Therapeutic Anticoagulation with Argatroban and Heparins Reduces Granulocyte Migration: Possible Impact on ECLS-Therapy?

Introduction. Anticoagulants such as argatroban and heparins (low-molecular-weight and unfractionated) play an immense role in preventing thromboembolic complications in clinical practice. Nevertheless, they can also have a negative effect on the immune system. This study is aimed at investigating the influence of these substances on polymorphonuclear neutrophils (PMNs), whose nonspecific defense mechanisms can promote thrombogenesis. Methods. Blood samples from 30 healthy volunteers were investigated, whereby PMNs were isolated by density gradient centrifugation and incubated with 0.8 mu g/mL of argatroban, 1.0 U/mL of low-molecular-weight heparin (LMWH), 1.0 U/mL of unfractionated heparin (UFH), or without drug (control). A collagen-cell mixture was prepared and filled into 3D mu-slide chemotaxis chambers (IBIDI (R) GmbH, Germany). Stimulation was initiated by using a chemokine gradient of n-formyl-methionine-leucyl-phenylalanine (fMLP), and microscopic observation was conducted for 4.5 hours. The cells' track length and track straightness, as well as the number of attracted granulocytes, level of ROS (reactive oxygen species) production, and NET (neutrophil extracellular traps) formation, were analyzed and categorized into migration distances and time periods. Results. All three anticoagulants led to significantly reduced PMN track lengths, with UFH having the biggest impact. The number of tracks observed in the UFH group were significantly reduced compared to the control group. Additionally, the UFH group demonstrated a significantly lower track straightness compared to the control. ROS production and NET formation were unaffected. Conclusion. Our data provide evidence that anticoagulants have an inhibitory effect on the extent of PMN migration and chemotactic migration efficiency, thus indicating their potential immune-modulatory and prothrombotic effects

Preeclampsia before 26 weeks of gestation: Obstetrical prognosis for the subsequent pregnancy

International audienceIntroduction: Gestational age at delivery seems to be a risk factor of recurrence of preeclampsia. The objective of this study was to analyze adverse pregnancy outcomes and recurrence of preeclampsia during the subsequent pregnancy in women with a history of pre-eclampsia delivered before 26 weeks of gestation.Material and method: We performed a retrospective study in two French tertiary care hospitals between 2000 and 2018. Patients with a history of pre-eclampsia delivered before 26 weeks of gestation were analyzed. Information on the immediate subsequent pregnancy was collected. Adverse composite outcome was defined as recurrent preeclampsia, HELLP syndrome, placental abruption, fetal growth restriction <3rd percentile or <10e percentile with Doppler abnormalities, maternal death and fetal death.Results: Among the 107 patients who met the criteria, 48 were analyzed for a subsequent pregnancy. Seventeen women (35.4 %) developed an adverse composite outcome, occurring for 15 women (31.2 %) before 34 weeks. Ten women (20.8 %) developed a recurrent preeclampsia occurring for 5 women (10.4 %) before 34 weeks. We related 3 HELLP syndromes, 1 placental abruption, 9 fetal growth restrictions, 3 fetal deaths and no maternal death. Compared to baseline normotensive women, chronic hypertension was significantly associated with an increased risk of adverse composite outcome (19.3 vs 58.8 %, p-value 0.014).Conclusion: In our population, preeclampsia with delivery before 26 weeks is associated with 35.4 % of adverse composite outcomes and 20.8 % of recurrent preeclampsia during the immediate subsequent pregnancy. These results justify the importance of an ongoing monitoring of these patients during subsequent pregnancy

In Vivo Effects of Neostigmine and Physostigmine on Neutrophil Functions and Evaluation of Acetylcholinesterase and Butyrylcholinesterase as Inflammatory Markers during Experimental Sepsis in Rats

Introduction. Recent studies have shown that acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) may serve as important diagnostic and therapeutic targets in sepsis. Since polymorphonuclear neutrophils (PMNs) play a pivotal role in the early phase of sepsis, we evaluated the potential therapeutic effects of cholinesterase inhibitors on PMN functions during cecal ligation and puncture- (CLP-) induced sepsis and investigated the roles of AChE and BChE as inflammatory markers under standardized experimental conditions. Methods. Sham surgery or CLP was performed in male Wistar rats (n=60). Animals were randomized into four groups: physostigmine, 100 μg/kg; neostigmine, 75 μg/kg; 0.9% saline (control group); and sham group, each applied four times over 24 h. The levels of reactive oxygen species (ROS) production and CD11b/CD62l expression were quantified by flow cytometry at t=0, 6, 15, 20, and 24 h. Blood gas analysis as well as AChE and BChE activity levels was measured by validated point-of-care measurements. Clinical scores and survival times were determined. Results. CLP induced a significant increase in ROS production and CD11b upregulation by rat PMNs. Treatment with physostigmine or neostigmine significantly reduced ROS production and CD11b upregulation by PMNs 20 h after CLP induction. In physostigmine-treated animals, survival times were significantly improved compared to the control animals, but not in neostigmine-treated animals. While AChE activity significantly decreased in the control animals at t>6 h, AChE activity did not change in the sham group. BChE activity decreased at t>20 h in the control animals. Conclusion. While AChE activity may serve as an acute inflammatory marker, BChE activity shows a delayed decrease. Administration of centrally acting physostigmine in CLP-induced sepsis in rats has protective effects on PMN functions and improves survival times, which may be of interest in clinical practice

