An organic O donor for biological hydroxylation reactions.

All biological hydroxylation reactions are thought to derive the oxygen atom from one of three inorganic oxygen donors, O2, H2O2, or H2O. Here, we have identified the organic compound prephenate as the oxygen donor for the three hydroxylation steps of the O2-independent biosynthetic pathway of ubiquinone, a widely distributed lipid coenzyme. Prephenate is an intermediate in the aromatic amino acid pathway and genetic experiments showed that it is essential for ubiquinone biosynthesis in Escherichia coli under anaerobic conditions. Metabolic labeling experiments with 18O-shikimate, a precursor of prephenate, demonstrated the incorporation of 18O atoms into ubiquinone. The role of specific iron-sulfur enzymes belonging to the widespread U32 protein family is discussed. Prephenate-dependent hydroxylation reactions represent a unique biochemical strategy for adaptation to anaerobic environments.

Première vague de la COVID-19 au Québec : motivation du personnel soignant à traiter des patients infectés.

INTRODUCTION: Retention of healthcare workers (HCWs) in the healthcare system during the COVID-19 pandemic could become a challenge. It is therefore important to better understand what are the motivational elements that could explain a greater or lesser motivation to care for infected patients. OBJECTIVES: To evaluate factors modulating HCWs' willingness to treat COVID-19 infected patients. METHODS: HCWs from Québec, Canada, were invited to complete an online survey during the first wave of the COVID-19 pandemic between the months of April and July 2020. The survey focused on the intention to avoid treating infected patients, prior experiences in treating COVID-19 patients and anxiety levels. Descriptive statistics and multiple regression analysis were used to assess which factors explained differences in HCWs intention to avoid treating patients. RESULTS: A total of 430 HCW completed the survey. A majority were women (87%) and nurses (50%). Of those, 12% indicated having considered measures to avoid working with COVID-19 infected patients and 5% indicated having taken actions to avoid working with infected patients. A further 18% indicated that they would use a hypothetical opportunity to avoid working with infected patients. Having previously treated infected patients was associated with a significant reduction in the intention to avoid work (OR: 0.56 CI 0.36-0.86). Amongst HCWs, physicians had a significantly reduced intention to avoid treating infected patients (OR: 0.47 CI 0.23-0.94). We also found that an increase in anxiety score was associated with a greater intention to avoid treating COVID-19 infected patients (OR: 1.06 CI 1.04-1.08). CONCLUSION: Study results suggest that previous experience in treating COVID-19 infected patients is protective in terms of work-avoidance intentions. We also found that amongst HCWs, physicians had a significantly lower intention to avoid working with COVID-19 infected patients. Finally, our results show that increase in anxiety is associated with a higher intention to avoid treating infected patients. Characterization of factors associated with low anxiety levels and low reluctance to work during the COVID-19 pandemic could be useful in staffing facilities during the present and future healthcare crisis.

Safety and tolerance of cidofovir as a 2% gel for local application in high-grade cervical intraepithelial neoplasia: A phase 1 investigation.

OBJECTIVES: The primary objective was to evaluate the safety and local tolerance of a topical 2% (w/w) cidofovir gel, applied directly to the cervices of women with high-grade cervical intraepithelial neoplasia (CIN 2+). The secondary objective was to evaluate the pharmacokinetics of cidofovir during the treatment. MATERIALS AND METHODS: Nine women with CIN 2+, were treated with a course of 3 g of cidofovir gel, applied locally once per week for 3 weeks in total (9 g). The treatment was administered in a cervical cap, applied to the cervix for 5 or 10 hours (n = 6 and 3 patients, respectively). Follow-up included a structured questionnaire, a gynecological examination, blood analysis for hematology, C-reactive protein (CRP), and renal function assessment plus pharmacokinetic analyses of cidofovir after each treatment and at the end of the full course. RESULTS: No clinically significant hematological/biochemical abnormalities or serious adverse events (SAE) were reported, although 6 mild to moderate adverse events (AE) occurred in relation to the study drug: 1 flu-like syndrome and 5 local AEs. Plasma concentrations of cidofovir were very low (mean Cmax of 103.0 and 99.2 ng/mL after 5 and 10 hours of exposure, respectively). CONCLUSION: Cidofovir, directly applied on CIN 2+, is reasonably well tolerated and the systemic exposure following topical application is much lower than that seen with intravenous administration, at the approved dose.
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Molecular Cobalt Complexes with Pendant Amines for Selective Electrocatalytic Reduction of Carbon Dioxide to Formic Acid.

