Culture Medium Factorial Design Optimization for Fibrinolytic Enzymes Production by Bionectria sp.

Thrombotic diseases can be clinically treated with fibrinolytic enzymes and many attempts have been made at laboratory level to increase fibrinolytic enzymes production from microbial sources and to reduce the process cost, including culture medium design, optimization of environmental conditions, and over expression with genetically modified strains. In this contribution we present the optimization of culture medium composition and incubation temperature for fibrinolytic enzyme production by Bionectria sp., a selected fungal strain from Las Yungas (Tucumán). Optimization was carried out at Erlenmeyer scale (100-mL working volume) via factorial design methodology. All trials included a common mineral base (%, w/v: NaCl 0.2, KH2PO4 0.05, MgSO4·7H2O 0.05). According to four factorial designs it could be demonstrated the convenience of using soy peptone as N-source, glucose as C-source, and the possibility to eliminate starch, meat peptone and meat extract from original medium composition, whilst 25°C was selected as the optimal incubation temperature. Results showed that culture medium could be successfully optimized by factorial design, achieving a reduction in the production process costs by means of a decrease in culture medium components, the improvement in culture broth rheology, mycelial morphology and mass/energy transfer, and the subsequent two-fold enhancement in productivity.Fil: Arnau, Victor Gonzalo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Rovati, Jose Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Figueroa, L. I. C.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Fariña, Julia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaXLVI Reunión Anual Sociedad Argentina de Investigaciones Bioquímicas y MolecularesPuerto Madryn, Chubut, ArgentinaArgentinaSociedad Argentina de Investigaciones Bioquímicas y Moleculare

Validity of Daily Physical Activity Measurements of Fitbit Charge 2

Physical activity monitors collect continuous data to provide a summary of daily activity. The Fitbit Charge 2 monitors heart rate as well as steps, calories, and active minutes throughout the day. There is currently no research validating the Fitbit Charge 2 at measuring daily physical activity levels in a real life setting. PURPOSE: To compare measures of daily steps and active minutes of Fitbit Charge 2 with a research-grade accelerometer. METHODS: Sixteen active college students (Mean±SD; 23±4.9yrs; 16.43±10.19%fat; 9 male) consented to be part of the study. Participants wore an ActiGraph GT3X accelerometer and Fitbit Charge 2 concurrently for seven consecutive days. Both devices were programed with each participant’s information and the participants were instructed to perform their daily activities wearing both devices and only remove them to shower and to sleep. Data were considered valid when participants wore both devices for at least 10 hours on 4 or more days of the week. Steps and active minutes (moderate-vigorous physical activity) were recorded by each device. Mean bias was calculated by subtracting ActiGraph steps and active minutes from those obtained from the Fitbit Charge 2 for each day and an average daily mean bias was calculated using values from all seven days. Absolute percentage error was also calculated [100(|Fitbit Charge 2 - ActiGraph|)/ActiGraph] to indicate the overall 7-day difference between the Fitbit Charge 2 and ActiGraph. Pearson correlations and paired sample t-test were performed to compare Fitbit Charge 2 measurements with the corresponding ActiGraph measurements with significance considered at p\u3c0.05. RESULTS: The Fitbit Charge 2 overestimated steps by 2,451.3±2085.4 compared to the ActiGraph using the daily average steps over the seven days. This was 32.2±40.7% above the ActiGraph measurement. Average mean bias for daily active minutes was -52.1±58.9 with the Fitbit Charge 2 underestimating compared to the ActiGraph. Active minutes for the Fitbit Charge 2 were an average of 69±26.1% away from the ActiGraph. Steps for the Fitbit Charge 2 were significantly correlated to ActiGraph steps (r=0.575, p=0.02) while active minutes were not significantly correlated (r= -0.255, p=0.34). Paired sample t-test results showed a significant difference between the Fitbit Charge 2 steps and active minutes compared with the ActiGraph (p\u3c0.01 for both). CONCLUSION: The Fitbit Charge 2 may be useful for measuring steps in a free-living environment, however active minutes are significantly underestimated

Accuracy of Fitbit Charge 2 at Estimating VO2max, Calories, and Steps on a Treadmill

