Huntington's disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the ...mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.
► TR-FRET assay development for duplex detection of soluble and aggregated huntingtin ► Detection of aggregates in brain and peripheral tissue of Huntington's disease models ► Inverse correlation between soluble and aggregated huntingtin upon disease progression
Objective:
Congenital adrenal hyperplasia results from 21-hydroxylase deficiency in more than ninety percent of cases. The classical form of 21-hydroxylase deficiency presents in the neonatal period ...with virilization or adrenal insufficiency, with or without concurrent salt wasting. We report on a rare case of classic 21-hydroxylase deficiency diagnosed in late adulthood.
Case report:
A 39-year-old male patient presented for workup of infertility. Urologic investigation revealed small testes, bilateral testicular masses, and asthenozoospermia. The patient's steroid metabolism showed markedly increased levels of adrenal androgens, in particular of 17-hydroxyprogesterone and 21-deoxycortisol. The gas chromatographic-mass spectrometric (GC-MS) urinary steroid profile was dominated by metabolites of 17-hydroxyprogesterone, while the endogenous glucocorticoid production was subnormally low. ACTH levels in plasma were elevated. These hormonal findings were consistent with 21-hydroxylase deficiency. Therapy with dexamethasone was initiated. The CYP21A2 gene analysis revealed the mutation I172N (ATC → AAC) in exon 4 of allele 1 and a large gene deletion in allele 2.
Conclusions:
Cases of 21-hydroxylase deficiency diagnosed in late adulthood are rare; however, clinicians should be alert of this possibility.
The adipokine adiponectin has been postulated to play an important role in the modulation of glucose and lipid metabolism in insulin-sensitive tissues. Up to now, adiponectin gene (
apM1
) expression ...was exclusively found in white adipose tissue. In our study, we show for the first time
apM1
expression in human myotubes and investigated its hormonal regulation. In vitro differentiated human myotubes were incubated with an adiponectin-containing HEK293 cell culture supernatant and agents were added as indicated. After 48 hours, adiponectin mRNA was assessed by RT-PCR. Adiponectin protein was visualized by immunoblotting and quantified by radioimmunoassay.
After exposure to an adiponectin containing HEK293 cell culture supernatant, adiponectin mRNA was significantly and consistently induced in human myotubes (90-fold over control, p<0.001, n=8). This increase was paralleled by a significant rise of intracellular adiponectin protein (8-fold over control, p<0.05, n=4). Addition of choleratoxin (1µg/ml) enhanced
apM1
induction significantly and isoproterenol (10µM) tended to enhance
apM1
induction as well. For TNF-α (5nM), IL-6 (5nM), insulin (5nM), and leptin (5nM), we found no regulatory impact on
apM1
expression in human myotubes.
In summary, we show an expression of
apM1
in human myotubes. Furthermore we observed consistent enhancement of the expression by choleratoxin and isoproterenol. Therefore, we conclude that the stimulatory G protein subunit Gsα is involved in the regulation of
apM1
expression and catecholamines might act as positive regulators. This would be compatible with the known modulating role of catecholamines on
apM1
expression in human white adipose tissue. However, other insulin resistance- associated hormones such as TNF-α, IL-6, insulin, and leptin, do not seem to regulate
apM1
expression in human myotubes.
Hospitalized patients with heart failure and decreased ejection fraction are at substantial risk for mortality and rehospitalization, yet no acute therapies are proven to decrease this risk. ...Therefore, in-hospital use of medications proved to decrease long-term mortality is a critical strategy to improve outcomes. Although endorsed in guidelines, predictors of initiation and continuation of angiotensin-converting enzyme (ACE) inhibitors/angiotensin receptor blockers (ARBs), β blockers, and aldosterone antagonists have not been well studied. We assessed noncontraindicated use patterns for the 3 medications using the Get With the Guidelines–Heart Failure (GWTG-HF) registry from February 2009 through March 2010. Medication continuation was defined as treatment on admission and discharge. Multivariable logistic regression using generalized estimating equations was used to determine factors associated with discharge use. In total 9,474 patients were enrolled during the study period. Of those treated before hospitalization, overall continuation rates were 88.5% for ACE inhibitors/ARBs, 91.6% for β blockers, and 71.9% for aldosterone-antagonists. Of patients untreated before admission, 87.4% had ACE inhibitors/ARBs and 90.1% had β blocker initiated during hospitalization or at discharge, whereas only 25.2% were started on an aldosterone antagonist. In multivariate analysis, admission therapy was most strongly associated with discharge use (adjusted odds ratios 7.4, 6.0, and 20.9 for ACE inhibitors/ARBs, β blockers, and aldosterone antagonists, respectively). Western region, younger age, and academic affiliation were also associated with higher discharge use. Although ACE inhibitor/ARB and β-blocker continuation rates were high, aldosterone antagonist use was lower despite potential eligibility. In conclusion, being admitted on evidence-based medications is the most powerful, independent predictor of discharge use.