We report here on a new series of CO2-reducing molecular catalysts based on Earth-abundant elements that are very selective for the production of formic acid in dimethylformamide (DMF)/water mixtures (Faradaic efficiency of 90 ± 10%) at moderate overpotentials (500-700 mV in DMF measured at the middle of the catalytic wave). The [CpCo(PR2NR'2)I]+ compounds contain diphosphine ligands, PR2NR'2, with two pendant amine residues that act as proton relays during CO2-reduction catalysis and tune their activity. Four different PR2NR'2 ligands with cyclohexyl or phenyl substituents on phosphorus and benzyl or phenyl substituents on nitrogen were employed, and the compound with the most electron-donating phosphine ligand and the most basic amine functions performs best among the series, with turnover frequency >1000 s-1. State-of-the-art benchmarking of catalytic performances ranks this new class of cobalt-based complexes among the most promising CO2-to-formic acid reducing catalysts developed to date; addressing the stability issues would allow further improvement. Mechanistic studies and density functional theory simulations confirmed the role of amine groups for stabilizing key intermediates through hydrogen bonding with water molecules during hydride transfer from the Co center to the CO2 molecule.

Treatment of Post-Traumatic Stress Disorders with the Alpha-1 Adrenergic Antagonist Prazosin.

OBJECTIVE: The present review aims to assess the clinical efficacy and safety of the α-1-adrenergic antagonist prazosin as primary pharmacologic treatment for post-traumatic stress disorder (PTSD). METHOD: A systematic review was performed using keywords (i.e., prazosin, α-1-adrenergic antagonist, α-1-blocker, post-traumatic stress disorder) in the databases PubMed/Medline (1966-May 2016), Embase (1966-May 2016), ScienceDirect (1823-May 2016), OvidSP (1946-May 2016) and Nature (1845-May 2016). To be considered for inclusion, studies had to test the efficacy of prazosin either alone or added to ongoing treatment in adults with PTSD, use validated tools to assess and monitor the disorders, allow comparisons on the basis of univariate analyses (i.e., p-values of t-tests and effect sizes) and list the identified adverse reactions. RESULTS: 12 studies were included: 5 randomized controlled trials, 4 open-label prospective trials and 3 retrospective file reviews. The evaluation concerned 276 patients exposed to civilian trauma (19%) or war trauma (81%). Prazosin significantly decreases trauma nightmares, avoidance, hypervigilance and improves patient status in all studies. No significant difference of blood pressure was observed at the end of trials. CONCLUSIONS: Beyond the methodological and clinical biases of these studies, the present review not only confirms the effectiveness and good tolerability of prazosin, but also suggests its possible use as primary pharmacologic treatment for PTSD. Uncertainties remain, however, regarding the prescription modalities and dosages.

Cu/Cu2 O Electrodes and CO2 Reduction to Formic Acid: Effects of Organic Additives on Surface Morphology and Activity.

