Scale-up from shake flasks to fermenters in batch and continuous mode with Corynebacterium glutamicum on lactic acid based on oxygen transfer and pH

Scale-up from shake flasks to fermenters has been hampered by the lack of knowledge concerning the influence of operating conditions on mass transfer, hydromechanics, and power input. However, in recent years the properties of shake flasks have been described with empirical models. A practical scale-up strategy for everyday use is introduced for the scale-up of aerobic cultures from shake flasks to fermenters in batch and continuous mode. The strategy is based on empirical correlations of the volumetric mass transfer coefficient (k(L) a) and the pH. The accuracy of the empirical k(L) a correlations and the assumptions required to use these correlations for an arbitrary biological medium are discussed. To determine the optimal pH of the culture medium a simple laboratory method based on titration curves of the medium and a mechanistic pH model, which is solely based on the medium composition, is applied. The effectiveness of the scale-up strategy is demonstrated by comparing the behavior of Corynebacterium glutamicum on lactic acid in shake flasks and fermenters in batch and continuous mode. The maximum growth rate (micro(max) = 0.32 h(-1)) and the oxygen substrate coefficient (Y O2 /S= 0.0174 mol/l) of C. glutamicum on lactic acid were equal for shake flask, fermenter, batch, and continuous cultures. The biomass substrate yield was independent of the scale, but was lower in batch cultures (Y(X/S) = 0.36 g/g) than in continuous cultures (Y(X/S) = 0.45 g/g). The experimental data (biomass, respiration, pH) could be described with a simple biological model combined with a mechanistic pH model.     

Extracting Objective Estimates of Sedentary Behavior from Accelerometer Data: Measurement Considerations for Surveillance and Research Applications

BackgroundAccelerometer-based activity monitors are widely used in research and surveillance applications for quantifying sedentary behavior (SB) and physical activity (PA). Considerable research has been done to refine methods for assessing PA, but relatively little attention has been given to operationalizing SB parameters (i.e., sedentary time and breaks) from accelerometer data - particularly in relation to health outcomes. This study investigated: (a) the accrued patterns of sedentary time and breaks; and (b) the associations of sedentary time and breaks in different bout durations with cardiovascular risk factors.MethodsAccelerometer data on 5,917 adults from the National Health Examination and Nutrition Survey (NHANES) 2003–2006 were used. Sedentary time and breaks at different bout durations (i.e., 1, 2–4, 5–9, 10–14, 15–19, 20–24, 25–29, and ≥30-min) were obtained using a threshold of <100 counts per minute. Sedentary time and breaks were regressed on cardiovascular risk factors (waist circumference, triglyceride, and high-density lipoprotein cholesterol) and body mass index across bout durations.ResultsThe results revealed that the majority of sedentary time occurred within relatively short bout durations (≈70% and ≈85% for <5-min and <10-min, respectively). The associations of sedentary time and breaks with health outcomes varied depending on how bout time was defined. Estimates of SB parameters based on bout durations of 5 min or shorter were associated with reduced cardiovascular risk factors while durations longer than 10-min were generally associated with increased risk factors.ConclusionsThe present study demonstrates that the duration of sedentary bouts should be further considered when operationalizing the SB parameters from accelerometer data. The threshold of 5 minutes to define a bout is defensible, but a 10 minute threshold would provide a more conservative estimate to clearly capture the prolonged nature of sedentary behavior. Additional research is needed to determine the relative sensitivity and specificity of these thresholds.     