Apical reabsorption of dibasic amino acids and cystine in kidney is mediated by the heteromeric amino acid antiporter rBAT/b super(0,+)AT (system b super(0,+)). Mutations in rBAT cause cystinuria ...type A, whereas mutations in b super(0,+)AT cause cystinuria type B, b super(0+)AT is the catalytic subunit, whereas it is believed that rBAT helps the routing of the rBAT/b super(0,+)AT heterodimeric complex to the plasma membrane. In the present study, we have functionally characterized the cystinuria-specific R365W (Arg super(365) arrow right Trp) mutation of human rBAT, which in addition to a trafficking defect, alters functional properties of the b super(0+) transporter. In oocytes, where human rBAT interacts with the endogenous b super(0,+)AT subunit to form an active transporter, the rBAT(R365W) mutation caused a defect of arginine efflux without altering arginine influx or apparent affinities for intracellular or extracellular arginine. Transport of lysine or leucine remained unaffected. In HeLa cells, functional expression of rBAT(R365W)/b super(0,+)AT was observed only at the permissive temperature of 33 degree C. Under these conditions, the mutated transporter showed 50% reduction of arginine influx and a similar decreased accumulation of dibasic amino acids. Efflux of arginine through the rBAT (R365W)/b super(0,+)AT holotransporter was completely abolished. This supports a two-translocation-pathway model for antiporter b super(0,+), in which the efflux pathway in the rBAT(R365W)/b super(0,+)AT holotransporter is defective for arginine translocation or dissociation. This is the first direct evidence that mutations in rBAT may modify transport properties of system b super(0,+).
Abstract Objective To examine the frequency and determinants of an assessment for rehabilitation during the hospitalization for acute stroke. Design Prospective cohort of patients admitted with acute ...stroke in the Get With The Guidelines–Stroke (GWTG-Stroke) program from January 8, 2008, to March 31, 2011. Setting Acute hospitals (n=1532) in the United States participating in the GWTG-Stroke program. Participants Adults with a stroke diagnosis (N=616,982) from a GWTG-Stroke–participating acute hospital. Interventions Not applicable. Main Outcome Measure Documentation of an assessment for rehabilitation services during the acute hospitalization. Results Overall, almost 90% of stroke patients had documentation of an acute assessment for rehabilitation. In multivariable analysis, patients significantly more likely to be assessed for rehabilitation were younger, male, black or of other nonwhite races (Asian, American Indian, Alaska Native, Native Hawaiian, or Pacific Islander) when compared with white, independently ambulating before admission, and admitted from the community. Patients who received a stroke consult, cared for in a stroke unit, and treated in the northeast region of the United States were also more likely to be assessed. Conclusions There is evidence that rehabilitation was considered for 90% of acute stroke patients in this sample. Future research is needed to examine what assessments are conducted and by whom, and how these are used to determine the appropriate level of rehabilitation care for their needs.
Objective:
We report on a rare case of 17α-hydroxylase/17,20-lyase deficiency caused by homocygosity of a novel mutation of the cytochrome P450c17 (CYP17) gene.
Case report:
A 20-year old female ...Turkish patient (46, XX), phenotypically female, presented with primary amenorrhea, sexual infantilism, and easy fatiguability. Hormonal data showed low or undetectable plasma concentrations of 17α-hydroxycorticoids, androgens, and estrogens, while ACTH, FSH, and LH levels were elevated. The plasma levels of the mineralocorticoid precursors were increased, whereas aldosterone concentration was within the lower normal range and the plasma renin activity was decreased. The hormonal findings are consistent with combined 17α-hydroxylase/17,20-lyase deficiency and therapy with hydrocortisone and conjugated estrogens was initiated. CYP17 gene analysis revealed homocygosity of the mutation Y27Stop (TAC to TAA) in exon 1; which has not been previously described.
Conclusions:
This is the first description of homocygosity of a mutation in the CYP17 gene leading to a stop codon in a patient with combined 17α-hydroxylase/17,20-lyase defiency.
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