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Article: Cloning, partial sequencing and 17β‐estradiol modulation of hepatic vitellogenin gene of the Neotropical catfish Rhamdia quelen

Fernandes, Letícia da Silva P. / Mathias, Francielle T. / Richardi, Vinicius S. / Cardoso, Cibele C. / Silva de Assis, Helena C.

Journal of applied ichthyology. 2021 Aug., v. 37, no. 4

2021

Abstract: ... to the lack of the two domains, the β’‐c and the Ct. Moreover, from the cloning and sequencing of vtg of R ...

Abstract	Expression of vitellogenin gene (vtg) is used as a biomarker for the evaluation of the exposure of estrogenic substances in fish. However, scientific information regarding this biomarker in Neotropical fish is limited. In this study, a 760 bp partial sequence of the vtg mRNA from the liver of the catfish Rhamdia quelen was cloned, representing almost 20% of the full‐length vtg. The phylogenetic tree analysis recovered the vtg of R. quelen inside the clade of Siluriformes. The alignment of the deduced amino acid sequence of R. quelen vtg with other species confirmed that the described sequence is gene specific. Also, this cloned sequence presents almost 70% of identity with the vtgI subtype, that is known to translate into the incomplete form of Vtg due to the lack of the two domains, the β’‐c and the Ct. Moreover, from the cloning and sequencing of vtg of R. quelen, a protocol for a RT‐qPCR was developed with the goal to be applied as a biomarker of the hypothalamic‐pituitary‐gonad‐liver (HPGL) axis, in order to evaluate the effects of endocrine disruptor exposure in Neotropical fish. Thus, this protocol was applied in the effects of a dose of 10 mg/kg of 17β‐estradiol (E2) on vtg expression in male and female fish. The results showed that after 17 days of the exposure injection the E2 treatment upregulated vtg expression in both sexes. No observed differences in the levels of gene expression between the male and female fish were observed, and no statistical interaction between E2‐treatment and sex were found. The results obtained from cloning add new information regarding vtg in Siluriformes fish, an order poorly studied in this aspect. Also, the vtg RT‐qPCR protocol stablished for vtg of R. quelen will expand the application of this animal model, a Neotropical fish, in investigation regarding the effects of endocrine disruptors.
Keywords	Neotropics ; Rhamdia quelen ; amino acid sequences ; animal models ; biomarkers ; catfish ; endocrine-disrupting chemicals ; females ; gene expression ; genes ; ichthyology ; liver ; males ; phylogeny ; tropical fish ; vitellogenin
Language	English
Dates of publication	2021-08
Size	p. 545-552.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	283875-8
ISSN	0175-8659
ISSN	0175-8659
DOI	10.1111/jai.14203
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Cloning, partial sequencing and expression analysis of the neural form of P450 aromatase (cyp19a1b) in the South America catfish Rhamdia quelen.

Silva de Assis, Helena C / Navarro-Martín, Laia / Fernandes, Letícia S P / Cardoso, Cibele C / Pavoni, Daniela Parada / Trudeau, Vance L

Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology

2018 Volume 221-222, Page(s) 11–17

Abstract: Brain aromatase is a key enzyme exclusively expressed in fish radial glial cells that convert androgens into estrogens, thus controlling neuroendocrine functions and neurogenesis. As an important step in characterizing the neuroendocrine systems of ... ...

