Multicenter Evaluation of the Clinical Performance and the Neutralizing Antibody Activity Prediction Properties of 10 High-Throughput Serological Assays Used in Clinical Laboratories.

As the coronavirus disease 2019 (COVID-19) pandemic second wave is emerging, it is of the upmost importance to screen the population immunity in order to keep track of infected individuals. Consequently, immunoassays for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with high specificity and positive predictive values are needed to obtain an accurate epidemiological picture. As more data accumulate about the immune responses and the kinetics of neutralizing-antibody (nAb) production in SARS-CoV-2-infected individuals, new applications are forecast for serological assays such as nAb activity prediction in convalescent-phase plasma from recovered patients. This multicenter study, involving six hospital centers, determined the baseline clinical performances, reproducibility, and nAb level correlations of 10 commercially available immunoassays. In addition, three lateral-flow chromatography assays were evaluated, as these devices can be used in logistically challenged areas. All assays were evaluated using the same patient panels in duplicate, thus enabling accurate comparison of the tests. Seven immunoassays examined in this study were shown to have excellent specificity (98 to 100%) and good to excellent positive predictive values (82 to 100%) when used in a low (5%)-seroprevalence setting. We observed sensitivities as low as 74% and as high as 95% at ≥15 days after symptom onset. The determination of optimized cutoff values through receiver operating characteristic (ROC) curve analyses had a significant impact on the diagnostic resolution of several enzyme immunoassays by increasing the sensitivity significantly without a large trade-off in specificity. We found that spike-based immunoassays seem to be better correlates of nAb activity. Finally, the results reported here will add to the general knowledge of the interlaboratory reproducibility of clinical performance parameters of immunoassays and provide new evidence about nAb activity prediction.

Evaluating age-related changes of some facial signs among men of four different ethnic groups.

BACKGROUND: The alterations of some facial characteristics, with age, in men of different ethnic groups are a scarcely studied topic. OBJECTIVE: To illustrate and grade, among men of four different ethnic descents (Asians, African-Americans, Caucasians, Indians), aged 18-80 year, the changes in the severity of some facial signs occurring with age. METHODS: Digital Photographs (full face, profile, 45°) of 1058 subjects were taken under standard conditions of lightning. These allowed us to zoom in on 15-20 different facial signs on all subjects. Their observations by 15 trained experts, under calibrated screens, allowed these signs to be graded, with the help of four referential skin Atlases, specific to the gender and ethnic groups studied. RESULTS: Most facial signs show a rather regular increase with age, with different kinetics according to the ethnic group. The major changes, in all four ethnic groups, mostly concern skin structural alterations (wrinkles, neck sagging or texture). The changes in the severity of some wrinkles (forehead, inter-ocular…) may be viewed as ethnic signatures. The modifications in pigmentation disorders seem more closely linked to the constitutive skin pigmentation (photo-types), among which pigmentary spots are confirmed as a more specific Asian trait. CONCLUSION: Men of four different ethnic origins present different and ethnic-related facial alterations with increasing age, of different amplitudes.

Influences of age, ethnic group, and skin sites on a provisory skin marking, experimentally induced, in vivo.

BACKGROUND: To study capacity of skin to being imprinted by a marking head rapidly applied (10 seconds) onto the skin under a standardized pressure (15 kPa). MATERIAL AND METHODS: Referential photographs of skin markings, resulting from different times of application serve at grading the extent of skin imprints. The imprinting tool, equipped with 5 linear bulbs of different sizes but same thickness (1 mm), was applied onto the cheeks of 641 women of different ages (18-80 years) of 3 different ethnic groups. In the cohort of 198 Caucasian women, the marking head was pressed onto 3 different sites (outer arm, cheek, and dorsal forearm) and the kinetics of skin recovery from cheek marking was recorded on a smaller and younger part of this Caucasian cohort, ie, 141 Caucasian women, aged 18-59 years. RESULTS: The recorded intensities of skin markings were found highly correlated with age in all 3 studied ethnic groups (Chinese, African-American, and Caucasian). Caucasian women seem more sensitive to the marking and the kinetics of recovery from such imprint was found highly dependent from its intensity, ie, the higher the grades, the slower the recovery. In Caucasian women, intensities were different between skin sites, where arm showed higher resistance to marking. Possible impact of photo-aging on the decreased skin resistance to such provoked deformation is likely of minor amplitude, as compared to the one induced by chronological aging. CONCLUSION: This method offers a new vision on skin aging, in its variable response to a standard deformation that appears mostly driven by chronological aging.

