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Background Septic shock is characterized by breakdown of the endothelial glycocalyx and endothelial damage, contributing to fluid extravasation, organ failure and death. Albumin has shown benefit in septic shock patients. Our aims were: (1) to identify the relations between circulating levels of syndecan-1 (SYN-1), sphingosine-1-phosphate (S1P) (endothelial glycocalyx), and VE-cadherin (endothelial cell junctions), severity of the disease, and survival; (2) to evaluate the effects of albumin supplementation on endothelial dysfunction in patients with septic shock. Methods This was a retrospective analysis of a multicenter randomized clinical trial on albumin replacement in severe sepsis or septic shock (the Albumin Italian Outcome Sepsis Trial, ALBIOS). Concentrations of SYN-1, S1P, soluble VE-cadherin and other biomarkers were measured on days 1, 2 and 7 in 375 patients with septic shock surviving up to 7 days after randomization. Results Plasma concentrations of SYN-1 and VE-cadherin rose significantly over 7 days. SYN-1 and VE-cadherin were elevated in patients with organ failure, and S1P levels were lower. SYN-1 and VE-cadherin were independently associated with renal replacement therapy requirement during ICU stay, but only SYN-1 predicted its new occurrence. Both SYN-1 and S1P, but not VE-cadherin, predicted incident coagulation failure. Only SYN-1 independently predicted 90-day mortality. Albumin significantly reduced VE-cadherin, by 9.5% (p = 0.003) at all three time points. Conclusion Circulating components of the endothelial glycocalyx and of the endothelial cell junctions provide insights into severity and progression of septic shock, with special focus on incident coagulation and renal failure. Albumin supplementation lowered circulating VE-cadherin consistently over time. Clinical Trial Registration: ALBIOS ClinicalTrials.gov number NCT00707122.

In this study natural-based complex polyphenols, obtained through a smart synthetic approach, have been evaluated for their ability to inhibit the formation of A? oligomers, the most toxic species causing synaptic dysfunction, neuroinflammation, and neuronal death leading to the onset and progression of Alzheimer's disease. In vitro neurotoxicity tests on primary hippocampal neurons have been employed to select nontoxic candidates. Solution NMR and molecular docking studies have been performed to clarify the interaction mechanism of A? with the synthesized polyphenol derivatives, and highlight the sterical and chemical requirements important for their antiaggregating activity. NMR results indicated that the selected polyphenolic compounds target A? oligomeric species. Combined NMR and docking studies indicated that the A? central hydrophobic core, namely, the 17-31 region, is the main interaction site. The length of the peptidomimetic scaffold and the presence of a guaiacol moiety were identified as important requirements for the antiaggregating activity. In vivo experiments on an A? oligomer-induced acute mouse model highlighted that the most promising polyphenolic derivative (PP04) inhibits detrimental effects of A? oligomers on memory and glial cell activation. NMR kinetic studies showed that PP04 is endowed with the chemical features of true inhibitors, strongly affecting both the A? nucleation and growth rates, thus representing a promising candidate to be further developed into an effective drug against neurodegenerative diseases of the amyloid type.

Neutralizing activity against primary human immunodeficiency virus type 1 (HIV-1) isolates from 17 persons who were long-term disease nonprogressors (LTNPs) and 13 persons who were fast progressors (FPs) was compared. Sera from LTNPs showed higher neutralizing activity both in titer and in host spectrum than did sera from FPs. However, LTNP sera had limited neutralizing activity against HIV-1 subtypes from different geographic areas. Sera collected 6 years earlier from both groups had limited neutralizing activity, indicating that early responses are not predictive for disease progression. LTNPs had very low virus loads, as reflected by only one positive isolation, which was an MT-2-negative phenotype. Virus was isolated from all FPs, and the isolates showed a phenotype switch from MT-2 negative to MT-2 positive. Development of high-titer, broadly cross-reactive neutralizing antibodies is associated with control of virus replication and low virus load in HIV-1-infected LTNPs.

