bluuebunny/crossref_metadata_2025 huggingface dataset:
Unusual multiple metastatic localisations in adult Burkitt’s lymphoma. A case report.
Authors: Nagy Georgiana , Chira Romeo , Crisan Doinita , Munteanu Mihai , Mircea Petru Adrian
Published: December 2016

Burkitt’s lymphoma is an aggressive B-cell non-Hodgkin lymphoma. It is less common in adults accounting for less than 5% of non-Hodgkin lymphoma cases. Radiological methods (ultrasonography, computed tomography) are indispensable for the initial evaluation and appreciation of organ extension; complete diagnosis is confirmed by the histopathological examination.We present the clinical case and ultrasound imaging particularities of a young patient diagnosed with multisystem involvement Burkitt’s lymphoma, with rapid progressive evolution towards exitus.
Evaluation of nasal fluid flow in fetuses by Doppler ultrasound
Authors: Zielinski Rafal , Respondek-Liberska Maria
Published: December 2016

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Sonographer preference for Knox versus ballistic gelatin for the creation of deep venous thrombosis ultrasound phantoms
Authors: Olivieri Patrick , Doctor Michael , Siadecki Sebastian , Rose Gabriel , Drake Aaran , Saul Turandot
Published: December 2016

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Metal-vapor integration/transportation based on metal-atom desorption from polymer surfaces with a low glass-transition temperature
Authors: Tsujioka Tsuyoshi , Shirakawa Riku , Matsumoto Saki , Nishimura Ryo , Uchida Kingo
Published: December 2016

Vacuum evaporation, which is widely used to obtain a metal-film coating on substrates and/or targets, results in a large loss of evaporation materials. The authors discovered a selective metal-vapor deposition phenomenon in which metal-vapor atoms can be deposited selectively without a shadow mask. The core phenomenon of selective deposition is the desorption of metal-vapor atoms from organic surfaces with a low-glass transition temperature (low-Tg). The authors investigated metal-atom desorption from low-Tg polymer [polydimethylsiloxane (PDMS)] surfaces during vacuum deposition and found that greater than 2-μm-thick PDMS film enables high desorption efficiency. The authors proposed two vacuum deposition methods: metal-vapor transportation and metal-vapor integration. The former method attained reduced substrate surface temperatures during vacuum evaporation and the latter enabled extremely high deposition efficiency and three-dimensional deposition in a one-batch process. These new methods can greatly widen applications of vacuum deposition.
Improvement of Ice Hockey Players’ On-Ice Sprint With Combined Plyometric and Strength Training
Authors: Dæhlin Torstein E. , Haugen Ole C. , Haugerud Simen , Hollan Ivana , Raastad Truls , Rønnestad Bent R.
Published: December 2016

Background:

Combined plyometric and strength training has previously been suggested as a strategy to improve skating performance in ice hockey players. However, the effects of combined plyometric and strength training have not previously been compared with the effects of strength training only.

Purpose:

To compare the effects of combined plyometric and strength training on ice hockey players’ skating sprint performance with those of strength training only.

Methods:

Eighteen participants were randomly assigned to 2 groups that completed 5 strength-training sessions/wk for 8 wk. One group included plyometric exercises at the start of 3 sessions/wk (PLY+ST), and the other group included core exercises in the same sessions (ST). Tests of 10- and 35-m skating sprints, horizontal jumping, 1-repetition-maximum (1 RM) squat, skating multistage aerobic test (SMAT), maximal oxygen consumption, repeated cycle sprints, and body composition were performed before and after the intervention.

Results:

The participants increased their 1RM squat, lean mass, and body mass (P < .05), with no difference between the groups. Furthermore, they improved their 3×broad jump, repeated cycle sprint, and SMAT performance (P < .05), with no difference between the groups. PLY+ST gained a larger improvement in 10-m on-ice sprint performance than ST (P < .025).

Conclusion:

Combining plyometric and strength training for 8 wk was superior to strength training alone at improving 10-m on-ice sprint performance in high-level ice hockey players.
Aerobic Evaluation in Elite Slalom Kayakers Using a Tethered Canoe System: A New Proposal
Authors: Ferrari Homero Gustavo , Messias Leonardo H.D. , Reis Ivan G.M. , Gobatto Claudio A. , Sousa Filipe A.B. , Serra Camila C.S. , Manchado-Gobatto Fúlvia B.
Published: December 2016

Background:

Among other aspects, aerobic fitness is indispensable for performance in slalom canoe.

Purpose:

To propose the maximal-lactate steady-state (MLSS) and critical-force (CF) tests using a tethered canoe system as new strategies for aerobic evaluation in elite slalom kayakers. In addition, the relationship between the aerobic parameters from these tests and the kayakers’ performances was studied.

Methods:

Twelve male elite slalom kayakers from the Brazilian national team participated in this study. All tests were conducted using a tethered canoe system to obtain the force records. The CF test was applied on 4 d and analyzed by hyperbolic (CFhyper) and linear (CFlin) mathematical models. The MLSS intensity (MLSSint) was obtained by three 30-min continuous tests. The time of a simulated race was considered the performance index.

Results:

No difference (P < .05) between CFhyper (65.9 ± 1.6 N) and MLSSint (60.3 ± 2.5 N) was observed; however, CFlin (71.1 ± 1.7 N) was higher than MLSSint. An inverse and significant correlation was obtained between MLSSint and performance (r = –.67, P < .05).