Copper/copper oxide (Cu/Cu2 O) electrodes are known to display interesting electrocatalytic performances for the reduction of CO2 , and thus, deserve further investigation for optimization. Here, we show that the addition of nitrogen-based organic additives greatly improves the activity of these electrodes (higher current densities, greater selectivity, and higher faradaic yields). The best effector is found to be tetramethyl cyclam. For example, electrolysis at -2.0 V versus Fc+ /Fc in CO2 -saturated DMF/H2 O (99:1, v/v) in the presence of this effector results in formic acid with almost 90 % faradaic yield. SEM and XPS analysis of the electrode surface reveals that the organic additive promotes the formation of active Cu0 nanoparticles from Cu2 O during electrolysis. This simple approach provides a straightforward strategy toward the optimization of Cu/Cu2 O electrodes.

Preoperative axillary lymph node staging by ultrasound-guided cytology using a four-level sonographic score.

BACKGROUND: The staging of axillary lymph nodes is critical to the management and prognosis of breast cancer, the most frequent cancer in females. Neoadjuvant therapy and lymph node dissection are recommended when malignant cells invade the lymph nodes. Therefore the pre-operative examination of these lymph nodes is crucial to treatment. METHODS: In this study, we examined the effectiveness of cytology through ultrasound-guided fine needle aspiration (USG-FNA) and ultrasound (US) imaging using an established classification system in correctly identifying lymph node status compared to the final histological results after surgery. RESULTS: Cytology by USG-FNA and US classification were found to be promising methods of axillary lymph node staging. CONCLUSIONS: US and CB offer minimally invasive techniques to pre-operatively examine these lymph nodes in patients with primary breast cancer.

Highly Pathogenic H5N1 and Novel H7N9 Influenza A Viruses Induce More Profound Proteomic Host Responses than Seasonal and Pandemic H1N1 Strains.

Influenza A viruses (IAV) are important human and animal pathogens with potential for causing pandemics. IAVs exhibit a wide spectrum of clinical illness in humans, from relatively mild infections by seasonal strains to acute respiratory distress syndrome during infections with some highly pathogenic avian influenza (HPAI) viruses. In the present study, we infected A549 human cells with seasonal H1N1 (sH1N1), 2009 pandemic H1N1 (pdmH1N1), or novel H7N9 and HPAI H5N1 strains. We used multiplexed isobaric tags for relative and absolute quantification to measure proteomic host responses to these different strains at 1, 3, and 6 h post-infection. Our analyses revealed that both H7N9 and H5N1 strains induced more profound changes to the A549 global proteome compared to those with low-pathogenicity H1N1 virus infection, which correlates with the higher pathogenicity these strains exhibit at the organismal level. Bioinformatics analysis revealed important modulation of the nuclear factor erythroid 2-related factor 2 (NRF2) oxidative stress response in infection. Cellular fractionation and Western blotting suggested that the phosphorylated form of NRF2 is not imported to the nucleus in H5N1 and H7N9 virus infections. Fibronectin was also strongly inhibited in infection with H5N1 and H7N9 strains. This is the first known comparative proteomic study of the host response to H7N9, H5N1, and H1N1 viruses and the first time NRF2 is shown to be implicated in infection with highly pathogenic strains of influenza.

Live birth after autograft of ovarian tissue cryopreserved during childhood.

Ovarian insufficiency is a major long-term adverse event, following the administration of a myeloablative conditioning regimen, and occurring in >80% of children and adolescents receiving such treatment for malignant or non-malignant disease. Cryopreservation of ovarian tissue is currently offered to preserve the fertility of these young patients. At least 35 live births have been reported after transplantation of cryopreserved ovarian tissue in adult patients, but the procedure remains unproven for ovarian tissue harvested at a prepubertal or pubertal age. We report here the first live birth after autograft of cryopreserved ovarian tissue in a woman with primary ovarian failure after a myeloablative conditioning regimen as part of a hematopoietic stem cell transplantation performed for homozygous sickle-cell anemia at age 14 years. This first report of successful fertility restoration after the graft of ovarian tissue cryopreserved before menarche offers reassuring evidence for the feasibility of the procedure when performed during childhood.

The CJIE1 prophage of Campylobacter jejuni affects protein expression in growth media with and without bile salts.

