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Viruses target mitochondria to promote their replication, and infection-induced stress during the progression of infection leads to the regulation of antiviral defenses and mitochondrial metabolism which are opposed by counteracting viral factors. The precise structural and functional changes that underlie how mitochondria react to the infection remain largely unclear. Here we show extensive transcriptional remodeling of protein-encoding host genes involved in the respiratory chain, apoptosis, and structural organization of mitochondria as herpes simplex virus type 1 lytic infection proceeds from early to late stages of infection. High-resolution microscopy and interaction analyses unveiled infection-induced emergence of rough, thin, and elongated mitochondria relocalized to the perinuclear area, a significant increase in the number and clustering of endoplasmic reticulum-mitochondria contact sites, and thickening and shortening of mitochondrial cristae. Finally, metabolic analyses demonstrated that reactivation of ATP production is accompanied by increased mitochondrial Ca2+ content and proton leakage as the infection proceeds. Overall, the significant structural and functional changes in the mitochondria triggered by the viral invasion are tightly connected to the progression of the virus infection.

Summary: Background: Although immunomodulators have established benefit against the new coronavirus disease (COVID-19) in general, it is uncertain whether such agents improve outcomes without increasing the risk of secondary infections in the specific subgroup of previously immunocompromised patients. We assessed the effect of immunomodulators on outcomes of immunocompromised patients hospitalized for COVID-19. Methods: The protocol was prospectively registered with PROSPERO (CRD42022335397). MEDLINE, Cochrane Central Register of Controlled Trials and references of relevant articles were searched up to 01-06-2022. Authors of potentially eligible randomized controlled trials were contacted to provide data on immunocompromised patients randomized to immunomodulators vs control (i.e., placebo or standard-of-care). Findings: Eleven randomized controlled trials involving 397 immunocompromised patients hospitalized for COVID-19 were included. Ten trials had low risk of bias. There was no difference between immunocompromised patients randomized to immunomodulators vs control regarding mortality [30/182 (16.5%) vs 41/215 (19.1%); RR 0.93, 95% CI 0.61–1.41; p = 0.74], secondary infections (RR 1.00, 95% CI 0.64–1.58; p = 0.99) and change in World Health Organization ordinal scale from baseline to day 15 (weighed mean difference 0.27, 95% CI -0.09–0.63; p = 0.15). In subgroup analyses including only patients with hematologic malignancy, only trials with low risk of bias, only trials administering IL-6 inhibitors, or only trials administering immunosuppressants, there was no difference between comparators regarding mortality. Interpretation: Immunomodulators, compared to control, were not associated with harmful or beneficial outcomes, including mortality, secondary infections, and change in ordinal scale, when administered to immunocompromised patients hospitalized for COVID-19. Funding: Hellenic Foundation for Research and Innovation.

Abstract Ultrasound-guided hydrodissection with 5% dextrose in water (DW5) creates a peri-nervous compartment, separating the nerve from the neighboring anatomical structures. The aim of this randomized study was to determine the minimum volume of lidocaine 2% with epinephrine 1:200,000 required when using this technique to achieve an effective median nerve block at the elbow in 95% of patients (MEAV95). Fifty-two patients scheduled for elective hand surgery received an ultrasound-guided circumferential perineural injection of 4 ml DW5 and an injection of local anesthetic (LA) following a biased coin up-and-down sequential allocation method. A successful block was defined as a light touch completely suppressed on the two distal phalanges of the index finger within a 30-min evaluation period. The MEAV95 of lidocaine 2% with epinephrine was 4 ml [IQR 3.5–4.0]. Successful median nerve block was obtained in 38 cases (82.6%) with median onset time of 20.0 [10.0–21.2] minutes (95% CI 15–20). The analgesia duration was 248 [208–286] minutes (95% CI 222–276). Using an ultrasound-guided hydrodissection technique with DW5, the MEAV95 to block the median nerve at the elbow with 2% lidocaine with epinephrine was 4 ml [IQR 3.5–4.0]. This volume is close to that usually recommended in clinical practice. Trial registration clinicaltrials.gov. NCT02438657, Date of registration: May 8, 2015.