A new tool to quantify the geometrical characteristics of facial skin pores. Changes with age and a making-up procedure in Caucasian women.

BACKGROUND: Facial skin pores (FSP) are common and benign signs that generate frequent esthetic concerns or complaints. Despite their worldwide prevalence, related literature remains scarce. Hence, a new device has been developed and applied to validating studies, aiming at best describing FSP as they are self-perceived, i.e. their anatomic features, their possible alterations with age and their appearance after application of a make-up product. METHODS: Dermascore+® is an imaging device dedicated to a direct observation and acquisition of various characteristics of the skin surface. Images are captured under a high magnification and under different lighting configurations, to highlight the skin relief, based upon parallel polarized images. Dedicated software allows FSP to being detected and their morphological parameters to being extracted and computed. By measuring each detected FSP in a given region of interest, a statistically significant impact of both age and an applied cosmetic product upon their morphological features can be observed and quantified. RESULTS: Although the size and density of FSP are not affected by aging, their shape becomes elongated. A few tested make up products show variable effects that closely correlate with visual assessments made by trained estheticians. CONCLUSION: The shape of FSP present on cheeks shows age-related changes, toward a more elongated aspect, likely linked to a progressively altered (more isotropic) skin surface micro-relief. The new tool Dermascore+® allows foundations to being rapidly differentiated and screened according to their variable effects upon the visual appearance through instrumental, objective depiction of FSP.

Influence of gravity upon some facial signs.

OBJECTIVES: Facial clinical signs and their integration are the basis of perception than others could have from ourselves, noticeably the age they imagine we are. Facial modifications in motion and their objective measurements before and after application of skin regimen are essential to go further in evaluation capacities to describe efficacy in facial dynamics. Quantification of facial modifications vis à vis gravity will allow us to answer about 'control' of facial shape in daily activities. METHODS: Standardized photographs of the faces of 30 Caucasian female subjects of various ages (24-73 year) were successively taken at upright and supine positions within a short time interval. All these pictures were therefore reframed - any bias due to facial features was avoided when evaluating one single sign - for clinical quotation by trained experts of several facial signs regarding published standardized photographic scales. RESULTS: For all subjects, the supine position increased facial width but not height, giving a more fuller appearance to the face. More importantly, the supine position changed the severity of facial ageing features (e.g. wrinkles) compared to an upright position and whether these features were attenuated or exacerbated depended on their facial location. Supine station mostly modifies signs of the lower half of the face whereas those of the upper half appear unchanged or slightly accentuated. These changes appear much more marked in the older groups, where some deep labial folds almost vanish. These alterations decreased the perceived ages of the subjects by an average of 3.8 years. CONCLUSION: Although preliminary, this study suggests that a 90° rotation of the facial skin vis à vis gravity induces rapid rearrangements among which changes in tensional forces within and across the face, motility of interstitial free water among underlying skin tissue and/or alterations of facial Langer lines, likely play a significant role.

Effect of fertilizer formulation and bioaugmentation on biodegradation and leaching of crude oils and refined products in soils.

The effects of soil characteristics and oil types as well as the efficacy of two fertilizer formulations and three bioaugmentation packages in improving the bioremediation of oil-contaminated soils were assessed as a means of ex situ treatment selection and optimization through seven laboratory microcosm studies. The influence of bioremediation on leaching of oil from the soil was also investigated. The studies demonstrated the benefits ofbiostimulation to overcome nutrient limitation, as most of the soils were nutrient depleted. The application of both liquid and pelleted slow-release N and P fertilizers increased both the hydrocarbon biodegradation rates (by a factor of 1.4 to 2.9) and the percentage of hydrocarbon mass degraded (by > 30% after 12 weeks and 80% after 37 weeks), when compared with the unamended soils. Slow-release fertilizers can be particularly useful when multiple liquid applications are not practical or cost-effective. Bioaugmentation products containing inoculum plus fertilizer also increased biodegradation by 20% to 37% compared with unamended biotic controls; however, there was no clear evidence of additional benefits due to the inocula, compared with fertilizer alone. Therefore biostimulation is seen as the most cost-effective bioremediation strategy for contaminated soils with the levels of crude oil and refined products used in this study. However, site-specific considerations remain essential for establishing the treatability of oil-contaminated soils.