BACKGROUND: The presence of Campylobacter jejuni temperate bacteriophages has increasingly been associated with specific biological effects. It has recently been demonstrated that the presence of the prophage CJIE1 is associated with increased adherence and invasion of C. jejuni isolates in cell culture assays. RESULTS: Quantitative comparative proteomics experiments were undertaken using three closely related isolates with CJIE1 and one isolate without CJIE1 to determine whether there was a corresponding difference in protein expression levels. Initial experiments indicated that about 2% of the total proteins characterized were expressed at different levels in isolates with or without the prophage. Some of these proteins regulated by the presence of CJIE1 were associated with virulence or regulatory functions. Additional experiments were conducted using C. jejuni isolates with and without CJIE1 grown on four different media: Mueller Hinton (MH) media containing blood; MH media containing 0.1% sodium deoxycholate, which is thought to result in increased expression of virulence proteins; MH media containing 2.5% Oxgall; and MHwithout additives. These experiments provided further evidence that CJIE1 affected protein expression, including virulence-associated proteins. They also demonstrated a general bile response involving a majority of the proteome and clearly showed the induction of almost all proteins known to be involved with iron acquisition. The data have been deposited to the ProteomeXchange with identifiers PXD000798, PXD000799, PXD000800, and PXD000801. CONCLUSION: The presence of the CJIE1 prophage was associated with differences in protein expression levels under different conditions. Further work is required to determine what genes are involved in causing this phenomenon.

Myofibroblastic stromal reaction and lymph node status in invasive breast carcinoma: possible role of the TGF-ß1/TGF-ßR1 pathway.

BACKGROUND: The microenvironment modulates tissue specificity in the normal breast and in breast cancer. The stromal loss of CD34 expression and acquisition of SMA myofibroblastic features may constitute a prerequisite for tumor invasiveness in breast carcinoma. The aim of the present study is to examine the stromal expression of CD34 and SMA in cases of invasive ductal carcinoma and to try to demonstrate the role played by the TGF-ß 1 et TGF-ß R1 pathway in the transformation of normal breast fibrocytes into myofibroblasts. METHODS: We carried out an immunohistochemical study of CD34, SMA, TGF-ß and TGF-ß R1 on a series of 155 patients with invasive ductal carcinoma. We also treated a breast fibrocytes cell line with TGF-ß1. RESULTS: We found a loss of stromal expression of CD34 with the appearance of a myofibroblastic reaction in almost 100% cases of invasive ductal carcinoma. The strong stromal expression of SMA correlates with the presence of lymph node metastases. We were also able to show a greater expression of TGF-ß in the tumor cells as well as a higher expression of TGF- ß R1 in the tumor stroma compared to normal breast tissue. Finally, we demonstrated the transformation of breast fibrocytes into SMA positive myofibroblasts after being treated with TGF-ß1. CONCLUSIONS: Our study demonstrated that a significant tumor myofibroblastic reaction is correlated with the presence of lymph node metastasis and that this myofibroblastic reaction can be induced by TGF-ß1. Future research on fibrocytes, myofibroblasts, TGF-ß and stromal changes mechanisms is essential in the future and may potentially lead to new treatment approaches.

Identification of novel silent KEL alleles causing KEL:-5 (Ko) phenotype or discordance between KEL:1,-2 phenotype/KEL*01/02 genotype.