Summary: The group B Coxsackieviruses (CVB), belonging to the Enterovirus genus, can establish persistent infections in human cells. These persistent infections have been linked to chronic diseases including type 1 diabetes. Still, the outcomes of persistent CVB infections in human pancreas are largely unknown. We established persistent CVB infections in a human pancreatic ductal-like cell line PANC-1 using two distinct CVB1 strains and profiled infection-induced changes in cellular protein expression and secretion using mass spectrometry-based proteomics. Persistent infections, showing characteristics of carrier-state persistence, were associated with a broad spectrum of changes, including changes in mitochondrial network morphology and energy metabolism and in the regulated secretory pathway. Interestingly, the expression of antiviral immune response proteins, and also several other proteins, differed clearly between the two persistent infections. Our results provide extensive information about the protein-level changes induced by persistent CVB infection and the potential virus-associated variability in the outcomes of these infections. : Biological Sciences; Microbiology; Virology; Proteomics Subject Areas: Biological Sciences, Microbiology, Virology, Proteomics

Summary: Dopaminergic (DA) neurons have been implicated as key targets in neurological disorders, notably those involving locomotor impairment, and are considered to be highly vulnerable to mitochondrial dysfunction, a common feature of such diseases. Here we investigated a Drosophila model of locomotor disorders in which functional impairment is brought about by pan-neuronal RNAi knockdown of subunit COX7A of cytochrome oxidase (COX). Despite minimal neuronal loss by apoptosis, the expression and activity of tyrosine hydroxylase was decreased by half. Surprisingly, COX7A knockdown specifically targeted to DA neurons did not produce locomotor defect. Instead, using various drivers, we found that COX7A knockdown in specific groups of cholinergic and glutamatergic neurons underlay the phenotype. Based on our main finding, the vulnerability of DA neurons to mitochondrial dysfunction as a cause of impaired locomotion in other organisms, including mammals, warrants detailed investigation.

Plants and many lower organisms, but not mammals, express alternative oxidases (AOXs) that branch the mitochondrial respiratory chain, transferring electrons directly from ubiquinol to oxygen without proton pumping. Thus, they maintain electron flow under conditions when the classical respiratory chain is impaired, limiting excess production of oxygen radicals and supporting redox and metabolic homeostasis. AOX from Ciona intestinalis has been used to study and mitigate mitochondrial impairments in mammalian cell lines, Drosophila disease models and, most recently, in the mouse, where multiple lentivector-AOX transgenes conferred substantial expression in specific tissues. Here, we describe a genetically tractable mouse model in which Ciona AOX has been targeted to the Rosa26 locus for ubiquitous expression. The AOXRosa26 mouse exhibited only subtle phenotypic effects on respiratory complex formation, oxygen consumption or the global metabolome, and showed an essentially normal physiology. AOX conferred robust resistance to inhibitors of the respiratory chain in organello; moreover, animals exposed to a systemically applied LD50 dose of cyanide did not succumb. The AOXRosa26 mouse is a useful tool to investigate respiratory control mechanisms and to decipher mitochondrial disease aetiology in vivo.

Mitochondria have been increasingly recognized as a central regulatory nexus for multiple metabolic pathways, in addition to ATP production via oxidative phosphorylation (OXPHOS). Here we show that inducing mitochondrial DNA (mtDNA) stress in Drosophila using a mitochondrially-targeted Type I restriction endonuclease (mtEcoBI) results in unexpected metabolic reprogramming in adult flies, distinct from effects on OXPHOS. Carbohydrate utilization was repressed, with catabolism shifted towards lipid oxidation, accompanied by elevated serine synthesis. Cleavage and translocation, the two modes of mtEcoBI action, repressed carbohydrate rmetabolism via two different mechanisms. DNA cleavage activity induced a type II diabetes-like phenotype involving deactivation of Akt kinase and inhibition of pyruvate dehydrogenase, whilst translocation decreased post-translational protein acetylation by cytonuclear depletion of acetyl-CoA (AcCoA). The associated decrease in the concentrations of ketogenic amino acids also produced downstream effects on physiology and behavior, attributable to decreased neurotransmitter levels. We thus provide evidence for novel signaling pathways connecting mtDNA to metabolism, distinct from its role in supporting OXPHOS.