Évaluation des expositions professionnelles et environnementales chez des patients présentant des troubles de la reproduction « Programme MATEREXPO-REPROTOXIF »

International audienceIntroductionLe programme MATEREXPO-REPROTOXIF est un programme pilote dédié à l’évaluation des expositions professionnelles ou environnementales chez des patients présentant des troubles de la reproduction. La mise en évidence d’une exposition à des facteurs de risque reprotoxiques doit permettre la mise en place de mesures de prévention visant à optimiser les futures grossesses.MéthodeCe programme a été mis en place en Île-de-France (IDF) au centre hospitalier intercommunal de Créteil (CHIC) et à l’hôpital Fernand-Widal, avec le soutien de l’Anses et de l’ARS IDF. Au CHIC, le programme a débuté en février 2018 par la prise en charge des patientes présentant des pathologies de grossesse et des malformations congénitales chez les nouveau-nés, le versant « troubles de la fertilité » a été développé à partir de septembre 2019. Les couples présentant l’un des critères de prise en charge se voient proposer un entretien. Le recueil d’informations est réalisé par une infirmière de recherche clinique dédiée. Le questionnaire, analogue à celui développé dans le cadre du programme ARTEMIS à Bordeaux, est expertisé par un ingénieur spécialisé en hygiène industrielle, ce qui permet d’identifier à partir des situations d’expositions en milieu professionnel ou dans l’environnement général, les expositions à des agents reprotoxiques.RésultatsSur les 15 premiers mois d’activité, 107 patientes ont pu bénéficier d’une consultation environnementale. L’âge moyen des patientes était de 32 ans.Globalement, 20 patientes (18,7 %) n’exerçaient pas d’activité professionnelle et 87 (81,3 %) étaient en activité professionnelle pendant leur grossesse. Au moins une exposition professionnelle ayant un retentissement potentiel sur la grossesse a été rapportée chez 48,3 % des patientes exerçant une activité professionnelle pendant leur grossesse.Concernant les expositions environnementales, au moins une circonstance d’exposition extraprofessionnelle ayant un retentissement potentiel sur la grossesse a été rapportée chez 102 patientes (95,3 %). Le niveau d’exposition est néanmoins considéré comme étant faible chez la plupart des patientes.ConclusionLes données préliminaires du programme MATEREXPO-REPROTOXIF montrent fréquemment des circonstances d’exposition à des facteurs de risque reprotoxiques dans l’environnement général et professionnel des patientes interrogées.Il est prévu une poursuite du programme dans le cadre du consortium PREVENIR « Prévention Environnement Reproduction » dédié à l’évaluation des expositions environnementales professionnelles et extra-professionnelles chez des patients souffrant de troubles de la reproduction dans 8 centres hospitaliers sur le territoire national

Recurrent necrotizing cellulitis, multi-organ autoimmune disease and humoral immunodeficiency due to a novel NFKB1 frameshift mutation

Background: Mutations in NFKB1(nuclear factor of kappa light polypeptide gene enhancer in B-cells 1) are associated with a variety of clinical symptoms, including lymphadenopathy, splenomegaly, hepatomegaly, autoimmune haemolytic anaemia, arthralgia, recurrent respiratory tract infections and post-operative necrotizing cellulitis. Case presentation: We describe a case of a 47-year-old man, who presented with deep necrotizing cellulitis after incision of a submucous abscess by a dentist. Surgical intervention led to a massive progress. Pyoderma gangraenosum (PG) was diagnosed clinically and confirmed histopathologically. High dose corticosteroids and intravenous immunoglobulins (IVIG) improved wound healing dramatically. Until now, immune mediated inflammation events not only affected the skin, but also multiple inner organs, i.e. the heart, lungs and gut. Sequencing of all coding exons of NFKB1 revealed a heterozygous 1bp deletion in exon 23 predicting a frameshift starting at codon Ala891 and resulting in a subsequent stop codon at position 6 in the new reading frame: NM_003998.4: c.2671del; p.(Ala891Glnfs*6) Acute episodes were always successfully treated with corticosteroids, IVIG and concomitant antibiotics. To prevent further exacerbations, the patient receives IVIG once a month, low-dose corticosteroids and methotrexate. Conclusion: This is the first case of a patient with recurrent necrotizing cellulitis and immune mediated multi organ involvement (heart, lungs, intestine) carrying the novel frameshift mutation c.2671del (p. Ala891Glnfs*6) in NFKB1 effectively treated with IVIG, low-dose corticosteroids and methotrexate