Current fitness activity trackers can account for steps, calories burned, heart rate, and distance traveled. A new feature has recently been introduced on the Fitbit Charge 2, “Cardio Fitness Level,” which is comparable to a VO2max score in that it allows consumers to be aware of their aerobic fitness level. PURPOSE: To assess the accuracy of the Fitbit Charge 2 at estimating VO2 score (“Cardio Fitness Level”), calories, and steps when compared to indirect calorimetry and video analyzed steps, respectively. METHODS: Twenty-two healthy adults (Mean±SD; 24.1±4.2yrs; 16.9±9.0%fat; 15 male) completed two separate visits. On the first visit, anthropometric measurements were taken followed by a 10-minute outdoor run. Participants ran for 10 minutes at their own pace on flat terrain as recommended by Fitbit to generate a Cardio Fitness score. On the second visit, participants came fasted, at least 8 hours, and completed a standardized VO2max protocol (Arizona State protocol) using a PARVO TrueOne2400 metabolic cart. The treadmill was set at 3mph for the first 3 minutes with 0% grade. Following the first stage, the speed was raised to the participant’s pre-selected speed (between 5-8mph) with 0% grade. After stage 2 the grade increased every minute by 1.5% and speed was kept constant until fatigue was reached. Calories and step counts from the Fitbits were correlated with the metabolic cart and tally counter respectively, using 2-tailed Pearson correlations. Significance was set at pRESULTS: Participants completed the VO2max test in an average of 11:05. Eight of the 22 estimated VO2max ranges given by Fitbit included the value given by the metabolic cart. Fitbit ranges for seven participants were below the metabolic cart values and the Fitbit ranges for the remaining seven participants were above the metabolic cart values. Calories were correlated between the Fitbit and metabolic cart (r = 0.874, pCONCLUSION: VO2 scores given by the Fitbit Charge 2 did not always match values given by the metabolic cart but may serve as a rough estimate of fitness level. Fitbit Charge 2 may also be useful in tracking calories and steps in a controlled setting, but results may differ in real world conditions

Thyroid hormone regulates distinct paths to maturation in pigment cell lineages

Thyroid hormone (TH) regulates diverse developmental events and can drive disparate cellular outcomes. In zebrafish, TH has opposite effects on neural crest derived pigment cells of the adult stripe pattern, limiting melanophore population expansion, yet increasing yellow/orange xanthophore numbers. To learn how TH elicits seemingly opposite responses in cells having a common embryological origin, we analyzed individual transcriptomes from thousands of neural crest-derived cells, reconstructed developmental trajectories, identified pigment cell-lineage specific responses to TH, and assessed roles for TH receptors. We show that TH promotes maturation of both cell types but in distinct ways. In melanophores, TH drives terminal differentiation, limiting final cell numbers. In xanthophores, TH promotes accumulation of orange carotenoids, making the cells visible. TH receptors act primarily to repress these programs when TH is limiting. Our findings show how a single endocrine factor integrates very different cellular activities during the generation of adult form

AAV2/8-hSMAD3 gene delivery attenuates aortic atherogenesis, enhances Th2 response without fibrosis, in LDLR-KO mice on high cholesterol diet

BACKGROUND: Inflammation is a key etiologic component in atherogenesis and transforming growth factor beta 1 (TGFβ1) is a well known anti-inflammatory cytokine which potentially might be used to limit it. Yet TGFβ1 is pleiomorphic, causing fibrosis, cell taxis, and under certain circumstances, can even worsen inflammation. SMAD3 is an important member of TGFβ1′s signal transduction pathway, but is a fully intracellular protein. OBJECTIVES: With the hope of attenuating TGFβ1′s adverse systemic effects (eg. fibrosis) and accentuating its anti-inflammatory activity, we proposed the use of human (h)SMAD3 as an intracellular substitute for TGFβ1. STUDY DESIGN: To test this hypothesis adeno-associated virus type 2/8 (AAV)/hSMAD3 or AAV/Neo (control) was tail vein injected into the low density lipoprotein receptor knockout (LDLR-KO) mice, then placed on a high-cholesterol diet (HCD). RESULTS: The hSMAD3 delivery was associated with significantly lower atherogenesis as measured by larger aortic cross sectional area, thinner aortic wall thickness, and lower aortic systolic blood velocity compared with Neo gene-treated controls. HSMAD3 delivery also resulted in fewer aortic macrophages by immunohistochemistry for CD68 and ITGAM, and quantitative reverse transcriptase polymerase chain reaction analysis of EMR and ITGAM. Overall, aortic cytokine expression showed an enhancement of Th2 response (higher IL-4 and IL-10); while Th1 response (IL-12) was lower with hSMAD3 delivery. While TGFβ1 is often associated with increased fibrosis, AAV/hSMAD3 delivery exhibited no increase of collagen 1A2 or significantly lower 2A1 expression in the aorta compared with Neo-delivery. Connective tissue growth factor (CTGF), a mediator of TGFβ1/SMAD3-induced fibrosis, was unchanged in hSMAD3-delivered aortas. In the liver, all three of these genes were down-regulated by hSMAD3 gene delivery. CONCLUSION: These data strongly suggest that AAV/hSMAD3 delivery gave anti-atherosclerosis therapeutic effect without the expected undesirable effect of TGFβ1-associated fibrosis