Abstract The alternative oxidase (AOX) from Ciona intestinalis was previously shown to be expressible in mice and to cause no physiological disturbance under unstressed conditions. Because AOX is known to become activated under some metabolic stress conditions, resulting in altered energy balance, we studied its effects in mice subjected to dietary stress. Wild‐type mice (Mus musculus, strain C57BL/6JOlaHsd) fed a high‐fat or ketogenic (high‐fat, low‐carbohydrate) diet show weight gain with increased fat mass, as well as loss of performance, compared with chow‐fed animals. Unexpectedly, AOX‐expressing mice fed on these metabolically stressful, fat‐rich diets showed almost indistinguishable patterns of weight gain and altered body composition as control animals. Cardiac performance was impaired to a similar extent by ketogenic diet in AOX mice as in nontransgenic littermates. AOX and control animals fed on ketogenic diet both showed wide variance in weight gain. Analysis of the gut microbiome in stool revealed a strong correlation with diet, rather than with genotype. The microbiome of the most and least obese outliers reared on the ketogenic diet showed no consistent trends compared with animals of normal body weight. We conclude that AOX expression in mice does not modify physiological responses to extreme diets.

Mycobacterium tuberculosis remains one of the most problematic infectious agents, owing to its highly developed mechanisms to evade host immune responses combined with the increasing emergence of antibiotic resistance. Host-directed therapies aiming to optimize immune responses to improve bacterial eradication or to limit excessive inflammation are a new strategy for the treatment of tuberculosis. In this study, we have established a zebrafish-Mycobacterium marinum natural host-pathogen model system to study induced protective immune responses in mycobacterial infection. We show that priming adult zebrafish with heat-killed Listeria monocytogenes (HKLm) at 1 day prior to M. marinum infection leads to significantly decreased mycobacterial loads in the infected zebrafish. Using rag1−/− fish, we show that the protective immunity conferred by HKLm priming can be induced through innate immunity alone. At 24 h post-infection, HKLm priming leads to a significant increase in the expression levels of macrophage-expressed gene 1 (mpeg1), tumor necrosis factor α (tnfa) and nitric oxide synthase 2b (nos2b), whereas superoxide dismutase 2 (sod2) expression is downregulated, implying that HKLm priming increases the number of macrophages and boosts intracellular killing mechanisms. The protective effects of HKLm are abolished when the injected material is pretreated with nucleases or proteinase K. Importantly, HKLm priming significantly increases the frequency of clearance of M. marinum infection by evoking sterilizing immunity (25 vs 3.7%, P=0.0021). In this study, immune priming is successfully used to induce sterilizing immunity against mycobacterial infection. This model provides a promising new platform for elucidating the mechanisms underlying sterilizing immunity and to develop host-directed treatment or prevention strategies against tuberculosis. This article has an associated First Person interview with the first author of the paper.

Abstract The machinery of mitochondrial DNA (mtDNA) maintenance is only partially characterized and is of wide interest due to its involvement in disease. To identify novel components of this machinery, plus other cellular pathways required for mtDNA viability, we implemented a genome‐wide RNAi screen in Drosophila S2 cells, assaying for loss of fluorescence of mtDNA nucleoids stained with the DNA‐intercalating agent PicoGreen. In addition to previously characterized components of the mtDNA replication and transcription machineries, positives included many proteins of the cytosolic proteasome and ribosome (but not the mitoribosome), three proteins involved in vesicle transport, some other factors involved in mitochondrial biogenesis or nuclear gene expression, > 30 mainly uncharacterized proteins and most subunits of ATP synthase (but no other OXPHOS complex). ATP synthase knockdown precipitated a burst of mitochondrial ROS production, followed by copy number depletion involving increased mitochondrial turnover, not dependent on the canonical autophagy machinery. Our findings will inform future studies of the apparatus and regulation of mtDNA maintenance, and the role of mitochondrial bioenergetics and signaling in modulating mtDNA copy number.

