ABSTRACT
Pubertal gynecomastia is a common condition observed in up to 65% of adolescent males. It is usually idiopathic and tends to regress within 1-2 years. In this descriptive cross-sectional study, we investigated 25 adolescent males with prominent (>B3) and/or persistent (>2 years) pubertal gynecomastia (P/PPG) to determine whether a hormonal/genetic defect might underline this condition. Endocrine investigation revealed the absence of hormonal disturbance for 18 boys (72%). Three patients presented Klinefelter syndrome and three a partial androgen insensitivity syndrome (PAIS) as a result of p.Ala646Asp and p.Ala45Gly mutations of the androgen receptor gene. The last patient showed a 17α-hydroxylase/17,20-lyase deficiency as a result of a compound heterozygous mutation of the CYP17A1 gene leading to p.Pro35Thr(P35T) and p.Arg239Stop(R239X) in the P450c17 protein. Enzymatic activity was analyzed: the mutant protein bearing the premature stop codon R239X showed a complete loss of 17α-hydroxylase and 17,20-lyase activity. The mutant P35T seemed to retain 15-20% of 17α-hydroxylase and about 8-10% of 17,20-lyase activity. This work demonstrates that P/PPG had an endocrine/genetic cause in 28% of our cases. PAIS may be expressed only by isolated gynecomastia as well as by 17α-hydroxylase/17,20-lyase deficiency. Isolated P/PPG is not always a 'physiological' condition and should thus be investigated through adequate endocrine and genetic investigations, even though larger studies are needed to better determine the real prevalence of genetic defects in such patients.
Subject(s)
Gynecomastia/genetics , Adolescent , Androgen-Insensitivity Syndrome/genetics , Androgen-Insensitivity Syndrome/metabolism , Cohort Studies , Cross-Sectional Studies , Gynecomastia/metabolism , Hormones/metabolism , Humans , Male , Mutation , Receptors, Androgen/genetics , Steroid 17-alpha-Hydroxylase/genetics , TranscriptomeABSTRACT
Primary emissions from a log wood burner and a pellet boiler were characterized by online measurements of the organic aerosol (OA) using a high-resolution time-of-flight aerosol mass spectrometer (HR-TOF-AMS) and of black carbon (BC). The OA and BC concentrations measured during the burning cycle of the log wood burner, batch wise fueled with wood logs, were highly variable and generally dominated by BC. The emissions of the pellet burner had, besides inorganic material, a high fraction of OA and a minor contribution of BC. However, during artificially induced poor burning BC was the dominating species with â¼80% of the measured mass. The elemental O:C ratio of the OA was generally found in the range of 0.2-0.5 during the startup phase or after reloading of the log wood burner. During the burnout or smoldering phase, O:C ratios increased up to 1.6-1.7, which is similar to the ratios found for the pellet boiler during stable burning conditions and higher than the O:C ratios observed for highly aged ambient OA. The organic emissions of both burners have a very similar H:C ratio at a given O:C ratio and therefore fall on the same line in the Van Krevelen diagram.
Subject(s)
Air Pollutants/analysis , Cooking/instrumentation , Particulate Matter/analysis , Soot/analysis , Air Pollution/statistics & numerical data , Biomass , Cooking/statistics & numerical data , Environmental Monitoring , WoodABSTRACT
This study reports the potential toxicological impact of particles produced during biomass combustion by an automatic pellet boiler and a traditional logwood stove under various combustion conditions using a novel profluorescent nitroxide probe, BPEAnit. This probe is weakly fluorescent but yields strong fluorescence emission upon radical trapping or redox activity. Samples were collected by bubbling aerosol through an impinger containing BPEAnit solution, followed by fluorescence measurement. The fluorescence of BPEAnit was measured for particles produced during various combustion phases: at the beginning of burning (cold start), stable combustion after refilling with the fuel (warm start), and poor burning conditions. For particles produced by the logwood stove under cold-start conditions, significantly higher amounts of reactive species per unit of particulate mass were observed compared to emissions produced during a warm start. In addition, sampling of logwood burning emissions after passing through a thermodenuder at 250 degrees C resulted in an 80-100% reduction of the fluorescence signal of the BPEAnit probe, indicating that the majority of reactive species were semivolatile. Moreover, the amount of reactive species showed a strong correlation with the amount of particulate organic material. This indicates the importance of semivolatile organics in particle-related toxicity. Particle emissions from the pellet boiler, although of similar mass concentration, were not observed to lead to an increase in fluorescence signal during any of the combustion phases.
