remote-magnetic-actuation-micrometric-probes-for-situ-3d-mapping

Remote Magnetic Actuation of Micrometric Probes for <em>in situ</em> 3D Mapping of Bacterial Biofilm Physical Properties

Bacterial adhesion and growth on interfaces lead to the formation of three-dimensional heterogeneous structures so-called biofilms. The cells dwelling in ...

Sorbonne Universités, UPMC

In order to study the acclimatization process over 14 days of exposure to tropical climate, 9 triathletes performed 4 outdoor indirect continuous multistage tests in both thermoneutral and tropical conditions. The thermoneutral test (TN, 14 degree C, 45% rh) was performed before traveling to the tropical area (Martinique, FWI). The tropical tests were performed 2, 8, and 14 days after arrival (32.9 degree C, 78% rh). During each trial, we measured tympanic temperature, sweat rate, body mass loss, heart rate (HR), and performance. The results showed that 1). the mean tympanic temperature was greater in T2 (P <.001), T8 (P <.01) and T14 (P <.01) than in TN and significantly lower in T14 than in T2 (P <.05); 2). the mean sweat rate was significantly greater (P <.001) in T2, T8 and T14 than in TN and significantly greater (P <.05) in T8 and T14 than in T2; 3). the body mass loss after trials was significantly greater (P <.001) in T2, T8 and T14 than in TN and significantly greater (P <.05) in T8 and T14 than in T2; 4). the mean HR and HR at rest were significantly higher (P <.005) in T2 than in TN, T8, T14 and the mean HR was significantly lower (P <.05) in T14 than in the other trials; and 5). the performance time was significantly lower in T2 (P < 0.02), T8 (P < 0.03) and T14 (P < 0.05) than in TN. We concluded that 14 days of exposure to tropical climate led to changes in physiological parameters but were still insufficient to ensure complete acclimatization in well-trained athletes. The hot/wet climate induced impairment of physiological responses and performance that were still evident on the 14th day.

Effect of fourteen days of acclimatization on athletic performance in tropical climate.

The aim of this study was to examine the heart rate (HR) response to Olympic windsurfing competition and to check if there was any correlation between racing HR, performance, and the variables measured during laboratory maximal exercise. Ten elite windsurfers [age: 20.93 (3.46) years; height: 178.10 (6.34) cm; body mass: 66.79 (5.90) kg] performed a laboratory maximal oxygen consumption (.VO(2max)) trial and national windsurf competitions wearing a HR monitor. One hundred and forty-three individual races were examined. Racing HR was expressed as a percentage of (1) HR(max) (maximal treadmill HR) and (2) HR(reserve) (HR(max)-HR(rest)). The performance (racing classification: RC, which is inversely proportional to performance) was significantly correlated to the racing HR response in both light wind (LW): LW-RC=-0.12(%HR(reserve))+13.03; r=-0.71, r(2)=0.50, p<0.001, and medium wind (MW): MW-RC=-0.11(%HR(reserve))+10.99; r=-0.66, r(2)=0.43, p<0.001. The results showed similar correlations between performance and %HR(max). Post racing lactate concentration was higher in LW compared to MW [7.14 (0.21) and 5.18 (2.02) mmol.l(-1), respectively]. There was a negative correlation between the highest racing HR (%HR(reserve)) of each athlete and the second ventilatory threshold expressed as a percentage of .VO(2max) ( r=-0.71, p<0.05). To summarize, this study showed that light and medium wind Olympic windsurfing performances are highly dependent on the capacity of the athlete to maintain a high HR for long periods of time. Furthermore, windsurfing is highly dependent on the athlete's physical fitness level as shown by the correlations between racing HRs and laboratory physiological variables.

Correlation between heart rate and performance during Olympic windsurfing competition.

To study the pathophysiological mechanisms involved in the decrease of post-triathlon diffusing capacity (DLco), blood rheologic properties (blood viscosity: eta(b); changes in plasma volume: deltaPV) and atrial natriuretic factor (ANF) were assessed in ten triathletes during cycle-run (CR) and run-cycle (RC) trials at a metabolic intensity of 75% of maximal oxygen consumption ( VO(2max)). The DLco was measured before and 10 min after trials. ANF and deltaPV were measured at rest, after the cycle and run of CR and RC trials, and at the end of and 10 min after exercise. RC led to a greater deltaDLco decrease, a lower ANF concentration and a lower deltaPV than did CR, whereas for both CR and RC eta(b) was increased throughout exercise and 10 min after. In addition, after CR the deltaDLco decrease was inversely correlated ( r=-0.764; P<0.01) with deltaPV. The association of decreased plasma volume, increased eta(b), and lower ANF concentrations after RC suggested that lower blood pulmonary volume may have caused the greater decrease in Dlco as compared with CR. The inverse correlation between deltaPV and deltaDLco reinforces the hypothesis that fluid shifts limit the post-exercise DLco decrease after the CR succession in triathletes. Lastly, cycling in the crouched position might increase intra-thoracic pressure, decrease thorax volume due to the forearm position on the handlebars, and weaken peripheral muscular pump efficacy, all of which would limit venous return to the heart, and thus result in low pulmonary blood volume. Compared with cycling, running appeared to induce the opposite effects.

