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UNLABELLED: Determination of 17α-nyaroxyprogesterone (17OHP) is used for the diagnosis and monitoring of Congenital Adrenal Hyperplasia (CAH). Problems associated with the specificity of antibodies used in direct immunoassays can yield high false results. OBJECTIVES: To analyze serum levels of direct 17OHP (17OHPd) and previous extraction (17OHPe) in the neonatal period, in order to establish reference values. To relate levels of 17OHPd and 17OHPe with other androgens in CAH patients. SUBJECTS AND METHODS: Serum 17OHPd and 17OHPe were measured via RIA-DPC in 400 healthy newborns and infants (aged 2-365 days), and 100 treated CAH patients (aged 1-18 years). The extraction was performed with 3% isopropanol/heptane. The influence of age and gender was assessed by ANOVA. RESULTS: The serum levels of 17OHP were significantly correlated with chronological age, but not with gestational age, sex or birth weight. The difference between 17OHPd and 17OHPe decreased with age. The correlation index between 17OHPd and 17OHPe in CAH patients was 0.93 (p < 0.01). CONCLUSION: The present results provide 17OHP reference values for infants from birth up to one year of life. The extraction method is necessary in the neonatal period up to 6 months of life. Our data might be useful to make an early CAH diagnosis and follow-up newborns with high 17OHP levels without adrenal pathology.
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17-alfa-Hidroxiprogesterona/sangue , Radioimunoensaio/métodos , Hiperplasia Suprarrenal Congênita/diagnóstico , Fatores Etários , Peso ao Nascer , Diagnóstico Precoce , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Masculino , Triagem Neonatal , Valores de Referência , Fatores SexuaisRESUMO
Glyphosate is the most widely used herbicide in the world with application in agriculture, forestry, industrial weed control, garden and aquatic environments. However, its use is highly controversial for the possible impact on not-target organisms, such as amphibians, which are vanishing at an alarming and rapid rate. Due to the high solubility in water and ionic nature, the glyphosate requires of surfactants to increase activity. In addition, for the control of coca (Erythroxylum coca) and agricultural weeds in Colombia, formulated glyphosate is mixed and sprayed with the adjuvant Cosmo-Flux 411F to increase the penetration and activity of the herbicide. This study evaluates the acute toxic and sublethal effects (embryonic development, tadpole body size, tadpole swimming performance) of the mixture of the formulated glyphosate Roundup Active and Cosmo-Flux 411F to anuran embryos and tadpoles of four Colombian species under 96h laboratory standard tests and microcosms, which are more similar to field conditions as they include soil, sand and macrophytes. In the laboratory, embryos and tadpoles of Engystomops pustulosus were the most tolerant (LC50 = 3904 microg a.e./L; LC50=2 799 pg a.e./L, respectively), while embryos and tadpoles of Hypsiboas crepitans (LC50=2 203 microg a.e./L; LC50=1424 microgg a.e./L, respectively) were the most sensitive. R. humboldti and R. marina presented an intermediate toxicity. Embryos were significantly more tolerant to the mixture than tadpoles, which could be likely attributed to the exclusion of chemicals by the embryonic membranes and the lack of organs, such as gills, which are sensitive to surfactants. Sublethal effects were observed for the tadpole body size, but not for the embryonic development and tadpole swimming performance. In microcosms, no toxicity (LC50 could not be estimated), or sublethal responses were observed at concentrations up to fourfold (14.76 kg glyphosate a.e./ha) the highest field application rate of 3.69 kg glyphosate a.e./ha. Thus, toxicity was less in the microcosms than in laboratory tests, which may be attributed to the presence of sediments and organic matter which rapidly adsorb glyphosate and surfactants such as POEA. It is concluded that the mixture of glyphosate (Roundup Active) and Cosmo-Flux*411F, as used in the field, has a negligible toxic effect to embryos and tadpoles of the species tested in this study.