Abstract	Brain aromatase is a key enzyme exclusively expressed in fish radial glial cells that convert androgens into estrogens, thus controlling neuroendocrine functions and neurogenesis. As an important step in characterizing the neuroendocrine systems of Rhamdia quelen (jundiá), a partial cDNA sequence (1045 bp) of brain aromatase (cyp19a1b) was cloned and sequenced. At the nucleotide level the cDNA sequence was found to be 88% identical to cyp19a1b of two species of catfish, Ictalurus punctatus and Silurus meridionalis. The predicted amino acid sequence was between 80 and 91% similar to other teleosts. Real-time RT-qPCR analysis revealed that cyp19a1b was detected in pituitary, hypothalamus, telencephalon, head and posterior kidneys, liver and gonads (testis and ovary) of both males and females. The effects of E2 on cyp19a1b expression are sexually dimorphic in R. quelen. The injection of 17β-estradiol (E2) decreased head kidney mRNA levels of cyp19a1b in males and increased cyp19a1b mRNA levels in the pituitary and head kidney of females. This study demonstrated that the R. quelen cyp19a1b gene is expressed in brain, pituitary and peripheral tissues in both males and females.
MeSH term(s)	Animals ; Aromatase/biosynthesis ; Aromatase/genetics ; Catfishes/genetics ; Catfishes/metabolism ; Cloning, Molecular ; Fish Proteins/biosynthesis ; Fish Proteins/genetics ; Gene Expression Regulation, Developmental/physiology ; Organ Specificity ; Sequence Analysis, DNA
Chemical Substances	Fish Proteins ; Aromatase (EC 1.14.14.1)
Language	English
Publishing date	2018-07
Publishing country	England
Document type	Journal Article
ZDB-ID	121247-3
ISSN	1879-1107 ; 0305-0491 ; 1096-4959
ISSN (online)	1879-1107
ISSN	0305-0491 ; 1096-4959
DOI	10.1016/j.cbpb.2018.04.001
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Article ; Online: PIMREG expression level predicts glioblastoma patient survival and affects temozolomide resistance and DNA damage response.

Serafim, Rodolfo Bortolozo / Cardoso, Cibele / Arfelli, Vanessa Cristina / Valente, Valeria / Archangelo, Leticia Fröhlich

Biochimica et biophysica acta. Molecular basis of disease

2022 Volume 1868, Issue 6, Page(s) 166382

Abstract: PIMREG expression strongly correlates with cellular proliferation in both malignant and normal cells. Throughout embryo development, PIMREG expression is prominent in the central nervous system. Recent studies have described elevated PIMREG expression in ...

Abstract	PIMREG expression strongly correlates with cellular proliferation in both malignant and normal cells. Throughout embryo development, PIMREG expression is prominent in the central nervous system. Recent studies have described elevated PIMREG expression in different types of tumors, which correlates with patient survival and tumor aggressiveness. Given the emerging significance of PIMREG in carcinogenesis and its putative role in the context of the nervous system, we investigated the expression and function of PIMREG in gliomas, the most common primary brain tumors. We performed an extensive analysis of PIMREG expression in tumors samples from glioma patients. We then assessed the effects of PIMREG silencing and overexpression on the sensitivity of glioblastoma cell lines treated with genotoxic agents commonly used for treating patients and assessed for treatment response, proliferation and migration. Our analysis shows that glioblastoma exhibits the highest levels of PIMREG expression among all cancers analyzed and that elevated PIMREG expression is a biomarker for glioma progression and patient outcome. Moreover, PIMREG is induced by genotoxic agents, and its silencing renders glioblastoma cells sensitive to temozolomide treatment and affects ATR- and ATM-dependent signaling. Our data demonstrate that PIMREG is involved in DNA damage response and temozolomide resistance of glioblastoma cells and further supports a role for PIMREG in tumorigenesis.
MeSH term(s)	Antineoplastic Agents, Alkylating/pharmacology ; Antineoplastic Agents, Alkylating/therapeutic use ; Cell Line, Tumor ; DNA Damage ; Glioblastoma/drug therapy ; Glioblastoma/genetics ; Glioblastoma/metabolism ; Humans ; Temozolomide/pharmacology ; Temozolomide/therapeutic use
Chemical Substances	Antineoplastic Agents, Alkylating ; Temozolomide (YF1K15M17Y)
Language	English
Publishing date	2022-03-14
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	60-7
ISSN	1879-260X ; 1879-2596 ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-260X ; 1879-2596 ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbadis.2022.166382
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Article ; Online: Factors Associated with Disagreement of Fibrosis Stages between 2D-Shear Wave Elastography and Transient Elastography in Chronic Hepatitis B.