Effects of the order of running and cycling of similar intensity and duration on pulmonary diffusing capacity in triathletes.

Marine microbiologists commonly assay lipase activities by using a synthetic fluorescent analog, 4-methylumbelliferyl (MUF)-oleate. The technique is convenient, but it is considered to be unspecific because of the structure of this analog. This study reports the design of a new specific and sensitive lipase assay based on the use of a radiolabeled triglyceride, [3H]triolein. Free fatty acids (FFA) resulting from its hydrolysis are isolated as a function of time in a one-step liquid-liquid extraction and then radioassayed. MUF-oleate and [3H]triolein techniques were compared by measuring lipase activities at similar substrate concentrations along a trophic gradient in the Southwest Lagoon of New Caledonia, near Nouméa. Hydrolysis rates decreased from the nearshore station to the offshore station and showed similar trends regardless of the technique used. Rates decreased from 5.83 to 0.88 nmol of FFA. liter-1. h-1 and from 0.76 to 0.23 nmol of 3H-FFA. liter-1. h-1, respectively. These results appeared to be consistent with bacterial production results, which also decreased similarly (from 0.59 to 0.26 micrograms of C. liter-1. h-1). However, the ratio of MUF-oleate activities to [3H]triolein activities, which was constant at the offshore stations (3.8 +/- 0.1), gradually increased at the nearshore stations (from 4.1 to 7.6). This result shows that the two assays respond in different ways to changes in environmental conditions and validates the need to set up more specific enzymatic assays.

Application of a specific and sensitive radiometric assay for microbial lipase activities in marine water samples from the lagoon of nouméa.

The purpose of this study was to estimate the exercise intensity from the competition heart rate (HR) of professional triathletes during a multi-triathlon race. Five internationally ranked professional triathletes completed incremental cycling and running tests to assess the first and second ventilatory thresholds (i.e., VT and RCT) and the HR at VT and RCT. HR was then monitored during a 5 d multi-triathlon race: a prologue time trial (PTT, 0.2 km swim -- 5 km cycle -- 1.2 km run) that opened the race; short-distance triathlons (SHD; 1.3 km swim -- 36 km cycle -- 8.4 km run) performed on the 2nd and 5th days; and sprint-distance triathlons (SPD; 0.75 km swim -- 20 km cycle -- 5 km run) performed on the 3rd and 4th days. All trials except the last (i.e., the second SHD) were performed above HR corresponding to RCT. PTT elicited significantly higher mean HR than the other trials (except for the first SPD trial). In contrast, the last SHD elicited significantly lower HR than the other trials. These responses were globally similar in the 3 segments (i.e., swim, cycle, and run). This study demonstrates that the triathletes performed at very high intensity during a drafting-permitted multi-triathlon race. However, as shown for multi-day cycling distances, the HR responses depended on (i) the distance covered and (ii) group behavior.

Exercise intensity during repeated days of racing in professional triathletes.

To study the relationship between performance and exercise-induced arterial hypoxemia (EIAH), 5 internationally ranked (INT) and 8 regionally ranked (REG) triathletes performed cycle-run successions (CR) and control runs (R) in competition-like conditions: at approximately 75% VO2max.

Influence of performance level on exercise-induced arterial hypoxemia during prolonged and successive exercise in triathletes.

Aflatoxin B(1) (AFB(1)) is a risk factor for hepatocellular carcinoma (HCC) in many low-resource countries. Although its metabolites bind at several positions in TP53, a mutation at codon 249 (AGG to AGT, arginine to serine, p.R249S) accounts for 90% of TP53 mutations in AFB(1)-related HCC. This specificity suggests that p.R249S confers a selective advantage during hepatocarcinogenesis. Using HCC cell lines, we show that p.R249S has lost the capacity to bind to p53 response elements and to transactivate p53 target genes. In p53-null Hep3B cells, stable transfection of p.R249S or of another mutant, p.R248Q, did not induce significant changes in cell proliferation and survival after cytotoxic stress. In contrast, in a cell line that constitutively expresses both p.R249S and the hepatitis B virus antigen HBx (PLC/PRF/5), silencing of either p.R249S or HBx by RNA interference slowed down proliferation, with no additive effects when both factors were silenced. Furthermore, the two proteins appear to form a complex. In human HCC samples, mutation at codon 249 did not correlate with p.R249S protein accumulation or HBx truncation status. We suggest that p.R249S may contribute to hepatocarcinogenesis through interaction with HBx, conferring a subtle growth advantage at early steps of the transformation process, but that this interaction is not required for progression to advanced HCC.