Conclusion:

In summary, MLSS and CF tests on a tethered canoe system may be used for aerobic assessment of elite slalom kayakers. In addition, CFhyper may be used as an alternative low-cost and noninvasive method to estimate MLSSint, which is related with slalom kayakers’ performance.
Movement Patterns and Metabolic Responses During an International Rugby Sevens Tournament
Authors: Couderc Anthony , Thomas Claire , Lacome Mathieu , Piscione Julien , Robineau Julien , Delfour-Peyrethon Rémi , Borne Rachel , Hanon Christine
Published: December 2016

Purpose:

To investigate the running demands and associated metabolic perturbations during an official rugby sevens tournament.

Methods:

Twelve elite players participated in 7 matches wearing GPS units. Maximal sprinting speed (MSS) and maximal aerobic speed (MAS) were measured. High-intensity threshold was individualized relative to MAS (>100% of MAS), and very-high-intensity distance was reported relative to both MAS and MSS. Blood samples were taken at rest and after each match.

Results:

Comparison of prematch and postmatch samples revealed significant (P < .01) changes in pH (7.41–7.25), bicarbonate concentration ([HCO3–]) (24.8–13.6 mmol/L), and lactate concentration ([La]) (2.4–11.9 mmol/L). Mean relative total distance covered was 91 ± 13 m/min with ~17 m/min at high-intensity. Player status (whole-match or interchanged players), match time, and total distance covered had no significant impact on metabolic indices. Relative distance covered at high intensity was negatively correlated with pH and [HCO3–] (r = .44 and r = .42, respectively; P < .01) and positively correlated with [La] (r = .36; P < .01). Total distance covered and distance covered at very high intensity during the 1-min peak activity in the last 3 min of play were correlated with [La] (r = .39 and r = .39, respectively; P < .01).

Conclusions:

Significant alterations in blood-metabolite indices from prematch to postmatch sampling suggest that players were required to tolerate a substantial level of acidosis related to metabolite accumulation. In addition, the ability to produce energy via the glycolytic energy pathway seems to be a major determinant in match-related running performance.
A Comparison of Isometric Midthigh-Pull Strength, Vertical Jump, Sprint Speed, and Change-of-Direction Speed in Academy Netball Players
Authors: Thomas Christopher , Comfort Paul , Jones Paul A. , Dos’Santos Thomas
Published: December 2016

Purpose:

To investigate the relationships between maximal isometric strength, vertical jump (VJ), sprint speed, and change-of-direction speed (CoDS) in academy netball players and determine whether players who have high performance in isometric strength testing would demonstrate superior performance in VJ, sprint speed, and CoDS measures.

Method:

Twenty-six young female netball players (age 16.1 ± 1.2 y, height 173.9 ± 5.7 cm, body mass 66.0 ± 7.2 kg) from a regional netball academy performed isometric midthigh pull (IMTP), squat jumps (SJs), countermovement jumps (CMJs), 10-m sprints, and CoDS (505).

Results:

IMTP measures displayed moderate to strong correlations with sprint and CoDS performance (r = –.41 to –.66). The VJs, which included SJs and CMJs, demonstrated strong correlations with 10-m sprint times (r = –.60 to –.65; P < .01) and CoDS (r = –.60 to –.71; P = .01). Stronger players displayed significantly faster sprint (ES = 1.1–1.2) and CoDS times (ES = 1.2–1.7) and greater VJ height (ES = 0.9–1.0) than weaker players.

Conclusion:

The results of this study illustrate the importance of developing high levels of lower-body strength to enhance VJ, sprint, and CoDS performance in youth netball players, with stronger athletes demonstrating superior VJ, sprint, and CoDS performances.
Importance of Various Training-Load Measures in Injury Incidence of Professional Rugby League Athletes
Authors: Thornton Heidi R. , Delaney Jace A. , Duthie Grant M. , Dascombe Ben J.
Published: December 2016

Purpose:

To investigate the ability of various internal and external training-load (TL) monitoring measures to predict injury incidence among positional groups in professional rugby league athletes.

Methods:

TL and injury data were collected across 3 seasons (2013–2015) from 25 players competing in National Rugby League competition. Daily TL data were included in the analysis, including session rating of perceived exertion (sRPE-TL), total distance (TD), high-speed-running distance (>5 m/s), and high-metabolic-power distance (HPD; >20 W/kg). Rolling sums were calculated, nontraining days were removed, and athletes’ corresponding injury status was marked as “available” or “unavailable.” Linear (generalized estimating equations) and nonlinear (random forest; RF) statistical methods were adopted.

Results:

Injury risk factors varied according to positional group. For adjustables, the TL variables associated most highly with injury were 7-d TD and 7-d HPD, whereas for hit-up forwards they were sRPE-TL ratio and 14-d TD. For outside backs, 21- and 28-d sRPE-TL were identified, and for wide-running forwards, sRPE-TL ratio. The individual RF models showed that the importance of the TL variables in injury incidence varied between athletes.

Conclusions:

Differences in risk factors were recognized between positional groups and individual athletes, likely due to varied physiological capacities and physical demands. Furthermore, these results suggest that robust machine-learning techniques can appropriately monitor injury risk in professional team-sport athletes.
Within-Season Distribution of External Training and Racing Workload in Professional Male Road Cyclists
Authors: Metcalfe Alan J. , Menaspà Paolo , Villerius Vincent , Quod Marc , Peiffer Jeremiah J. , Govus Andrew D. , Abbiss Chris R
Published: December 2016

Purpose:

To describe the within-season external workloads of professional male road cyclists for optimal training prescription.