Miodownik, Fernanda Gdalevici / Cardoso, Ana Carolina / Cancella Nabuco, Leticia / Franz, Cibele / Perez, Renata / Alves Villela-Nogueira, Cristiane

Viruses

2023 Volume 15, Issue 4

Abstract: Introduction and objectives: The agreement of elastography techniques in chronic Hepatitis B (CHB) needs evaluation. We aimed to evaluate, in CHB, the agreement between transient elastography (TE) and two-dimensional shear wave elastography (2D-SWE), ... ...

Abstract	Introduction and objectives: The agreement of elastography techniques in chronic Hepatitis B (CHB) needs evaluation. We aimed to evaluate, in CHB, the agreement between transient elastography (TE) and two-dimensional shear wave elastography (2D-SWE), analyzing the factors related to the disagreement of measures. Materials and methods: CHB patients underwent liver stiffness measures with both TE and 2D-SWE on the same day. For concordance analysis, we defined liver fibrosis as F0/1 vs. F ≥ 2, F0/1-F2 vs. F ≥ 3 and F0/1-F2-F3 vs. F4 for both methods. Logistic regression analysis was used to identify the variables independently associated with the disagreement between methods. Results: A total of 150 patients were enrolled. Liver fibrosis categorization according to TE was: F0-F1 = 73 (50.4%), F ≥ 2 = 40 (27.6%), F ≥ 3 = 21 (14.5%) and F4 = 11 (7.6%), and according to 2D-SWE was: F0/F1 = 113 (77.9%), F ≥ 2 = 32 (22.1%), F≥ 3 = 25 (17.2%) and F4 = 11 (7.6%). It was observed that 20.0% of the sample had steatosis (CAP≥ 275 dB/m). TE and SD-SWE estimated equal fibrosis stages in 79.3% of cases. Spearman's correlation coefficient was 0.71 ( Conclusions: In CHB, there is strong correlation and good agreement between TE and 2D-SWE in identifying fibrosis stages. Diabetes mellitus and antiviral therapy may impact the agreement of stiffness measures obtained with these elastographic methods.
MeSH term(s)	Humans ; Hepatitis B, Chronic/complications ; Elasticity Imaging Techniques/methods ; Liver Cirrhosis/pathology ; Fatty Liver ; Antiviral Agents ; Liver/diagnostic imaging ; Liver/pathology
Chemical Substances	Antiviral Agents
Language	English
Publishing date	2023-03-26
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15040846
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Article ; Online: HJURP is recruited to double-strand break sites and facilitates DNA repair by promoting chromatin reorganization.

Serafim, Rodolfo B / Cardoso, Cibele / Storti, Camila B / da Silva, Patrick / Qi, Hongyun / Parasuram, Ramya / Navegante, Geovana / Peron, Jean Pierre S / Silva, Wilson A / Espreafico, Enilza M / Paçó-Larson, Maria L / Price, Brendan D / Valente, Valeria

Oncogene

2024 Volume 43, Issue 11, Page(s) 804–820

Abstract: HJURP is overexpressed in several cancer types and strongly correlates with patient survival. However, the mechanistic basis underlying the association of HJURP with cancer aggressiveness is not well understood. HJURP promotes the loading of the histone ... ...

