approach. The search for alternative and autologous methods provides assisted reproduction patients with a better chance of a successful pregnancy. In this study, mesenchymal cells derived from menstrual blood resembled the outside uterine surface and could potentially be employed for improving embryo outgrowth. Our protocol enriches the embryonic microenvironment and facilitates high-quality single-embryo transfer.
Flavia Fonseca Bloise, Anderson Teixeira Santos, Juliana de Brito, Cherley Borba Vieira de Andrade, Thamires Siqueira Oliveira, Aline Fonseca Pereira de Souza, Klaus Novaes Fontes, Johnatas D Silva, Natália Blanco, Pedro Leme Silva,et al.
Mary Ann Liebert Inc
BACKGROUND
Sepsis can cause the nonthyroidal illness syndrome (NTIS), resulting in perturbed thyroid hormone (TH) signaling and reduced thyroxine (T4) levels. TH is a major regulator of muscle function, via its influence on mitochondria. The present study aimed to evaluate the relationship among TH signaling, mitochondrial function and the antioxidant defense system in the diaphragms of septic mice.
METHODS
Male C57Bl/6 mice were divided into two groups: cecal ligation and puncture (CLP) and sham. Twenty-four hours after surgery, plasma, diaphragms and livers were collected. TH metabolism and responses were analyzed by measuring mRNA expression of Dio1 in the liver, and Thra, Thrb, Dio2, Slc16a10 and Slc16a2 (encodes MCT 10 and 8), in the diaphragm. T4 plasma levels were measured by radioimmunoassay. Damage to diaphragm mitochondria was assessed by electron microscopy and qPCR, and function with oxygraphy. The diaphragm anti-oxidative defense system was examined by qPCR, analyzing Sod1, Sod2, Sod3, Gpx1 and Cat expression. The effect of TH replacement was tested by treating the mice with T4 and tri-iodothyronine (T3) (CLP+TH) after surgery.
RESULTS
CLP mice presented reduced total plasma T4 concentrations, as well as downregulated Dio1 and upregulated Il1b mRNA expression in the liver. CLP mice also displayed downregulated Thra, Thrb, Slc16a10 and Slc16a2 expression in the diaphragm, suggesting that TH signaling was compromised. The expression of Ppargc1a (encodes PGC1a) was downregulated, which correlated with the decrease in the number of total mitochondria, increase in the percentage of injured mitochondria, downregulation of respiratory chain complex 2 and 3 mRNA expression and reduced maximal respiration. Additionally, septic animals presented a 3-fold increase in Ucp3 and G6pdh expression, downregulated Sod3, Gpx1 and Cat expression; and upregulated Sod2 expression, potentially due to elevated ROS levels. The mitochondrial number and the percentage of injured mitochondrial were similar between sham and CLP+TH mice.
CONCLUSIONS
Sepsis induced responses consistent with NTIS, resulted in mitochondrial damage and functional impairment, and modulated the expression of key antioxidant enzymes in the diaphragm. Thus, impaired diaphragm function during sepsis seems to involve altered local TH signaling, mitochondrial dysfunction and oxidative stress defense.
Rhayanna B Gaglianone, F. Bloise, T. Ortiga-Carvalho, T. Quirico-Santos, M. Costa and C. Mermelstein
Sarcolemma instability and increased calcium influx in muscle fibers are characteristics of the Duchenne muscular dystrophy. Excessive calcium activates calcium-dependent enzymes, such as calpains (CAPN) and matrix metalloproteases (MMP). Here, we analyzed calcium deposits, the activity of CAPN and MMP and the expression of Myh, SERCA and myogenic regulatory factors in different skeletal muscles during myonecrosis (4-weeks) and regeneration (12-weeks) phases of the mdx muscular pathology. Alizarin red staining was used to assess calcium deposits, casein and gelatin zymography were performed to evaluate CAPN and MMP activity, and qPCR was used to evaluate the expression of Myh, Capn, Atp2a1 and Atp2a2, Myod1 and Myog. We observed the following characteristics in mdx muscles: (i) calcium deposits almost exclusively in mdx muscles, (ii) lower CAPN1 activity in mdx muscles, (iii) higher CAPN2 activity in mdx muscles (only at 12 wks), (iv) autolyzed CAPN activity exclusively in mdx muscles, (v) lower expression of Capn1 and higher expression of Capn2 in mdx muscles; (vi) lower expression of Atp2a1 and Atp2a2 in mdx muscles, (vii) higher MMP (pre pro MMP2, pro MMP2, MMP2 and MMP9) activity in mdx muscles, (viii) MMP2 activity exclusively in mdx muscles at 12 wks, (ix) MMP9 activity exclusively in mdx muscles, (x) higher expression of Myog in mdx muscles at 12 wks, and (xi) lower expression of Myh (Myh7, Myh2, Myh1, Myh4) in mdx muscles, particularly Myh7 and Myh2. The collection of our results provides valuable information for a better characterization of mdx pathology phenotype.
K. N. Fontes, M. W. Reginatto, N. L. Silva, C. B. V. Andrade, F. F. Bloise, V. R. S. Monteiro, J. L. Silva-Filho, G. E. Imperio, P. M. Pimentel-Coelho, A. A. S. Pinheiro,et al.
Springer Science and Business Media LLC
Rhayanna B. Gaglianone, Anderson Teixeira Santos, Flavia Fonseca Bloise, Tania Maria Ortiga-Carvalho, Manoel Luis Costa, Thereza Quirico-Santos, Wagner Seixas da Silva, and Claudia Mermelstein
Springer Science and Business Media LLC