Development of a submaximal test to predict elliptical cross-trainer VO2max

The purpose of this study was to develop an equation to predict VO2max from a submaximal elliptical cross-trainer test. Fifty-four apparently healthy subjects (25 men and 29 women, mean +/- SD age: 29.5 +/- 7.1 years, height: 173.3 +/- 12.6 cm, weight: 72.3 +/- 7.9 kg, percent body fat: 17.3 +/- 5.0%, and elliptical cross-trainer VO2max: 43.9 +/- 7.2 ml x kg(-1) x min(-1)) participated in the study and were randomly assigned to an original sample group (n = 40) and a cross-validation group (n = 14). Each subject completed an elliptical cross-trainer submaximal (3 5-minute submaximal stages) and a VO2max test on the same day, with a 15-minute rest period in between. Stepwise multiple regression analyses were used to develop an equation for estimating elliptical cross-trainer VO2max from the data of the original sample group. The accuracy of the equation was tested by using data from the cross-validation group. Because there was no shrinkage in R2 between the original sample group and the cross-validation group, data were combined in the final prediction equation (R2 = 0.732, standard error of the estimate = 3.91 ml x kg(-1) x min(-1), p < 0.05): VO2max = 73.676 + 7.383(gender) - 0.317(weight) + 0.003957(age x cadence) - 0.006452(age x heart rate at stage 2). The correlation coefficient between the predicted and measured VO2max values was r = 0.86. Dependent t-tests resulted in no significant differences (p > 0.05) between predicted (43.8 ml x kg(-1) x min(-1)) and measured (43.9 ml x kg(-1) x min(-1)) VO2max measurements. Results indicate that the protocol and equation developed in the current study can be used by exercise professionals to provide acceptably accurate estimates of VO2max in non-laboratory-based settings.     

Prediction of maximal VO2 from a submaximal StairMaster test in young women

The StairMaster 4000 PT is a popular step ergometer which provides a submaximal test protocol (SM Predicted VO(2)max) for the prediction of VO(2)max (ml.kg(-1).min(-1)). The purpose of this study was to evaluate the SM Predicted VO(2)max protocol by comparing it to results from a VO(2)max treadmill test in 20 young healthy women aged 20-25 years. Subjects were 10 step-trained (ST) women who had performed aerobic activities and exercised on a step ergometer for 20-30 minutes at least 3 times per week for the past 3 months, and 10 non-step-trained (NST) women who had performed aerobic activities no more than twice a week during the past 3 months and had no previous experience on a step ergometer. The SM Predicted VO(2)max protocol used 2 steady state heart rates between approximately 115-150 b.min(-1) to estimate VO(2)max. The Bruce maximal treadmill protocol (Actual VO(2)max) was used to measure VO(2)max by open circuit spirometry. Each subject performed both tests within a 7-day period. The means and standard deviations for the Actual VO(2)max tests were 39.8 +/- 6.1 ml.kg(-1).min(-1) for the ST group, 37.6 +/- 6.3 ml.kg(-1).min(-1) for the NST group, and 38.7 +/- 6.2 ml.kg(-1).min(-1) for the Total group (N = 20); and for the SM Predicted VO(2)max tests, means and standard deviations were 40.78 +/- 14.0 ml.kg(-1).min(-1), 30.9 +/- 4.8 ml.kg(-1).min(-1) and 35.9 +/- 11.4 ml.kg(-1).min(-1). There was no significant difference (p > 0.05) between the means of the Actual VO(2)max and SM Predicted VO(2)max test for the Total group (N = 20) or the ST group (n = 10), but a significant difference (p < 0.05) was shown for the NST group. The coefficient of determination (R(2)) and standard error of estimate (SEE) for the SM Predicted VO(2)max and Actual VO(2)max tests were R(2) = 0.18, SEE = 5.72 ml.kg(-1).min(-1) for the Total group; R(2) = 0.00, SEE = 6.68 ml.kg(-1).min(-1) for the NST group; and R(2) = 0.33, SEE = 5.32 ml.kg(-1).min(-1) for ST group. In conclusion, the SM Predicted VO(2)max test has acceptable accuracy for the ST group, but significantly underpredicted the NST group by almost 7 ml; and, as demonstrated by the high SEEs, it has a low level of precision for both ST and NST subjects.     