Abstract	HJURP is overexpressed in several cancer types and strongly correlates with patient survival. However, the mechanistic basis underlying the association of HJURP with cancer aggressiveness is not well understood. HJURP promotes the loading of the histone H3 variant, CENP-A, at the centromeric chromatin, epigenetically defining the centromeres and supporting proper chromosome segregation. In addition, HJURP is associated with DNA repair but its function in this process is still scarcely explored. Here, we demonstrate that HJURP is recruited to DSBs through a mechanism requiring chromatin PARylation and promotes epigenetic alterations that favor the execution of DNA repair. Incorporation of HJURP at DSBs promotes turnover of H3K9me3 and HP1, facilitating DNA damage signaling and DSB repair. Moreover, HJURP overexpression in glioma cell lines also affected global structure of heterochromatin independently of DNA damage induction, promoting genome-wide reorganization and assisting DNA damage response. HJURP overexpression therefore extensively alters DNA damage signaling and DSB repair, and also increases radioresistance of glioma cells. Importantly, HJURP expression levels in tumors are also associated with poor response of patients to radiation. Thus, our results enlarge the understanding of HJURP involvement in DNA repair and highlight it as a promising target for the development of adjuvant therapies that sensitize tumor cells to irradiation.
MeSH term(s)	Humans ; Centromere/metabolism ; Centromere Protein A/genetics ; Centromere Protein A/metabolism ; Chromatin/genetics ; Chromosomal Proteins, Non-Histone/genetics ; Chromosomal Proteins, Non-Histone/metabolism ; DNA Repair/genetics ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Glioma/genetics
Chemical Substances	Centromere Protein A ; Chromatin ; Chromosomal Proteins, Non-Histone ; DNA-Binding Proteins ; HJURP protein, human
Language	English
Publishing date	2024-01-26
Publishing country	England
Document type	Journal Article
ZDB-ID	639046-8
ISSN	1476-5594 ; 0950-9232
ISSN (online)	1476-5594
ISSN	0950-9232
DOI	10.1038/s41388-024-02937-1
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Zs.A 2218: Show issues

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Article ; Online: A survey of management practices that influence performance and welfare of dairy calves reared in southern Brazil.

Hötzel, Maria J / Longo, Cibele / Balcão, Lucas F / Cardoso, Clarissa S / Costa, João H C

PloS one

2014 Volume 9, Issue 12, Page(s) e114995

Abstract: Here we report dairy calf management practices used by 242 smallholder family farmers in the South of Brazil. Data were collected via a semi-structured questionnaire with farmers, inspection of the production environment and an in-depth interview with a ... ...

Abstract	Here we report dairy calf management practices used by 242 smallholder family farmers in the South of Brazil. Data were collected via a semi-structured questionnaire with farmers, inspection of the production environment and an in-depth interview with a sample of 26 farmers. Herds had an average of 22.3 lactating cows and an average milk production of 12.7 L/cow/day. Calves were dehorned in 98% of the farms, with a hot iron in 95%. Male calves were castrated in 71% of the farms; methods were surgery (68%), emasculator (29%), or rubber rings (3%). No pain control was used for these interventions. In 51% of the farms all newborn male calves were reared, sold or donated to others; in 35% all newborn males were killed on the farm. Calves were separated from the dam up to 12 h after birth in 78% of the farms, and left to nurse colostrum from the dam without intervention in 55% of the farms. The typical amount of milk fed to calves was 4 L/day until a median age of 75 days. In 40% of the farms milk was provided in a bucket, in 49% with bottles, and in 11% calves suckled from a cow. Solid feeding in the milk-feeding period started at a median age of 10 days. Calves were housed individually in 70% of the farms; in 81% of the farms calves were housed in indoor pens, in 6% in outdoor hutches and in 13% they were kept on pasture. Diarrhoea was reported as the main cause of calf mortality in 71% of the farms. Farmers kept no records of calf disease, mortality, or use of medicines. Changing the scenario identified in this survey is essential to support the sustainable development of dairy production, an activity of great economic and social relevance for the region.
MeSH term(s)	Animal Husbandry/methods ; Animals ; Brazil ; Breeding ; Cattle/physiology ; Dairying/methods ; Female ; Lactation ; Male
Language	English
Publishing date	2014
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0114995
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Article ; Online: Optimizing orthotopic cell transplantation in the mouse adrenal gland.

Cardoso, Cibele C / Bornstein, Stefan R / Hornsby, Peter J

Cell transplantation

2010 Volume 19, Issue 5, Page(s) 565–572

Abstract: Orthotopic cell transplantation models are important for a complete understanding of cell-cell interactions as well as tumor biology. In published studies of orthotopic transplantation in the mouse adrenal gland, human neuroblastoma cells have been shown ...