BACKGROUND: The Kell system, encoded by the KEL gene, is one of the most clinically important blood group systems. Molecular defects may lead to the absence of Kell antigen expression. The very rare KEL:5 results from silent KEL genes, also called KELnull alleles. In a few cases, the rare KEL:1,-2 phenotype may be associated with silent KEL*02 alleles. STUDY DESIGN AND METHODS: The aim of this study was to perform DNA investigations to identify silent KEL alleles among 10 KEL:-5 patients and 121 individuals presenting the rare KEL:1,-2 phenotype. Serologic investigations were performed on patients' red blood cells and serum. The KEL gene analysis was done by using a BeadChip assay (HEA Version, 1.2, Immucor), real-time polymerase chain reaction, and/or sequencing of all 19 exons of the KEL gene. RESULTS: In KEL:-5 patients, two novel KELnull alleles were described: 821G>A being the second described KELnull allele on a KEL*01 backbone and 184Tdel. In the 121 KEL:1,-2 individuals, nine (7.4%) were found to display a discordant KEL:1,-2 phenotype and KEL*01/KEL*02 genotype. Three novel silent KEL*02 alleles were described: 1084C>A, 1708G>A, and IVS11+5g>a. CONCLUSION: The number of silent KEL alleles and the notion that KEL null alleles are on a KEL*02 background may evolve in the coming years. Systematic DNA analysis showed that the number of discordant phenotype/genotype results, related to silent KEL*02 alleles was higher than expected in France. These data emphasize that clinical practice based on DNA analysis for blood group antigens requires caution and should improve the performance of the blood group phenotype prediction.

[Alzheimer's disease, amyloid-b peptides and ubiquitin-proteasome system: Therapeutic perspectives]. / Maladie d'Alzheimer, peptides ß-amyloïdes et système ubiquitine-protéasome - Perspectives thérapeutiques.

The Alzheimer's disease - an age-related neurodegenerative disorder leading to a progressive cognitive impairment - is characterized by an intracerebral accumulation of soluble ß-amyloid (Aß) oligomers, followed by the appearance of abnormally ubiquitinylated neurofibrillary tangles - a process associated with a chronic inflammation. The systematic presence of ubiquitinylated inclusions reflects a decrease in the proteasome activity due to (and contributing to) the presence of Aß oligomers - a central dysfunction in the etiology of the disease. The involvement of the ubiquitin-proteasome system opens new therapeutic perspectives for both prevention and treatment.

Title: Maladie d'Alzheimer, peptides ß-amyloïdes et système ubiquitine-protéasome - Perspectives thérapeutiques. Abstract: La maladie d'Alzheimer ­ une maladie neurodégénérative liée à l'âge entraînant une altération progressive des fonctions cognitives ­ se caractérise par une accumulation intracérébrale d'oligomères ß-amyloïdes (Aß) solubles, suivie d'une apparition d'enchevêtrements neurofibrillaires anormalement ubiquitinylés ­ un processus associé à une inflammation chronique. La présence systématique d'inclusions ubiquitinylées traduit une baisse d'activité du protéasome due (et concourant) à la présence d'oligomères Aß ­ un dysfonctionnement central dans l'étiologie de la maladie. L'implication du système ubiquitine-protéasome ouvre de nouvelles perspectives thérapeutiques tant préventives que curatives.

Sonographic Aeration Scoring Indicates Disease Severity in Critically Ill Patients with COVID-19.

AIMS AND METHODS: We evaluated an ultrasound score from 0 to 32 points in eight pulmonary regions to monitor critically ill COVID-19 patients. The score was correlated to surrogate parameters of disease severity, i.e., the oxygenation index, respiratory support, mortality, plasma interleukin-6, and WHO and ARDS classifications. RESULTS: A total of 27 patients were repeatedly examined, and 71 examinations were evaluated. Patients with severe COVID-19 scored higher (median 17) than those with moderate disease (median 11, p < 0.01). The score did not differentiate between stages of ARDS as defined by the Berlin criteria (p = 0.1) but could discern ARDS according to the revised ESICM definition (p = 0.002). Non-survivors had higher ultrasound scores than survivors (median 18.5 vs. 14, p = 0.04). The score correlated to the oxygenation index (ρ = -0.56, p = 0.03), and changes in the score between examinations correlated to changes in oxygenation (ρ = -0.41, p = 0.16). The correlation between the score and interleukin-6 was ρ = 0.35 (p < 0.001). The interrater reliability for the score was ICC = 0.87 (p < 0.001). CONCLUSIONS: The ultrasound score is a reliable tool that might help monitor disease severity and may help stratify the risk of mortality.