Subject(s)
Fires , Fluorescent Dyes/chemistry , Nitrogen Oxides/chemistry , Particulate Matter/chemistry , Wood/metabolism , Molecular Weight , Oxidation-Reduction , Reactive Oxygen Species/metabolism , Spectrometry, Fluorescence , TemperatureABSTRACT
In the biosynthesis of steroid hormones the neutral lipid cholesterol, a normal constituent of lipid bilayers is transformed via a series of hydroxylation, oxidation, and reduction steps into a vast array of biologically active compounds: mineralocorticoids, glucocorticoids, and sex hormones. Glucocorticoids regulate many aspects of metabolism and immune function, whereas mineralocorticoids help maintain blood volume and control renal excretion of electrolytes. Sex hormones are essential for sex differentiation in male and support reproduction. They include androgens, estrogens, and progestins. A block in the pathway of steroid biosynthesis leads to the lack of hormones downstream and accumulation of the upstream compounds that can activate other members of the steroid receptor family. This review deals with the clinical consequences of these blocks.
Subject(s)
Gonadal Steroid Hormones/biosynthesis , Gonadal Steroid Hormones/deficiency , Steroids/biosynthesis , Animals , Cholesterol/metabolism , Disorders of Sex Development/etiology , Endocrine System Diseases/etiology , Endocrine System Diseases/genetics , Endocrine System Diseases/metabolism , Female , Gonadal Steroid Hormones/genetics , Humans , Lipid Metabolism/genetics , Male , Models, Biological , Water-Electrolyte Imbalance/etiologyABSTRACT
BACKGROUND: P450c17 has two distinct activities: 17alpha-hydroxylase activity and 17,20-lyase activity. Combined 17alpha-hydroxylase/17,20-lyase deficiency leads to a severe defect in the production of cortisol and sex steroids. In affected males this results in impaired masculinization with ambiguous or female external genitalia. Female patients have normal genitalia but show a lack of pubertal development in adolescence. An increased production of mineralocorticoids often leads to hypertension and hypokalemia in both sexes. METHODS: To better understand the mechanisms of P450c17 deficiency, we studied 2 patients (both 46,XY) with combined 17alpha-hydroxylase/17,20-lyase deficiency of different severity: one with complete lack of masculinization and one with ambiguous genitalia. RESULTS: Four mutations were identified by sequencing of the CYP17A1 gene: I332T and A355T in the less severely affected patient; G111S and R440H in the patient with complete lack of masculinization. The three novel mutations were expressed in COS1 cells and all mutant proteins except I332T showed a complete loss of both enzymatic activities. I332T retained some residual 17alpha-hydroxylase as well as 17,20-lyase activity. CONCLUSION: We identified 2 patients with the phenotypical spectrum of P450c17 deficiency. Three novel mutations in the CYP17A1 gene were identified and their functional characterization provided a good phenotype-genotype correlation. The location of the mutated residues in the three-dimensional model of P450c17 gave some additional insights into its structure-function relationship.
Subject(s)
Adrenal Hyperplasia, Congenital/genetics , Gonadal Dysgenesis, 46,XY/genetics , Steroid 17-alpha-Hydroxylase/genetics , Animals , COS Cells , Chlorocebus aethiops , Female , Genetic Association Studies , Humans , Infant , Male , Mutation , Steroid 17-alpha-Hydroxylase/metabolismABSTRACT
Although factors involved in male sexual differentiation have been well studied, the pathways regulating female sexual differentiation remain incompletely defined. To date, no genes have been identified to play a similar role in ovarian development as was shown for the SRY or SOX9 genes in testicular development. In mice, Wnt4 regulates the development of the female reproductive tract, antagonizes the production of testosterone, and is important for oocyte development. The recent demonstration of heterozygous WNT4 defects in patients with Mullerian agenesis and signs of ovarian hyperandrogenism added WNT4 to the growing list of genes such as SRY, SOX9, WT1, DAX1, and SF-1 contributing to human sexual development. In particular, WNT4 was the first human gene to be identified to direct development of the bipotential gonad towards ovaries. From a more clinical point of view, it seems that the absence of a uterus (and not other Müllerian abnormalities) and the androgen excess are the pathognomonic signs of WNT4 defects, suggesting that WNT4 deficiency might be a clinical entity distinct from the typical Mayer-Rokitansky-Kuster-Hauser syndrome.