Abstract	Orthotopic cell transplantation models are important for a complete understanding of cell-cell interactions as well as tumor biology. In published studies of orthotopic transplantation in the mouse adrenal gland, human neuroblastoma cells have been shown to invade and occupy the adrenal, but in these investigations a true orthotopic model was not established. Here we show an orthotopic model in which transplanted cells are retained within the adrenal gland by formation of a fibrin clot. To establish an appropriate technique, we used brightly fluorescent 10 microm polystyrene microspheres injected into the mouse adrenal gland. In the absence of fibrinogen/thrombin for clot formation, much of the injected material was extruded to the outside of the gland. When the microspheres were injected in a fibrinogen/thrombin mixture, fluorescence was confined to the adrenal gland. As a model neoplastic cell originating from the cortex of the gland, we used a tumorigenic bovine adrenocortical cell line. When 3 x 10(5) cells were implanted orthotopically, by 16 days the cell mass had expanded and had invaded the cortex, whereas when 1 x 10(5) cells were used, tumor masses were much smaller. We therefore subsequently used 3 x 10(5) cells. When mice were sacrificed at different time points, we found that tumor growth resulting was progressive and that by 26 days cells there was extensive invasion into the cortex or almost complete replacement of the cortex with tumor cells. As a model neoplastic cell of neural crest origin, we used SK-N-AS human neuroblastoma cells. Orthotopic transplantation of 3 x 10(5) cells resulted in extensive invasion and destruction of the gland by 26 days. In summary, the present orthotopic model for intra-adrenal cell transplantation is valuable for investigation of growth of neoplastic cells of both cortical and medullary origin and should be useful for future studies of cortex-medulla interactions.
MeSH term(s)	Adrenal Glands/cytology ; Adrenal Glands/metabolism ; Animals ; Cattle ; Cell Line, Tumor ; Cell Transplantation/methods ; Cells, Cultured ; Child ; Disease Models, Animal ; Female ; Fibrin/metabolism ; Fibrinogen/administration & dosage ; Humans ; Mice ; Mice, Transgenic ; Microspheres ; Neoplasm Transplantation/methods ; Neuroblastoma/metabolism ; Neuroblastoma/pathology ; Thrombin/administration & dosage ; Transplantation, Heterologous/methods
Chemical Substances	Fibrin (9001-31-4) ; Fibrinogen (9001-32-5) ; Thrombin (EC 3.4.21.5)
Language	English
Publishing date	2010-06-03
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1135816-6
ISSN	1555-3892 ; 0963-6897
ISSN (online)	1555-3892
ISSN	0963-6897
DOI	10.3727/096368910X509077
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Zs.A 3556: Show issues

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Article ; Online: The lncRNA RMEL3 protects immortalized cells from serum withdrawal-induced growth arrest and promotes melanoma cell proliferation and tumor growth.

Cardoso, Cibele / Serafim, Rodolfo B / Kawakami, Akinori / Gonçalves Pereira, Cristiano / Roszik, Jason / Valente, Valeria / Vazquez, Vinicius L / Fisher, David E / Espreafico, Enilza M

Pigment cell & melanoma research

2018 Volume 32, Issue 2, Page(s) 303–314

Abstract: ... C > T substitutions at dipyrimidine sites including CC > TT, typical of UV signature. RMEL3 mutation ...