Flavin conjugates for delivery of peptide nucleic acids.

Oligonucleotides and their analogues, such as peptide nucleic acids (PNAs), can be used in chemical strategies to artificially control gene expression. Inefficient cellular uptake and inappropriate cellular localization still remain obstacles in biological applications, however, especially for PNAs. Here we demonstrate that conjugation of PNAs to flavin resulted in efficient internalization into cells through an endocytic pathway. The flavin-PNAs exhibited antisense activity in the sub-micromolar range, in the context of a treatment facilitating endosomal escape. Increased endosomal release of flavin conjugates into the cytoplasm and/or nucleus was shown by chloroquine treatment and also--when the flavin-PNA was conjugated to rhodamine, a mild photosensitizer--upon light irradiation. In conclusion, an isoalloxazine moiety can be used as a carrier and attached to a cargo biomolecule, here a PNA, for internalization and functional cytoplasmic/nuclear delivery. Our findings could be useful for further design of PNAs and other oligonucleotide analogues as potent antisense agents.

A steric blocker of translation elongation inhibits IGF-1R expression and cell transformation.

The insulin-like growth factor 1 receptor (IGF-1R) is involved in transformation, survival, mitogenesis and differentiation. It is overexpressed in many tumors and a validated target for anticancer therapy. In cell-free systems, polypyrimidic peptide nucleic acids (PNAs) can form triplex-like structures with messenger RNAs and halt the ribosomal machinery during the translation elongation. A 17-mer PNA that formed a PNA(2):mRNA complex with a purine-rich sequence located in the coding region of IGF-1R mRNA induced the synthesis of a truncated IGF-1R in vitro. This PNA down-regulated expression of the receptor by 70-80% in prostate cancer cells without affecting insulin receptor expression that exhibits high homology with IGF-1R. Inhibition occurs at the translational level, since the IGF-1R mRNA level measured by quantitative RT-PCR was not affected by PNA treatment. In addition, IGF-1R knockdown by PNA led to an attenuation of phosphorylation of downstream signaling pathways, PI3K/AKT and MAPK, involved in survival and mitogenesis and also to a decrease in cell transformation. Of the steric blockers tested, which included phosphorodiamidate morpholino oligomers and locked nucleic acids, PNA was unique in its ability to form triplex structures with mRNA and to arrest translation elongation.

Detection of high-grade lesions on cell blocks from residual fluids of Pap smears diagnosed as low-grade abnormalities: a preliminary pilot study.

OBJECTIVE: To investigate the feasibility of the technique of cell blocks (CBs) from residual fluids of Papanicolaou (Pap) smears diagnosed as low-grade abnormalities in the detection of high-grade lesions on biopsies. STUDY DESIGN: In the present pilot study, we evaluated the sensitivity and specificity of 70 CBs from liquid-based cervicovaginal smears of women with atypical squamous cells (ASCs) of undetermined significance (ASCUS; n = 39), ASCs that cannot exclude high-grade squamous intraepithelial lesion (ASC-H; n = 17) and low-grade squamous intraepithelial lesions (LSILs; n = 14) in the detection of cervical intraepithelial neoplasia (CIN) type 2+ lesions. RESULTS: Of the 70 CBs, 22 were diagnosed as negative, 27 as CIN1 and 21 as CIN2+. The sensitivity and specificity of CB preparation for the diagnosis of CIN2+ lesions were 50 and 100%, respectively, in ASCUS, 92 and 100%, respectively, in ASC-H, 100 and 100%, respectively, in LSILs. CONCLUSIONS: Our study confirms that CB preparation is a simple and reproducible technique with a good specificity that could be added advantageously to Pap smears to detect CIN2+ lesions in women with ASCs and LSILs.