Subject(s)
Sexual Development/genetics , Wnt Proteins/genetics , Animals , Female , Humans , Mice , Mullerian Ducts/abnormalities , Mullerian Ducts/metabolism , Ovary/abnormalities , Ovary/metabolism , Wnt4 ProteinABSTRACT
The pathways leading to female sexual determination in mammals are incompletely defined. Loss-of-function mutations in the WNT4 gene appear to cause developmental abnormalities of sexual differentiation in women and mice. We recruited six patients with different degrees of Müllerian abnormalities, with or without renal aberrations and a normal female 46,XX karyotype. A clear androgen excess was found only in one patient. This 19-year-old woman was affected by primary amenorrhoea, absence of Müllerian ducts derivatives, clinical (acne and hirsutism) and biochemical (repeatedly high levels of testosterone) signs of androgen excess. Direct sequencing of her WNT4 gene followed by functional studies in human ovarian cells (OVCAR3) was performed. This patient carried the novel R83C loss-of-function dominant negative mutation in her WNT4, confirming the role of WNT4 in the development and maintenance of the female phenotype in women. Our study can also help refine the phenotype of WNT4 deficiency in humans. In fact, it appears that at least in this limited casuistic small group of patients, the absence of a uterus (and not other Müllerian abnormalities) and the androgen excess are the pathognomonic signs of WNT4 defects, suggesting that this might be a clinical entity distinct from the classic Mayer-Rokitansky-Kuster-Hauser syndrome.
Subject(s)
Abnormalities, Multiple/diagnosis , Proto-Oncogene Proteins/deficiency , Wnt Proteins/deficiency , Adolescent , Adult , Amino Acid Sequence , Amino Acid Substitution , Child , Child, Preschool , Female , Gonadal Dysgenesis/diagnosis , Humans , Infant , Kidney/abnormalities , Molecular Sequence Data , Mullerian Ducts/abnormalities , Ovary/abnormalities , Phenotype , Proto-Oncogene Proteins/genetics , Sequence Alignment , Syndrome , Tumor Cells, Cultured , Uterus/abnormalities , Wnt Proteins/genetics , Wnt4 ProteinABSTRACT
CONTEXT: The characteristics of P450c17 deficiency include 46,XY disorder of sex development, hypertension, hypokalemia, and lack of pubertal development. OBJECTIVE: To better understand this rare enzymatic deficiency, we analyzed the CYP17A1 gene in six affected patients. DESIGN AND PATIENTS: We examined six patients, five 46,XY, and one 46,XX (age 9-29 yr) with complete lack of masculinization (female infantile external genitalia, no uterus) and delayed puberty, respectively, and different degrees of hypertension. MAIN OUTCOME MEASUREMENTS: Genotype-phenotype correlation was measured. RESULTS: Four homozygote mutations were identified by direct sequencing of the CYP17A1 gene corresponding to an alanin 302-proline (A302P) exchange; the loss of lysine 327 (K327del); the deletion of glutamate 331 (E331del); and the replacement of arginine 416 with a histidine (R416H). Both P450c17 activities were abolished in all the mutant proteins, except one, when expressed in COS1 cells. The E331del-mutated P450c17 retained 17alpha-hydroxylase activity. The mutant proteins were normally expressed, suggesting that the loss of enzymatic activity is not due to defects of synthesis, stability, or localization of P450c17 proteins. CONCLUSION: These studies confirm lack of masculinization in 46,XY individuals as the pathognomic sign of the complete P450c17 deficiency. In XX individuals P450c17 deficiency should be considered in cases of delayed puberty. Age of onset and the severity of hypertension do not seem to be constant. Careful examination of long-term follow-ups in two of our patients suggested to us that estrogen treatment in P450c17-deficient patients might worsen the enzymatic defect, leading to aggravation of the hypertension.