Methods:

Training and racing of 4 international competitive professional male cyclists (age 24 ± 2 y, body mass 77.6 ± 1.5 kg) were monitored for 12 mo before the world team-time-trial championships. Three within-season phases leading up to the team-time-trial world championships on September 20, 2015, were defined as phase 1 (Oct–Jan), phase 2 (Feb–May), and phase 3 (June–Sept). Distance and time were compared between training and racing days and over each of the various phases. Times spent in absolute (<100, 100–300, 400–500, >500 W) and relative (0–1.9, 2.0–4.9, 5.0–7.9, >8 W/kg) power zones were also compared for the whole season and between phases 1–3.

Results:

Total distance (3859 ± 959 vs 10911 ± 620 km) and time (240.5 ± 37.5 vs 337.5 ± 26 h) were lower (P < .01) in phase 1 than phase 2. Total distance decreased (P < .01) from phase 2 to phase 3 (10911 ± 620 vs 8411 ± 1399 km, respectively). Mean absolute (236 ± 12.1 vs 197 ± 3 W) and relative (3.1 ± 0 vs 2.5 ± 0 W/kg) power output were higher (P < .05) during racing than training, respectively.

Conclusion:

Volume and intensity differed between training and racing over each of 3 distinct within-season phases.
Modern Techniques and Technologies Applied to Training and Performance Monitoring
Authors: Sands William A. , Kavanaugh Ashley A. , Murray Steven R. , McNeal Jeni R. , Jemni Monèm
Published: December 2016

Athlete preparation and performance continue to increase in complexity and costs. Modern coaches are shifting from reliance on personal memory, experience, and opinion to evidence from collected training-load data. Training-load monitoring may hold vital information for developing systems of monitoring that follow the training process with such precision that both performance prediction and day-to-day management of training become adjuncts to preparation and performance. Time-series data collection and analyses in sport are still in their infancy, with considerable efforts being applied in “big data” analytics, models of the appropriate variables to monitor, and methods for doing so. Training monitoring has already garnered important applications but lacks a theoretical framework from which to develop further. As such, we propose a framework involving the following: analyses of individuals, trend analyses, rules-based analysis, and statistical process control.
Countermovement-Jump-Phase Characteristics of Senior and Academy Rugby League Players
Authors: McMahon John J. , Murphy Shannon , Rej Sophie J.E. , Comfort Paul
Published: December 2016

Purpose:

Gross measures of countermovement-jump (CMJ) performance are commonly used to track maturational changes in neuromuscular function in rugby league (RL). The purpose of this study was to conduct both a gross and a more detailed temporal-phase analysis of the CMJ performances of senior and academy RL players, to provide greater insight into how neuromuscular function differs between these groups.

Methods:

Twenty senior and 14 academy (under-19) male RL players performed 3 maximal-effort CMJs on a force platform, with forward dynamics subsequently employed to allow gross performance measures and entire kinetic– and kinematic–time curves to be compared between groups.

Results:

Jump height (JH), reactive strength index modified, concentric displacement, and relative concentric impulse (C-IMP) were the only gross measures that were greater for senior players (d = 0.58–0.91) than for academy players. The relative force- and displacement–time curves were similar between groups, but the relative power– and velocity–time curves were greater (d = 0.59–0.97) for the senior players at 94–96% and 89–100% of the total movement time, respectively.

Conclusions:

The CMJ distinguished between senior and academy RL players, with seniors demonstrating greater JH through applying a larger C-IMP and thus achieving greater velocity throughout the majority of the concentric phase and at takeoff. Therefore, academy RL players should train to improve triple (ie, ankle, knee, and hip) extension velocity during the CMJ to bring their JH scores in line with those attained by senior players.
Is There an Optimal Pole Length for Double Poling in Cross Country Skiing?
Authors: Onasch Franziska , Killick Anthony , Herzog Walter
Published: December 2016

The aim of this study was to determine the effects of pole length on energy cost and kinematics in cross country double poling. Seven sub-elite male athletes were tested using pole sets of different lengths (ranging between 77% and 98% of participants’ body height). Tests were conducted on a treadmill, set to a 2% incline and an approximate racing speed. Poling forces, contact times, and oxygen uptake were measured throughout the testing. Pole length was positively correlated with ground contact time (r = .57, p < .001) and negatively correlated with poling frequency (r = −.48, p = .003). Pole length was also positively correlated with pole recovery time and propulsive impulse produced per poling cycle (r = .36, p = .031; r = .35, p = .042, respectively). Oxygen uptake and pole length were negatively correlated (r = −.51, p = .004). This acute study shows that increasing pole length for double poling in sub-elite cross country skiers under the given conditions seems to change the poling mechanics in distinct ways, resulting in a more efficient poling action by decreasing an athlete’s metabolic cost.
The Validity and Reliability of an iPhone App for Measuring Running Mechanics
Authors: Balsalobre-Fernández Carlos , Agopyan Hovannes , Morin Jean-Benoit
Published: December 2016

