Fabrication and Application of Isotopically Labelled Gold Arrays for Multiplexed Peptide Analysis

A new array-based technology for the simultaneous capture, chemical labelling and mass spectrometry analysis of peptides is presented. Isotopically labelled self-assembled monolayer (SAM) gold arrays are constructed and used simultaneously to capture and label a range of peptides. The array-immobilised, labelled peptides were released by MALDI ablation, analysed by MALDI mass spectrometry and readily identified as labelled peptides from their characteristic isotope pattern. This new solid-phase array platform has the advantage of minimal sample manipulation and is suitable for multiple analyses of single protein digests on a single MALDI target plate

The risk stratification of adverse neonatal outcomes in women with gestational diabetes (STRONG) study

Aims: To assess the risk of adverse neonatal outcomes in women with gestational diabetes (GDM) by identifying subgroups of women at higher risk to recognize the characteristics most associated with an excess of risk. Methods: Observational, retrospective, multicenter study involving consecutive women with GDM. To identify distinct and homogeneous subgroups of women at a higher risk, the RECursive Partitioning and AMalgamation (RECPAM) method was used. Overall, 2736 pregnancies complicated by GDM were analyzed. The main outcome measure was the occurrence of adverse neonatal outcomes in pregnancies complicated by GDM. Results: Among study participants (median age 36.8 years, pre-gestational BMI 24.8 kg/m2), six miscarriages, one neonatal death, but no maternal death was recorded. The occurrence of the cumulative adverse outcome (OR 2.48, 95% CI 1.59–3.87), large for gestational age (OR 3.99, 95% CI 2.40–6.63), fetal malformation (OR 2.66, 95% CI 1.00–7.18), and respiratory distress (OR 4.33, 95% CI 1.33–14.12) was associated with previous macrosomia. Large for gestational age was also associated with obesity (OR 1.46, 95% CI 1.00–2.15). Small for gestational age was associated with first trimester glucose levels (OR 1.96, 95% CI 1.04–3.69). Neonatal hypoglycemia was associated with overweight (OR 1.52, 95% CI 1.02–2.27) and obesity (OR 1.62, 95% CI 1.04–2.51). The RECPAM analysis identified high-risk subgroups mainly characterized by high pre-pregnancy BMI (OR 1.68, 95% CI 1.21–2.33 for obese; OR 1.38 95% CI 1.03–1.87 for overweight). Conclusions: A deep investigation on the factors associated with adverse neonatal outcomes requires a risk stratification. In particular, great attention must be paid to the prevention and treatment of obesity

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

Synthèse des aloisines immobilisées

Des résultats récents ont montré que l aloisine A (7-nButyl-6-(4-hydroxyphenyl) [5H] pyrrolo[2,3-b]pyrazine) est un puissant inhibiteur des kinases dépendantes des cyclines. Une méthode permettant d identifier d éventuelles cibles cellulaires secondaires d un inhibiteur consiste à l immobiliser sur un support solide, afin d identifier les protéines ayant une forte affinité pour l inhibiteur par chromatographie d affinité d un extrait cellulaire. Dans ce but, l aloisine A portant un bras espaceur, ainsi que son analogue N-méthylé servant de contrôle négatif, ont été synthétisés. Nous rapportons ici la préparation de ces nouveaux dérivés de l aloisine conjugués à une chaîne de triéthylèneglycol en diverses positions de la molécule et terminés par une fonction amine permettant son immobilisation sur une matrice d agarose. Par ailleurs, la stratégie d immobilisation de la molécule via une réaction de cycloaddition [3+2] offre la perspective de synthèses de dérivés de l aloisine conjugués à diverses molécules d intérêt biologique telles que la biotine ou encore la tyrosine facilitant son transport à travers la barrière hémato-encéphaliqueRecent results have shown that aloisine A (7-nButyl-6-(4-hydroxyphenyl)[5H]pyrrolo[2,3-b] pyrazine) is a potent inhibitor of CDK. An original methodology aiming at identifying potential secondary inhibition targets relies on the immobilization of the inhibitor on solid matrix, followed by identification of proteins with high affinity for the inhibitor by affinity chromatography of a cellular extract. To this end, both aloisine A and the negative control N-methyl aloisine bearing extended linker chain have been synthesized. We present here the preparation of the new aloisine analogues bearing a triethylene glycol chain at different positions of the molecule, terminated by an amine suitable for the immobilisation on an agarose-based matrix. Furthemore, we show that conjugation of the molecule with the linker via a cycloaddition [3+2] allows efficient coupling to the matrix as well as access to aloisine derivatives conjugated with biologically relevent molecules such as biotine, or a tyrosine residue targeting LAT1 for transport of drugs through the blood-brain barrierLYON1-BU.Sciences (692662101) / SudocSudocFranceF

Synthesis of C-Linked immobized analogs of aloisine A by click chemistry

International audienceAn efficient approach for the immobilization of a series of analogs of aloisine A, an in vitro inhibitor of protein kinases, to polymeric supports via a [3+2] cycloaddition reaction is reported

Synthesis of C-Linked immobized analogs of aloisine A by click chemistry

International audienceAn efficient approach for the immobilization of a series of analogs of aloisine A, an in vitro inhibitor of protein kinases, to polymeric supports via a [3+2] cycloaddition reaction is reported

Synthèse des aloisines immobilisées

Des résultats récents ont montré que l aloisine A (7-nButyl-6-(4-hydroxyphenyl) [5H] pyrrolo[2,3-b]pyrazine) est un puissant inhibiteur des kinases dépendantes des cyclines. Une méthode permettant d identifier d éventuelles cibles cellulaires secondaires d un inhibiteur consiste à l immobiliser sur un support solide, afin d identifier les protéines ayant une forte affinité pour l inhibiteur par chromatographie d affinité d un extrait cellulaire. Dans ce but, l aloisine A portant un bras espaceur, ainsi que son analogue N-méthylé servant de contrôle négatif, ont été synthétisés. Nous rapportons ici la préparation de ces nouveaux dérivés de l aloisine conjugués à une chaîne de triéthylèneglycol en diverses positions de la molécule et terminés par une fonction amine permettant son immobilisation sur une matrice d agarose. Par ailleurs, la stratégie d immobilisation de la molécule via une réaction de cycloaddition [3+2] offre la perspective de synthèses de dérivés de l aloisine conjugués à diverses molécules d intérêt biologique telles que la biotine ou encore la tyrosine facilitant son transport à travers la barrière hémato-encéphaliqueRecent results have shown that aloisine A (7-nButyl-6-(4-hydroxyphenyl)[5H]pyrrolo[2,3-b] pyrazine) is a potent inhibitor of CDK. An original methodology aiming at identifying potential secondary inhibition targets relies on the immobilization of the inhibitor on solid matrix, followed by identification of proteins with high affinity for the inhibitor by affinity chromatography of a cellular extract. To this end, both aloisine A and the negative control N-methyl aloisine bearing extended linker chain have been synthesized. We present here the preparation of the new aloisine analogues bearing a triethylene glycol chain at different positions of the molecule, terminated by an amine suitable for the immobilisation on an agarose-based matrix. Furthemore, we show that conjugation of the molecule with the linker via a cycloaddition [3+2] allows efficient coupling to the matrix as well as access to aloisine derivatives conjugated with biologically relevent molecules such as biotine, or a tyrosine residue targeting LAT1 for transport of drugs through the blood-brain barrierLYON1-BU.Sciences (692662101) / SudocSudocFranceF