Instrumental and clinical studies of the facial skin tone and pigmentation of Shanghaiese women. Changes induced by age and a cosmetic whitening product.

The pigmentation patterns of facial skin of 354 healthy Chinese women aged 18-80 years were investigated clinically and instrumentally. Chromasphere(®) was used to acquire pictures from the cheeks of subjects. Facial skin tone was described by L* parameter from the L,a,b system as well as Individual Typology Angle (ITA). Results show that skin tone becomes significantly darker along the life span. Both size of hyper-pigmented spots and their contrast with surrounding skin were found increased with age. As additional study, 40 women from these 354 subjects were asked to apply daily a whitening cosmetic product for a 2-month period. Such application led to a significantly lighter skin tone, although this study was not vehicle controlled and we cannot exclude that the increase in L* observed was in some part because of cumulative effects of previously used whitening products, there was an association with lighter skin tone as assessed through both instrumental measurements and self-perception by most subjects.

Neutralization of mitogenic lectins by intravenous immunoglobulin (IVIg) prevents T cell activation: does IVIg really have a direct effect on T cells?

Intravenous immunoglobulin (IVIg) is used for the treatment of an increasing number of autoimmune diseases. Clinical observations on IVIg-treated patients have revealed a modulation of T cell populations and functions in these patients. In vitro studies aimed at understanding the mechanisms underlying the effects of IVIg on T cells led to the conclusion that IVIg directly affected lectin-activated T cell functions. However, more recent studies have suggested the absence of a direct effect of IVIg on T cells. In the present work, we revisited the effect of IVIg on T cells using lectin-stimulated human T cells and showed that IVIg inhibited T cell functions only when added simultaneously with the activating lectin. Further, we showed that IVIg depleted from lectin-reactive IgG was no longer inhibitory, suggesting that the effect of IVIg on T cells was the consequence of lectin neutralization, possibly by interaction with glycans present in F(ab')(2) portion of IgG molecules. Our results challenge the previously widely accepted notion that IVIg exerts its anti-inflammatory effects by acting directly on T cells and suggest that effects of IVIg observed in treated patients are rather a consequence of the recently reported inhibitory effect of IVIg on antigen presentation.

Influence of season on some skin properties: winter vs. summer, as experienced by 354 Shanghaiese women of various ages.

The facial skin of 354 women, aged 18-80, living in Shanghai, was investigated over two successive 6-month periods, summer and winter. Results from clinical assessments indicate that aging signs, such as wrinkling and sagging, are unaffected over such period. However, physical measurements revealed alterations in some functional criteria of the skin, such as sebum output, skin colour, melanin content of pigmented spots, skin hydration, all being increased during summer. The relationships between all criteria, as well as technical or applied inferences/consequences from this study, are discussed.

Immunomodulatory effects of therapeutic preparations of human albumin.

BACKGROUND AND OBJECTIVES: Albumin is the most abundant protein in plasma and is considered to be immunologically inert. However, we recently observed that therapeutic human albumin preparations, used as protein control in studies involving high doses of IVIg, modulated the MHC II-restricted activation of antigen-specific T cells. In the present work, we characterized this effect in more details. MATERIALS AND METHODS: An in vitro antigen presentation assay using mouse cells was used to evaluate the effect of therapeutic human albumin preparations on the activation of ovalbumin-specific T cells. Flow cytometry and quantitative real-time PCR were used to monitor the expression of genes involved in this process. RESULTS: Therapeutic human albumin preparations increased T cell activation in a dose-dependent manner. The effect was explained by an increase in the expression of MHC II and of two other genes (CIITA and H2-M) involved in antigen presentation by murine monocytic cells. Similarly, the expression of HLA-DR on the surface of human monocytic cells was increased following incubation with therapeutic human albumin preparations. CONCLUSION: Altogether, these results reveal a possible physiological role of albumin in immunological processes, leading to an increased ability of antigen presenting cells to trigger T cell activation. This immunomodulatory effect needs to be considered, at least in studies in which albumin is used as a presumably inert control protein.