Improvements to the Chemical Synthesis of Biologically Active RNA Using 2′-O-Fpmp Chemistry

Abstract

The synthetic cycle protocol for the solid phase synthesis of RNA using 5′-O-(DMTr)-2′-O-(Fpmp)-ribonucleoside phosphoramidites is optimised. A simple and reliable two step deprotection procedure is developed to isolate biologically active RNA. It is demonstrated that fully deprotected RNA is completely stable under the deprotection conditions and that it does not undergo internucleotide cleavage and/or migration. Ribozymes and substrate RNAs synthesized using this chemistry were found to be catalytically active.     

Using a Simple Apparatus to Measure Direct and Diffuse Photosynthetically Active Radiation at Remote Locations

Plant canopy interception of photosynthetically active radiation (PAR) drives carbon dioxide (CO₂), water and energy cycling in the soil-plant-atmosphere system. Quantifying intercepted PAR requires accurate measurements of total incident PAR above canopies and direct beam and diffuse PAR components. While some regional data sets include these data, e.g. from Atmospheric Radiation Measurement (ARM) Program sites, they are not often applicable to local research sites because of the variable nature (spatial and temporal) of environmental variables that influence incoming PAR. Currently available instrumentation that measures diffuse and direct beam radiation separately can be cost prohibitive and require frequent adjustments. Alternatively, generalized empirical relationships that relate atmospheric variables and radiation components can be used but require assumptions that increase the potential for error. Our goal here was to construct and test a cheaper, highly portable instrument alternative that could be used at remote field sites to measure total, diffuse and direct beam PAR for extended time periods without supervision. The apparatus tested here uses a fabricated, solar powered rotating shadowband and other commercially available parts to collect continuous hourly PAR data. Measurements of total incident PAR had nearly a one-to-one relationship with total incident radiation measurements taken at the same research site by an unobstructed point quantum sensor. Additionally, measurements of diffuse PAR compared favorably with modeled estimates from previously published data, but displayed significant differences that were attributed to the important influence of rapidly changing local environmental conditions. The cost of the system is about 50% less than comparable commercially available systems that require periodic, but not continual adjustments. Overall, the data produced using this apparatus indicates that this instrumentation has the potential to support ecological research via a relatively inexpensive method to collect continuous measurements of total, direct beam and diffuse PAR in remote locations.     

Familial aggregation of VO(2max) response to exercise training: results from the HERITAGE Family Study.

The aim of this study was to test the hypothesis that individual differences in the response of maximal O(2) uptake (VO(2max)) to a standardized training program are characterized by familial aggregation. A total of 481 sedentary adult Caucasians from 98 two-generation families was exercise trained for 20 wk and was tested for VO(2max) on a cycle ergometer twice before and twice after the training program. The mean increase in VO(2max) reached approximately 400 ml/min, but there was considerable heterogeneity in responsiveness, with some individuals experiencing little or no gain, whereas others gained >1.0 l/min. An ANOVA revealed that there was 2.5 times more variance between families than within families in the VO(2max) response variance. With the use of a model-fitting procedure, the most parsimonious models yielded a maximal heritability estimate of 47% for the VO(2max) response, which was adjusted for age and sex with a maternal transmission of 28% in one of the models. We conclude that the trainability of VO(2max) is highly familial and includes a significant genetic component.     

Linear Tuning of Gamma Amplitude and Frequency to Luminance Contrast: Evidence from a Continuous Mapping Paradigm

Individual differences in the visual gamma (30–100Hz) response and their potential as trait markers of underlying physiology (particularly related to GABAergic inhibition) have become a matter of increasing interest in recent years. There is growing evidence, however, that properties of the gamma response (e.g., its amplitude and frequency) are highly stimulus dependent, and that individual differences in the gamma response may reflect individual differences in the stimulus tuning functions of gamma oscillations. Here, we measured the tuning functions of gamma amplitude and frequency to luminance contrast in eighteen participants using MEG. We used a grating stimulus in which stimulus contrast was modulated continuously over time. We found that both gamma amplitude and frequency were linearly modulated by stimulus contrast, but that the gain of this modulation (as reflected in the linear gradient) varied across individuals. We additionally observed a stimulus-induced response in the beta frequency range (10–25Hz), but neither the amplitude nor the frequency of this response was consistently modulated by the stimulus over time. Importantly, we did not find a correlation between the gain of the gamma-band amplitude and frequency tuning functions across individuals, suggesting that these may be independent traits driven by distinct neurophysiological processes.     