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Anuros , Glicina/análogos & derivados , Herbicidas/toxicidade , Larva/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Xantonas/toxicidade , Animais , Anuros/classificação , Colômbia , Glicina/toxicidade , Dose Letal Mediana , Testes de Toxicidade Aguda , GlifosatoRESUMO
INTRODUCTION: Individuals with chronic pain often receive prescription opioid medication, and they may use cannabis to treat pain as well, although the risks of cannabis-opioid co-use are significant. This study aimed to investigate whether two transdiagnostic factors, emotion regulation and distress tolerance, had significant indirect effects in the relationship between pain and cannabis use in adults with chronic pain and an opioid prescription. METHODS: Participants (n = 450; mean age = 38.6 ± 11.09) were recruited using Qualtrics panel service and were 75 % female and 79 % White, non-Hispanic. Participants completed a 30-minute self-report survey capturing three-month cannabis use, the Difficulties in Emotional Regulation Scale (DERS), and the Distress Tolerance Scale (DTS). The Graded Pain Scale (GCPS) assessed pain severity/intensity and disability. Analyses used the SPSS PROCESS macro, with both single (i.e., one transdiagnostic factor) and parallel indirect effects (i.e., both the DERS and DTS) examined. RESULTS: There were statistically significant indirect effects for both the DERS and DTS in the relationship between pain intensity or disability and three-month cannabis use in single factor models. In the parallel indirect effect model, only the DERS was statistically significant (intensity indirect effect coefficient = 0.0195 % confidence interval [95 %CI] = 0.0065, 0.390; disability indirect effect coefficient = 0.0147, 95 %CI = 0.0055, 0.0274). CONCLUSIONS: When examining parallel indirect effects, only emotional regulation and not distress tolerance mediated the relationship between chronic pain and cannabis use among those with an opioid prescription. Clinically, interventions aimed at improving emotional regulation in individuals with chronic pain can help limit cannabis and opioid co-use.
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Cannabis , Dor Crônica , Regulação Emocional , Alucinógenos , Transtornos Relacionados ao Uso de Opioides , Adulto , Humanos , Feminino , Pessoa de Meia-Idade , Masculino , Analgésicos Opioides/uso terapêutico , Dor Crônica/tratamento farmacológico , Transtornos Relacionados ao Uso de Opioides/psicologiaRESUMO
Osteosarcoma (OS) is the most common primary bone tumour in the paediatric age group. Treatment-refractory pulmonary metastasis continues to be the major complication of OS, reducing the 5-year survival rate for these patients to 10-20%. The mechanisms underlying the metastatic process in OS are still unclear, but undoubtedly, a greater understanding of the factors and interactions involved in its regulation will open new and much needed opportunities for therapeutic intervention. Recent published data have identified a new role for bone-specific macrophages (osteoclasts) and tumour-associated macrophages (TAMs), in OS metastasis. In this review we discuss the contribution of TAMs and osteoclasts in the establishment and maintenance of secondary metastatic lesions, and their novel role in the prevention of metastatic disease in a primary bone cancer such as osteosarcoma.
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Neoplasias Ósseas/patologia , Macrófagos/fisiologia , Osteoclastos/fisiologia , Osteossarcoma/secundário , Animais , HumanosRESUMO
A facile vacuum-assisted vapor deposition process has been developed to control the pore size of ordered mesoporous silica materials in a stepwise manner with angstrom precision, providing an unprecedented paradigm for screening a designer hydrophobic drug nanocarrier with optimized pore diameter to maximize drug solubility.