The effects of UV radiation on photosynthesis estimated as chlorophyll fluorescence in Zygnemopsis decussata (Chlorophyta) growing in a high mountain lake (Sierra Nevada, Southern Spain)

The effect of increased UV radiation on photosynthesis estimated as in vivo chlorophyll fluorescence i.e. optimal quantum yield (Fv/Fm) and electron transport rate (ETR) in the green filamentous alga Zygnemopsis decussata (Streptophyta, Zygnematales) growing in the high mountain lake "La Caldera" (Sierra Nevada, Spain) at 3050 m altitude was evaluated. Two sets of in situ experiments were conducted: (1) On July 2006, Fv/Fm was measured throughout the day at different depths (0.1, 0.25, 0.5 and 1 m) and in the afternoon, ETR and phenolic compounds were determined. In addition, in order to analyze the effect of UV radiation, Fv/Fm was determined in algae incubated for 3 days at 0.5m under three different light treatments: PAR+UVA+UVB (PAB), PAR+UVA (PA) and PAR (P). (2) On August 2007, Fv/Fm was determined under PAB, PA and P treatments and desiccation/rehydration conditions. Fv/Fm decreased in algae growing in surface waters (0.1 m) but also at 1 m depth compared to that at 0.5 m depth. The decrease of Fv/Fm at noon due to photoinhibition was small (less than 10%) except in algae growing at 1 m depth (44%). The maximal electron transport rate was 3.5-5 times higher in algae growing at 0.25-0.5 m respectively than that at 0.1 and 1 m depth. These results are related to the accumulation of phenolic compounds: i.e. the algae at 0.25-0.5 m presented respectively about a 3-5 times higher concentration of phenolic compounds than that of algae at 0.1-1 m depth. The protection mechanisms seem to be stimulated by UVB radiation, since Fv/Fm was higher in the presence of UVB (PAB treatment) compared to PA or P treatments. UVA exerts the main photoinhibitory effect, not only at midday, but also in the afternoon. UVB radiation also had a protective effect in algae grown under desiccation conditions for three days. During re-hydration, the rapid increase of Fv/Fm (after 1 h) was higher in the UVB-grown algae than in algae grown under UVA radiation. After 5 h, Fv/Fm values were similar in algae submitted to desiccation/rehydration under PAB and P treatments as they were in the control (submerged algae). The combined effect of desiccation and UVA produced the greatest decrease of photosynthesis in Z. decussata. Thus UVB, in contrast to other species, may support the recovery process. Z. decussata can acclimate to severe stress conditions in this high mountain lake by the photoprotection mechanism induced by UVB radiation through dynamic photoinhibition and the accumulation of phenolic compounds (UV screen and antioxidant substances).This research was supported by the Spanish Ministries of Environment (PN2003/25) and Education and Science (CGL2005/01564, AGL2005/02655, CGL 2008/01127, CGL 2008/05407) and Consejería de Innovación, Ciencia y Empresa, Junta de Andalucía (Excelencia project P07-CVI-02598)