Clinical study on the effects of a cosmetic product on dermal extracellular matrix components using a high-resolution multiphoton tomograph.

BACKGROUND/PURPOSE: The aim of this study was to demonstrate the effects of selected plant extracts in a cosmetic cream on the dermal network components after a 3-month treatment using an in vivo multiphoton tomographic device. METHODS: Twenty-four Caucasian women aged between 45 and 65 applied randomly a cosmetic emulsion B containing active ingredients (soy and jasmine) twice a day on one arm and its vehicle A (without active ingredients) on the other arm during 3 months. Measurements were performed on the internal side of the forearm before starting the treatment (T0), after 4 week (T4) and 12 weeks (T12) treatment. Measurements consisted of a multi-layers acquisitions using a multiphoton tomograph with subcellular resolution. Optical sections (about 6 microm thick) were recorded from 0 to about 200 microm using two different wavelengths: 760 and 820 nm. To compare the series of images and obtain an objective quantification of the signal of second harmonic generation (SHG) and autofluorescence, the method used consisted of taking the integrated brightness of an image (same rectangular area for all images) as a measure of the signal. Following this step, a ratio between brightness of images from the area treated with cream A or B and brightness of untreated area was calculated and used as an assessment of treatment efficacy. The parameter used for statistical analysis (variance analysis) is the difference before and after 12 weeks of treatment by either cream A or B of the signal ratios calculated in the upper dermis (118-130 microm) and those from a deeper region of the upper dermis (165-178 microm). RESULTS: Signals (autofluorescence+SHG) of extracellular matrix do not change significantly with time (weeks 0, 4 and 12) when cream A (vehicle with no active ingredient) is applied. Treatment with cream B results in an enhancement in the signal level of extracellular matrix at week 12. The comparison of signals, in both areas (118-130 microm and 160-178 microm), show an higher increase in the deeper region than in the more superficial one for product B while we do not notice any change with product A. CONCLUSION: The multiphoton tomograph provided excellent high-resolution images, which describe clearly the different skin layers, single cells and extracellular matrix components in all the 24 volunteers. Statistic analyses reveal a real effect for product B with selected plant extracts, known to increase collagen synthesis. Changes observed are characteristics of modifications in dermal collagen and elastin content. To our knowledge, it is the first time that it was possible to demonstrate in vivo the effect of a cosmetic product on the superficial dermal layer, in a non-invasive and non-destructive process, i.e. without cutting the skin.

In vitro and in vivo confocal Raman study of human skin hydration: assessment of a new moisturizing agent, pMPC.

The hydration capacities of a biomimetic polymer, 2-methacryloyloxethylphosphorylcholine polymer (pMPC), alone and microencapsulated, in association with another well known hydrating polymer, Hyaluronic acid, were investigated in vitro on skin models and in vivo on volunteers by using confocal Raman microspectroscopy. The hydration impact and the relative water content in the Stratum corneum were calculated from the Raman spectra using the OH (water)/CH3 (protein) ratio. Moreover, the follow-up of the presence of pMPC through the Stratum corneum was possible with confocal Raman microspectroscopy, using a characteristic vibration of pMPC, different from that of the encapsulating material. From our in vitro measurements, the improved hydration of the Stratum corneum was confirmed by the use of the encapsulated form of pMPC, which was higher when combined with Hyaluronic acid. On the basis of these in vitro findings, we validated this trend in in vivo measurements on 26 volunteers, and found a good correlation with the in vitro results. Mechanical and ultrastructural studies have been carried out to demonstrate the positive effects of the pMPC on the Stratum corneum function, namely the interaction with lamellar lipids and the plasticizing effects, which are both supposed to spell out the moisturizing effect. This study demonstrates the efficiency of a original hydrating agent, pMPC, entrapped with Hyaluronic acid in a new type of microcapsules by the use of a novel tool developed for both in vitro and in vivo approaches. This indicates a new step to evaluate and improve new moisturizers in response to the cosmetics or dermatologic demands.

Equivalence of face and volar forearm for the testing of moisturizing and firming effect of cosmetics in hydration and biomechanical studies.