Herbivore and pathogen effects on tree growth are additive,but mediated by tree diversity and plant traits

Herbivores and fungal pathogens are key drivers of plant community composition and functioning. The effects of herbivores and pathogens are mediated by the diversity and functional characteristics of their host plants. However, the combined effects of herbivory and pathogen damage, and their consequences for plant performance, have not yet been addressed in the context of biodiversity–ecosystem functioning research. We analyzed the relationships between herbivory, fungal pathogen damage and their effects on tree growth in a large‐scale forest‐biodiversity experiment. Moreover, we tested whether variation in leaf trait and climatic niche characteristics among tree species influenced these relationships. We found significant positive effects of herbivory on pathogen damage, and vice versa. These effects were attenuated by tree species richness—because herbivory increased and pathogen damage decreased with increasing richness—and were most pronounced for species with soft leaves and narrow climatic niches. However, herbivory and pathogens had contrasting, independent effects on tree growth, with pathogens decreasing and herbivory increasing growth. The positive herbivory effects indicate that trees might be able to (over‐)compensate for local damage at the level of the whole tree. Nevertheless, we found a dependence of these effects on richness, leaf traits and climatic niche characteristics of the tree species. This could mean that the ability for compensation is influenced by both biodiversity loss and tree species identity—including effects of larger‐scale climatic adaptations that have been rarely considered in this context. Our results suggest that herbivory and pathogens have additive but contrasting effects on tree growth. Considering effects of both herbivory and pathogens may thus help to better understand the net effects of damage on tree performance in communities differing in diversity. Moreover, our study shows how species richness and species characteristics (leaf traits and climatic niches) can modify tree growth responses to leaf damage under real‐world conditions.     

The mitogen-activated protein kinase signaling pathway stimulates mos mRNA cytoplasmic polyadenylation during Xenopus oocyte maturation

The Mos protein kinase is a key regulator of vertebrate oocyte maturation. Oocyte-specific Mos protein expression is subject to translational control. In the frog Xenopus, the translation of Mos protein requires the progesterone-induced polyadenylation of the maternal Mos mRNA, which is present in the oocyte cytoplasm. Both the Xenopus p42 mitogen-activated protein kinase (MAPK) and maturation-promoting factor (MPF) signaling pathways have been proposed to mediate progesterone-stimulated oocyte maturation. In this study, we have determined the relative contributions of the MAPK and MPF signaling pathways to Mos mRNA polyadenylation. We report that progesterone-induced Mos mRNA polyadenylation was attenuated in oocytes expressing the MAPK phosphatase rVH6. Moreover, inhibition of MAPK signaling blocked progesterone-induced Mos protein accumulation. Activation of the MAPK pathway by injection of RNA encoding Mos was sufficient to induce both the polyadenylation of synthetic Mos mRNA substrates and the accumulation of endogenous Mos protein in the absence of MPF signaling. Activation of MPF, by injection of cyclin B1 RNA or purified cyclin B1 protein, also induced both Mos protein accumulation and Mos mRNA polyadenylation. However, this action of MPF required MAPK activity. By contrast, the cytoplasmic polyadenylation of maternal cyclin B1 mRNA was stimulated by MPF in a MAPK-independent manner, thus revealing a differential regulation of maternal mRNA polyadenylation by the MAPK and MPF signaling pathways. We propose that MAPK-stimulated Mos mRNA cytoplasmic polyadenylation is a key component of the positive-feedback loop, which contributes to the all-or-none process of oocyte maturation.     

Clinical relevance of matrix metalloproteinase-13 determined with a new highly specific and sensitive ELISA in ascitic fluid of advanced ovarian carcinoma patients

Matrix metalloproteinases (MMPs) are involved in many physiological and pathophysiological processes, including tumor cell invasion and metastasis. For one member of this family, MMP-13 (collagenase-3), a new, highly specific ELISA with a sensitivity of 0.5 ng MMP-13/ml was established. The protein levels of MMP-13 in ascitic fluids of 30 patients with advanced ovarian cancer FIGO stage III (n = 19) and IV (n = 11) were measured with this ELISA. Using a cut-off value of 0.5 ng MMP-13/mg total protein, two patient subpopulations with short (median 16 months) and long (median 36 months) overall survival were identified. Together with other prognostic markers, determination of MMP-13 in ascitic fluid may help to identify patients at risk for early death and help to individualize adjuvant therapy.     