The purpose of this investigation was to analyze the validity of an iPhone application (Runmatic) for measuring running mechanics. To do this, 96 steps from 12 different runs at speeds ranging from 2.77–5.55 m·s−1 were recorded simultaneously with Runmatic, as well as with an opto-electronic device installed on a motorized treadmill to measure the contact and aerial time of each step. Additionally, several running mechanics variables were calculated using the contact and aerial times measured, and previously validated equations. Several statistics were computed to test the validity and reliability of Runmatic in comparison with the opto-electronic device for the measurement of contact time, aerial time, vertical oscillation, leg stiffness, maximum relative force, and step frequency. The running mechanics values obtained with both the app and the opto-electronic device showed a high degree of correlation (r = .94–.99, p < .001). Moreover, there was very close agreement between instruments as revealed by the ICC (2,1) (ICC = 0.965–0.991). Finally, both Runmatic and the opto-electronic device showed almost identical reliability levels when measuring each set of 8 steps for every run recorded. In conclusion, Runmatic has been proven to be a highly reliable tool for measuring the running mechanics studied in this work.
Comparison of 3D Joint Angles Measured With the Kinect 2.0 Skeletal Tracker Versus a Marker-Based Motion Capture System
Authors: Guess Trent M. , Razu Swithin , Jahandar Amirhossein , Skubic Marjorie , Huo Zhiyu
Published: December 2016

The Microsoft Kinect is becoming a widely used tool for inexpensive, portable measurement of human motion, with the potential to support clinical assessments of performance and function. In this study, the relative osteokinematic Cardan joint angles of the hip and knee were calculated using the Kinect 2.0 skeletal tracker. The pelvis segments of the default skeletal model were reoriented and 3-dimensional joint angles were compared with a marker-based system during a drop vertical jump and a hip abduction motion. Good agreement between the Kinect and marker-based system were found for knee (correlation coefficient = 0.96, cycle RMS error = 11°, peak flexion difference = 3°) and hip (correlation coefficient = 0.97, cycle RMS = 12°, peak flexion difference = 12°) flexion during the landing phase of the drop vertical jump and for hip abduction/adduction (correlation coefficient = 0.99, cycle RMS error = 7°, peak flexion difference = 8°) during isolated hip motion. Nonsagittal hip and knee angles did not correlate well for the drop vertical jump. When limited to activities in the optimal capture volume and with simple modifications to the skeletal model, the Kinect 2.0 skeletal tracker can provide limited 3-dimensional kinematic information of the lower limbs that may be useful for functional movement assessment.
Changes in the Force-Velocity Mechanical Profile After Short Resistance Training Programs Differing in Set Configurations
Authors: Iglesias-Soler Eliseo , Fernández-del-Olmo Miguel , Mayo Xián , Fariñas Juan , Río-Rodríguez Dan , Carballeira Eduardo , Carnero Elvis A. , Standley Robert A. , Giráldez-García Manuel A. , Dopico-Calvo Xurxo , Tuimil Jose Luis
Published: December 2016

The main aim of this study was to analyze the effect of resistance training programs differing in set configuration on mechanical force-velocity profiles. Thirteen participants performed 10 unilateral knee extension training sessions over 5 weeks. Each limb was randomized to one of the following set configurations: traditional (4 sets of 8 repetitions at maximum intended velocity, 10RM load, 3-min pause between sets) or interrepetition rest (32 maximum intended velocity repetitions, 10RM load, 17.4 s of rest between each repetition). Velocity of each repetition was recorded throughout the program. Before and after training, individual linear force velocities were calculated, and the following parameters were obtained: force and velocity axis intercept, slope, and estimated maximum power. Mean velocity was higher throughout the program for interrepetition rest configuration (0.54 ± 0.01 vs. 0.48 ± 0.01 m∙s−1 for interrepetition rest, and traditional configuration respectively; main effect of set configuration: P < .001). There was a significant increase in force and velocity intercepts, but a steeper negative slope after both training protocols (main effect of time: P < .001 for every variable). Differences in resistance training velocity did not affect the adaptations. Our results suggest that, in a short-term program, maximum intended rather than actual velocity is a key factor to modulate strength adaptations.
The Associations Between Urbanicity and Physical Activity and Sitting Time in Mexico
Authors: Hermosillo-Gallardo Maria E. , Jago Russell , Sebire Simon J.
Published: December 2016

Background:

Approximately 17.4% of people in Mexico self-report physical activity levels below the World Health Organization’s guidelines and an average sedentary time of 16 hours per day.1 Low physical activity has been associated with noncommunicable disease risk factors and previous research suggests that urbanicity might be an important determinant of physical activity. The aim of this study was to measure urbanicity in Mexico and assess if it is associated with physical activity and sitting time.

Methods:

A sample of 2880 men and 4211 women aged 20 to 69 was taken from the 2012 Mexico National Health and Nutrition Survey and multivariable linear regression models were used to examine the association between physical activity, sitting time and urbanicity; adjusting for sex, education level, socioeconomic status and Body Mass Index. The urbanicity score and the 7 urbanicity subscores were estimated from the CENSUS 2010.

Results:

The subscores of demographic, economic activity, diversity and communication were negatively associated with physical activity. Sitting time was positively associated with the overall urbanicity, and the demographic and health subscores.

Conclusions:

There was evidence of associations between urbanicity and physical activity in Mexico.
Are Immigrants More Physically Active Than Native-Born Australians and Does it Changes Over Time? Evidence From a Nationally Representative Longitudinal Survey
Authors: Joshi Suresh , Jatrana Santosh , Paradies Yin
Published: December 2016

Background:

We investigated the differences and over time changes in recommended physical activity among foreign-born (FB) from English speaking countries (ESC) and non-English speaking countries (NESC) relative to native-born (NB) Australians, and whether the association between nativity and duration of residence (DoR) and physical activity is mediated by English language proficiency, socioeconomic status and social engagement/membership.