This article analyzes the beginnings of the reception theory in German philosophy, then its developments. It argues that the logical sense of this theory lays in a new definition of the use, with the example of the reception of Brian de Palma’s Scarface in the French suburbs.

INTRODUCTION:Diabetes Associated Protein in Insulin-sensitive Tissues (DAPIT) is a subunit of mitochondrial ATP synthase and has also been found to associate with the vacuolar H+-ATPase. Its expression is particularly high in cells with elevated aerobic metabolism and in epithelial cells that actively transport nutrients and ions. Deletion of DAPIT is known to induce loss of mitochondrial ATP synthase but the effects of its over-expression are obscure. RESULTS:In order to study the consequences of high expression of DAPIT, we constructed a transgenic cell line that constitutively expressed DAPIT in human embryonal kidney cells, HEK293T. Enhanced DAPIT expression decreased mtDNA content and mitochondrial mass, and saturated respiratory chain by decreasing H+-ATP synthase activity. DAPIT over-expression also increased mitochondrial membrane potential and superoxide level, and translocated the transcription factors hypoxia inducible factor 1α (Hif1α) and β-catenin to the nucleus. Accordingly, cells over-expressing DAPIT used more glucose and generated a larger amount of lactate compared to control cells. Interestingly, these changes were associated with an epithelial to mesenchymal (EMT)-like transition by changing E-cadherin to N-cadherin and up-regulating several key junction/adhesion proteins. At physiological level, DAPIT over-expression slowed down cell growth by G1 arrest and migration, and enhanced cell detachment. Several cancers also showed an increase in genomic copy number of Usmg5 (gene encoding DAPIT), thereby providing strong correlative evidence for DAPIT possibly having oncogenic function in cancers. CONCLUSIONS:DAPIT over-expression thus appears to modulate mitochondrial functions and alter cellular regulations, promote anaerobic metabolism and induce EMT-like transition. We propose that DAPIT over-expression couples the changes in mitochondrial metabolism to physiological and pathophysiological regulations, and suggest it could play a critical role in H+-ATP synthase dysfunctions.

Recent epidemiological and laboratory-based studies suggest that the anti-diabetic drug metformin prevents cancer progression. How metformin diminishes tumor growth is not fully understood. In this study, we report that in human cancer cells, metformin inhibits mitochondrial complex I (NADH dehydrogenase) activity and cellular respiration. Metformin inhibited cellular proliferation in the presence of glucose, but induced cell death upon glucose deprivation, indicating that cancer cells rely exclusively on glycolysis for survival in the presence of metformin. Metformin also reduced hypoxic activation of hypoxia-inducible factor 1 (HIF-1). All of these effects of metformin were reversed when the metformin-resistant Saccharomyces cerevisiae NADH dehydrogenase NDI1 was overexpressed. In vivo, the administration of metformin to mice inhibited the growth of control human cancer cells but not those expressing NDI1. Thus, we have demonstrated that metformin's inhibitory effects on cancer progression are cancer cell autonomous and depend on its ability to inhibit mitochondrial complex I.