Effects of the TP53 p.R249S mutant on proliferation and clonogenic properties in human hepatocellular carcinoma cell lines: interaction with hepatitis B virus X protein.

The objective of this work was to develop an economically relevant enzymatic process of butyl ester production using crude high-oleic sunflower oil. Novozym 435, a non-regiospecific biocatalyst, provided the best compromise between activity and butyl-ester yield. The inhibition caused by the presence of phopholipids in crude oil was eliminated by using tert-butanol. It demonstrates the key role of the medium polarity in order to insure the stability of a process. Initial substrate concentrations and their molar ratio were optimized in a continuous packed-bed reactor to maximize product yield and productivity. The best compromise was obtained for an initial oil concentration of 500 mM and a molar ratio of 5. It enabled a high productivity of 13.8 tons year(-1)kg Novozym 435(-1) with a butyl-ester purity of 96.5% to be obtained. Experiments with the continuous reactor were performed over 50 days without any loss of enzyme activity.

Continuous lipase-catalyzed production of esters from crude high-oleic sunflower oil.

Hepatocellular carcinoma (HCC) may develop according to two major pathways, one involving HBV infection and TP53 mutation and the other characterized by HCV infection and CTNNB1 mutation. We have investigated HBV/HCV infections and TP53/CTNNB1 mutations in 26 HCC patients from Thailand. HBV DNA (genotype B or C) was detected in 19 (73%) of the cases, including 5 occult infections and 3 coinfections with HCV. TP53 and CTNNB1 mutations were not mutually exclusive, and most of TP53 mutations were R249S, suggesting a significant impact of aflatoxin-induced mutagenesis in HCC development.

Mutations in TP53 and CTNNB1 in Relation to Hepatitis B and C Infections in Hepatocellular Carcinomas from Thailand.

Enzymatic transesterification of triglycerides in a continuous way is always a great challenge with a large field of applications for biodiesel, bio-lubricant, bio-surfactant, etc. productions. The lipase B from Candida antarctica (CalB) is the most appreciated enzyme because of its high activity and its non-regio-selectivity toward positions of fatty acid residues on glycerol backbone of triglycerides. Nevertheless, in the field of heterogeneous catalysis, we demonstrated that the medium hydrophilic nature of the support used for its commercial form (Lewatit VPOC1600) is a limitation. Glycerol is adsorbed onto support inducing drastic decrease in enzyme activity. Glycerol would form a hydrophilic layer around the enzyme resulting in diffusional limitations during triglyceride transfer to the enzyme. Accurel MP, a very hydrophobic macroporous polymer of propylene, was found not to adsorb glycerol. Immobilization conditions using this support were optimized. The best support was Accurel MP1001 (particle size<1000 μm) and a pre-treatment of the support with acetone instead of ethanol enables the adsorption rate and the immobilized enzyme quantity to be maximized. An economical approach (maximization of the process net present value) was expanded in order to explore the impact of immobilization on development of an industrial packed bed reactor. The crucial ratio between the quantity of lipase and the quantity of support, taking into account enzyme, support and equipped packed bed reactor costs was optimized in this sense. The biocatalyst cost was found as largely the main cost centre (2-10 times higher than the investments for the reactor vessel). In consequence, optimal conditions for immobilization were a compromise between this immobilization yield (90% of lipase immobilized), biocatalyst activity, reactor volume and total investments.

Selection of CalB immobilization method to be used in continuous oil transesterification: analysis of the economical impact.