Vanadyl binding to a testicular S-100-like protein and to calmodulin: Electron paramagnetic resonance spectra of VO2+-protein complexes

The binding of vanadyl to a porcine and bovine testicular S-100-like protein and to calmodulin was demonstrated using X-band (9.2 gHz) electron paramagnetic resonance (EPR) spectroscopy in aqueous solution at pH 7.4. In liquid solutions at 22°C, the vanadyl-protein complexes yielded VO²⁺ near rigid limit spectra. At 122 K, each of the three high-field resonances (i.e., 3/2, 5/2, and 7/2 parallel components) splits into two components indicating the presence of two classes of vanadyl-binding sites in each protein. The spectra of the frozen solutions were simulated to give parallel and perpendicular components of the hyperfine coupling constant and g factors similar to other vanadyl-protein complexes.     

Time to exhaustion and time spent at a high percentage of VO2max in severe intensity domain in children and adults

The aim of the study was to compare time spent at a high percentage of VO2max (>90% of VO2max) (ts90%), time to achieve 90% of VO2max (ta90%), and time to exhaustion (TTE) for exercise in the severe intensity domain in children and adults. Fifteen prepubertal boys (10.3 ± 0.9 years) and 15 men (23.5 ± 3.6 years) performed a maximal graded exercise to determine VO2max, maximal aerobic power (MAP) and power at ventilatory threshold (PVTh). Then, they performed 4 constant load exercises in a random order at PVTh plus 50 and 75% of the difference between MAP and PVTh (PΔ50 and PΔ75) and 100 and 110% of MAP (P100 and P110). VO2max was continuously monitored. The P110 test was used to determine maximal accumulated oxygen deficit (MAOD). No significant difference was found in ta90% between children and adults. ts90% and TTE were not significantly different between children and adults for the exercises at PΔ50 and PΔ75. However, ts90% and TTE during P100 (p < 0.05 and p < 0.01, respectively) and P110 (p < 0.001) exercises were significantly shorter in children. Children had a significantly lower MAOD than adults (34.3 ± 9.4 ml · kg vs. 53.6 ± 11.1 ml · kg). A positive relationship (p < 0.05) was obtained between MAOD and TTE values during the P100 test in children. This study showed that only for intensities at, or higher than MAP, lower ts90% in children was linked to a reduced TTE, compared to adults. Shorter TTE in children can partly be explained by a lower anaerobic capacity (MAOD). These results give precious information about exercise intensity ranges that could be used in children's training sessions. Moreover, they highlight the implication of both aerobic and anaerobic processes in endurance performances in both populations.     

Crude Oil Treatment Leads to Shift of Bacterial Communities in Soils from the Deep Active Layer and Upper Permafrost along the China-Russia Crude Oil Pipeline Route

The buried China-Russia Crude Oil Pipeline (CRCOP) across the permafrost-associated cold ecosystem in northeastern China carries a risk of contamination to the deep active layers and upper permafrost in case of accidental rupture of the embedded pipeline or migration of oil spills. As many soil microbes are capable of degrading petroleum, knowledge about the intrinsic degraders and the microbial dynamics in the deep subsurface could extend our understanding of the application of in-situ bioremediation. In this study, an experiment was conducted to investigate the bacterial communities in response to simulated contamination to deep soil samples by using 454 pyrosequencing amplicons. The result showed that bacterial diversity was reduced after 8-weeks contamination. A shift in bacterial community composition was apparent in crude oil-amended soils with Proteobacteria (esp. α-subdivision) being the dominant phylum, together with Actinobacteria and Firmicutes. The contamination led to enrichment of indigenous bacterial taxa like Novosphingobium, Sphingobium, Caulobacter, Phenylobacterium, Alicylobacillus and Arthrobacter, which are generally capable of degrading polycyclic aromatic hydrocarbons (PAHs). The community shift highlighted the resilience of PAH degraders and their potential for in-situ degradation of crude oil under favorable conditions in the deep soils.     