Subject(s)
Gonadal Dysgenesis, 46,XX/genetics , Gonadal Dysgenesis, 46,XY/genetics , Steroid 17-alpha-Hydroxylase/genetics , Adolescent , Adult , Animals , Blood Pressure , COS Cells , Child , Chlorocebus aethiops , DNA Mutational Analysis , Genes, Reporter , Gonadal Dysgenesis, 46,XX/diagnostic imaging , Gonadal Dysgenesis, 46,XX/physiopathology , Gonadal Dysgenesis, 46,XY/diagnostic imaging , Gonadal Dysgenesis, 46,XY/physiopathology , Humans , Mutation , Progesterone/metabolism , Radiography , Steroid 17-alpha-Hydroxylase/metabolism , TransfectionABSTRACT
UNLABELLED: Anterior pituitary agenesis (APA) has very rarely been reported. Therefore, its phenotypical and genotypical features are not well known. The aim of this study was to ascertain whether the clinical picture in 4 subjects with APA and multiple pituitary hormone deficiencies (MPHD) was different compared to the one observed in a selected control group consisting of 7 MPHD individuals with hypoplastic (and not aplastic) adenohypophysis and pituitary stalk interruption syndrome. Another goal was to investigate genetic basis of APA by analyzing for the first time in this condition many of the transcriptional factors which are required for both structural development and cellular differentiation of hypophysis. Age at diagnosis was significantly lower in APA children than in controls (1.5+/-2.3 vs 11.1+/-7.6 yr, p<0.0005). Microphallus and neonatal cholestasis were observed only in APA subjects (chi-squared=4.3, p<0.05) and also neonatal hypoglycemia was more frequent in APA patients than in controls (X2=4.05, p<0.05). Molecular analyses of the genes of the transcriptional factors POU1F1, PROP1, LHX3, LHX4, ISL1 and HESX1 detected no mutations in APA patients. CONCLUSIONS: a) if compared with a selected cohort of MPHD patients with both adenohypophysis hypoplasia and pituitary stalk interruption syndrome, the ones with APA show an earlier and more severe picture of hypopituitarism; b) mutations in several transcription factors that are known to be essential for the development of Rathke's pouch are not necessarily found in humans with APA.
Subject(s)
Hypopituitarism/etiology , Pituitary Gland, Anterior/abnormalities , Pituitary Gland, Anterior/embryology , Transcription Factors/genetics , Child, Preschool , Female , Homeodomain Proteins/genetics , Humans , Hypopituitarism/diagnosis , Hypopituitarism/genetics , Infant , Infant, Newborn , LIM-Homeodomain Proteins , Magnetic Resonance Imaging , Male , Mutation , Pituitary Gland/abnormalities , Pituitary Hormones/deficiency , Retrospective Studies , Transcription Factor Pit-1/geneticsABSTRACT
AIMS/HYPOTHESIS: Loss of pancreatic beta cells is the crucial event in the development of type 1 diabetes. It is the result of an imbalance between autoimmune destruction and insufficient regeneration of islet cells. To study the role of islet cell regeneration in the pathogenesis of type 1 diabetes, we focused on PAX4, a paired homeodomain transcriptional repressor that is involved in islet cell growth. METHODS: The study included 379 diabetic children and 1,070 controls from two distinct populations, and a cohort of children who had not developed type 1 diabetes, despite the presence of islet cell antibodies. Genomic DNA analysis of PAX4 was carried out via direct sequencing of PCR-amplified fragments and allelic discrimination. We compared the transrepression potential of the PAX4 variants in betaTC3 cells and analysed their influence on beta cell growth. RESULTS: The type 1 diabetic subjects are different from the normal individuals in terms of the genotype distribution of the A1168C single nucleotide polymorphism in PAX4. The C/C genotype is frequent among type 1 diabetic children (73%) and rare among the control population (32%). Conversely, the A/C genotype is prevalent among control subjects (62%) and antibody-positive children without type 1 diabetes (73.6%), but uncommon among subjects with type 1 diabetes (17.5%). The combination of PAX4A and PAX4C is functionally more active than PAX4C alone (the "diabetic" variant). Beta cells expressing PAX4A and PAX4C efficiently proliferate when stimulated with glucose, whereas cells expressing the PAX4C variant alone do not. CONCLUSIONS/INTERPRETATION: We have identified a link between beta cell regenerative capacity and susceptibility to type 1 diabetes. This finding could explain the fact that not all of the individuals who develop autoimmunity against beta cells actually contract the disease. The C/C genotype of the A1168C polymorphism in PAX4 can be viewed as a predisposition marker that can help to detect individuals prone to develop type 1 diabetes.