Methods:

This study applies multilevel group-meancentered mixed (hybrid) logistic regression models to 12 waves of longitudinal data (12,634 individuals) from the Household, Income and Labor Dynamics in Australia survey with engagement in physical activities for more than 3 times a week as the outcome variable.

Results:

Immigrants from ESC had higher odds of physical activity, while immigrants from NESC had significantly lower odds of physical activity than NB Australians, after adjusting for covariates. There was no evidence that these differences changed by DoR among immigrants from NESC, whereas ESC immigrants had higher odds of physical activity when their DoR was more than 20 years. We also found a mediating role of English language proficiency on immigrants physical activities.

Conclusion:

Appropriate health promotion interventions should be implemented to foster physical activities among NESC immigrants, considering English language proficiency as an important factor in designing interventions.
Psychosocial and Friend Influences on Objective Sedentary Behavior and Screen Time: A Mixed Methods Analysis
Authors: Garcia Jeanette M. , Agaronov Alen , Sirard John R. , Whaley Diane , Rice David J. , Weltman Arthur
Published: December 2016

Background:

Sedentary behavior (SB) increases throughout adolescence, and is associated with adverse health outcomes.

Purpose:

Examine psychosocial and friend influences on SB and screen time in adolescents using a mixed-methods design.

Methods:

108 middle and high school students wore accelerometers to measure objective SB, completed screen time and psychosocial questionnaires, and nominated friends to complete activity questionnaires. Focus groups centered around influences on SB behavior. Regression analyses and NVivo software analyzed quantitative and qualitative data.

Results:

Screen time was associated with greater screen time enjoyment, lower self-efficacy, and friends’ screen time (r2 = .21, P < .0001). Friends influenced whether adolescents engaged in screen time behaviors, with active friends encouraging less screen time.

Conclusion:

Active friends influenced adolescents to engage in less SB. Interventions should place an emphasis on encouraging less screen time, and providing opportunities for adolescents and their friends to engage in activities that promote physical activity rather than SB.
Patterns and Predictors of Change in Moderate-to-Vigorous Physical Activity Over Time in Multiple Sclerosis
Authors: Klaren Rachel E. , Sasaki Jeffer E. , McAuley Edward , Motl Robert W.
Published: December 2016

Background:

Physical inactivity is common in persons with multiple sclerosis (MS), but there is very little known about the pattern and predictors of changes in physical activity over time.

Purpose:

This study examined changes in moderate-to-vigorous physical activity (MVPA) over a 30-month time period and the demographic and clinical predictors of such changes in relapsing-remitting MS (RRMS).

Methods:

269 persons with MS wore an accelerometer for a 7-day period and completed a demographic/clinical scale every 6 months over a 30-month period. Data were analyzed using latent class growth modeling (LCGM).

Results:

LCGM identified a two-class model for changes in levels of MVPA over time. Class 1 involved higher initial levels of MVPA and linear decreases in MVPA over time, whereas Class 2 involved lower initial levels of MVPA and linear increases in MVPA over time. LCGM further indicated that males were more likely (OR = 5.8, P < .05) and those with higher disability status were less likely (OR = 0.51, P < .05) to belong to Class 1 than Class 2.

Conclusion:

Levels of MVPA change over time in persons with RRMS and the pattern of change suggests that behavioral physical activity interventions for persons with MS might target men and those with lower disability.
Cotranscription of genes encoding indoleacetic acid production in Pseudomonas syringae subsp. savastanoi
Authors: Palm C J , Gaffney T , Kosuge T
Published: December 2016

Indoleacetic acid (IAA) production by the plant pathogen Pseudomonas syringae subsp. savastanoi is essential for tumor formation on olive and oleander. The bacterium produces IAA from tryptophan in reactions catalyzed by tryptophan monooxygenase and indoleacetamide hydrolase. The genetic determinants are, respectively, iaaM and iaaH. In oleander isolates, the genes encoding the IAA biosynthetic enzymes are located on a plasmid; in olive isolates, the genes occur on the chromosome. The IAA genes from the oleander isolate strain EW2009 are located within a 4-kilobase (kb) segment of the 52-kb plasmid pIAA1. Escherichia coli strains harboring a recombinant plasmid, pCJP3, which contains this 4-kb fragment, excreted IAA into culture media, and crude cell extracts had both tryptophan monooxygenase and indoleacetamide hydrolase activity. In vitro coupled transcription-translation of pCJP3 demonstrated that this fragment coded for proteins of 62 and 47 kilodaltons which correspond to tryptophan monooxygenase and indoleacetamide hydrolase, respectively. Expression of these genes was dependent upon a vector promoter in pCJP3. However, in the absence of a vector promoter, E. coli containing recombinant plasmids with additional pIAA1 DNA in front of iaaM had high levels of tryptophan monooxygenase. Northern (RNA) hybridization experiments verified that iaaM and iaaH are cotranscribed as a portion of a ca. 4- to 5-kb transcript in vivo. Southern hybridization experiments with IAA plasmids from different oleander strains of P. syringae subsp. savastanoi revealed that all IAA plasmids contained a region of at least 10 kb of homology, with the IAA genes at one end. Repetitive DNA and a copy of IS51 were found at the end of this region of homology.
Multiple and tandemly arranged promoters of the cell wall protein gene operon in Bacillus brevis 47
Authors: Adachi T , Yamagata H , Tsukagoshi N , Udaka S
Published: December 2016