Abstract	RMEL3 is a recently identified lncRNA associated with BRAFV600E mutation and melanoma cell survival. Here, we demonstrate strong and moderate RMEL3 upregulation in BRAF and NRAS mutant melanoma cells, respectively, compared to melanocytes. High expression is also more frequent in cutaneous than in acral/mucosal melanomas, and analysis of an ICGC melanoma dataset showed that mutations in RMEL3 locus are preponderantly C > T substitutions at dipyrimidine sites including CC > TT, typical of UV signature. RMEL3 mutation does not correlate with RMEL3 levels, but does with poor patient survival, in TCGA melanoma dataset. Accordingly, RMEL3 lncRNA levels were significantly reduced in BRAFV600E melanoma cells upon treatment with BRAF or MEK inhibitors, supporting the notion that BRAF-MEK-ERK pathway plays a role to activate RMEL3 gene transcription. RMEL3 overexpression, in immortalized fibroblasts and melanoma cells, increased proliferation and survival under serum starvation, clonogenic ability, and xenografted melanoma tumor growth. Although future studies will be needed to elucidate the mechanistic activities of RMEL3, our data demonstrate that its overexpression bypasses the need of mitogen activation to sustain proliferation/survival of non-transformed cells and suggest an oncogenic role for RMEL3.
MeSH term(s)	Animals ; Apoptosis/drug effects ; Carcinogenesis/drug effects ; Carcinogenesis/genetics ; Carcinogenesis/pathology ; Cell Line, Transformed ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Clone Cells ; Cytoprotection/drug effects ; Fibroblasts/drug effects ; Fibroblasts/pathology ; GTP Phosphohydrolases/genetics ; Gene Expression Regulation, Neoplastic/drug effects ; Humans ; Melanoma/genetics ; Melanoma/pathology ; Membrane Proteins/genetics ; Mice ; Mitogen-Activated Protein Kinases/antagonists & inhibitors ; Mitogen-Activated Protein Kinases/metabolism ; NIH 3T3 Cells ; Protein Kinase Inhibitors/pharmacology ; Proto-Oncogene Proteins B-raf/genetics ; RNA, Long Noncoding/genetics ; RNA, Long Noncoding/metabolism ; Serum/metabolism
Chemical Substances	Membrane Proteins ; Protein Kinase Inhibitors ; RNA, Long Noncoding ; Proto-Oncogene Proteins B-raf (EC 2.7.11.1) ; Mitogen-Activated Protein Kinases (EC 2.7.11.24) ; GTP Phosphohydrolases (EC 3.6.1.-) ; NRAS protein, human (EC 3.6.1.-)
Language	English
Publishing date	2018-12-16
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2409570-9
ISSN	1755-148X ; 1600-0749 ; 0893-5785 ; 1755-1471
ISSN (online)	1755-148X ; 1600-0749
ISSN	0893-5785 ; 1755-1471
DOI	10.1111/pcmr.12751
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Article: Impression Cytology Is a Non-invasive and Effective Method for Ocular Cell Retrieval of Zika Infected Babies: Perspectives in OMIC Studies.

Barbosa, Raquel Hora / Dos Santos, Maria Luiza B / Silva, Thiago P / Rosa-Fernandes, Liva / Pinto, Ana M V / Spínola, Pricila S / Bonvicino, Cibele R / Fernandes, Priscila V / Lucena, Evandro / Palmisano, Giuseppe / Melo, Rossana C N / Cardoso, Claudete Aparecida Araújo / Lemos, Bernardo

Frontiers in molecular neuroscience

2019 Volume 12, Page(s) 279

Abstract: Importance: Non-invasive techniques for retrieving ocular surface cells from babies infected by zika virus (ZIKV) during the gestational period remain to be determined.: Objectives: The aim of this study was to describe an optimized impression ... ...

