Natália Galvão Garcia

Differences in neonatal exposure to estradiol or testosterone on ovarian function and hormonal levels

Exposure to an excess of androgen or estrogen can induce changes in reproductive function in adult animals that resemble polycystic ovary syndrome in humans. However, considerable differences exist among several types of animal models. Little is known about the molecular features of steroidogenesis and folliculogenesis in the ovaries of rats exposed to different sex steroids as neonates. Here, we evaluated the impact of androgen and estrogen exposure on the ovaries of adult female rats during their neonatal period in the gene expression of Lhr and Cyp17a1, two key players of steroidogenesis. We also assessed hormone levels, folliculogenesis and the theca-interstitial cell population. The study was performed on the second postnatal day in thirty female Wistar rats that were sorted into the following three intervention groups: testosterone, estradiol and vehicle (control group). The animals were euthanized 90 days after birth. The main outcomes were hormone serum levels, ovary histomorphometry and gene expression of Lhr and Cyp17a1 as analyzed via quantitative real-time PCR. We found that exposure to excess testosterone in early life increased the LH and testosterone serum levels, the LH/FSH ratio, ovarian theca-interstitial area and gene expression of Lhr and Cyp17a1 in adult rats. Estrogen induced an increase in the ovarian theca-interstitial area, the secondary follicle population and gene expression of Lhr and Cyp17a1. All animals exposed to the sex steroids presented with closed vaginas. Our data suggest that testosterone resulted in more pronounced reproductive changes than did estrogen exposure. Our results might provide some insight into the role of different hormones on reproductive development and on the heterogeneity of clinical manifestations of conditions such as polycystic ovary syndrome.

Necrotizing Sialometaplasia of Palate: A Case Report

Background. Necrotizing sialometaplasia (NS) is an uncommon benign reactive necrotizing inflammatory process involving minor salivary gland that often mimics malignancy both clinically and histopathologically. Case Report. We report the case of a healthy 26-year-old man with a painless swelling in the hard palate near the middle raphe, asymptomatic, well limited, and raised edges. The patient was submitted to incisional biopsy and histopathological examination. The histological diagnosis was necrotizing sialometaplasia. Discussion. The clinical and histological similarity between this entity and a malignant lesion implies a risk of unnecessary or mistreatment. Therefore, clinicians and pathologists should be aware of this lesion as to avoid errors in the diagnosis and treatment of this benign pathologic condition.

Idiopathic bone cavity: case series involving children and adolescents.

AIM This study aimed at reviewing a series of clinical cases of children and adolescents treated for idiopathic bone cavity (IBC). METHODS Ten lesions diagnosed as IBC in nine patients treated between February 2000 and December 2008 were reviewed, focusing on clinical and radiographic aspects, as well the therapeutic approach. RESULTS Most of the patients were Caucasian with no sex predilection (55.5% males and 45.5% females), and all were in the second decade of life at the time of IBC diagnosis. The lesions were asymptomatic and detected during routine radiographic examinations. All the lesions were located in the mandible, and the anterior region was the most affected. The most adopted therapeutic approach was simple curettage of the bone cavity, and all surgically-treated patients presented complete bone healing after 6 months, postoperatively. CONCLUSIONS Clinical and radiographic diagnoses associated with surgical exploration are the most important treatment options in order to avoid radical surgery or unnecessary endodontic treatment, mainly when idiopathic bone cavities are detected in children and adolescents. Further studies aimed at elucidating the etiopathogenesis of this condition and defining the most appropriate treatment should be conducted.

Peripheral compound odontoma erupting in the gingiva.

BackgroundPeripheral odontoma arising in the extraosseous soft tissues is rare and if not removed early, may enlarge over time and eventually erupt in the oral cavity.Case presentationA 15-year-old girl presented with “denticles on the gingiva”. During the intraoral examination, seven small tooth-like structures were found. These were exposed in the anterior left gingiva between the permanent maxillary lateral incisor and canine teeth, and the left first premolar was absent. Radiographic examination revealed irregular tooth-like structures without evidence of bone involvement.ConclusionThe lesion was surgically removed, and the specimens were analyzed histopathologically. The diagnosis of compound odontoma was established.Clinical significanceThis is the twelfth reported case of peripheral odontoma in the gingiva and the first one that erupted in the oral cavity.

Extensive cutaneous lesions in paracoccidioidomycosis successfully treated with itraconazole and β-glucan.

