3D-Printed Models Applied in Medical Research Studies

The aim of this chapter is to show experiments in cardiology and fetal medicine, two specialties of medicine, through the development of three dimensional (3D) physical models produced on additive manufacturing (AM) technologies, also known as 3D printing, from files acquired on noninvasive-imaging technologies (NITs) as 3D ultrasound (3DUS), magnetic resonance imaging (MRI), computed tomography (CT), and micro-computed tomography (micro-CT). The presentation of eight different experiments demonstrates that the combination of AM technologies and files obtained from NITs may improve our understanding of medical anatomical characteristics for medical research, simulation procedures, and educational purposes

Discordant Zika Virus Findings in Twin Pregnancies Complicated by Antenatal Zika Virus Exposure: A Prospective Cohort.

BACKGROUND: There are limited data on the natural history of antenatal Zika virus (ZIKV) exposure in twin pregnancies, especially regarding intertwin concordance of prenatal, placental, and infant outcomes. METHODS: This prospective cohort study included twin pregnancies referred to a single institution from September 2015 to June 2016 with maternal ZIKV. Polymerase chain reaction (PCR) testing of maternal, placental, and neonatal samples was performed. Prenatal ultrasounds were completed for each twin, and histomorphologic analysis was performed for each placenta. Abnormal neonatal outcome was defined as abnormal exam and/or abnormal imaging. Two- to three-year follow-up of infants included physical exams, neuroimaging, and Bayley-III developmental assessment. RESULTS: Among 244 pregnancies, 4 twin gestations without coinfection were identified. Zika virus infection occurred at 16-33 weeks gestation. Zika virus PCR testing revealed discordance between dichorionic twins, between placentas in a dichorionic pair, between portions of a monochorionic placenta, and between a neonate and its associated placenta. Of the 8 infants, 3 (38%) had an abnormal neonatal outcome. Of 6 infants with long-term follow-up, 3 (50%) have demonstrated ZIKV-related abnormalities. CONCLUSIONS: Neonatal PCR testing, placental findings, and infant outcomes can be discordant between co-twins with antenatal ZIKV exposure. These findings demonstrate that each twin should be evaluated independently for vertical transmission

The role of TP53 and p21(WAF1) gene polymorphisms and immunohistochemical expression of p53, p21 (WAF1), p16 (INK4a) and cyclin D1 and their importance in the development and / or severity of cervical neoplasias