Abstract	Importance: Non-invasive techniques for retrieving ocular surface cells from babies infected by zika virus (ZIKV) during the gestational period remain to be determined. Objectives: The aim of this study was to describe an optimized impression cytology method for the isolation of viable cells from Zika infected babies with and without Congenital Zika Syndrome (CZS) in satisfactory amount and quality to enable easy adoption in the field and application in the context of genomic and molecular approaches. Design settings and participants: Ocular surface samples were obtained with a hydrophilic nitrocellulose membrane (through optimized impression cytology method) from twelve babies referred to the Pediatric Service of the Antonio Pedro Hospital, Universidade Federal Fluminense (UFF), Niteroi, Rio de Janeiro, Brazil. After an authorized written informed consent from the parents, samples were collected from both eyes of 12 babies (4 babies with maternal ZIKV exposure during gestation and presence of clinical signs which included ocular abnormalities and microcephaly; 4 babies with maternal ZIKV exposure during gestation but no clinical signs; and 4 unaffected control babies with negative PCR for Zika virus and without clinical signs). Cells were used for microscopy analyses and evaluated for their suitability for downstream molecular applications in transcriptomic and proteomic experiments. Results: Our optimized impression cytology protocol enabled the capture of a considerable number of viable cells. The microscopic features of the conjunctival epithelial cells were described by both direct analysis of the membrane-attached cells and analysis of cytospinned captured cells using several staining procedures. Epithelial basal, polyhedral and goblet cells were clearly identified in all groups. All cases of ZIKV infected babies showed potential morphological alterations (cell keratinization, pyknosis, karyolysis, anucleation, and vacuolization). Molecular approaches were also performed in parallel. Genomic DNA and RNA were successfully isolated from all samples to enable the establishment of transcriptomic and proteomic studies. Conclusions and relevance: Our method proved to be a suitable, fast, and non-invasive tool to obtain ocular cell preparations from babies with and without Zika infection. The method yielded sufficient cells for detailed morphological and molecular analyses of samples. We discuss perspectives for the application of impression cytology in the context of ZIKV studies in basic and clinical research.
Language	English
Publishing date	2019-12-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2452967-9
ISSN	1662-5099
ISSN	1662-5099
DOI	10.3389/fnmol.2019.00279
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Article: New methods for investigating experimental human adrenal tumorigenesis.

Cardoso, Cibele C / Bornstein, Stefan R / Hornsby, Peter J

Molecular and cellular endocrinology

2008 Volume 300, Issue 1-2, Page(s) 175–179

Abstract: Adenomas and nodules of the human adrenal cortex are common, whereas adrenocortical carcinomas are rare. Genes such as IGF2 have been suggested to be important in human adrenocortical tumorigenesis but their role has not been directly investigated. We ... ...

Abstract	Adenomas and nodules of the human adrenal cortex are common, whereas adrenocortical carcinomas are rare. Genes such as IGF2 have been suggested to be important in human adrenocortical tumorigenesis but their role has not been directly investigated. We describe here elements of a system in which hypotheses concerning the molecular basis for the formation of benign and malignant adrenocortical lesions can be experimentally tested. Various viral vectors have been employed in the study of adrenocortical cell biology. Because of the low proliferative rate of primary human adrenocortical (pHAC) cells, a lentiviral system is ideal for transducing these cells with genes that may alter their characteristics or cause them to acquire benign or malignant tumorigenicity. Cultures of pHAC cells were highly infectible with lentiviruses and showed a higher proliferative potential when transduced with a lentivirus encoding IGF2. For tumorigenesis studies of genetically modified adrenocortical cells, we use RAG2(-/-), gammac(-/-) mice. Using this immunodeficient mouse model, we established an orthotopic intra-adrenal cell transplantation technique for adrenocortical cells that should be of value for future studies of the experimental conversion of human adrenocortical cells to a benign or malignant tumorigenic state.
MeSH term(s)	Adrenal Gland Neoplasms/etiology ; Adrenal Gland Neoplasms/pathology ; Adrenal Gland Neoplasms/physiopathology ; Animals ; Cell Transformation, Neoplastic ; Cell Transplantation/methods ; Genetic Vectors ; Humans ; Lentivirus/genetics ; Lentivirus/metabolism
Language	English
Publishing date	2008-11-13
Publishing country	Ireland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	187438-x
ISSN	1872-8057 ; 0303-7207
ISSN (online)	1872-8057
ISSN	0303-7207
DOI	10.1016/j.mce.2008.10.046
Database	MEDical Literature Analysis and Retrieval System OnLINE

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