We performed a cross-sectional and partly retrospective virological evaluation of 31 long-term responders (LTRs) to zidovudine (ZDV) (persistent increase in the CD4+ cell counts without progression of HIV infection throughout a period of ZDV therapy3 years) and 17 well-matched controls who developed a marked immunological deterioration over a 24-month period of ZDV therapy. The biological phenotype of HIV-1 was assessed by testing the capacity of the isolates to replicate in the MT-2, HUT-78, C-8166, and U-937 T cell lines, and mutations at codons 215 and 41 of RT were checked in proviral DNA from uncultured PBMCs. Show/low non-syncytium-inducing (S/L-NSI) and rapid/high syncytium-inducing (R/H-SI) variants were detected in 25 (81%) and 2 (6%) LTRs, respectively. HIV-1 could not be isolated in the remaining four LTRs (13%). Conversely, 12 of 17 (71%) controls yielded R/H-SI variants. Conversion from the S/L-NSI to R/H to R/H-SI phenotype occurred in 5 controls but in none of the 18 LTRs tested. Mutant sequences in proviral DNA from control PBMCs were consistently detected (94%), while a wild-type sequence of the residues investigated was found in the majority of LTRs (77%). In our series, patients who received immunological and clinical benefits even after prolonged ZDV treatment had S/L-NSI viruses and a low risk to develop ZDV resistance. Conversely, subjects who demonstrated an immunological and clinical deterioration yielded R/H-SI variants or shifted from S/L-NSI to R/H-SI phenotypes and were at higher risk to develop mutations indicating ZDV resistance.

Conduct of clinical trials in babies, children and young people is often hindered by issues that could have been foreseen before the trial opened; that is, some clinical trials are often underprepared. In order to identify a good approach to trial preparedness, the European Network of Paediatric Research at the European Medicines Agency formed a working group. The Working Group included representation from regulators, industry, academics, paediatric clinical research networks and parents. The Working Group consulted widely about how to prepare for paediatric clinical trials. The Group’s detailed recommendations have been published (https://www.ema.europa.eu/en/documents/other/preparedness-medicines-clinical-trials-paediatrics-recommendations-enpr-ema-working-group-trial_en.pdf). This paper is a summary of the key recommendations including the following: start early, preferably in parallel to designing the medicine’s development plan and individual protocols; identify the rationale and clinical need; listen to the perspectives of children and families, and of patient advocacy groups; identify how many people will be eligible for the trial; identify the resources needed, such as clinical facilities (including play therapy) and out-of-pocket expenditure by participants and their families; use all available data to estimate what is possible; present information about preparedness in a structured way; deploy proportionate resources to support the preparation of trials. A well-prepared, well-designed trial is likely to require fewer changes during its course, be run in a shorter time frame and achieve expected objectives., Clinical trials in children require careful preparation, or they risk avoidable delays. This report provides a structured approach to clinical trial preparation for paediatric populations, and includes practical suggestions to improve the process based on learning from industry, regulators, sites, and patient advocates.

Here we describe the identification and preliminary characterization of a new class of pyrrolo(imidazo)quinoxaline hydrazones as flurescent probes for Aβ 1-42 fibrils. All the newly developed compounds were able to bind amyloid fibrils formed in vitro and some of them displayed an increase of their fluorescence upon binding. When tested on brain tissue preparations presenting Aβ deposits, the described hydrazones selectively stained amyloid structures and did not display aspecific binding. The hydrazones did not show antifibrillogenic activity and electron microscopy analysis revealed that they do not interfere with fibrils structure. The described pyrrolo(imidazo) quinoxalines could be useful for studying amyloid structures in vitro. Moreover, their experimentally proven ability to cross the blood-brain barrier in mouse opens the possibility of developing these compounds as potential amyloid imaging agents for in vivo applications.

We investigated the pharmacological profile of a novel series of quinoxaline-based 5-HT(3) receptor ligands bearing an extra basic moiety oil the piperazine N-4. High affinity and selectivity were dependent on the electronic properties of the substituents, and at cardiac level 3a and 3c modulated chronotropy but not inotropy. In von Bezold-Jarisch reflex test 3a-c were partial agonists while 3i was a full agonist. Preliminary pharmacokinetic studies indicated that 3a is it brain penetrating agent.