The prevalence of intragenic deletions in patients with idiopathic hypogonadotropic hypogonadism and Kallmann syndrome

Idiopathic hypogonadotropic hypogonadism (IHH) and Kallmann syndrome (KS) are clinically and genetically heterogeneous disorders caused by a deficiency of gonadotrophin-releasing hormone (GnRH). Mutations in three genes—KAL1, GNRHR and FGFR1—account for 15–20% of all causes of IHH/KS. Nearly all mutations are point mutations identified by traditional PCR-based DNA sequencing. The relatively new method of multiplex ligation-dependent probe amplification (MLPA) has been successful for detecting intragenic deletions in other genetic diseases. We hypothesized that MLPA would detect intragenic deletions in ∼15–20% of our cohort of IHH/KS patients. Fifty-four IHH/KS patients were studied for KAL1 deletions and 100 were studied for an autosomal panel of FGFR1, GNRH1, GNRHR, GPR54 and NELF gene deletions. Of all male and female subjects screened, 4/54 (7.4%) had KAL1 deletions. If only anosmic males were considered, 4/33 (12.1%) had KAL1 deletions. No deletions were identified in any of the autosomal genes in 100 IHH/KS patients. We believe this to be the first study to use MLPA to identify intragenic deletions in IHH/KS patients. Our results indicate ∼12% of KS males have KAL1 deletions, but intragenic deletions of the FGFR1, GNRH1, GNRHR, GPR54 and NELF genes are uncommon in IHH/KS

Predictors of Placebo Response to Local (Intra-Articular) Therapy In Osteoarthritis:An Individual Participant Data Meta-Analysis

Objective: We undertook this study to evaluate potential predictors of placebo response with intra-articular (IA) injections for knee/hip osteoarthritis (OA) using individual participant data (IPD) from existing trials.Methods: Randomized placebo-controlled trials evaluating IA glucocorticoid or hyaluronic acid published to September 2018 were selected. IPD for disease characteristics and outcome measures were acquired. Potential predictors of placebo response included participant characteristics, pain severity, intervention, and trial design. Placebo response was defined as at least a 20% reduction in baseline pain. Logistic regression models and odds ratios were computed as effect measures to evaluate patient and pain mechanisms and then pooled using a random effects model. Generalized mixed-effect models were applied to intervention and trial characteristics. Results: Of 56 eligible trials, 6 shared data, and these were combined with the existing 4 OA Trial Bank studies, yielding 10 studies with IPD of 621 placebo participants for analysis. In the total placebo population, at short-term follow-up, the use of local anesthetic and ultrasound guidance were associated with reduced odds of placebo response. At midterm follow-up, mid- to long-term trial duration was associated with increased odds of placebo response, and worse baseline function scores were associated with reduced odds of a placebo response. Conclusion: The administration of local anesthetics or ultrasound guidance may reduce IA placebo response at short-term follow-up. At midterm follow-up, participants with worse baseline function scores may be less likely to respond to IA placebo, and mid- to long-term trial duration may enhance the placebo response. Further studies are required to corroborate these potential predictors of IA placebo response.</p

Targeted next generation sequencing approach identifies eighteen new candidate genes in normosmic hypogonadotropic hypogonadism and Kallmann Syndrome

The genetic basis is unknown for ∼60% of normosmic hypogonadotropic hypogonadism (nHH)/Kallmann syndrome (KS). DNAs from (17 male and 31 female) nHH/KS patients were analyzed by targeted next generation sequencing (NGS) of 261 genes involved in hypothalamic, pituitary, and/or olfactory pathways, or suggested by chromosome rearrangements. Selected variants were subjected to Sanger DNA sequencing, the gold standard. The frequency of Sanger-confirmed variants was determined using the ExAC database. Variants were classified as likely pathogenic (frameshift, nonsense, and splice site) or predicted pathogenic (nonsynonymous missense). Two novel FGFR1 mutations were identified, as were 18 new candidate genes including: AMN1, CCKBR, CRY1, CXCR4, FGF13, GAP43, GLI3, JAG1, NOS1, MASTL, NOTCH1, NRP2, PALM2, PDE3A, PLEKHA5, RD3, and TRAPPC9, and TSPAN11. Digenic and trigenic variants were found in 8/48 (16.7%) and 1/48 (2.1%) patients, respectively. NGS with confirmation by Sanger sequencing resulted in the identification of new causative FGFR1 gene mutations and suggested 18 new candidate genes in nHH/KS