Cyanide-resistant non-phosphorylating respiration is known in mitochondria from plants, fungi, and microorganisms but is absent in mammals. It results from the activity of an alternative oxidase (AOX) that conveys electrons directly from the respiratory chain (RC) ubiquinol pool to oxygen. AOX thus provides a bypath that releases constraints on the cytochrome pathway and prevents the over-reduction of the ubiquinone pool, a major source of superoxide. RC dysfunctions and deleterious superoxide overproduction are recurrent themes in human pathologies, ranging from neurodegenerative diseases to cancer, and may be instrumental in ageing. Thus, preventing RC blockade and excess superoxide production by means of AOX should be of considerable interest. However, because of its energy-dissipating properties, AOX might produce deleterious effects of its own in mammals. Here we show that AOX can be safely expressed in the mouse (MitAOX), with major physiological parameters being unaffected. It neither disrupted the activity of other RC components nor decreased oxidative phosphorylation in isolated mitochondria. It conferred cyanide-resistance to mitochondrial substrate oxidation and decreased reactive oxygen species (ROS) production upon RC blockade. Accordingly, AOX expression was able to support cyanide-resistant respiration by intact organs and to afford prolonged protection against a lethal concentration of gaseous cyanide in whole animals. Taken together, these results indicate that AOX expression in the mouse is innocuous and permits to overcome a RC blockade, while reducing associated oxidative insult. Therefore, the MitAOX mice represent a valuable tool in order to investigate the ability of AOX to counteract the panoply of mitochondrial-inherited diseases originating from oxidative phosphorylation defects.

Abstract  The objective of this preliminary study was to evaluate the usefulness of front face fluorescence spectroscopy to predict some chemical parameters [pH, fat, dry matter (DM), fat in DM, total nitrogen (TN) and water soluble nitrogen (WSN)] and cheese meltability of semi-hard and hard cheeses. Dynamic testing rheology was used to determine the melting point of cheeses corresponding to the temperature at which loss tangent (tan δ) = 1. Tryptophan and vitamin A fluorescence spectra were, also, recorded on cheese samples at 20 °C. The partial least squares (PLS) regression with the leave one-out cross-validation technique was used to build up calibration models. Excellent predictions were obtained from the tryptophan and vitamin A models for fat (R 2 = 0.99 and 0.97, respectively), DM (R 2 = 0.94 and 0.96, respectively), fat in DM (R 2 = 0.92 and 0.99, respectively), TN (R 2 = 0.91 and 0.91, respectively). Excellent predictions were also obtained for WSN (R 2 = 0.96) and melting point (R 2 = 0.97) from vitamin A spectra, while only good predictions for these two parameters (R 2 = 0.90 and R 2 = 0.87, respectively) were obtained from tryptophan spectra. The results for pH were good (R 2 = 0.82) and approximate (R 2 = 0.76) with tryptophan and vitamin A, respectively. [ABSTRACT FROM AUTHOR]

Abstract  Ten traditional M1 (n = 5) and M2 (n = 5) soft cheeses produced from raw milk, and five other stabilised M3 (n = 5) cheeses manufactured from pasteurised milk, were studied using mid infrared (MIR) and front face fluorescence (FFFS) spectroscopies. MIR (3000–900 cm−1), tryptophan (excitation: 290 nm, emission: 305-450 nm), 400-640 emission spectra (excitation: 380 nm) and vitamin A (excitation: 280–350 nm, emission: 410 nm) spectra were recorded at two sampling zones (external (E) and central (C)) of the investigated cheeses. When the factorial discriminant analysis (FDA) was applied to the MIR spectra, the classification was not satisfactory. With tryptophan fluorescence spectra, correct classification of 94.4 and 69.4% was observed for the calibration and validation spectra, respectively. Better classification was obtained using vitamin A fluorescence spectra, since 91.8 and 80.6% of the calibration and validation spectra, respectively, were correctly classified. When the first five principal components (PCs) of the PCA extracted from each data set were pooled into a single matrix and analysed by FDA, the classification was considerably improved, obtaining a percentage of correct classification of 100 and 91.7% for the calibration and validation samples, respectively. It was concluded that concatenation of the physico-chemical and spectroscopic data sets is an efficient technique for the identification of soft cheese varieties. [ABSTRACT FROM AUTHOR]

