Two histologically different tumours in a neonate born from an assisted reproductive technology pregnancy

The first case of a female neonate born from an in vitro fertilization with embryo transfer (IVF-ET) and intracytoplasmic sperm injection (ICSI) (IVF-ET (ICSI) with two histologically different tumours (craniopharyngioma and hepatoblastoma) is described. Anti-neoplasmatic therapy was abandoned due to the significant extent of the disease (craniopharyngioma, 15×12 cm in diameter with active internal hydrocephalus; and right liver lobe hepatoblastoma, 5 cm in diameter) and the severely impaired general condition of the neonate. The neonate died on the 30th day of life due to cerebellar and brainstem herniation, followed by circulatory and respiratory failure

Ocena częstości występowania mutacji genów BRAF, KRas oraz metylacji genu RASSF1A w wolu guzkowym na podstawie badania materiału cytologicznego uzyskanego drogą biopsji aspiracyjnej cienkoigłowej

  Introduction: Standard pre-operative diagnosis of nodular goitre is not always conclusive. The decision about nodular goitre surgery is increasingly based on molecular methods. The aim of the study was to determine BRAF T1799A mutation and KRas proto-oncogene mutation, and the analysis of RASSF1A promoter methylation level in cytological material obtained from FNAB specimens of thyroid nodules. Material and methods: The study population consisted of 85 women and 12 men. The study material was genomic DNA isolated from peripheral blood and thyroid bioptates. Pyrosequencing was used for the evaluation of RASSF1 methylation level. KRas mutation was investigated with Sanger sequencing. BRAF mutation was analysed by standard methods of real-time amplification detection (real-time PCR) with the use of specific starters surrounding the mutated site. Results: A significant positive correlation was demonstrated between mean methylation of four CpG islands of RASSF1A gene and thyroid tumour volume and its largest diameter (p < 0.05). KRas mutation was not detected in any of the 97 patients. In 7/85 subjects (8.2%) BRAF mutation was observed. In 6/7 patients with BRAF mutation, FNAB of thyroid nodules confirmed a benign nature of the lesions; the material was non-diagnostic in one patient, and papillary thyroid cancer was diagnosed on the basis of postoperative histopathology assessment. Conclusions: The results of genetic tests reported in our study indicate that the presence of BRAF mutation or higher RASSF1A methylation levels in FNAB cytology specimens of benign lesions may be useful in the assessment of oncological risk, while the evaluation of KRas proto-oncogene mutation is not a valuable test in pre-operative diagnosis of nodular goitre. (Endokrynol Pol 2015; 66 (5): 384–393)    Wstęp: Na podstawie standardowej przedoperacyjnej diagnostyki wola guzkowego nie zawsze uzyskuje się jednoznaczne rozpoznanie. Coraz częściej w kwalifikacji wola guzkowego do zabiegu operacyjnego wykorzystywane są metody badań molekularnych. Celem pracy było oznaczenie mutacji T1799A genu BRAF i mutacji protoonkogenu KRas oraz analiza stopnia metylacji promotora genu RASSF1A w materiale komórkowym uzyskanym z guzków tarczycy na drodze biopsji aspiracyjnej cienkoigłowej. Materiały i metody: Badaniami objęto 85 kobiet i 12 mężczyzn. Materiał do badań stanowił genomowy DNA wyizolowany z krwi obwodowej pacjentów oraz z bioptatów tarczycy. Do oceny stopnia metylacji genu RASSF1 wykorzystano metodę pirosekwencjonowania. Mutacje genu KRas badano metodą sekwencjonowania Sangera. Do oznaczania mutacji BRAF użyto standardowej metodologii detekcji amplifikacji w czasie rzeczywistym (real-time PCR) z zastosowaniem specyficznych starterów otaczających miejsce zmutowane. Wyniki: Wykazano, że średnia metylacji czterech wysp CpG w genie RASSF1A znamiennie, dodatnio koreluje z objętością guza tarczycy i największym wymiarem guza (p < 0,05). U żadnej z 97 osób nie stwierdzono mutacji Kras. U 7/85 badanych (8,2%) stwierdzono obecność mutacji genu BRAF. U 6/7 osób z obecnością mutacji BRAF, BAC guzków tarczycy wykazała łagodny charakter tych zmian, u jednej osoby otrzymano materiał niediagnostyczny, a na podstawie pooperacyjnego badania histopatologicznego rozpoznano raka brodawkowatego tarczycy. Wnioski: Otrzymane wyniki badań genetycznych wskazują, że obecność mutacji genu BRAF lub wyższego odsetka metylacji genu RASSF1A w materiale cytologicznym z biopsji aspiracyjnej cienkoigłowej zmiany cytologicznie łagodnej może mieć znaczenie w ocenie zagrożenia onkologicznego, podczas gdy ocena mutacji protonkogenu KRas nie jest przydatna w diagnostyce przedoperacyjnej wola guzkowego. (Endokrynol Pol 2015; 66 (5): 384–393)