Georg Stary, MD Alexandra Geusau, MD Division of Immunology Allergy and Infectious Diseases Departmentof Dermatology Medical University of Vienna Vienna Austria E-mail: alexandra.geusau@meduniwien.ac.at References 1 Workowski KA, Berman SM. Sexually Transmitted Diseases Treatment Guidelines 2006. MMWR Morb Mortal Wkly Rep 2006; 55: 1–94. 2 David G, Perpoint T, Boibieux A, et al. Secondary pulmonary syphilis: report of a likely case and literature review. Clin Infect Dis 2006; 42: 11–15. 3 Woehrl S, Geusau A. Clinical update: syphilis in adults. Lancet 2007; 369: 1912–1914. 4 Mahto M, Mohammed F, Wilkins E, et al. Pseudohepatic tumor associated with secondary syphilis in an HIV-positive male. Int J STD AIDS 2006; 17: 139–141. 5 Hamlyn E, Taylor C. Sexually transmitted proctitis. Postgrad Med J 2006; 82: 733–736. 6 Geusau A, Mayerhofer S, Schmidt B, et al. The year 2002 re-emergence of syphilis in Austria. Int J STD AIDS 2004; 15: 496–497. 7 Gerbase AC, Rowley JT, Mertens TE. Global epidemiology of sexually transmitted diseases. Lancet 1998; 351(Suppl. 3): 2–4. 8 Müller H, Eisendle K, Bräuninger W, et al. Comparative analysis of immunohistochemistry, polymerase chain reaction and focus-floating microscopy for the detection of Treponema pallidum in mucocutaneous lesions of primary, secondary and tertiary lesions. Br J Dermatol 2011; 165: 50–60. 9 Guarner J, Greer PW, Bartlett J, et al. Congenital syphilis in a newborn: an immunopathologic study. Mod Pathol 1999; 12: 82–87.

May Sonic Hedgehog proteins be markers for malignancy in uterine smooth muscle tumors

Several studies have demonstrated that the Sonic Hedgehog signaling pathway (SHH) plays an important role in tumorigenesis and cellular differentiation. We analyzed the protein expression of SHH pathway components and evaluated whether their profile could be useful for the diagnosis, prognosis, or prediction of the risk of malignancy for uterine smooth muscle tumors (USMTs). A total of 176 samples (20 myometrium, 119 variants of leiomyoma, and 37 leiomyosarcoma) were evaluated for the protein expression of the SHH signaling components, HHIP1 (SHH inhibitor), and BMP4 (SHH target) by immunohistochemistry. Western blot analysis was performed to verify the specificity of the antibodies. We grouped leiomyoma samples into conventional leiomyomas and unusual leiomyomas that comprise atypical, cellular, mitotically active leiomyomas and uterine smooth muscle tumors of uncertain malignant potential. Immunohistochemical analysis showed that SMO, SUFU, GLI1, GLI3, and BMP4 expression gradually increased depending on to the histologic tissue type. The protein expression of SMO, SUFU, and GLI1 was increased in unusual leiomyoma and leiomyosarcoma samples compared to normal myometrium. The inhibitor HHIP1 showed higher expression in myometrium, whereas only negative or basal expression of SMO, SUFU, GLI1, and GLI3 was detected in these samples. Strong expression of SHH was associated with poorer overall survival. Our data suggest that the expression of SHH proteins can be useful for evaluating the potential risk of malignancy for USMTs. Moreover, GLI1 and SMO may serve as future therapeutic targets for women with USMTs.

Oral Verrucous Carcinoma Mimicking a Chronic Candidiasis: A Case Report

Verrucous carcinoma has a special propensity to mimic benign lesions of the oral cavity. A case of the oral verrucous carcinoma in maxillary alveolar ridge, extending to buccal vestibule, cheek, and labial mucosa, which was diagnosed and initially treated as chronic candidiasis, is presented. Clinical, histopathological, and therapeutic considerations related to diagnosis of the verrucous carcinoma in the oral cavity are discussed.

Impaired branched-chain amino acid metabolism may underlie the nonalcoholic fatty liver disease-like pathology of neonatal testosterone-treated female rats.