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Nanoestruturas/química , Compostos de Organossilício/química , Porosidade , Interações Hidrofóbicas e Hidrofílicas , Oxirredução , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
BACKGROUND: Overexpression of CEACAM6 has been reported for a number of malignancies. However, the mechanism of how CEACAM6 contributes to cancer formation and its role in head and neck squamous cell carcinoma (HNSCC) remains unclear. Therefore, we examined the role of CEACAM6 in head and neck squamous cell carcinoma (HNSCC). METHODS: CEACAM6 expression was examined in normal squamous epithelia as well as a number of patient HNSCC samples and tumours derived from HNSCC cell lines injected into NOD/SCID mice. CEACAM6 expression was manipulated in HNSCC cell lines by shRNA-mediated CEACAM6 knockdown or virally-delivered overexpression of CEACAM6. The role of CEACAM6 in tumour growth and chemotherapeutic sensitivity was then assessed in vivo and in vitro respectively. RESULTS: CEACAM6 expression was significantly increased in highly tumourigenic HNSCC cell lines when compared to poorly tumourigenic HNSCC cell lines. Moreover, HNSCC patient tumours demonstrated focal expression of CEACAM6. Functional investigation of CEACAM6, involving over-expression and knock down studies, demonstrated that CEACAM6 over-expression could enhance tumour initiating activity and tumour growth via activation of AKT and suppression of caspase-3 mediated cell death. CONCLUSION: We report that CEACAM6 is focally overexpressed in a large fraction of human HNSCCs in situ. We also show that over-expression of CEACAM6 increases tumour growth and tumour initiating activity by suppressing PI3K/AKT-dependent apoptosis of HNSCC in a xenotransplant model of HNSCC. Finally, our studies indicate that foci of CEACAM6 expressing cells are selectively ablated by treatment of xenotransplant tumours with pharmacological inhibitors of PI3K/AKT in vivo.
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Antígenos CD/genética , Apoptose/genética , Carcinoma de Células Escamosas/genética , Moléculas de Adesão Celular/genética , Transformação Celular Neoplásica/genética , Expressão Gênica , Neoplasias de Cabeça e Pescoço/genética , Animais , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Proteínas Ligadas por GPI/genética , Técnicas de Silenciamento de Genes , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Imidazóis/administração & dosagem , Imidazóis/farmacologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Quinolinas/administração & dosagem , Quinolinas/farmacologia , Carcinoma de Células Escamosas de Cabeça e PescoçoRESUMO
BACKGROUND: Iodide organification defect (IOD) is characterized by a reduced ability of the thyroid gland to retain iodide resulting in hypothyroidism. Mutations in thyroid peroxidase (TPO) gene appear to be the most common cause of IOD and are commonly inherited in an autosomal recessive fashion. The TPO gene is located on the chromosome 2p25. It comprises 17 exons, covers approximately 150 kb of genomic DNA and codes 933 amino acids. OBJECTIVES: In this study, we characterize the clinical and molecular basis of seven patients from four unrelated families with congenital hypothyroidism (CH) because of IOD. DESIGN AND METHODS: All patients underwent clinical, biochemical and imaging evaluation. The promoter and the complete coding regions of the human TPO along with the flanking intronic regions were analysed by single-strand conformation polymorphism analysis and direct DNA sequencing. Segregation analysis of mutations was carried out, and the effect of the novel missense identified mutations was investigated by 'in silico' studies. RESULTS: All subjects had congenital and persistent primary hypothyroidism. Three novel mutations: c.796C>T [p.Q266X], c.1784G>A [p.R595K] and c.2000G>A [p.G667D] and a previously reported mutation: c.1186_1187insGGCC [p.R396fsX472] have been identified. Four patients were compound heterozygous for p.R396fsX472/p.R595K mutations, two patients were homozygous for p.R595K, and the remaining patient was a compound heterozygous for p.Q266X/p.G667D. CONCLUSIONS: Our findings confirm the genetic heterogeneity of TPO defects and the importance of the implementation of molecular studies to determinate the aetiology of the CH with dyshormonogenesis.