The objective of the study was to compare measurements of skin hydration and of biomechanical properties performed on different zones of face and volar forearm. Two short-term (1 h) and two long-term (3 weeks) studies were conducted with a moisturizing and a firming product, respectively, on groups of female volunteers with dry skin. Measurements (Corneometer) and Dermal Torque Meter or DTM were performed on different zones of the face and of the volar forearm, 1 h after product application, and after 1, 2 and 3 weeks of repeated twice daily application. While the sebaceous-gland rich T-zone behaves differently, probably due to sebum/skin, sebum/product and/or sebum/measuring device interactions, there are no statistically significant differences between measurements made on temple, cheek, maxilla and volar forearm. The volar forearm is representative of the face for measuring skin hydration and biomechanical properties, and relevant for the assessment of the efficacy of cosmetic products destined for facial use.

[Comparison of eight deep breaths and tidal volume breathing preoxygenation techniques in morbid obese patients]. / Comparaison de la technique de préoxygénation à huit capacités vitales et à volume courant chez les patientes ayant une obésité morbide.

OBJECTIVE: To compare two techniques of preoxygenation, eight deep breaths (8DB) and tidal volume breathing in obese patients by measuring end-tidal fractional oxygen concentration (FETO2) and apnea time from 100% of hemoglobin saturation to 95% (T95%). STUDY DESIGN: Prospective randomized study. METHODS: Twenty obese patients (BMI >40 kg/m2) without cardiorespiratory disease nor difficult intubation criteria were randomized into two groups of ten. One group received preoxygenation with eight deep breaths in one minute (8DB) and the other with three minutes tidal volume preoxygenation (3TV) both under FIO2 100%. FETO2 every minute of preoxygenation and T95% were measured. Data were analyzed with Mann and Whitney test. A p <0.05 was considered significant. RESULT: There was no significant difference between the groups regarding FETO2 values [84 +/- 4% (8DB) and 88 +/- 5% (3TV)] and T95% [176 +/- 23 s (8DB) and 181 +/- 35 s (3TV)]. The PETCO2 was significantly inferior in the 8DB group at the end of preoxygenation [PETCO2 =29 +/- 1 mmHg (8DB) and PETCO2 =36 +/- 5 mmHg (3TV)]. CONCLUSION: 8DB and 3TV preoxygenation techniques in morbid obese patients induce similar FETO2 and T95%. However hyperventilation effects in the 8DB group are unknown.

Adipocyte functions are modulated by cell size change: potential involvement of an integrin/ERK signalling pathway.

OBJECTIVES: Adipocyte is the only cell whose size may vary dramatically in physiological conditions. We hypothesized that increase in fat cell size per se could modulate several signalling pathways by changing the relationships between the cell and the extracellular matrix. The aim of the current study was (i). to examine whether within the same fat depot, metabolic functions of adipocyte were modified by cell size and (ii). if such an adaptation exists, to look for an integrin/extracellular-signal-regulated kinases (ERKs) signalling pathway. RESULTS: We isolated two populations of adipocytes with different volumes (67 and 22 x 10(3) microm(3)) within the same adipose location. In large compared to small fat cells, fatty acid synthase and lipoprotein lipase activities were increased two- and seven-fold, respectively; GLUT4 protein concentration and leptin expression were increased three-fold; lipolytic capacity was increased four-fold. The integrin/ERK signalling pathway could be the one responsible for the adaptation of adipose functions to cell size. In large compared with small adipocytes, we showed that beta(1)-integrins are present in adipose membranes and at a higher concentration in large than in small cells. In isolated adipocytes, stimulation of beta(1)-integrins with a specific monoclonal antibody results in ERK(1) and ERK(2) activation. In large compared to small cells, cytoplasmic concentrations of these two mitogen-activated protein kinases were increased two-fold, whereas their activities were increased 10-fold. CONCLUSION: A beta(1)-integrin/ERKs signalling pathway is present in mature adipocyte. Increase in cell size, by modifying the relationships between cell and extracellular matrix, could turn on this pathway. Since ERKs can modulate transcription factors and subsequently modulate gene expression important for adipose function, this pathway could play an important role in the adaptation of adipose functions to cell size.