Polycystic ovary syndrome (PCOS) is frequently associated with non-alcoholic fatty liver disease (NAFLD), but the mechanisms involved in the development of NAFLD in PCOS are not well known. We investigated histological changes and metabolomic profile in the liver of rat models of PCOS phenotype induced by testosterone or estradiol. Two-day old female rats received sc injections of 1.25 mg testosterone propionate (Testos; n = 10), 0.5 mg estradiol benzoate (E2; n = 10), or vehicle (control group, CNT; n = 10). Animals were euthanized at 90–94 d of age and the liver was harvested for histological and metabolomic analyses. Findings showed only Testos group exhibited fatty liver morphology and higher levels of ketogenic and branched-chain amino acids (BCAA). Enrichment analysis showed effects of testosterone on BCAA degradation pathway and mitochondrial enzymes related to BCAA metabolism. Testos group also had a decreased liver fatty acid elongase 2 (ELOVL2) activity. E2 group had reduced lipid and acylcarnitine metabolites in the liver. Both groups had increased organic cation transporters (SLC22A4 and SLC16A9) activity. These findings indicate that neonatal testosterone treatment, but not estradiol, produces histological changes in female rat liver that mimic NAFLD with testosterone-treated rats showing impaired BCAA metabolism and dysfunctions in ELOVL2, SLC22A4 and SLC16A9 activity.

Oncomirs Expression Profiling in Uterine Leiomyosarcoma Cells

MicroRNAs (miRNAs) are small non-coding RNAs that act as regulators of gene expression at the post-transcriptional level. They play a key role in several biological processes. Their abnormal expression may lead to malignant cell transformation. This study aimed to evaluate the expression profile of 84 miRNAs involved in tumorigenesis in immortalized cells of myometrium (MM), uterine leiomyoma (ULM), and uterine leiomyosarcoma (ULMS). Specific cell lines were cultured and qRT-PCR was performed. Thirteen miRNAs presented different expression profiles in ULM and the same thirteen in ULMS compared to MM. Eight miRNAs were overexpressed, and five were underexpressed in ULM. In ULMS cells, five miRNAs exhibited an overexpression and eight were down-regulated. Six miRNAs (miR-1-3p, miR-130b-3p, miR-140-5p, miR-202-3p, miR-205-5p, and miR-7-5p) presented a similar expression pattern in cell lines compared to patient samples. Of these, only three miRNAs showed significant expression in ULM (miR-1-3p, miR-140-5p, and miR-7-5p) and ULMS (miR-1-3p, miR-202-3p, and miR-7-5p). Our preliminary approach identified 24 oncomirs with an altered expression profile in ULM and ULMS cells. We identified four differentially expressed miRNAs with the same profile when compared with patients’ samples, which strongly interacted with relevant genes, including apoptosis regulator (BCL2), epidermal growth factor receptor (EGFR), vascular endothelial growth factor A (VEGFA), insulin like growth factor 1 receptor (IGF1R),serine/threonine kinase (RAF1), receptor tyrosine kinase (MET), and bHLH transcription factor (MYCN). This led to alterations in their mRNA-target.

Loss of cytokeratin 10 indicates malignant transformation in actinic cheilitis

ObjectivesThe aim of this study was to investigate the relationship the expression of cytokeratins (CK10 and CK13) and the cell proliferation index determined by Ki-67 of lip squamous cell carcinoma and actinic cheilitis with different degrees of dysplasia.Materials and methodsForty-five paraffin-embedded actinic cheilitis with and without dysplasia and 20 lip squamous cell carcinoma were analyzed by immunohistochemistry using anti-human anti-CK10, anti-CK13, and anti-Ki-67 antibodies.ResultsThe majority of actinic cheilitis showed immunopositivity for CK10 and CK13 with decrease or loss of expression in dysplastic areas. In lip squamous cell carcinoma of the lip, heterogeneous expression of CK13 and immunonegativity for CK10 were observed. There was a statistically significant difference between CK10 expression in lip squamous cell carcinoma and in actinic cheilitis with or without dysplasia (p < 0.001). The cell proliferation index was higher in actinic cheilitis with dysplasia and lip squamous cell carcinoma than in actinic cheilitis without epithelial dysplasia. A significant correlation was found between the intensity of the epithelial dysplasia and the cell proliferation index (p < 0.001).ConclusionThese results provide evidence that there is a downregulation of CK10 expression in dysplastic areas of patients with actinic cheilitis and in those with lip squamous cell carcinoma (LSCC) and that the index of cell proliferation, determined by Ki-67, is directly correlated with the intensity of the epithelial dysplasia.Clinical relevanceAltogether, these results suggest that CK10 expression and the epithelial cell proliferation index can help to identify malignant transformation in the lip region.