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Hipotireoidismo Congênito/genética , Iodeto Peroxidase/genética , Adolescente , Criança , Análise Mutacional de DNA , Feminino , Humanos , Masculino , MutaçãoRESUMO
BACKGROUND: Turner syndrome (TS) patients usually have low bone mineral density (BMD) and increased risk of osteoporotic fractures. We have previously demonstrated an association of bb (BsmI polymorphic site) and ff (FokI polymorphic site) vitamin D receptor (VDR) genotypes with reduced BMD in TS patients. AIM: To analyze the relationship between VDR-Cdx2 polymorphism and BMD as well as bone metabolic variables in TS patients. METHODS: Fifty-five TS patients and 59 control women were studied. VDR-Cdx2 genotypes were determined using TaqMan probes in a real time thermocycler. Lumbar and femoral BMD were determined by dual-energy X-ray absorptiometry (DEXA) and serum intact parathyroid hormone, osteocalcin and beta3-CrossLaps were determined by electrochemiluminescence. RESULTS: Patients with genotype GG had higher levels of both osteocalcin and beta-CrossLaps as compared to patients with genotype GA (p < 0.01 and p < 0.05, respectively). CONCLUSION: Patients carrying genotype GG have higher levels of bone formation and resorption markers. This indicates a more active bone turnover that could impact on their future bone mineral density.
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Densidade Óssea/genética , Polimorfismo de Nucleotídeo Único , Receptores de Calcitriol/genética , Síndrome de Turner/genética , Absorciometria de Fóton , Adolescente , Adulto , Biomarcadores/análise , Biomarcadores/metabolismo , Estudos de Casos e Controles , Feminino , Fraturas Ósseas/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Genótipo , Humanos , Polimorfismo de Nucleotídeo Único/fisiologia , Regiões Promotoras Genéticas/genética , Síndrome de Turner/metabolismo , Síndrome de Turner/fisiopatologia , Adulto JovemRESUMO
Fungal infections have increased in recent decades with considerable morbidity and mortality, mainly in immunosuppressed or admitted-to-the-ICU patients. The fungal resistance to conventional antifungal treatments has become a public health problem, especially with Candida that presents resistance to several antifungals. Therefore, generating new alternatives of antifungal therapy is fundamental. One of these possibilities is the use of antimicrobial peptides, such as LL-37, which acts on the disruption of the microorganism membrane and promotes immunomodulatory effects in the host. In this study, we evaluated the in vitro antifungal activity of the LL-37 analogue peptides (AC-1, LL37-1, AC-2, and D) against different Candida spp. and clinical isolates obtained from patients with vulvovaginal candidiasis. Our results suggest that the peptides with the best ranges of MICs were LL37-1 and AC-2 (0.07 µM) against the strains studied. This inhibitory effect was confirmed by analyzing the yeast growth curves that evidenced a significant decrease in the fungal growth after exposure to LL-37 peptides. By the XTT technique we observed a significant reduction in the biofilm formation process when compared to yeasts untreated with the analogue peptides. In conclusion, we suggest that LL-37 analogue peptides may play an important antimicrobial role against Candida spp.
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Background: Congenital iodide transport defect (ITD) is an uncommon cause of dyshormonogenic congenital hypothyroidism characterized by the absence of active iodide accumulation in the thyroid gland. ITD is an autosomal recessive disorder caused by loss-of-function variants in the sodium/iodide symporter (NIS)-coding SLC5A5 gene. Objective: We aimed to identify, and if so to functionally characterize, novel ITD-causing SLC5A5 gene variants in a cohort of five unrelated pediatric patients diagnosed with dyshormonogenic congenital hypothyroidism with minimal to absent 99mTc-pertechnetate accumulation in the thyroid gland. Methods: The coding region of the SLC5A5 gene was sequenced using Sanger sequencing. In silico analysis and functional in vitro characterization of a novel synonymous variant were performed. Results: Sanger sequencing revealed a novel homozygous synonymous SLC5A5 gene variant (c.1326A>C in exon 11). In silico analysis revealed that the c.1326A>C variant is potentially deleterious for NIS pre-mRNA splicing. The c.1326A>C variant was predicted to lie within a putative exonic splicing enhancer reducing the binding of splicing regulatory trans-acting protein SRSF5. Splicing minigene reporter assay revealed that c.1326A>C causes exon 11 or exon 11 and 12 skipping during NIS pre-mRNA splicing leading to the NIS pathogenic variants p.G415_P443del and p.G415Lfs*32, respectively. Significantly, the frameshift variant p.G415Lfs*32 is predicted to be subjected to degradation by nonsense-mediated decay. Conclusions: We identified the first exonic synonymous SLC5A5 gene variant causing aberrant NIS pre-mRNA splicing, thus expanding the mutational landscape of the SLC5A5 gene leading to dyshormonogenic congenital hypothyroidism.