Novel Electrochemically Active Bacterium Phylogenetically Related to Arcobacter butzleri,Isolated from a Microbial Fuel Cell

Exoelectrogenic bacteria are organisms that can transfer electrons to extracellular insoluble electron acceptors and have the potential to be used in devices such as microbial fuel cells (MFCs). Currently, exoelectrogens have been identified in the Alpha-, Beta-, Gamma- and Deltaproteobacteria, as well as in the Firmicutes and Acidobacteria. Here, we describe use of culture-independent methods to identify two members of the genus Arcobacter in the Epsilonproteobacteria that are selectively enriched in an acetate-fed MFC. One of these organisms, Arcobacter butzleri strain ED-1, associates with the electrode and rapidly generates a strong electronegative potential as a pure culture when it is supplied with acetate. A mixed-community MFC in which ∼90% of the population is comprised of the two Arcobacter species generates a maximal power density of 296 mW/liter. This demonstration of exoelectrogenesis by strain ED-1 is the first time that this property has been shown for members of this genus.A microbial fuel cell (MFC) is a mimic of a biological system in which microorganisms transfer electrons from organic compounds to a conductive external electron acceptor under anaerobic conditions (6). In an MFC, the electron acceptor is provided by an artificial anode, which is connected to an electric circuit. Although the basic processes involved in the generation of electricity by bacteria have been known for many years, recent interest in MFC development has been stimulated by the need to find alternative, carbon-neutral sources of energy generation. MFCs are particularly useful for breakdown of organic matter in wastewater treatment plants, in which production of electricity as a by-product can be used to power the process or can be sold to offset the cost of operation (6). At present, although the key principles of MFC design and operation are well understood (19), the technical aspects and particularly the microbiological aspects (18) are still in development. Further optimization of the design and microbial composition of these devices is desirable as current MFCs achieve power densities of no more than 1,550 mW/liter (7), which limits their real-world applications (6).The basic microbiological characteristics which influence the efficiency of an MFC are bacterial metabolism and bacterial electron transfer. Although most current MFCs perform optimally when they contain a mixed microbial community, some pure cultures that exhibit strong electrogenic activity in the MFC environment have been characterized (19). The electrogenic properties and some aspects of extracellular electron transfer have been defined for pure cultures of organisms such as Geobacter sulfurreducens (2, 3), Escherichia coli (27), Shewanella putrefaciens (15, 16), Rhodoferax ferrireducens (5), Rhodopseudomonas palustris DX-1 (40), and Ochrobactrum anthropi YZ-1 (41). The current list of confirmed exoelectrogens includes representatives of four of the five classes of Proteobacteria (only the Epsilonproteobacteria are not represented), as well as representatives of the Firmicutes and Acidobacteria (18). However, it is likely that novel electrogenic bacteria remain to be discovered.The metabolic characteristics required for an electrogenic bacterium depend upon the specific application for which an MFC is used, because not all electrogenic bacteria are able to fully oxidize several substrates. For example, Shewanella oneidensis oxidizes lactate to acetate under anaerobic conditions, while G. metallireducens oxidizes acetate but not glucose (20). R. ferrireducens can oxidize acetate, lactate, and glucose but does not degrade ethanol, another common fermentation end product (11). For this reason, MFCs which are employed in wastewater treatment when complex compounds have to be degraded are often inoculated with a diverse microbial community (for example, methanogenic sludge [30]). Degradation of acetate is a key bacterial characteristic because acetate is a primary organic intermediate in the degradation of organic matter in anoxic aquatic sediments. Moreover, the ability to use artificial electron acceptors (anodic electrodes) provides bacteria such as Geobacteraceae with a competitive advantage over other microorganisms under such conditions. Analysis of the microbial community firmly attached to anodes harvesting electricity from a variety of sediments demonstrated that microorganisms in the family Geobacteraceae were highly enriched on these anodes (2, 35). Moreover, it was shown that an MFC initially inoculated with methanogenic sludge failed to consume acetate in the absence of anodic electrodes over a 1-year period (8).Arcobacter spp. are inhabitants of human and animal hosts (14, 37) and also occur in various environments, including wastewater (24), surface water (21), seawater (9), and groundwater (32). Arcobacter spp. belong to the Epsilonproteobacteria, which includes pathogens (e.g., Campylobacter jejuni and Helicobacter pylori), opportunistic pathogens, and nonpathogenic environmental isolates (4). Typically, these bacteria have genomes with low G+C contents (27 to 30%), although some Epsilonproteobacteria, such as Wolinella spp. and Campylobacter curvus, have higher G+C contents. The environmental bacteria group into four clusters: Nautiliales, Arcobacter, Sulfurospirillum, and Thiovulgaceae. The genus Arcobacter comprises Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius, all of which have been isolated from animals or food (particularly poultry), as well as Arcobacter halophilus, Arcobacter nitrofigilis, “Candidatus Arcobacter sulfidicus,” and a number of species characterized so far only at the 16S rRNA gene level (4). A feature of both “Ca. Arcobacter sulfidicus” and Arcobacter sp. strain FWKBO is autotrophic metabolism under microaerobic conditions, in contrast to the heterotrophic growth of A. butzleri. Both of these organisms use oxidation of sulfide to sulfur and are obligate autotrophs. Some Arcobacter spp. may be capable of Mn and Fe reduction; isolates from Black Sea sediments (36) oxidized acetate in the presence of Mn oxide. This was the first evidence of Mn or Fe reduction in nitrate-reducing Arcobacter microaerophiles and nitrate reducers; previously, the only other epsilonproteobacterium identified with this ability was Sulfurospirillum barnesii. Thus, organisms related to Arcobacter comprise an ecologically significant new group of dissimilatory Fe- and Mn-reducing bacteria.In the present study we isolated and characterized two strains phylogenetically related to Arcobacter spp. which are selectively enriched in an acetate-fed MFC. One of these strains, A. butzleri strain ED-1, which specifically associates with the MFC electrode, shows electrochemical activity when it is grown on acetate, and hence it is the first example of an exoelectrogenic epsilonproteobacterium.     

