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This study aims to evaluate the effect of diclofenac addition to the preservation solution Celsior on liver graft preservation. Liver from Wistar rats were cold flushed in situ, harvested, and then stored in Celsior solution (24 h, 4[degrees]C) supplemented or not with 50 mg/L of diclofenac sodium salt. Reperfusion was performed (120 min, 37[degrees]C) using the isolated perfusion rat liver model. Perfusate samples were collected to evaluate transaminases' activities after cold storage and by the end of reperfusion. To evaluate liver function, bile flow, hepatic clearance of bromosulfophthalein, and vascular resistance were assessed. Diclofenac scavenging property (DPPH assay) as well as oxidative stress parameters (SOD and MPO activities and the concentration of glutathione, conjugated dienes, MDA, and carbonylated proteins) were measured. Transcription factors (PPAR-[gamma] and NF-kB), inflammation (COX-2, IL-6, HMGB- 1, and TLR-4), as well as apoptosis markers (Bcl-2 and Bax) were determined by quantitative RT-PCR. Enriching the preservation solution Celsior with diclofenac sodium salt attenuated liver injuries and improved graft function. Oxidative stress, inflammation, and apoptosis were significantly reduced in Celsior + Diclo solution. Also, diclofenac activated PPAR-[gamma] and inhibited NF-kB transcription factors. To decrease graft damage and improve transplant recovery, diclofenac sodium salt may be a promising additive to preservation solution. Key words: diclofenac, organ preservation solution, cold ischemia/reperfusion injury, inflammation, 1. Introduction Organ preservation is a crucial feature for a successful transplantation procedure (Petrenko et al. 2019). To ensure optimal graft preservation, two requirements are needed: hypothermia and a preservation [...]

BackgroundRecurrent bacterial and fungal infections, eczema, and increased serum IgE levels characterize patients with the hyper-IgE syndrome (HIES). Known genetic causes for HIES are mutations in signal transducer and activator of transcription 3 (STAT3) and dedicator of cytokinesis 8 (DOCK8), which are involved in signal transduction pathways. However, glycosylation defects have not been described in patients with HIES. One crucial enzyme in the glycosylation pathway is phosphoglucomutase 3 (PGM3), which catalyzes a key step in the synthesis of uridine diphosphate N-acetylglucosamine, which is required for the biosynthesis of N-glycans.ObjectiveWe sought to elucidate the genetic cause in patients with HIES who do not carry mutations in STAT3 or DOCK8.MethodsAfter establishing a linkage interval by means of SNPchip genotyping and homozygosity mapping in 2 families with HIES from Tunisia, mutational analysis was performed with selector-based, high-throughput sequencing. Protein expression was analyzed by means of Western blotting, and glycosylation was profiled by using mass spectrometry.ResultsMutational analysis of candidate genes in an 11.9-Mb linkage region on chromosome 6 shared by 2 multiplex families identified 2 homozygous mutations in PGM3 that segregated with disease status and followed recessive inheritance. The mutations predict amino acid changes in PGM3 (p.Glu340del and p.Leu83Ser). A third homozygous mutation (p.Asp502Tyr) and the p.Leu83Ser variant were identified in 2 other affected families, respectively. These hypomorphic mutations have an effect on the biosynthetic reactions involving uridine diphosphate N-acetylglucosamine. Glycomic analysis revealed an aberrant glycosylation pattern in leukocytes demonstrated by a reduced level of tri-antennary and tetra-antennary N-glycans. T-cell proliferation and differentiation were impaired in patients. Most patients had developmental delay, and many had psychomotor retardation.ConclusionImpairment of PGM3 function leads to a novel primary (inborn) error of development and immunity because biallelic hypomorphic mutations are associated with impaired glycosylation and a hyper-IgE-like phenotype.