O câncer de colo do útero é o terceiro tipo de câncer mais frequente em mulheres no mundo, e a infecção persistente pelo papilomavirus humano (HPV) oncogênico é condição necessária, mas não suficiente para seu desenvolvimento. As oncoproteínas virais E6 e E7 interferem direta ou indiretamente na ação de várias proteínas celulares. Entretanto, as variantes proteicas, resultantes de polimorfismos genéticos, podem apresentar comportamento distinto mediante a infecção pelo HPV. O objetivo deste estudo foi avaliar possíveis associações entre polimorfismos nos genes TP53 (p53 PIN3, p53 72C>G) e p21 (p21 31C>A) e o desenvolvimento de neoplasias cervicais, considerando os níveis de expressão das proteínas p53, p21, p16 e ciclina D1, e fatores de risco clássicos para o câncer cervical. Foram selecionadas 466 mulheres residentes no Rio de Janeiro, 281 com diagnóstico histopatológico de neoplasia cervical de baixo (LSIL) e alto grau (HSIL) e câncer (grupo de casos) e 185 sem história atual ou pregressa de alteração citológica do colo uterino (grupo controle). A técnica de PCR-RFLP (reação em cadeia da polimerase - polimorfismo de comprimento de fragmento de restrição), foi empregada na análise dos polimorfismos p53 72C>G e p21 31C>A, usando as enzimas de restrição BstUI e BsmaI, respectivamente. A avaliação do polimorfismo p53 PIN3 (duplicação de 16 pb) foi feita por meio da análise eletroforética direta dos produtos de PCR. A expressão das proteínas p53, p21, p16, ciclina D1 e Ki-67 e a pesquisa de anticorpos anti-HPV 16 e HPV pool foram avaliadas por imunohistoquímica (Tissue Microarray - TMA) em 196 biópsias do grupo de casos. O grupo controle se mostrou em equilíbrio de Hardy-Weinberg em relação aos três polimorfismos avaliados. As distribuições genotípicas e alélicas relativas a p53 PIN3 e p53 72C>G nos grupos controles e de casos não apresentaram diferenças significativas, embora o genótipo p53 72CC tenha aumentado o risco atribuído ao uso de contraceptivos das pacientes apresentarem lesões mais severas (OR=4,33; IC 95%=1,19-15,83). O genótipo p21 31CA(Ser/Arg) conferiu proteção ao desenvolvimento de HSIL ou câncer (OR=0,61, IC 95%=0,39-0,97), e modificou o efeito de fatores de risco associados à severidade das lesões. A interação multiplicativa de alelos mostrou que a combinação p53 PIN3A1, p53 72C(Pro) e p21 31C(Ser), representou risco (OR=1,67, IC95%=1,03-2,72) e a combinação p53 PIN3A1, p53 72C(Pro) e p21 31A(Arg) conferiu efeito protetor (OR=0,26, IC95%=0,08-0,78) para o desenvolvimento de HSIL e câncer cervical. Observou-se correlação positiva da expressão de p16 e p21 e negativa da ciclina D1 com o grau da lesão. A distribuição epitelial de p16, Ki-67, p21 e p53 se mostrou associada à severidade da lesão. Os polimorfismos analisados não apresentaram associação com a expressão dos biomarcadores ou positividade para HPV. Nossos resultados sugerem a importância do polimorfismo p21 31C>A para o desenvolvimento das neoplasias cervicais e ausência de correlação dos polimorfismos p53 PIN3 e p53 72C>G com a carcinogênese cervical, embora alguns genótipos tenham se comportado como modificadores de risco. Nossos resultados de TMA corroboram o potencial de uso de biomarcadores do ciclo celular para diferenciar as lesões precursoras do câncer cervical.Cervical cancer is the third most common female cancer worldwide, and persistent infection by the Human Papillomavirus (HPV) is a necessary but not sufficient condition to cause it. The viral oncoproteins E6 and E7 interfere directly or indirectly with the action of various cellular proteins. However, the protein variants, resulting from genetic polymorphisms, may act differently when encountering HPV infection. The aim of this study was to evaluate possible associations between polymorphisms in the TP53 (p53 PIN3, p53 72C>G) and p21 (p21 31C>A) genes, and the development of cervical neoplasia, considering the expression levels of p53, p21, p16 and cyclin D1 proteins, together with classic risk factors for cervical cancer. A total of 466 women resident in Rio de Janeiro were selected, being 281 with histopathological diagnosis of low (LSIL) or high grade (HSIL) cervical neoplasia or cancer (test group), and 185 with no current or previous history of alteration of cervical cytology (control group). The PCR-RFLP technique (polymerase chain reaction restriction fragment length polymorphism) was used to analyze the p53 72C>G and p21 31C>A polymorphisms, using BstUI and BsmaI restriction enzymes, respectively. Genotyping of the p53 PIN3 (duplication of 16 pb) polymorphism was performed by direct electrophoretic analysis of the PCR products. The expression of p53, p21, p16, cyclin D1 and Ki-67 proteins and the study of anti-HPV 16 and anti-HPV pool positivities were evaluated by immunohistochemisty (Tissue Microarray - TMA) in 196 biopsies of cases. The control group obeyed the Hardy-Weinberg principle in relation to the three polymorphisms analysed. The genotypic and allelic frequencies regarding p53 PIN3 and p53 72C>G in the control and test groups were not significantly different, although the p53 72CC genotype has increased the risk of more severe lesions attributed to the use of contraceptives (OR=4.33; IC 95%=1.19-15.83). The p21 31CA(Ser/Arg) genotype showed to protect against the development of HSIL or cancer (OR=0,61, IC 95%=0,39-0,97), and modified the effect of risk factors associated to the lesion severity. The multiplicative interaction of alleles showed that the combination p53 PIN3A1, p53 72C(Pro) and p21 31C(Ser) represented risk (OR=1,67, IC95%=1,03-2,72) and the combination p53 PIN3A1, p53 72C(Pro) and p21 31A(Arg) conferred protection (OR=0,26, IC95%=0,08-0,78) against the development of HSIL and cervical cancer. It was observed positive and negative correlations of, respectively, p16 and p21, and cyclin D1 expression with the cervical lesion grade. The epithelial distribution of p16, Ki-67, p21 and p53 was associated with the lesion severity. The polymorphisms analyzed showed neither association with the expression of the biomarkers nor positivity for HPV. Our results suggest the importance of polymorphism p21 31C>A in the development of cervical neoplasia and the lack of correlation between the polymorphisms p53 PIN3 and p53 72C>G with cervical carcinogenesis, although some genotypes acted as risk modifiers. Our TMA results corroborated the potential use of cell cycle biomarkers as an adjunctive tool to differentiate cervical precursor lesions