Characterization of a Novel β-Galactosidase from Bifidobacterium adolescentis DSM 20083 Active towards Transgalactooligosaccharides

This paper reports on the effects of both reducing and nonreducing transgalactooligosaccharides (TOS) comprising 2 to 8 residues on the growth of Bifidobacterium adolescentis DSM 20083 and on the production of a novel β-galactosidase (β-Gal II). In cells grown on TOS, in addition to the lactose-degrading β-Gal (β-Gal I), another β-Gal (β-Gal II) was detected and it showed activity towards TOS but not towards lactose. β-Gal II activity was at least 20-fold higher when cells were grown on TOS than when cells were grown on galactose, glucose, and lactose. Subsequently, the enzyme was purified from the cell extract of TOS-grown B. adolescentis by anion-exchange chromatography, adsorption chromatography, and size-exclusion chromatography. β-Gal II has apparent molecular masses of 350 and 89 kDa as judged by size-exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively, indicating that the enzyme is active in vivo as a tetramer. β-Gal II had an optimal activity at pH 6 and was not active below pH 5. Its optimum temperature was 35°C. The enzyme showed highest V_max values towards galactooligosaccharides with a low degree of polymerization. This result is in agreement with the observation that during fermentation of TOS, the di- and trisaccharides were fermented first. β-Gal II was active towards β-galactosyl residues that were 1→4, 1→6, 1→3, and 1↔1 linked, signifying its role in the metabolism of galactooligosaccharides by B. adolescentis.     

Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein (GST-RhoA(G17A)) from Epithelial Cell Lysates

Proteins of the Rho family of small GTPases are central regulators of the cytoskeleton, and control a large variety of cellular processes, including cell migration, gene expression, cell cycle progression and cell adhesion ¹. Rho proteins are molecular switches that are active in GTP-bound and inactive in GDP-bound state. Their activation is mediated by a family of Guanine-nucleotide Exchange Factor (GEF) proteins. Rho-GEFs constitute a large family, with overlapping specificities ². Although a lot of progress has been made in identifying the GEFs activated by specific signals, there are still many questions remaining regarding the pathway-specific regulation of these proteins. The number of Rho-GEFs exceeds 70, and each cell expresses more than one GEF protein. In addition, many of these proteins activate not only Rho, but other members of the family, contributing further to the complexity of the regulatory networks. Importantly, exploring how GEFs are regulated requires a method to follow the active pool of individual GEFs in cells activated by different stimuli. Here we provide a step-by-step protocol for a method used to assess and quantify the available active Rho-specific GEFs using an affinity precipitation assay. This assay was developed a few years ago in the Burridge lab ^3,4 and we have used it in kidney tubular cell lines ^5,6,7. The assay takes advantage of a "nucleotide free" mutant RhoA, with a high affinity for active GEFs. The mutation (G17A) renders the protein unable to bind GDP or GTP and this state mimics the intermediate state that is bound to the GEF. A GST-tagged version of this mutant protein is expressed and purified from E. coli, bound to glutathione sepharose beads and used to precipitate active GEFs from lysates of untreated and stimulated cells. As most GEFs are activated via posttranslational modifications or release from inhibitory bindings, their active state is preserved in cell lysates, and they can be detected by this assay⁸. Captured proteins can be probed for known GEFs by detection with specific antibodies using Western blotting, or analyzed by Mass Spectrometry to identify unknown GEFs activated by certain stimuli.     

Modeling the 3-D Structure of a Recombinant Laccase from Trametes trogii Active at a pH Close to Neutrality

A cDNA encoding a novel laccase from the white-rot fungus Trametes trogii was cloned and expressed in Pichia pastoris. The recombinant protein (Lcc2) exhibited kinetic parameters for both phenolic and non phenolic substrates that were different from the previously described Lcc1, the main laccase isoform expressed by T. trogii; in addition, the pH/activity profiles for phenolic substrates of Lcc2 were shifted upward by 1–1.5 pH units towards neutrality as compared to Lcc1. Comparative modeling of the two laccases (69.2% identity) showed that the overall fold of Lcc2 is very similar to Lcc1 and other laccases. The substrate cavity of Lcc2 contains the Asp residue which is thought to mediate the laccase activity at acidic pHs, whereas two hydrophobic residues (Phe, Ile) on the cavity orifice of Lcc2 replace the two polar residues (Thr, Ser) of Lcc1. These structural differences may be responsible for the unique kinetic performances of Lcc2.     

Assessing the Relative Contributions of Active Ankle and Knee Assistance to the Walking Mechanics of Transfemoral Amputees Using a Powered Prosthesis

Powered knee-ankle prostheses are capable of providing net-positive mechanical energy to amputees. Yet, there are limitless ways to deliver this energy throughout the gait cycle. It remains largely unknown how different combinations of active knee and ankle assistance affect the walking mechanics of transfemoral amputees. This study assessed the relative contributions of stance phase knee swing initiation, increasing ankle stiffness and powered plantarflexion as three unilateral transfemoral amputees walked overground at their self-selected walking speed. Five combinations of knee and ankle conditions were evaluated regarding the kinematics and kinetics of the amputated and intact legs using repeated measures analyses of variance. We found eliminating active knee swing initiation or powered plantarflexion was linked to increased compensations of the ipsilateral hip joint during the subsequent swing phase. The elimination of knee swing initiation or powered plantarflexion also led to reduced braking ground reaction forces of the amputated and intact legs, and influenced both sagittal and frontal plane loading of the intact knee joint. Gradually increasing prosthetic ankle stiffness influenced the shape of the prosthetic ankle plantarflexion moment, more closely mirroring the intact ankle moment. Increasing ankle stiffness also corresponded to increased prosthetic ankle power generation (despite a similar maximum stiffness value across conditions) and increased braking ground reaction forces of the amputated leg. These findings further our understanding of how to deliver assistance with powered knee-ankle prostheses and the compensations that occur when specific aspects of assistance are added/removed.     