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Hipotireoidismo Congênito , Simportadores , Criança , Hipotireoidismo Congênito/genética , Éxons , Humanos , Iodetos/metabolismo , Precursores de RNA , Simportadores/genéticaRESUMO
OBJECTIVE: Idiopathic short stature (ISS) describes short children with normal GH secretion. Although GH treatment increases their heights, growth response to the therapy differs among patients. Thyroid hormones (TH) are essential for longitudinal growth acting mainly through TH receptors (TR) α and ß. We have previously reported that GH treatment reduced peripheral TH action in Turner Syndrome by TR down-regulation. The aims of the study were to assess the effect of GH treatment to ISS on peripheral TH action and the correlation between thyroid status and growth response to the therapy. SUBJECTS, DESIGN AND MEASUREMENTS: Eighteen normal (control) and twenty-five ISS children were enrolled and evaluated before and after 12 months of life time (control) or 12 months of GH therapy (ISS). Fasting blood was used for serum biochemical evaluations, peripheral blood mononuclear cells for TR mRNA determination by QRT-PCR and growth parameters by standard methods. RESULTS: GH treatment modified neither TR mRNA levels nor serum markers of TH action in ISS evaluated as a whole group. However, the individual change in TRß mRNA levels correlated to the change in sex hormone-binding globulin (SHBG) levels after GH therapy. The growth response to GH correlated positively with the change in TRα mRNA level and negatively with that in TRß mRNA, TSH and SHBG levels. The change in each TR mRNA isoform after GH treatment correlated negatively with its own basal level. CONCLUSIONS: GH therapy induced individual changes in TR expression in ISS that correlated with their growth response. The basal TR mRNA level could predetermine the change in TR expression and therefore the sensitivity to GH treatment.
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Transtornos do Crescimento/sangue , Transtornos do Crescimento/tratamento farmacológico , Hormônio do Crescimento Humano/uso terapêutico , Criança , Transtornos do Crescimento/genética , Humanos , Imunoensaio/métodos , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/sangue , Fator de Crescimento Insulin-Like I/metabolismo , Osteocalcina/sangue , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Globulina de Ligação a Hormônio Sexual/metabolismo , Receptores beta dos Hormônios Tireóideos/genética , Hormônios Tireóideos/sangue , Tireotropina/sangue , Fatores de Tempo , Resultado do TratamentoRESUMO
BACKGROUND: Turner syndrome (TS) patients present low bone mineral density (BMD) and increased fracture risk, probably due to a genetic defect aggravated by hormonal deficiency. AIM: To study the relationship between vitamin D receptor (VDR) gene polymorphisms and BMD and bone parameters in TS patients. METHODS: DNA from 65 TS patients and 110 controls was amplified by PCR and digested with FokI, BsmI and ApaI restrictases. Lumbar and femoral BMD were determined by DEXA and serum intact parathyroid hormone, osteocalcin and beta-CrossLaps by electrochemiluminescence. RESULTS: Genotype distribution within the ApaI site was different in both groups: genotype Aa was more abundant in TS (63.8% vs. 41.3%; p<0.01), whereas AA predominated in controls (33.9% vs. 15.5%; p<0.01). Patients carrying genotype bb (BsmI) or ff (FokI) had lower BMD than those with other genotypes (p<0.01 and p<0.05, respectively). CONCLUSION: BsmI and FokI polymorphic sites of VDR could be genetic determinants of BMD in TS patients.