Abstract  Near infrared (NIR) spectroscopy was used to predict colour of European Emmental cheese samples. Colour values (L, a and b) were measured on 20 Emmental cheese samples using a Hunter-lab D25-D-2 optical head in the system according to Hunter to determine L (brightness), a (green-red component) and b (blue-yellow component). The diffuse reflectance of the investigated cheeses was also determined by a B�chi NIR Lab N-200 spectrometer using a rotating measuring cell in the range of 1000–2500 nm. The best results for L-value (squared correlation coefficient (R 2) = 0.56, root mean square error of cross-validation (RMSECV) = 0.76, ratio of prediction deviation (RPD) = 1.89 and range error ratio (RER) = 7.91), a-value (R 2 = 0.72, RMSECV = 0.15, RPD = 1.98 and RER = 7.6) and b-value (R 2 = 0.82, RMSECV = 0.52, RPD = 2.56 and RER = 9.42) were obtained when the first 12 principal components (PCs) of the principal component analysis (PCA) applied on normalised NIR spectra were used. It can be concluded that NIR spectroscopy could be used to predict b-value. The a- and L-values can also be predicted from NIR technique with approximate quantitative prediction. [ABSTRACT FROM AUTHOR]

The main objective of this study was to characterise the know-how diversity of small-scale facilities manufacturing traditional dry sausages and to investigate the relations between the know-how and the sensory diversity of the traditional dry sausages. First, a description of the know-how (typology) was observed on 108 out of 129 registered small-scale facilities in the Massif Central. The typology led to six groups with specific know-how differentiated mainly by the raw materials and drying. From these 6 groups, 12 small-scale facilities were selected (2 per group of know-how) in order to perform a Flash profile on the manufactured dry sausages with a panel of six experts. The Flash profile results showed discrimination between the dry sausages by the raw materials used. In addition, the canonical correlation analysis (CCA) showed a high correlation between the typology data and the sensory data. The similarity map 1–2 from the CCA illustrated the proximity between dry sausages from the same group of know-how and discriminated between dry sausages from different groups of know-how. Thus, it can be concluded that each group of know-how corresponded to traditional dry sausages with specific sensory characteristics. [ABSTRACT FROM AUTHOR]

The certification of aeronautical composite structures is based on a pragmatic approach, which is intended to be safe and essentially experimental but with a strong test/calculation dialogue called the “Test Pyramid”. However, this has proved to be extremely expensive and it appears necessary to reduce its cost either by developing Virtual testing, or by developing richer tests on an intermediate scale between coupon specimens and structural parts. It was in the aim of meeting this objective that the VERTEX program (French acronym for “Experimental modeling and Validation of compositE strucTures under complEX loading”) was launched in 2012. After positioning the VERTEX program in relation to the literature, this review will explain the methodology and present the measurement methods specifically developed for this scale. Then, three scientific themes that have been studied will be detailed (large notches, impact and wrinkling case studies). Finally, a proposal for validating the structures using envelope curves will be put forward, an assessment made, and perspectives presented.

The development of experimental conditions for rapid bacterial discrimination using fluorescence spectroscopy fingerprinting is presented. Colonies of Pseudomonas and related reference strains on agar plates were analyzed directly using an optic fiber coupled to a laboratory spectrofluorimeter. Spectra were collected using either classic fluorescence spectroscopy after excitation at 250 nm and 340 nm for aromatic amino acids and nucleic acids (AAA + NA) and nicotinamide adenine dinucleotide (NADH) respectively, or synchronous scanning in the excitation wavelength range 250-500 nm. Factorial discriminant analysis (FDA) showed 100% correct classification at the genus and species level from NADH spectra and 100% correct classification at the genus and species level for λ = 30, 70, 90 and 110 nm (cross-validation). Analysis of variance (ANOVA) confirmed that culture time (48 or 72 h) colony and optic fiber positioning had non-significant impacts on differences between species. The use of optical fiber-fluorescence spectroscopy for bacterial discrimination directly on colonies is fast, simple and reliable. The results are independent of culture growth phase and neither need reagent addition nor prior manual preparation of cells, thus eliminating all risk of human error or contamination during sample work-up.