Subject(s)
Diabetes Mellitus, Type 1/genetics , Genetic Variation , Homeodomain Proteins/genetics , Polymorphism, Single Nucleotide , Transcription Factors/genetics , Amino Acid Substitution , Animals , Binding Sites , Blood Glucose/metabolism , Child , DNA/blood , DNA/genetics , DNA/isolation & purification , Diabetes Mellitus, Type 1/blood , Female , Gene Frequency , Genetic Markers , Homeodomain Proteins/metabolism , Humans , Insulin/genetics , Male , Mutation, Missense , Paired Box Transcription Factors , Promoter Regions, Genetic , Reference Values , Switzerland , Transcription Factors/metabolismABSTRACT
OBJECTIVE: in diabetic patients with critical limb ischaemia (CLI) an inferior success rate following infrainguinal bypass surgery is quite often suggested. The aim of this retrospective analysis was, therefore, to evaluate the graft patency and, particularly, the clinical outcome at 1 year in diabetic compared with non-diabetic patients. MATERIAL AND METHODS: two hundred and eleven patients (diabetics 94; non-diabetics 117) with femorodistal reconstruction for CLI were studied. Groups were comparable with regard to the Fontaine classification, the distribution of vascular risk factors, graft material, distal anastomosis site, and the angiographic runoff grading. RESULTS: diabetes did not adversely affect graft function. For diabetics and non-diabetics primary cumulative patency rate at 1 year was found to be 66 and 56%, respectively (p=0.10) and a virtually identical limb salvage rate of 85 and 83% was achieved (p=0.76). With regard to healing of ischaemic foot ulcers a trend against diabetics was noted with a healing rate of 81% compared to 96% in non-diabetics at 1 year (p=0.067); gangrenous foot lesions could be equally remedied in 94% and in 87% among patients with and without diabetes (p=0.44). The survival rate of diabetics, however, was significantly lower with 78% at 1 year compared with 95% in non-diabetic patients (p=0.0004). CONCLUSIONS: our preliminary results support the view that infrainguinal bypass grafting can be safely done even in diabetics. Despite increased mortality in this group, liberal indication for reconstructive vascular surgery seems to be justified by favourable patency rates and clinical outcome in selected patients.
Subject(s)
Arterial Occlusive Diseases/surgery , Blood Vessel Prosthesis Implantation , Diabetic Angiopathies/surgery , Femoral Artery/surgery , Limb Salvage/methods , Vascular Patency , Aged , Anastomosis, Surgical/methods , Arterial Occlusive Diseases/mortality , Blood Vessel Prosthesis , Diabetic Angiopathies/mortality , Female , Foot Ulcer/surgery , Graft Survival , Humans , Male , Middle Aged , Postoperative Complications , Retrospective Studies , Risk Factors , Survival Rate , Treatment OutcomeABSTRACT
AIMS/HYPOTHESIS: In this nationwide prospective study we wanted to verify the trend of increasing diabetes incidence data from our earlier retrospective analysis of the military registry of Swiss men. SUBJECTS AND METHODS: The data collection of newly diagnosed children in Switzerland at an age younger than 15 years started in 1991. The countrywide survey used a small questionnaire which was sent back to the study centre. The questionnaire was anonymous and contained: hospital of diagnosis, initials, sex, birth date, date of diagnosis, residence, country of citizenship, and responsible physician. General data on the population were taken from publications of the Swiss Federal Statistical Office. RESULTS: A total of 941 children below the age of 15 years with newly diagnosed Type I (insulin-dependent) diabetes mellitus were collected (434 girls, 507 boys). The incidence in children aged 0 to 14 years rose significantly between 1991 and 1999 with a yearly average increase of 5.1%. In the age group 0 to 4 years a more than four-fold increase in incidence from 2.4/100,000 per year to 10.5/100,000 per year (p = 0.0002) was recorded, whereas the age-specific incidence in the 5 to 9-year-old and 10 to 14-year-old children did not change during the data collection period. The incidence was significantly higher in boys than in girls, whereas no difference was found between rural and urban populations. CONCLUSION/INTERPRETATION: The incidence of Type I diabetes is rising in children living in Switzerland but only the youngest age group of under 5 years of age is affected showing a large annual average increase of 23.8%.