Predicting timing of foot strike during running,independent of striking technique,using principal component analysis of joint angles

As 3-dimensional (3D) motion-capture for clinical gait analysis continues to evolve, new methods must be developed to improve the detection of gait cycle events based on kinematic data. Recently, the application of principal component analysis (PCA) to gait data has shown promise in detecting important biomechanical features. Therefore, the purpose of this study was to define a new foot strike detection method for a continuum of striking techniques, by applying PCA to joint angle waveforms. In accordance with Newtonian mechanics, it was hypothesized that transient features in the sagittal-plane accelerations of the lower extremity would be linked with the impulsive application of force to the foot at foot strike. Kinematic and kinetic data from treadmill running were selected for 154 subjects, from a database of gait biomechanics. Ankle, knee and hip sagittal plane angular acceleration kinematic curves were chained together to form a row input to a PCA matrix. A linear polynomial was calculated based on PCA scores, and a 10-fold cross-validation was performed to evaluate prediction accuracy against gold-standard foot strike as determined by a 10 N rise in the vertical ground reaction force. Results show 89–94% of all predicted foot strikes were within 4 frames (20 ms) of the gold standard with the largest error being 28 ms. It is concluded that this new foot strike detection is an improvement on existing methods and can be applied regardless of whether the runner exhibits a rearfoot, midfoot, or forefoot strike pattern.     

Geographical pattern of genetic diversity in Capsella bursa‐pastoris (Brassicaceae)—A global perspective

We analyzed the global genetic variation pattern of Capsella bursa‐pastoris (Brassicaceae) as expressed in allozymic (within‐locus) diversity and isozymic (between‐locus) diversity. Results are based on a global sampling of more than 20,000 C. bursa‐pastoris individuals randomly taken from 1,469 natural provenances in the native and introduced range, covering a broad spectrum of the species’ geographic distribution. We evaluated data for population genetic parameters and F‐statistics, and Mantel tests and AMOVA were performed. Geographical distribution patterns of alleles and multilocus genotypes are shown in maps and tables. Genetic diversity of introduced populations is only moderately reduced in comparison with native populations. Global population structure was analyzed with structure, and the obtained cluster affiliation was tested independently with classification approaches and macroclimatic data using species distribution modeling. Analyses revealed two main clusters: one distributed predominantly in warm arid to semiarid climate regions and the other predominantly in more temperate humid to semihumid climate regions. We observed admixture between the two lineages predominantly in regions with intermediate humidity in both the native and non‐native ranges. The genetically derived clusters are strongly supported in macroclimatic data space. The worldwide distribution patterns of genetic variation in the range of C. bursa‐pastoris can be explained by intensive intra‐ and intercontinental migration, but environmental filtering due to climate preadaption seems also involved. Multiple independent introductions of genotypes from different source regions are obvious. “Endemic” genotypes might be the outcome of admixture or of de novo mutation. We conclude that today's successfully established Capsella genotypes were preadapted and found matching niche conditions in the colonized range parts.     

New evidence for a postglacial homoploid hybrid origin of the widespread Central European Scabiosa columbaria L. s. str. (Dipsacaceae)

We have tested whether Scabiosa columbaria s. str. could have originated by recent homoploid hybrid speciation because it is of intermediate morphology and is distributed in areas under glacial influence. Existing morphological and new RAPD data for 11 ingroup populations were analysed using multivariate statistics. Most methods indicated that S. columbaria s. str. could be the first example for a homoploid hybrid origin from Central Europe although a rigorous proof was missing partly because of ongoing introgression. Sequence variation was absent for 5,000 bp from 11 regions supporting a postglacial origin of S. columbaria s. str. An analysis of realized macroclimatic niches using GIS data showed that the niches of S. columbaria s. str. were partly intermediate between its putative parental species rather than extreme as often found in other homoploid hybrids. This could be due to geographical preconditions: large new habitats were available in postglacial Europe while in the case of most other homoploid hybrids only ecologically challenging habitats were not occupied by parental taxa.