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Densidade Óssea/genética , Receptores de Calcitriol/genética , Síndrome de Turner/genética , Síndrome de Turner/metabolismo , Adolescente , Alelos , Sequência de Bases , Densidade Óssea/efeitos dos fármacos , Doenças Ósseas Metabólicas/etiologia , Doenças Ósseas Metabólicas/genética , Doenças Ósseas Metabólicas/metabolismo , Estudos de Casos e Controles , Primers do DNA/genética , Estradiol/uso terapêutico , Feminino , Frequência do Gene , Estudos de Associação Genética , Hormônio do Crescimento Humano/uso terapêutico , Humanos , Cariotipagem , Mosaicismo , Polimorfismo de Fragmento de Restrição , Proteínas Recombinantes/uso terapêutico , Síndrome de Turner/complicações , Síndrome de Turner/tratamento farmacológicoRESUMO
Clinical, biochemical and genetic analysis related to bone mineral density (BMD) were carried out in children born small for gestational age (SGA) that failed to achieve postnatal catch-up growth (CUG), SGA children that completed CUG and adequate for gestational age (AGA) children. Serum IGF-I, IGF-II, IGF binding protein-3 and acid-labile subunit were lower in the SGA-CUG children as compared with the other groups. Frequencies of polymorphic variants of vitamin D receptor, estrogen receptor and collagen genes were similar among groups. The genotype 194-192 of the IGF-I gene was higher in the SGA-CUG and 196-192 was higher in the SGA+CUG group. In the SGA-CUG group, the genotype SS of the COLIA1 gene was associated with lower BMD. Therefore, IGF system and COLIA1 polymorphism distinguish prepubertal SGA-CUG children from the SGA+CUG children of the same age. Furthermore, COLIA1 polymorphism could be useful to predict osteopenia in SGA-CUG children.
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Densidade Óssea , Doenças Ósseas Metabólicas/diagnóstico , Doenças Ósseas Metabólicas/genética , Desenvolvimento Infantil , Transtornos do Crescimento/sangue , Transtornos do Crescimento/genética , Recém-Nascido Pequeno para a Idade Gestacional , Polimorfismo Genético , Doenças Ósseas Metabólicas/etiologia , Proteínas de Transporte/sangue , Criança , Pré-Escolar , Colágeno Tipo I/genética , Cadeia alfa 1 do Colágeno Tipo I , Diagnóstico Precoce , Feminino , Estudos de Associação Genética , Glicoproteínas/sangue , Transtornos do Crescimento/fisiopatologia , Humanos , Recém-Nascido , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/sangue , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like II/análise , MasculinoRESUMO
The E2F transcription factor family is traditionally associated with cell cycle control. However, recent data has shown that activating E2Fs (E2F1-3a) are potent activators of apoptosis. In contrast, the recently cloned inhibitory E2Fs (E2F7 and 8) appear to antagonize E2F-induced cell death. In this review we will discuss (i) the potential role of E2Fs in UV-induced cell death and (ii) the implications of this to the development of UV-induced cutaneous malignancies.
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Apoptose , Fatores de Transcrição E2F/metabolismo , Neoplasias Cutâneas/metabolismo , Raios Ultravioleta/efeitos adversos , Animais , Fatores de Transcrição E2F/química , Fatores de Transcrição E2F/genética , Humanos , Queratinócitos/metabolismo , Queratinócitos/patologia , Queratinócitos/fisiologia , Neoplasias Cutâneas/etiologiaRESUMO
CONTEXT: Iodide transport defect (ITD) (Online Mendelian Inheritance in Man No. 274400) is an uncommon cause of dyshormonogenic congenital hypothyroidism due to loss-of-function variants in the SLC5A5 gene, which encodes the sodium/iodide symporter (NIS), causing deficient iodide accumulation in thyroid follicular cells. OBJECTIVE: This work aims to determine the molecular basis of a patient's ITD clinical phenotype. METHODS: The propositus was diagnosed with dyshormonogenic congenital hypothyroidism with minimal 99mTc-pertechnetate accumulation in a eutopic thyroid gland. The propositus SLC5A5 gene was sequenced. Functional in vitro characterization of the novel NIS variant was performed. RESULTS: Sanger sequencing revealed a novel homozygous missense p.G561E NIS variant. Mechanistically, the G561E substitution reduces iodide uptake, because targeting of G561E NIS to the plasma membrane is reduced. Biochemical analyses revealed that G561E impairs the recognition of an adjacent tryptophan-acidic motif by the kinesin-1 subunit kinesin light chain 2 (KLC2), interfering with NIS maturation beyond the endoplasmic reticulum, and reducing iodide accumulation. Structural bioinformatic analysis suggests that G561E shifts the equilibrium of the unstructured tryptophan-acidic motif toward a more structured conformation unrecognizable to KLC2. Consistently, knockdown of Klc2 causes defective NIS maturation and consequently decreases iodide accumulation in rat thyroid cells. Morpholino knockdown of klc2 reduces thyroid hormone synthesis in zebrafish larvae leading to a hypothyroid state as revealed by expression profiling of key genes related to the hypothalamic-pituitary-thyroid axis. CONCLUSION: We report a novel NIS pathogenic variant associated with dyshormonogenic congenital hypothyroidism. Detailed molecular characterization of G561E NIS uncovered the significance of KLC2 in thyroid physiology.