TP53 and p21 (CDKN1A) polymorphisms and the risk of systemic lupus erythematosus

Abstract Background The p53 and p21 proteins are important regulators of cell cycle and apoptosis and may contribute to autoimmune diseases, such as systemic lupus erythematosus (SLE). As genetic polymorphisms may cause changes in protein levels and functions, we investigated associations of TP53 and p21 (CDKN1A) polymorphisms (p53 72 G > C—rs1042522; p53 PIN3—rs17878362; p21 31 C > A—rs1801270; p21 70 C > T—rs1059234) with the development of systemic lupus erythematosus (SLE) in a Southeastern Brazilian population. Methods Genotyping of 353 female volunteers (cases, n = 145; controls, n = 208) was performed by polymerase chain reaction, restriction fragment length polymorphism and/or DNA sequencing. Associations between TP53 and p21 polymorphisms and SLE susceptibility and clinical manifestations of SLE patients were assessed by logistic regression analysis. Results Protective effect was observed for the genotype combinations p53 PIN3 A1/A1-p21 31 C/A, in the total study population (OR 0.45), and p53 PIN3 A1/A2-p21 31 C/C, in non-white women (OR 0.28). In Whites, p53 72 C-containing (OR 3.06) and p53 PIN3 A2-containing (OR 6.93) genotypes were associated with SLE risk, and higher OR value was observed for the combined genotype p53 72 G/C-p53 PIN3 A1/A2 (OR 9.00). Further, p53 PIN3 A1/A2 genotype was associated with serositis (OR 2.82), while p53 PIN3 A2/A2 and p53 72 C/C genotypes were associated with neurological disorders (OR 4.69 and OR 3.34, respectively). Conclusions Our findings showed that the TP53 and p21 polymorphisms included in this study may have potential to emerge as SLE susceptibility markers for specific groups of patients. Significant interactions of the TP53 polymorphisms with serositis and neurological disorders were also observed in SLE patients

Antenatal diagnosis of a large immature abdominal wall teratoma by 2d-3d ultrasound using hdlive and magnetic resonance imaging

We describe the first case of prenatally detected teratoma of the fetal abdomen wall using ultrasound and fetal magnetic resonance imaging (MRI). A heterogeneous mass, partly solid and cystic, originating from the anterior abdominal wall of the fetus close to an omphalocele sac was detected by means of 2D/3D ultrasound and MRI. Amniodrainage was performed and due to sign of impending fetal risk, an emergency Cesarean section was performed. A bulky, crumbly and bleeding tumoral mass was confirmed at delivery. Ligation of the supplying artery to the tumor was complicated by uncontrollable hemorrhage and early neonatal death. Pathology identified the tumor as an immature teratoma of the anterior fetal abdominal wall. 2D/3D ultrasound, especially using HDlive application and MRI demonstrated accurate detection and characterization of this congenital tumor.CDPI, Radiol, Rio De Janeiro, BrazilHosp Fed Servidores Estado Rio de Janeiro, Obstet, Rio De Janeiro, BrazilClin Perinatal Laranjeiras, Obstet & Gynecol, Rio De Janeiro, BrazilGuastalla Civil Hosp, Obstet & Gynecol, Reggio Emilia, ItalyUniv Fed Sao Paulo, Obstet, Paulista Sch Med, Rua Belchior Azevedo,156 Apto 111 Torre Vitoria, BR-05089030 Sao Paulo, BrazilFundacao Oswaldo Cruz, Pathol, Inst Fernandes Figueira, Rio De Janeiro, BrazilArcispedale S Maria Nuova, Pathol, Reggio Emilia, ItalyUniversidade Federal de Sao Paulo, Obstet, Paulista Sch Med, Rua Belchior Azevedo,156 Apto 111 Torre Vitoria, BR-05089030 Sao Paulo, Brazil.Web of Scienc

Practical considerations in the use of a porcine model (Sus scrofa domesticus) to assess prevention of postoperative peritubal adhesions.

Infertility has been a common postoperative problem caused by peritoneal adhesions. Since several prophylactic agents have recently shown promising preliminary results, more complete studies comparing their real efficacy and safety are needed urgently. The aim of this study was to investigate and describe practical considerations of a porcine model that can be used to assess such prophylactic agents. First, 10 healthy 5½ months old female pigs (24.3-31.3 Kg) underwent a standardized laparoscopy to provoke peritubal adhesion formation without prophylactic agents. After 30 days, a second-look laparoscopy was performed to evaluate adhesions and perform adnexectomy for histopathological evaluation. Adhesions at different sites were classified by grade, for which the scores range from 0 (no adhesion) to 3 (very strong vascularized adhesions), and also by area, with scores ranging from 0 (no adhesion) to 4 (>75% of the injured area). The histopathological evaluation of the distal uterine horns, oviducts and ovaries were compared withthose from a control group of six healthy pigs with no previous surgery. Biological samples were collected to assess vitality, inflammation and renal, hepatic and hematopoietic systems. There were small (but significant) changes in serum albumin (P = 0.07), globulin (P = 0.07), C-reactive protein (P = 0.011), fibrinogen (P = 0.023) and bilirubin (P0) and of strong / very strong adhesion (scores >1) was 75% (95% CI: 55-94.9) and 65% (95% CI: 45-85), respectively. The porcine model represents a useful animal platform that can be used to test the efficacy and safety of candidate prophylactic agents intended to prevent postoperative peritubal adhesions formation. We present several practical considerations and measures that can help to minimize animal suffering and avoid problems during such experiments