The genes encoding the major cell wall proteins, middle wall protein and outer wall protein, of Bacillus brevis 47 constitute a cotranscriptional unit (cwp [cell wall protein gene] operon). Primer extension assay of cwp operon transcripts showed the existence of six different 5' ends. This confirmed the results of the previous S1 nuclease protection assay and suggested the existence of several tandemly arranged promoters in the 5' region of the cwp operon. Promoter probe vectors carrying the Bacillus licheniformis alpha-amylase gene were constructed and used for deletion analysis of the 5' region. Three (P1, P2, and P3) of the six suggested promoters were shown to be located within three distinct fragments derived from the 5' region. The -35 and -10 regions of the P1 and P3 promoters resemble the consensus sequence recognized by the sigma-43-type RNA polymerase of Bacillus subtilis. The P2 promoter resembles only the consensus sequence in the -10 region. The P1 and P3 promoters were used to the same extents in Bacillus subtilis as in B. brevis, whereas the P2 promoter was used much less frequently in B. subtilis than in B. brevis. The P2 promoter is used constitutively in B. brevis 47 at all stages of growth, whereas P3 is used only at the exponential phase of growth. P2 could be a promoter of an unknown type that is preferentially used in B. brevis and might be responsible for the constitutive synthesis and secretion of the cell wall proteins into the medium at the stationary phase of growth.
Evolutionary relationship between the TonB-dependent outer membrane transport proteins: nucleotide and amino acid sequences of the Escherichia coli colicin I receptor gene
Authors: Nau C D , Konisky J
Published: December 2016

The nucleotide sequence of the Escherichia coli colicin I receptor gene (cir) has been determined. The predicted mature protein consists of 599 amino acids and has a molecular weight of 67,169. Several previously noted characteristics of other E. coli outer membrane protein sequences were also identified in the sequence of Cir. These include an overall acidic nature, the absence of long hydrophobic stretches of amino acids, and a lack of predicted alpha-helical secondary structure. Because two classes of outer membrane proteins (the TonB-dependent transport proteins and the porins) share some structural features, protein sequences from both of these groups were aligned pairwise and scored for sequence similarity. Statistical evidence suggested that the porins were not related to the proteins in the TonB-dependent group; however, there was a significant relationship between the proteins in the TonB-dependent group. On the basis of the multiple progressive sequence alignment and the similarity scores derived from it, a tree representing evolutionary distance between five TonB-dependent outer membrane transport proteins was generated.
Mechanism for iron-regulated transcription of the Escherichia coli cir gene: metal-dependent binding of fur protein to the promoters
Authors: Griggs D W , Konisky J
Published: December 2016

The molecular basis for the greatly elevated expression of the cir gene (encoding the colicin I receptor) in cells unable to maintain a critical supply of intracellular iron was investigated by genetic and biochemical means. Deletion analysis of the cloned promoter region allowed delineation of sequences necessary for control of transcription initiating at the two promoters, P1 and P2. Gel retardation assays were used to demonstrate both binding of purified Fur (ferric uptake regulation) protein to the iron control region and lack of binding to DNA fragments which are not involved in cir regulation. An operator sequence spanning 43 to 47 base pairs and completely encompassing the two promoters was identified by DNase I protection experiments (footprinting), with binding occurring in a metal-dependent fashion. Thus, during iron-replete growth, Fur appears to act as a repressor of transcription by blocking formation of a DNA-RNA polymerase complex, analogous to the mechanism previously described for regulation of the aerobactin operon (V. de Lorenzo, S. Wee, M. Herrero, and J.B. Neilands, J. Bacteriol. 169:2624-2630, 1987). Characterized and putative Fur recognition sites from several genes were analyzed and classified by statistical methods.
Modulation of alcohol dehydrogenase isoenzyme levels in Zymomonas mobilis by iron and zinc
Authors: Mackenzie K F , Eddy C K , Ingram L O
Published: December 2016

Zymomonas mobilis is an unusual microorganism which utilizes both iron-containing alcohol dehydrogenase (ADHII) and zinc-containing alcohol dehydrogenase (ADHI) isoenzymes during fermentative growth. This organism is obligately ethanologenic, and alcohol dehydrogenase activity is essential. The activities of ADHI and ADHII were altered by supplementing growth medium with iron or zinc salts and by iron starvation. Growth under iron-limiting conditions (chelators, minimal medium) reduced ADHII activity but did not prevent the synthesis of the ADHII protein. The inactive form of this enzyme appeared quite stable, was not renatured by iron addition, and persisted in the cell. The iron-induced increase in ADHII activity required de novo synthesis which was blocked by antibiotic additions. The ability of Z. mobilis to synthesize ADHII and ADHI may be advantageous in nature.
A recB recC sbcB recJ host prevents recA-independent deletions in recombinant cosmid DNA propagated in Escherichia coli
Authors: Ishiura M , Hazumi N , Koide T , Uchida T , Okada Y
Published: December 2016