Comparison of Smartphone Pedometer Apps on a Treadmill versus Outdoors

Previous research has focused on the accuracy of smartphone pedometer apps in laboratory settings, however less information is available in outdoor (free living) environments. PURPOSE: Determine the accuracy of 5 smartphone apps at recording steps at a walking speed in a laboratory versus an outdoor setting. METHODS: Twenty-three healthy college students consented (Mean±SD; 22±3.8yrs; BMI 24.9±4.13kg/m2) to participate in 2 separate visits. During the first visit participants walked 500 steps at 3mph on a treadmill while wearing a pedometer and a smartphone placed in the pocket using 5 pedometer apps concurrently (Moves, Google Fit (G-Fit), Runtastic, Accupedo, S-Health). During the second visit, participants walked 400 meters at 3mph on a sidewalk outside. Actual steps for each visit were recorded using a hand tally counter device. Zero and negative values were replaced with the mean value for that trial. Statistical analyses were performed using IBM SPSS 23.0. Mean bias scores were calculated between the step count for each app and the respective tally count for each trial. Mean bias scores were correlated between trials for each app using Pearson correlations and significance was set at p\u3c0.05. Mean Absolute Percent Error (MAPE) values were also calculated for each app for both trials. RESULTS: G-Fit recorded 2 zero values and 2 negative values and Moves recorded 1 zero value. Mean bias scores were significantly correlated between the indoor and outdoor protocols for the pedometer (r=0.67, p\u3c0.01) and S-Health (r=0.46, p\u3c0.5). The remaining apps were not correlated between protocols. The outdoor protocol producing a greater mean bias for the outdoor protocol for G-Fit, Runtastic, and Accupedo (mean bias ± SD indoor, outdoor; -4.3±53.1, -19.3±120.0; -10.7±63.3, -33.4±118.7; 16.0±143.6, 79.0±75.0; respectively) and a greater mean bias for the indoor protocol for the pedometer, Moves, and S-Health (mean bias indoor, outdoor; -1.4±41.5, 0.0±34.1; -117.4±196.7, -42.2±209.6; 11.3±28.4, 0.0±58.7; respectively). MAPE was below 5% for the pedometer and S-Health for both trials. CONCLUSION: Apps with the lowest error in a controlled setting may be less affected when used in other settings, while apps with greater variation in a controlled setting may be affected when used in a different environment

Accuracy of Fitbit Activity Trackers During Walking in a Controlled Setting

Activity trackers are widely used to measure daily physical activity. Many devices have been shown to measure steps more accurately at higher intensities, however, it is also important to determine the accuracy of these new devices at measuring steps while walking at a pace similar to that used during most daily activities. PURPOSE: To assess the accuracy of 6 popular activity trackers at measuring steps while walking on a treadmill. METHODS: Twenty-six college students (Mean±SD; 22.1±3.7yrs; 25.1±4.0kg/m2; 13 male) walked 500 steps at 3mph on a treadmill while wearing 6 different activity trackers (Pedometer, Fitbit Blaze, Charge HR, Alta, Flex, Zip, One). The Charge HR was placed two fingers above the right wrist while the Flex was next to the wrist bone. The Blaze was placed two fingers above the left wrist while the Alta was next to the wrist bone. The Fitbit Zip and the One were aligned with the hipbone on the left and right waistband respectively. Steps were counted by a trained researcher using a hand tally counter. Missing values were replaced with the mean value for that device. Step counts were correlated between Fitbit devices and the pedometer and tally counter using Pearson correlations. Significance was set at p\u3c0.05. Mean bias scores were calculated between the step counts for each device and the tally counter. Mean Absolute Percent Error (MAPE) values were also calculated for each device relative to the tally counter. RESULTS: Fitbit Zip and One were significantly correlated with the tally counter (r=0.50, p\u3c0.05; r=0.68, p\u3c0.01, respectively) while the other devices were not significantly correlated. Mean bias and MAPE values were as follows: Device (Mean Bias/MAPE) Pedometer (-0.2±39.2/3.8±6.8), Blaze (34.5±67.1/9.9±11.3), Charge HR (-12.6±61.5/7.0±10.3), Alta (-85.0±70.8/17.1±14.1), Flex (49.5±242.4/19.7±45.3), Zip (1.8±3.4/0.4±0.6), One (0.2±2.1/0.3±0.3). Fitbit Zip and One were within one half percent of actual steps while wrist-worn Fitbits ranged from 7.0-19.7% from actual step counts. CONCLUSION: Consistent with previous research, activity trackers worn at the waist provide the most accurate step counts compared to wrist-worn models. Differences found in wrist-worn models may result in significant over- or underestimation of activity levels when worn for long periods of time