Most bacteria live in the form of adherent communities forming three-dimensional material anchored to artificial or biological surfaces, with profound impact on many human activities. Biofilms are recognized as complex systems but their physical properties have been mainly studied from a macroscopic perspective. To determine biofilm local mechanical properties, reveal their potential heterogeneity, and investigate their relation to molecular traits, we have developed a seemingly new microrheology approach based on magnetic particle infiltration in growing biofilms. Using magnetic tweezers, we achieved what was, to our knowledge, the first three-dimensional mapping of the viscoelastic parameters on biofilms formed by the bacterium Escherichia coli. We demonstrate that its mechanical profile may exhibit elastic compliance values spread over three orders of magnitude in a given biofilm. We also prove that heterogeneity strongly depends on external conditions such as growth shear stress. Using strains genetically engineered to produce well-characterized cell surface adhesins, we show that the mechanical profile of biofilm is exquisitely sensitive to the expression of different surface appendages such as F pilus or curli. These results provide a quantitative view of local mechanical properties within intact biofilms and open up an additional avenue for elucidating the emergence and fate of the different microenvironments within these living materials.

Mapping of bacterial biofilm local mechanics by magnetic microparticle actuation.

The authors investigated the anthropometric and physiological characteristics of young Guadeloupian competitive swimmers in relation to swimming performance and compared the abilities of these children with those of the young white swimmers reported in the literature. All 2004 competitive swimmers between 10 and 14 y old (126 children, 61 boys and 65 girls, 12.0 ± 1.3 y) from Guadeloupe underwent anthropometric measurements and physiological and performance testing. Six boys on the French national swimming team are referred to hereafter as the 2011 elite subgroup. Anthropometric parameters, a jump-and-reach test, glide, and estimated aerobic power (eVO2max) were assessed in terms of swimming-performance analysis through a 400-m test. This study demonstrated that the Guadeloupian swimmers had more body fat than most age-matched white swimmers but had very poor hydrostatic lift; they had higher peak jump height and they swam as well as their white counterparts. The variability in 400-m performance between subjects was best described by glide, age, and eVO2max. Compared with the group of boys with the same age, the 2011 elite subgroup was significantly better for arm span, peak jump height, glide, and 400-m and 15-m performances. Further research is needed to investigate motor organization and energy cost of swimming in Afro-Caribbean swimmers.

Anthropometric and physiological characteristics in young afro-Caribbean swimmers. A preliminary study.

Postexercise alveolar-capillary membrane-diffusing capacity (DLco) often decreases in highly trained endurance athletes and seems linked to their training status. To test the hypothesis that training status influences postexercise DLco, 13 male and 2 female triathletes (20.2 ± 4.4 y old, 175.2 ± 6.7 cm tall; weight in a range of 66.6 ± 7.4 kg to 67.4 ± 7.8 kg during the 1-y study) were randomized into experimental (n = 7) and control (n = 8) groups and performed VO(2max) cycle tests and simulated cycle-run successions (CR) of 30 min + 20 min after 3 periods in the competitive season.

Aggravation of pulmonary diffusing capacity in highly trained athletes by 6 weeks of low-volume, low-intensity training.

We tested whether the silicone swim caps (SC) worn by young swimmers in a tropical climate negatively influence aerobic performance. Nine trained pre- adolescents [11.8 (± 0.8) years] swam randomized 800-m trials (water: 32.9°C, outdoors: shade, 29.2 ± 0.2 °C, 74 ± 0.3 % rh) with a SC or a nude head (NH). Performance times and heart rate (HR) were monitored continuously. Rectal temperature (Trec) was measured before and after trials. The rating of perceived exertion (RPE) was assessed. Stroke frequency (SF), stroke length (SL) and stroke index (SI) were measured every 50-m. The SC trial was significantly longer than NH (799 ± 16 and 781 ± 16 seconds, respectively). Mean delta Trec was significantly greater in SC (0.2 ± 0.1°C vs. -0.1 ± 0.1°C in SC vs. NH), mean SI was significantly different in SC versus NH (1.83 ± 0.07 vs 1.73 ± 0.06); but RPE and mean HR, SF and SL showed no change. We conclude that a silicone swim cap worn in tropical environment significantly decreased 800-m crawl performance without affecting HR or RPE. Silicone swim caps worn by young swimmers in a tropical environment may also have negative effects on training capacity. Key pointsSwimming in tropical climate represents a physiological stressSwimming with swim cap in warm water could induce thermal stressThermoregulation processes have to be used in order to make training in tropical climate safer.

Olivier Galy

Physicochime Curie,
CNRS UMR 168,
Institut Curie,
Physicochime Curie, CNRS UMR 168, Institut Curie

Sorbonne Universités, UPMC

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties

Olivier Galy

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Sorbonne Universités, UPMC

Remote Magnetic Actuation of Micrometric Probes for in situ 3D Mapping of Bacterial Biofilm Physical Properties

Physicochime Curie,
CNRS UMR 168,
Institut Curie,
Physicochime Curie, CNRS UMR 168, Institut Curie