Segments of DNA are deleted from recombinant cosmid DNAs with high frequency during propagation in standard recA Escherichia coli hosts. An attempt has been made to derive an appropriate strain of E. coli, suitable for cosmid cloning, in which such deletions do not occur. We examined the effects of a series of host recombinational mutations on the deletion process, using six independent recombinant cosmids that carry inserts of mouse, Chinese hamster, or human DNA. Various E. coli host cells carrying the recombinant cosmids were cultured serially in liquid medium, and the recombinant cosmid DNAs were extracted from the host cells and analyzed by agarose gel electrophoresis and by gene transfer of the DNAs into cultured mammalian cells. Of the mutations examined, only a recB recC sbcB recJ (or recN) quadruple combination of host mutations prevented the deletion of DNA segments. The recombinant cosmid DNAs propagated in E. coli hosts that carried this combination of mutations were functionally as well as structurally intact. We propose that the recJ (and/or recN) gene is involved in some aspect of the events that lead to deletions of cosmid DNA in a recB recC sbcB genetic background.
Nucleotide sequence and mutational analysis of the structural genes (anfHDGK) for the second alternative nitrogenase from Azotobacter vinelandii
Authors: Joerger R D , Jacobson M R , Premakumar R , Wolfinger E D , Bishop P E
Published: December 2016

The nucleotide sequence of a region of the Azotobacter vinelandii genome exhibiting sequence similarity to nifH has been determined. The order of open reading frames within this 6.1-kilobase-pair region was found to be anfH (alternative nitrogen fixation, nifH-like gene), anfD (nifD-like gene), anfG (potentially encoding a protein similar to the product of vnfG from Azotobacter chroococcum), anfK (nifK-like gene), followed by two additional open reading frames. The 5'-flanking region of anfH contains a nif promoter similar to that found in the A. vinelandii nifHDK gene cluster. The presumed products of anfH, anfD, and anfK are similar in predicted Mr and pI to the previously described subunits of nitrogenase 3. Deletion plus insertion mutations introduced into the anfHDGK region of wild-type strain A. vinelandii CA resulted in mutant strains that were unable to grow in Mo-deficient, N-free medium but grew in the presence of 1 microM Na2MoO4 or V2O5. Introduction of the same mutations into the nifHDK deletion strain CA11 resulted in strains that grew under diazotrophic conditions only in the presence of vanadium. The lack of nitrogenase 3 subunits in these mutant strains was demonstrated through two-dimensional gel analysis of protein extracts from cells derepressed for nitrogenase under Mo and V deficiency. These results indicate that anfH, anfD, and anfK encode structural proteins for nitrogenase 3.
Cascading regulation of histidase activity in Streptomyces griseus
Authors: Kroening T A , Kendrick K E
Published: December 2016

Mutants of Streptomyces griseus unable to utilize histidine as the sole nitrogen source have been isolated and characterized. Using a mutant defective in the production of histidase, we have demonstrated that urocanate functions as the inducer of the histidine utilization system. Another mutant produced histidase that was locked in an inactive form but could be activated by treatment with an extract from the wild-type strain or the histidase-negative strain. This mutant was deficient in the activity of a protein of Mr ca. 90,000 to 100,000 that is required for the activation of histidase. Histidase was synthesized constitutively but was maintained in an inactive form until after histidine or urocanate was added to the medium. At least four components were implicated in the activation of histidase: histidase, the activation protein, urocanate, and a phosphatase that is apparently inactive in cells grown without inducer. The functions of the last three factors could be supplanted in vitro by incubation of histidase with snake venom phosphodiesterase or 5' nucleotidase. The results suggest that histidine utilization by S. griseus is controlled posttranslationally by an activation cascade that involves at least two regulatory proteins.
Isolation and characterization of mutants of Rhizobium leguminosarum bv. viciae 248 with altered lipopolysaccharides: possible role of surface charge or hydrophobicity in bacterial release from the infection thread
Authors: de Maagd R A , Rao A S , Mulders I H , Goosen-de Roo L , van Loosdrecht M C , Wijffelman C A , Lugtenberg B J
Published: December 2016

Effects of alterations in lipopolysaccharide (LPS) structure of Rhizobium leguminosarum bv. viciae on effective symbiosis and on a number of cell surface characteristics were studied. Tn5 mutants with altered LPSs were screened for their inability to bind monoclonal antibody 3, one of three monoclonal antibodies to the tentative O-antigenic part of the wild-type LPS of strain 248. Ten class I LPS mutants completely lacked the O-antigen-containing LPS species. The class II LPS mutant had a severely diminished amount of an antigenically altered O-antigen-containing LPS. The class III LPS mutant had normal amounts of an altered, O-antigen-containing LPS. Class I and II mutants, but not the class III mutant, showed abnormal nodule development (i.e., blocked in the stage of bacterial release from the infection thread) resulting in nodules in which very few, at the most, plant cells contained bacteroids and which were unable to fix nitrogen. Class I and II mutants were nonmotile and were more sensitive to hydrophobic compounds than the parent strain. The most striking difference between the symbiotically defective class I and II LPS mutants on one hand and the wild-type strain and the class III mutant on the other hand was that the class I and II mutants have a more hydrophobic cell surface and a higher electrophoretic mobility. A role for an O-antigen-containing LPS in bacterial release from the infection thread, through its effects on general physicochemical cell surface characteristics, is proposed.
Point mutations in the regulatory region of the ilvGMEDA operon of Escherichia coli K-12
Authors: Lawther R P
Published: December 2016