Combined deformation and solidification-driven porosity formation in aluminum alloys

In die-casting processes, the high cooling rates and pressures affect the alloy solidification and deformation behavior, and thereby impact the final mechanical properties of cast components. In this study, isothermal semi-solid compression and subsequent cooling of aluminum die-cast alloy specimens were characterized using fast synchrotron tomography. This enabled the investigation and quantification of gas and shrinkage porosity evolution during deformation and solidification. The analysis of the 4D images (3D plus time) revealed two distinct mechanisms by which porosity formed; (i) deformation-induced growth due to the enrichment of local hydrogen content by the advective hydrogen transport, as well as a pressure drop in the dilatant shear bands, and (ii) diffusion-controlled growth during the solidification. The rates of pore growth were quantified throughout the process, and a Gaussian distribution function was found to represent the variation in the pore growth rate in both regimes. Using a one-dimensional diffusion model for hydrogen pore growth, the hydrogen flux required for driving pore growth during these regimes was estimated, providing a new insight into the role of advective transport associated with the deformation in the mushy region

Familial 46,XY sex reversal without campomelic dysplasia caused by a deletion upstream of the SOX9 gene

BACKGROUND: 46,XY sex reversal is a rare disorder and familial cases are even more rare. The purpose of the present study was to determine the molecular basis for a family with three affected siblings who had 46,XY sex reversal. METHODS: DNA was extracted from three females with 46,XY sex reversal, two normal sisters, and both unaffected parents. All protein coding exons of the SRY and NR5A1 genes were subjected to PCR-based DNA sequencing. In addition, array comparative genomic hybridization was performed on DNA from all seven family members. A deletion was confirmed using quantitative polymerase chain reaction. Expression of SOX9 gene was quantified using reverse transcriptase polymerase chain reaction. RESULTS: A 349kb heterozygous deletion located 353kb upstream of the SOX9 gene on the long arm of chromosome 17 was discovered in the father and three affected siblings, but not in the mother. The expression of SOX9 was significantly decreased in the affected siblings. Two of three affected sisters had gonadoblastomas. CONCLUSION: This is the first report of 46,XY sex reversal in three siblings who have a paternally inherited deletion upstream of SOX9 associated with reduced SOX9 mRNA expression

Spatio-temporal variability of surface-layer turbulent fluxes over the bay of bengal and arabian sea during the ICARB field experiment

We report the spatio-temporal variability of surface-layer turbulent fluxes of heat, moisture and momentum over the Bay of Bengal (BoB) and the Arabian Sea (AS) during the Integrated Campaign for Aerosols, gases Radiation Budget (ICARB) field experiment. The meteorological component of ICARB conducted during March - May 2006 onboard the oceanic research vessel Sagar Kanya forms the database for the present study. The bulk transfer coefficients and the surface-layer fluxes are estimated using a modified bulk aerodynamic method, and then the spatio-temporal variability of these air-sea interface fluxes is discussed in detail. It is observed that the sensible and latent heat fluxes over the AS are marginally higher than those over the BoB, which we attribute to differences in the prevailing meteorological conditions over the two oceanic regions. The values of the wind stress, sensible and latent heat fluxes are compared with those obtained for the Indian Ocean Experiment (INDOEX) period. The variation of drag coefficient (C<SUB>D</SUB> ), exchange coefficients of sensible heat and moisture (C<SUB>H</SUB> = C<SUB>E</SUB> ) and neutral drag coefficient (C <SUB>DN</SUB> ) with wind speed is also discussed