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Hipotireoidismo Congênito/genética , Erros Inatos do Metabolismo/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Simportadores/genética , Hormônios Tireóideos/metabolismo , Animais , Humanos , Recém-Nascido , Iodetos/metabolismo , Cinesinas , Masculino , Mutação de Sentido Incorreto , Fenótipo , Ratos , Glândula Tireoide/metabolismoRESUMO
Tumor initiation (TI) in xenotransplantation models of head and neck squamous cell carcinoma (HNSCC) is an inefficient process. Poor TI could be due to (1) posttransplant cell loss, (2) a rare sub-population of cancer stem cells or (3) a requirement for specific cellular interactions, which rely on cell number. By tracking GFP-expressing HNSCC cells, we conclude that the posttransplant loss of cancer cells is minimal in the xenotransplant model. Furthermore, an examination of putative cancer stem cell markers (such as CD133, CD44, SP and label retention) in HNSCC cell lines revealed no correlation between marker expression and tumorigenicity. In addition, single-cell clones randomly isolated from HNSCC cell lines and then transplanted into mice were all capable of initiating tumors with efficiencies varying almost 34-fold. As the observed variation in the clones was both more and less tumorigenic than the parental cells, a combination of two clones, at suboptimal cell numbers for TI, was implanted into mice and was found to modulate the tumor-initiating activity, thus indicating that TI is dependent on a 'critical' number of cells and, for the first time, that interactions between clonal variants within tumors can modulate the overall tumor-initiating activity. Put in context with previous literature on tumorigenic activity, we believe that interactions between clonal variants within a tumor as well as (1) stromal interactions, (2) angiogenic activity, (3) immunocompetence and (4) cancer stem cells may all contribute to tumorigenic potential and the propensity for tumor growth and recurrence.
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Carcinoma de Células Escamosas/patologia , Neoplasias de Cabeça e Pescoço/patologia , Antígeno AC133 , Animais , Antígenos CD/análise , Carcinoma de Células Escamosas/etiologia , Moléculas de Adesão Celular/análise , Linhagem Celular Tumoral , Proteínas Ligadas por GPI , Glicoproteínas/análise , Neoplasias de Cabeça e Pescoço/etiologia , Humanos , Receptores de Hialuronatos/análise , Camundongos , Camundongos SCID , Peptídeos/análiseRESUMO
Children born small for gestational age (SGA) are prone to developing obesity, insulin resistance and type 2 diabetes. Adiponectin and leptin are adipocytokines associated with insulin sensitivity parameters. We aimed to relate serum adiponectin and leptin levels with insulin sensitivity parameters in prepuberal SGA children with and without catch-up growth (SGA+CUG; SGA-CUG, respectively) and to analyze the usefulness of these adipocytokines as early markers of insulin resistance. We analysed adiponectin, proinsulin, leptin, growth factors, insulin, HOMA IR and HOMA beta(cell) in 23 SGA+CUG, 26 SGA-CUG children compared with 48 prepuberal appropiate for gestational age (AGA). SGA children had adiponectin levels comparable to AGA children. Leptin levels were different between sexes, showed to be higher in SGA+CUG group (p=0.040) and these were significantly correlated with insulin sensitivity parameters. These results suggest leptin resistance as an adaptive mechanism to increase energy balance, but an altered functional response of adipocytes cannot be discarded.