The ilvGMEDA operon of Escherichia coli K-12 is preceded by a regulatory region containing a promoter, a leader, and an attenuator. This region has been extensively characterized biochemically. In this note point mutations of the regulatory region are reported. The effect of these mutations on expression from the ilv regulatory region supports the previous biochemical analysis.
Induction of the alkylation-inducible aidB gene of Escherichia coli by anaerobiosis
Authors: Volkert M R , Hajec L I , Nguyen D C
Published: December 2016

Induction of the adaptive response to alkylation damage results in the expression of four genes arranged in three transcriptional units: the ada-alkB operon and the alkA and aidB genes. Adaptive-response induction requires the ada gene product and occurs when cells are treated with methylating agents. In previous studies we noted that aidB, but not alkA or ada-alkB, was induced in the absence of alkylation damage as cells were grown to stationary phase. In this note we present evidence that aidB is induced by anaerobiosis. Thus, aidB is subject to dual regulation by ada-dependent alkylation induction and ada-independent anaerobic induction.
Cloning and expression in Escherichia coli of an extremely thermostable oligo-1,6-glucosidase gene from Bacillus thermoglucosidasius
Authors: Watanabe K , Iha H , Ohashi A , Suzuki Y
Published: December 2016

The gene for an extremely thermostable oligo-1,6-glucosidase (dextrin-6-alpha-D-glucanohydrolase; EC 3.2.1.10) of obligately thermophilic Bacillus thermoglucosidasius KP1006 was cloned within a 4.2-kilobase HindIII-PvuII fragment of DNA by using the plasmid pUC19 as a vector and Escherichia coli C600 as a host. The gene was transcribed, presumably from its own promoter, in E. coli. E. coli with the hybrid plasmid accumulated oligo-1,6-glucosidase mainly in the cytoplasm. The level of enzyme production was comparable to that observed for B. thermoglucosidasius. The enzyme coincided absolutely with the B. thermoglucosidasius enzyme in its molecular weight (60,000), in its electrophoretic behavior on denaturing and nondenaturing polyacrylamide gels, in the temperature dependency of its stability and activity, and in its antigenic determinants.
Fragments of prochymosin produced in Escherichia coli form insoluble inclusion bodies
Authors: McCaman M T
Published: December 2016

Bovine prochymosin produced in Escherichia coli has been used as a model system to investigate factors which may cause a recombinant protein to accumulate as insoluble inclusion bodies. A series of plasmids was constructed to investigate the effect of deletions within the prochymosin-coding sequence on protein inclusion body formation. The results demonstrated that as much as 70% of the prochymosin-coding sequence could be deleted with no significant reduction in the accumulation of insoluble protein. The smallest deletion product identified (11,000 molecular weight) retained only one cysteine, yet this product still accumulated as an insoluble product in E. coli.
Bacillus subtilis phenylalanyl-tRNA synthetase genes: cloning and expression in Escherichia coli and B. subtilis
Authors: Brakhage A A , Putzer H , Shazand K , Röschenthaler R J , Grunberg-Manago M
Published: December 2016

The genes that encode the two subunits of Bacillus subtilis phenylalanyl-tRNA synthetase were cloned from alpha lambda library of chromosomal B. subtilis DNA by specific complementation of a thermosensitive Escherichia coli pheS mutation. Both genes (we named them pheS and pheT, analogous to the corresponding genes of E. coli) are carried by a 6.6-kilobase-pair PstI fragment which also complements E. coli pheT mutations. This fragment directs the synthesis of two proteins identical in size to the purified alpha and beta subunits of the phenylalanyl-tRNA synthetase of B. subtilis with Mrs of 42,000 and 97,000, respectively. A recombinant shuttle plasmid carrying the genes caused 10-fold overproduction of functional phenylalanyl-tRNA synthetase in B. subtilis.
Expression and nucleotide sequence of the Lactobacillus bulgaricus beta-galactosidase gene cloned in Escherichia coli
Authors: Schmidt B F , Adams R M , Requadt C , Power S , Mainzer S E
Published: December 2016

The Lactobacillus bulgaricus beta-galactosidase gene was cloned on a ca. 7-kilobase-pair HindIII fragment in the vector pKK223-3 and expressed in Escherichia coli by using its own promoter. The nucleotide sequence of the gene and approximately 400 bases of 3'- and 5'-flanking sequences was determined. The amino acid sequence of the beta-galactosidase, deduced from the nucleotide sequence of the gene, yielded a monomeric molecular mass of ca. 114 kilodaltons, slightly smaller than the E. coli lacZ and Klebsiella pneumoniae lacZ enzymes but larger than the E. coli evolved (ebgA) beta-galactosidase. The cloned beta-galactosidase was found to be indistinguishable from the native enzyme by several criteria. From amino acid sequence alignments, the L. bulgaricus beta-galactosidase has a 30 to 34% similarity to the E. coli lacZ, E. coli ebgA, and K. pneumoniae lacZ enzymes. There are seven regions of high similarity common to all four of these beta-galactosidases. Also, the putative active-site residues (Glu-461 and Tyr-503 in the E. coli lacZ beta-galactosidase) are conserved in the L. bulgaricus enzyme as well as in the other two beta-galactosidases mentioned above. The conservation of active-site amino acids and the large regions of similarity suggest that all four of these beta-galactosidases evolved from a common ancestral gene. However, these enzymes are quite different from the thermophilic beta-galactosidase encoded by the Bacillus stearothermophilus bgaB gene.