The role of C1 inhibitor and complement as acute phase reactants: are we missing the diagnosis of hereditary angioedema?

Abstract Background C1 inhibitor (C1-INH) and complement 4 (C4) have historically been referred to as positive acute phase reactants, however this has never been evaluated in hereditary angioedema (HAE) patients. Low function of C1-INH and low levels of C4 are important in the diagnosis of HAE type 1 and 2. If C1-INH and/or C4 are significant acute phase reactants, their levels may be falsely “normal” in patients with HAE when measured during times of infection or inflammation resulting in missed or delayed diagnosis. Case presentation We present a case series of four HAE patients who had C4, C1-INH, c-reactive protein (CRP) and ferritin measured at baseline and again during a self-reported upper respiratory tract infection (URTI) or flu-like illness. We did not identify any HAE patients who had a significant change in their C1-INH functional level in the context of a mild infection. However, the C4 level did increase into the normal range on three occasions (2 patients, with 1 patient having elevation during two separate illnesses). Conclusions C1 inhibitor may not be a clinically significant acute phase protein and appears to still be a reliable diagnostic marker of hereditary angioedema, even in times of modest acute inflammation, unlike complement C4 which can be elevated in this setting. </jats:sec

The role of C1 inhibitor and complement as acute phase reactants: are we missing the diagnosis of hereditary angioedema?

Background C1 inhibitor (C1-INH) and complement 4 (C4) have historically been referred to as positive acute phase reactants, however this has never been evaluated in hereditary angioedema (HAE) patients. Low function of C1-INH and low levels of C4 are important in the diagnosis of HAE type 1 and 2. If C1-INH and/or C4 are significant acute phase reactants, their levels may be falsely “normal” in patients with HAE when measured during times of infection or inflammation resulting in missed or delayed diagnosis. Case presentation We present a case series of four HAE patients who had C4, C1-INH, c-reactive protein (CRP) and ferritin measured at baseline and again during a self-reported upper respiratory tract infection (URTI) or flu-like illness. We did not identify any HAE patients who had a significant change in their C1-INH functional level in the context of a mild infection. However, the C4 level did increase into the normal range on three occasions (2 patients, with 1 patient having elevation during two separate illnesses). Conclusions C1 inhibitor may not be a clinically significant acute phase protein and appears to still be a reliable diagnostic marker of hereditary angioedema, even in times of modest acute inflammation, unlike complement C4 which can be elevated in this setting.Medicine, Faculty ofNon UBCAllergy and Immunology, Division ofHematology, Division ofMedicine, Department ofReviewedFacult