Impaired beta-adrenergic signaling pathway in white adipocytes of suckling fa/fa Zucker rats: a defect in receptor coupling.

BACKGROUND: In fa/fa Zucker rats, leptin receptor deficiency is responsible for both a deficit of energy expenditure and hyperphagia which lead to massive obesity and insulin resistance in adulthood. This obesity is also characterised by alterations of the beta-adrenergic signaling pathway. OBJECTIVE: To determine whether alterations in beta-adrenergic pathway could occur at the onset of obesity when fa/fa rats are not yet hyperinsulinemic. ANIMALS: Fourteen-day-old suckling fa/fa and Fa/fa littermates (from heterozygous lean (Fa/fa) female and homozygous obese (fa/fa) male mating). MEASUREMENTS: Membranes were prepared from isolated adipocytes after collagenase treatment of inguinal adipose tissue. The response of adenylyl-cyclase activity to stimulation by isoprenaline, GTPgamma-S or forskolin was studied. Bmax and Kd of (beta1+beta2) and of beta3 adrenoceptors were measured using 3H-CGP saturation binding experiments. mRNA concentration of beta1- and beta3-AR was determined by semi-quantitative RT-PCR. G(s)alpha protein was quantified by Western blotting and Gi protein by ADP-ribosylation. RESULTS: Despite an almost normal body weight, inguinal fat pad weight was increased two-fold by the expression of fa mutation. This increase was entirely accounted for by fat cell hypertrophy (x2.5 in volume). In fa/fa compared to Fa/fa pups, response of adenylyl cyclase to isoprenaline was decreased two-fold but responses to GTPgammaS or forskolin were unchanged. Density of (beta1+beta2) and beta3-AR was not affected by the fa/fa genotype, as well as G(s)alpha and Gi concentration. CONCLUSION: Response of inguinal fat cells to catecholamines was decreased without any quantitative modifications of the different elements of the adenylyl cyclase cascade. This suggests an alteration in the coupling between beta-AR and G proteins. Due to the important increase in fat cell volume we hypothesize that changes in the physical properties of plasma membranes and/or changes in cytoskeleton-extracellular-matrix interactions could disturb the beta-adrenergic pathway responsiveness. In addition to the excess of lipid storage, which occurs very early at the onset of obesity, the impairment of the responsiveness to catecholamines reported in this study might worsen the obesity syndrome.

Bilateral severe keratoconus after laser in situ keratomileusis in a patient with forme fruste keratoconus.

We report a case of bilateral keratoconus after laser in situ keratomileusis (LASIK). Before surgery, the patient had a forme fruste keratoconus, which evolved rapidly to a severe form of keratoconus in the months following LASIK. From this case, we conclude that forme fruste keratoconus is a contraindication to LASIK.

Conjugated dienes: a critical trait of lipoprotein oxidizability in renal fibrosis.

BACKGROUND: We assessed whether a differential oxidizability of apolipoprotein B (apo B)-containing lipoproteins (LDL and VLDL) may explain the oxidative stress that we had observed at the onset of renal fibrosis in Zucker obese (ZO) rats (Nephrol Dial Transplant 2000, 15: 467--476). METHODS: Ex vivo copper-induced oxidation of lipoproteins was performed in 1-, 3-, and 9-month-old ZO and age-matched lean (ZL) rats. LDL/VLDL oxidizability was determined by spectrophotometry at 234 nm by monitoring the formation of conjugated diene hydroperoxides. RESULTS: A significant increase in lag time (reflecting the resistance to oxidation) was observed in ZO rats at 3 months while the maximal diene production (reflecting the amount of hydroperoxides formed during oxidation) was higher in ZO than in ZL rats as early as 1 month. Lipoproteins were larger in ZO than in ZL rats, as shown by their core to surface component ratio. Furthermore, ZO lipoproteins had increased vitamin E and polyunsaturated fatty acid (PUFA) content, with no change in vitamin E/PUFA ratio. CONCLUSIONS: Rather than oxidizability of apo B-containing lipoproteins, the ability of these molecules to produce high levels of conjugated dienes, which can act as toxic tissue messengers, appears to be a critical trait in the development of renal fibrosis in this rat model of obesity and renal fibrosis.