Liver ischemia-reperfusion (IR) injury is inevitable in surgical procedures such as hepatic resection and liver transplantation. It represents a leading cause of liver graft dysfunction and primary nonfunction after transplantation. Phosphodiesterase (PDE) inhibitors are emerging as effective drugs able to reduce IR damage. The aim of this study was to investigate the effect of selective PDE-3 inhibitor olprinone (Olp) against liver IR injury. Male Wistar rats were subjected to 1 h of partial warm ischemia (70%) followed by 6 h of reperfusion. Before ischemia, rats were treated with saline (IR group), Olp (Olp group), or Olp with Akt inhibitor LY294002 (Olp plus LY group). After reperfusion, hepatic injury (transaminase activities), mitochondrial damage (glutamate dehydrogenase activity), oxidative stress (malondialdehyde and glutathione concentrations and catalase and superoxide dismutase activities), and protein kinase Akt activation were evaluated. Rat treatment with Olp reduced liver injury, prevented mitochondrial damage, decreased lipid peroxidation, and enhanced antioxidant enzymes. Also, Olp induced a significant activation in protein kinase Akt. Inhibition of Akt with LY294002 abolished all of the protective effects of Olp. In conclusion, Olp treatment may be an effective strategy in reducing liver IR injury through oxidative stress prevention and Akt activation. Key words: ischemia reperfusion injury, liver, olprinone, oxidative stress, Akt. Les lesions d'ischemie-reperfusion (IR) dans le foie sont inevitables dans le cadre d'interventions chirurgicales comme la resection et la greffe hepatiques. Ces lesions representent une cause premiere de dysfonctionnement du greffon hepatique et d'absence primitive de fonctionnement apres une greffe. Les inhibiteurs de la phosphodiesterase (PDE) se revelent etre des medicaments efficaces permettant d'attenuer les dommages lies a l'IR. Ces travaux avaient pour but d'etudier les effets de l'olprinone (Olp), un inhibiteur selectif de la PDE-3, contre les lesions d'IR. Nous avons soumis des rats Wistar males a une ischemie partielle (70 %) tiede de 1 h suivie d'une reperfusion de 6 h. Avant l'ischemie, nous avons administre aux rats une solution saline (groupe IR), de l'Olp (groupe Olp) ou de l'Olp avec du LY294002, un inhibiteur de l'Akt (groupe Olp plus LY). Apres la reperfusion, nous avons evalue les lesions hepatiques (activite des transaminases), les dommages mitochondriaux (activite de la glutamate deshydrogenase), le stress oxydatif (concentrations de malondialdehyde et de glutathion ainsi qu'activite de la catalase et de la superoxyde dismutase) ainsi que l'activation de la proteine kinase Akt. L'administration d'Olp aux rats permettait d'attenuer les lesions hepatiques, de prevenir les dommages mitochondriaux, de diminuer la peroxydation lipidique et de favoriser les enzymes anti-oxydants. De plus, l'Olp provoquait une activation marquee de la proteine kinase Akt. L'inhibition de l'Akt avec le LY294002 reduisait a neant tous les effets protecteurs de l'Olp. En conclusion, le traitement par l'Olp pourrait constituer une strategie efficace en vue d'attenuer les lesions d'IR hepatiques par la prevention du stress oxydatif et l'activation de l'Akt. [Traduit par la Redaction] Mots-cles : lesions d'ischemie-reperfusion, foie, olprinone, stress oxydatif, Akt., Introduction Ischemia-reperfusion (IR) injury remains an inevitable and unresolved problem in liver surgery; it determines the viability of the hepatic tissue after resection, repair of trauma, and liver transplantation (LT). [...]

Pyruvate Kinase (PK) deficiency is the most frequent red cell enzymatic defect responsible for hereditary non-spherocytic hemolytic anemia. The disease has been studied in several ethnic groups. However, it is yet an unknown pathology in Tunisia. We report here, the phenotypic and molecular investigation of PK deficiency in a Tunisian family. This study was carried out on two Tunisian brothers and members of their family. Hematological, biochemical analysis and erythrocyte PK activity were performed. The molecular characterization was carried out by gene sequencing technique. The first patient died few hours after birth by hydrops fetalis, the second one presented with neonatal jaundice and severe anemia necessitating urgent blood transfusion. This severe clinical picture is the result of a homozygous mutation of PKLR gene at exon 8 (c.1079G>A; p.Cys360Tyr). Certainly, this research allowed us to correlate the clinical phenotype severity with the identified mutation. Moreover, this will help in understanding the etiology of unknown anemia in our country.

Hyper IgE syndromes (HIES) is a heterogeneous group of Inborn Errors of Immunity characterized by eczema, recurrent skin and lung infections associated with eosinophilia and elevated IgE levels. Autosomal dominant HIES caused by loss of function mutations in Signal transducer and activator of transcription 3 (STAT3) gene is the prototype of these disorders. Over the past two decades, advent in genetic testing allowed the identification of ten other etiologies of HIES. Although Dedicator of Cytokinesis 8 (DOCK8) deficiency is no more classified among HIES etiologies but as a combined immunodeficiency, this disease, characterized by severe viral infections, food allergies, autoimmunity, and increased risk of malignancies, shares some clinical features with STAT3 deficiency. The present study highlights the diagnostic challenge in eleven patients with the clinical phenotype of HIES in a resource-limited region. Candidate gene strategy supported by clinical features, laboratory findings and functional investigations allowed the identification of two heterozygous STAT3 mutations in five patients, and a biallelic DOCK8 mutation in one patient. Whole Exome Sequencing allowed to unmask atypical presentations of DOCK8 deficiency in two patients presenting with clinical features reminiscent of STAT3 deficiency. Our study underlies the importance of the differential diagnosis between STAT3 and DOCK8 deficiencies in order to improve diagnostic criteria and to propose appropriate therapeutic approaches. In addition, our findings emphasize the role of NGS in detecting mutations that induce overlapping phenotypes. [ABSTRACT FROM AUTHOR]