RARE-58. CONGENITAL METASTATIC CHORDOMA OF THE CLIVUS

Abstract Chordomas are rare midline axial skeletal neoplasms that typically present in adults. They are infrequent in childhood with typical localization in the spheno-occipital skull base. They are derived from remnants of the embryonic notochord. We present the case of 4 months old girl, who was born with „blueberry muffin” syndrome and was first negatively diagnosed for neuroblastoma and leukemia (two negative skin biopsies were performed) was admitted with axial laxity. In imaging testes there was a tumor of the scull base, metastases in the lungs and kidneys (that were not seen at previous assessments) and a small lesion in the heart. The third biopsy of skin lesion was performed and pathological examination revealed a neoplasm composed of cords, clusters, and chains of multivacuolated cells embedded within a myxoid matrix and separated by fibrous septa. No atypical and dedifferentiated features were present. Mitotic activity was not observed. Neoplastic cells showed the typical cytoplasmic immunostaining for EMA, S100 and cytokeratin AE1/AE3, strong nuclear brachyury expression, and retention of nuclear INI-1 expression. The diagnosis of chordoma was established. Neoplastic tissue and blood samples were obtained for molecular analysis using next generation sequencing, including germline mutations assessment (are ongoing). Chemotherapy as for soft tissue sarcomas was undertaken. Currently a patient is on treatment with improvement of neurological status.</jats:p

Diagnostic Utility of Genetic and Immunohistochemical H3-3A Mutation Analysis in Giant Cell Tumour of Bone

To validate the reliability and implementation of an objective diagnostic method for giant cell tumour of bone (GCTB). H3-3A gene mutation testing was performed using two different methods, Sanger sequencing and immunohistochemical (IHC) assays. A total of 214 patients, including 120 with GCTB and 94 with other giant cell-rich bone lesions, participated in the study. Sanger sequencing and IHC with anti-histone H3.3 G34W and G34V antibodies were performed on formalin-fixed, paraffin-embedded tissues, which were previously decalcified in EDTA if needed. The sensitivity and specificity of the molecular method was 100% (95% CI: 96.97–100%) and 100% (95% CI: 96.15–100%), respectively. The sensitivity and specificity of IHC was 94.32% (95% CI: 87.24–98.13%) and 100% (95% CI: 93.94–100.0%), respectively. P.G35 mutations were discovered in 2/9 (22.2%) secondary malignant GCTBs and 9/13 (69.2%) GCTB after denosumab treatment. We confirmed in a large series of patients that evaluation of H3-3A mutational status using direct sequencing is a reliable tool for diagnosing GCTB, and it should be incorporated into the diagnostic algorithm. Additionally, we discovered IHC can be used as a screening tool. Proper tissue processing and decalcification are necessary. The presence of the H3-3A mutation did not exclude malignant GCTB. Denosumab did not eradicate the neoplastic cell population of GCTB.</jats:p

Diagnostic Utility of Genetic and Immunohistochemical H3-3A Mutation Analysis in Giant Cell Tumour of Bone

To validate the reliability and implementation of an objective diagnostic method for giant cell tumour of bone (GCTB). H3-3A gene mutation testing was performed using two different methods, Sanger sequencing and immunohistochemical (IHC) assays. A total of 214 patients, including 120 with GCTB and 94 with other giant cell-rich bone lesions, participated in the study. Sanger sequencing and IHC with anti-histone H3.3 G34W and G34V antibodies were performed on formalin-fixed, paraffin-embedded tissues, which were previously decalcified in EDTA if needed. The sensitivity and specificity of the molecular method was 100% (95% CI: 96.97&ndash;100%) and 100% (95% CI: 96.15&ndash;100%), respectively. The sensitivity and specificity of IHC was 94.32% (95% CI: 87.24&ndash;98.13%) and 100% (95% CI: 93.94&ndash;100.0%), respectively. P.G35 mutations were discovered in 2/9 (22.2%) secondary malignant GCTBs and 9/13 (69.2%) GCTB after denosumab treatment. We confirmed in a large series of patients that evaluation of H3-3A mutational status using direct sequencing is a reliable tool for diagnosing GCTB, and it should be incorporated into the diagnostic algorithm. Additionally, we discovered IHC can be used as a screening tool. Proper tissue processing and decalcification are necessary. The presence of the H3-3A mutation did not exclude malignant GCTB. Denosumab did not eradicate the neoplastic cell population of GCTB