Subject(s)
Diabetes Mellitus, Type 1/epidemiology , Adolescent , Age Distribution , Child , Child, Preschool , Ethnicity , Female , Humans , Incidence , Infant , Male , Sex Factors , Surveys and Questionnaires , Switzerland/epidemiologyABSTRACT
UNLABELLED: Persistent müllerian duct syndrome is a relatively rare inherited defect of sexual differentiation characterised by failure of regression of the müllerian ducts in males. In affected individuals, uterus and tubes are present because of defects of synthesis or action of anti-müllerian hormone (AMH), normally produced by the Sertoli cells of the testis. Patients are normally virilised, although mono- or bilateral cryptorchidism may be present. We observed two brothers (chromosomes 46 XY), aged 11 years and 2 months and 8 years and 3 months respectively, with bilateral cryptorchidism. The diagnosis of persistent müllerian duct syndrome was made on the basis of laparoscopic evidence of uterus and tubes, undetectable plasma levels of AMH and a 23 base pair duplicative insertion in exon 5 of the AMH gene, causing the introduction of a premature stop codon, homozygous in the two brothers. The surgical correction of the genital abnormalities was successfully carried out by laparoscopic orchidopexy according to Fowler-Stephens. CONCLUSION: Persistent müllerian duct syndrome should be taken into consideration in all cases of bilateral cryptorchidism. Laparoscopy is the elective procedure for diagnosis of this disease and laparoscopic surgery for orchidopexy of intra-abdominal testes. Mutation analysis of the anti-müllerian hormone gene in these patients helps to understand the structure-function relationship of the anti-müllerian hormone protein, although it is not clear at present whether anti-müllerian hormone is necessary to maintain normal testicular function.
Subject(s)
Cryptorchidism/genetics , Glycoproteins , Growth Inhibitors/genetics , Mullerian Ducts/abnormalities , Mutation , Testicular Hormones/genetics , Anti-Mullerian Hormone , Base Sequence , Child , Cryptorchidism/surgery , Growth Inhibitors/blood , Growth Inhibitors/deficiency , Homozygote , Humans , Laparoscopy , Male , Nuclear Family , Phenotype , Syndrome , Testicular Hormones/blood , Testicular Hormones/deficiencyABSTRACT
Steroidogenic factor 1 (NR5A1/SF-1) plays an essential role in the development of the hypothalamic-pituitary-adrenal and hypothalamic-pituitary-gonadal axes, controlling expression of their many important genes. The recent description of a 46,XY patient bearing a mutation in the NR5A1 gene, causing male pseudohermaphroditism and adrenal failure, demonstrated the crucial role of SF-1 in male gonadal differentiation. The role of SF-1 in human ovarian development was, until now, unknown. We describe a phenotypically and genotypically normal girl, with signs and symptoms of adrenal insufficiency and no apparent defect in ovarian maturation, bearing a heterozygote G-->T transversion in exon 4 of the NR5A1 gene that leads to the missense R255L in the SF-1 protein. The exchange does not interfere with protein translation and stability. Consistent with the clinical picture, R255L is transcriptionally inactive and has no dominant-negative activity. The inability of the mutant (MUT) NR5A1/SF-1 to bind canonical DNA sequences might offer a possible explanation for the failure of the mutant protein to transactivate target genes. This is the first report of a mutation in the NR5A1 gene in a genotypically female patient, and it suggests that NR5A1/SF-1 is not necessary for female gonadal development, confirming the crucial role of NR5A1/SF-1 in adrenal gland formation in both sexes.