Dipeptide Synthesis by an Aminopeptidase from Streptomyces septatus TH-2 and Its Application to Synthesis of Biologically Active Peptides

Dipeptide synthesis by aminopeptidase from Streptomyces septatus TH-2 (SSAP) was demonstrated using free amino acid as an acyl donor and aminoacyl methyl ester as an acyl acceptor in 98% methanol (MeOH). SSAP retained its activity after more than 100 h in 98% MeOH, and in the case of phenylalanyl-phenylalanine methyl ester synthesis, the enzyme reaction reached equilibrium when more than 50% of the free phenylalanine was converted to the product. In an investigation of the specificity of SSAP toward acyl donors and acyl acceptors, SSAP showed a broad specificity toward various free amino acids and aminoacyl methyl esters. Furthermore, we applied SSAP to the synthesis of several biologically active peptides, such as aspartyl-phenylalanine, alanyl-tyrosine, and valyl-tyrosine methyl esters.     

Active site plasticity revealed from the structure of the enterobacterial N-ribohydrolase RihA bound to a competitive inhibitor

Background  

Pyrimidine-preferring N-ribohydrolases (CU-NHs) are a class of Ca²⁺-dependent enzymes that catalyze the hydrolytic cleavage of the N-glycosidic bond in pyrimidine nucleosides. With the exception of few selected organisms, their physiological relevance in prokaryotes and eukaryotes is yet under investigation.     

Estimating 3D Movements from 2D Observations Using a Continuous Model of Helical Swimming

Helical swimming is among the most common movement behaviors in a wide range of microorganisms, and these movements have direct impacts on distributions, aggregations, encounter rates with prey, and many other fundamental ecological processes. Microscopy and video technology enable the automated acquisition of large amounts of tracking data; however, these data are typically two-dimensional. The difficulty of quantifying the third movement component complicates understanding of the biomechanical causes and ecological consequences of helical swimming. We present a versatile continuous stochastic model—the correlated velocity helical movement (CVHM) model—that characterizes helical swimming with intrinsic randomness and autocorrelation. The model separates an organism’s instantaneous velocity into a slowly varying advective component and a perpendicularly oriented rotation, with velocities, magnitude of stochasticity, and autocorrelation scales defined for both components. All but one of the parameters of the 3D model can be estimated directly from a two-dimensional projection of helical movement with no numerical fitting, making it computationally very efficient. As a case study, we estimate swimming parameters from videotaped trajectories of a toxic unicellular alga, Heterosigma akashiwo (Raphidophyceae). The algae were reared from five strains originally collected from locations in the Atlantic and Pacific Oceans, where they have caused Harmful Algal Blooms (HABs). We use the CVHM model to quantify cell-level and strain-level differences in all movement parameters, demonstrating the utility of the model for identifying strains that are difficult to distinguish by other means.     

Identification and Characterization of a Specific Receptor for Cholecystokinin on Isolated Fundic Glands from Guinea Pig Gastric Mucosa Using a Biologically Active 125I-CCK-8 Probe

Abstract

Specific binding sites for cholecystokinin (CCK) have been identified and characterized in fundic glands isolated by collagenase treatment from guinea pig gastric mucosa using a biologically active ¹²⁵I-labeled derivative of the C-terminal octa-peptide of CCK (¹²⁵IIE-CCK-8). The time course of binding to these glands was rapid, temperature dependent and saturable. At 24, 30 and 37° C, half-maximal binding was reached at 5 min and full binding at 30 min. The addition of a large excess of CCK-8 after 15 and 30 min of binding at 24° C caused a prompt and rapid decline in radioligand bound showing that the interaction was reversible. There was a progressive decline in the amount of ¹²⁵IIE-CCK-8 bound to fundic glands with increasing concentrations of CCK-8 and other structurally related peptides. Gastrin II displaced 50% of the radioligand at 1.6nM, CCK-8 at 3.2nM, gastrin I at 16nM, and desulfated-CCK-8 and pentagastrin at 59nM. Secretin did not displace the radioligand from fundic glands at 1.0uM. The binding was also tissue specific as glands isolated from the antral mucosa did not contain specific binding sites for ¹²⁵IIE-CCK-8. This data provides evidence for specific receptors for CCK on gastric fundic glands that may be involved in the control of acid and pepsinogen secretion.