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Adiponectina/sangue , Recém-Nascido Pequeno para a Idade Gestacional/sangue , Resistência à Insulina/fisiologia , Leptina/sangue , Puberdade/sangue , Glicemia/análise , Criança , Feminino , Homeostase , Humanos , Recém-Nascido , Recém-Nascido Pequeno para a Idade Gestacional/crescimento & desenvolvimento , Insulina/sangue , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Masculino , Proinsulina/sangue , Puberdade/fisiologiaRESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
RESUMO
Macrocystis pyrifera and Lessonia spicata are economically and ecologically relevant brown seaweeds that recently have been classified as members of two separated families within Laminariales (kelps). Here we describe for the first time the Macrocystis pyrifera x Lessonia spicata hybridization in the wild (Chiloe Island, Southeastern Pacific), where populations of the two parents exist sympatrically. Externally, this hybrid exhibited typical features of its parents M. pyrifera (cylindrical and flexible distal stipes, serrate frond margins and presence of sporophylls) and L. spicata (rigid and flat main stipe and first bifurcation), as well as intermediate features between them (thick unfused haptera in the holdfast). Histological sections revealed the prevalence of mucilage ducts within stipes and fronds (absent in Lessonia) and fully developed unilocular sporangia in the sporophylls. Molecular analyses confirmed the presence of the two parental genotypes for ITS1 nrDNA and the M. pyrifera genotype for two predominantly maternally inherited cytoplasmic markers (COI and rbcLS spacer) in the tissue of the hybrid. A metabolome-wide approach revealed that this hybrid is more chemically reminiscent to M. pyrifera. Nevertheless, several hits were identified as Lessonia exclusive or more remarkably, not present in any of the parent. Meiospores developed into apparently fertile gametophytes, which gave rise to F1 sporophytes that reached several millimeters before suddenly dying. In-vitro reciprocal crossing of Mar Brava gametophytes from both species revealed that although it is rare, interfamilial hybridization between the two species is possible but mostly overcome by pseudogamy of female gametophytes.
Assuntos
Técnicas de Genotipagem/métodos , Laminaria/fisiologia , Macrocystis/fisiologia , Metabolômica/métodos , DNA de Algas/genética , Genótipo , Hibridização Genética , Melhoramento Vegetal , Esporângios/fisiologia , SimpatriaRESUMO
Osteosarcoma is the most common paediatric primary bone malignancy. The major cause of death in osteosarcoma is drug-resistant pulmonary metastasis. Previous studies have shown that thioredoxin reductase 2 is a driver of metastasis in osteosarcoma and can be inhibited by auranofin (AF). Moreover, studies have shown that AF significantly reduces pulmonary metastases in xenotransplant models. Here, we describe a phase I/II study of AF in canine osteosarcoma, a well-recognized spontaneous model of human osteosarcoma. We performed a single-arm multicentre pilot study of AF in combination with standard of care (SOC) (amputation + carboplatin). We recruited 40 dogs to the trial and used a historical SOC-only control group (n = 26). Dogs >15 kg received 9 mg AF q3d PO and dogs <15 kg received 6 mg q3d. Follow-up occurred over at least a 3-year period. Auranofin plus SOC improved overall survival (OS) (P = .036) in all dogs treated. The improved outcome was attributable entirely to improved OS in male dogs (P = .009). At the time of writing, 10 dogs (25%) survive without measurable disease in the treatment group with survival times ranging between 806 and 1525 days. Our study shows that AF improves OS in male dogs when combined with SOC. Our findings have translational relevance for the management of canine and human osteosarcoma. Our data justify a larger multicentre phase 2 trial in dogs and a phase I/II trial in human patients with refractory disease at the time of initial surgery.