Subject(s)
Adrenal Gland Diseases/genetics , DNA-Binding Proteins/metabolism , Gene Expression Regulation , Mutation, Missense/genetics , Ovary/growth & development , Puberty , Transcription Factors/metabolism , Adrenal Gland Diseases/metabolism , Amino Acid Sequence , Base Sequence , Child, Preschool , DNA/genetics , DNA/metabolism , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , Female , Fushi Tarazu Transcription Factors , Genes, Reporter/genetics , Heterozygote , Homeodomain Proteins , Humans , Infant , Ovary/metabolism , Phenotype , Protein Binding , Receptors, Cytoplasmic and Nuclear , Steroidogenic Factor 1 , Transcription Factors/chemistry , Transcription Factors/geneticsABSTRACT
Lhx3 is a LIM-homeobox protein essential for pituitary development in mice. The human homologue gene spans 7.2 kb and contains 7 exons, including two alternatively spliced first exons. This structure encodes two distinct protein isoforms, LHX3a and LHX3b, that differ exclusively in their amino-terminus. The LHX3 gene was localized at 9q34.2-34.3. The predicted protein sequence is highly homologous to other known Lhx3 proteins, the highest degree of homology being in the conserved domains. The highest expression of LHX3 was found in pituitary gland, spinal cord, and lung. Among different pituitary cell types, corticotrophs appear to express preferentially LHX3b isoform, suggesting a distinct role of the b-form in the development of this cell lineage. Although the human LHX3 gene structure would provide a ground for clarification of the molecular basis of complete anterior pituitary deficiency, we were unable to identify any mutation in the LHX3 gene of 46 such patients.
Subject(s)
Chromosomes, Human, Pair 9 , Homeodomain Proteins/biosynthesis , Homeodomain Proteins/genetics , Alternative Splicing , Animals , Cell Line , Chromosome Mapping , DNA, Complementary/metabolism , Exons , Expressed Sequence Tags , Homeodomain Proteins/chemistry , Humans , Hypopituitarism/genetics , In Situ Hybridization, Fluorescence , LIM-Homeodomain Proteins , Lung/metabolism , Mice , Models, Genetic , Mutation , Physical Chromosome Mapping , Pituitary Gland/metabolism , Protein Isoforms , Protein Structure, Tertiary , Rats , Reverse Transcriptase Polymerase Chain Reaction , Spinal Cord/metabolism , Tissue Distribution , Transcription FactorsABSTRACT
CYP17 is a microsomal enzyme embodying two distinct activities, 17alpha-hydroxylase and 17,20-lyase, essential for the synthesis of cortisol and sex hormone precursors, respectively. The two activities are differentially regulated in a tissue and developmental stage-dependent fashion. Leptin might play a role in such differential control. Low dose leptin caused a significant increase in 17,20-lyase activity in adrenal NCI-H295R cells expressing leptin (OB) receptor (OB-R), without significant sustained influence on the 17alpha-hydroxylase activity. To analyze the time dependence of this leptin effect, the impact of long and short-term leptin treatment was studied. To assess the relationship with the OB-R signal transduction pathway, the same experiments were performed in intact cells and in a reconstituted system. The long- and short-term studies in intact cells and in microsomes suggest that the 17alpha-hydroxylase activity of CYP17 can be promptly stimulated by leptin, but that the effect is transient. In contrast, physiological doses of leptin steadily enhance 17,20-lyase activity. This influence is direct, OB-R specific and dependent on the integrity of the signal transduction pathway. The 17,20-lyase activity stimulation relies on phosphate incorporation, as demonstrated by the loss of leptin-dependent 17,20-lyase stimulation after phosphate removal, and by the fact that the DHEA production appears to be related exclusively to the presence of phosphorylated CYP17, independently from novel protein synthesis. The mechanism underlying the observed events seems to involve CYP17 phosphorylation, a feature of the OBR signal transduction pathway, and a process already shown to be crucial for 17,20-lyase activity.
Subject(s)
Adrenal Glands/enzymology , Leptin/pharmacology , Steroid 17-alpha-Hydroxylase/metabolism , Adrenal Cortex/metabolism , Adrenal Glands/cytology , Adrenal Glands/growth & development , Dose-Response Relationship, Drug , Humans , Puberty/metabolism , Recombinant Proteins/pharmacology , Tumor Cells, CulturedABSTRACT
Cytochrome P450 17alpha-hydroxylase (CYP17) is a single gene-encoded protein with two activities: 17alpha-hydroxylase and 17,20-lyase. The two catalytic activities are differentially regulated in health and disease. We took advantage of naturally occurring human mutations to understand the molecular bases of this differential regulation. We identified eight novel mutations in the CYP17 gene, different in nature and spread throughout the gene. As posttranslational modifications appear to be important for activity control, we investigated the phosphorylation state of wild-type and mutant CYP17 proteins. Although phospholabeled protein was seen when the wild-type and most mutant proteins were expressed, no phosphorylation was detected for the F417C mutant. F417C is the only 17,20-lyase deficiency case confirmed at the molecular level and represents the first phosphorylation CYP17-deficient mutant. In search of the physiological agents involved in this process, the effect of cAMP was tested on activity and phosphorylation state of our mutant CYP17 proteins. cAMP stimulates activity and phosphorylation in all cases, except in the F417C and R35L mutants. The lack of response to the physiological second messenger might explain the different phenotypes. The F417C mutant protein, which is already shown to be associated with the lack of electron transfer, provides for the first time a link between the electron transfer system and the phosphorylation state of the CYP17 enzyme in the control of 17,20-lyase activity.
Subject(s)
Adrenal Hyperplasia, Congenital , Phosphorylation , 8-Bromo Cyclic Adenosine Monophosphate/metabolism , Animals , COS Cells , Chlorocebus aethiops , DNA Mutational Analysis , Electron Transport/genetics , Heterozygote , Homozygote , Humans , Precipitin Tests , Protein Processing, Post-Translational/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Steroid 17-alpha-Hydroxylase/geneticsABSTRACT
A 55-year-old woman presented with androgenetic alopecia which had started at age 40. Her clinical history revealed that, unlike her younger sister, she was unable to conceive and was diagnosed as being sterile at age 30. At age 45, 21-hydroxylase deficiency (late-onset CAH) was assumed and glucocorticoid treatment suggested, but not initiated. There was slight hirsutism, but no other sign of virilization. Retesting of plasma steroids revealed elevated 17-OH-progesterone and free testosterone. Treatment with prednisone, cyproterone acetate, and spironolactone was started with significant clinical success. Surprisingly, the analyses of urinary steroid metabolites revealed a pattern that did not support the diagnosis of 21-hydroxylase deficiency (pregnanetriolone absent, pregnanediol, 17-OH-pregnanolone and pregnanetriol not increased). Abdominal CT showed bilateral adrenal hyperplasia and masses in both ovaries. Bilateral adnexectomy was performed, and cystic teratomas were diagnosed. Postoperative urinary steroid analyses showed a decreased tetrahydrocortisol/tetrahydrocortisone ratio (values around 0.08 as compared to age- and sex-matched controls with a ratio of about 0.5-0.8). Plasma cortisol appeared to be repeatedly elevated with exogenous sources excluded. Mass spectrometry showed that, while the tetrahydro metabolites were mainly cortisone-derived, the metabolites not reduced in A ring were mostly cortisol derivatives. This constellation clearly indicates cortisone reductase deficiency, a defect of hepatic 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD1). This enzyme catalyzes the oxidation of cortisol to cortisone and the reduction of cortisone to cortisol. In contrast to the corresponding kidney enzyme (11 beta-HSD2), its primary activity is, however, reductive. Although this is only the fifth reported case of that defect, more attention should be paid to this condition in hyperandrogenic women, even if elevated 17-OH-progesterone and testosterone suggest a more frequent cause.
Subject(s)
Androgens/blood , Cortisone Reductase/deficiency , Hydrocortisone/blood , Adult , Female , Gas Chromatography-Mass Spectrometry , Hormones/blood , Humans , Steroids/urineABSTRACT
The adrenal cortex encloses the neuroendocrine medulla and is itself subdivided into three distinct zones, each having a specific function and regulation. While the glomerulosa and the fasciculata control vital systems of mineral and energy supply, which are stringently regulated by higher control factors, the function of the reticularis is less clear, beyond supplying a pool of weak androgens, and consequently we do not understand its redundant regulation. The following questions need to be answered: 1. How is the formation of dehydroepiandrosterone (DHEA) and androstenedione differently regulated from glucocorticoid synthesis in normally functioning adrenals? 2. How might growth factors, which increase prepubertally, prime the adrenarche? 3. The regulation of the 17/20-lyase enzyme activity is one of the key factors of adrenal androgen secretion (review [2]). How can the two activities of the P450c17 enzyme be differently regulated in the same cell in a developmentally dependent fashion? This review focuses on the intra-adrenal growth factor system and on the role of 17/20-lyase regulation, as well as on their possible interactions. The increase of activity of the 17/20-lyase enzymatic activity is necessary for the rise of C19 steroids, while the relative increase of formation of DHEA is only possible in the presence of a low 3beta-hydroxysteroid-dehydrogenase (3betaHSD) activity.