bluuebunny/crossref_metadata_2025_split huggingface dataset:
Translating Stem Cell Research: Challenges at the Research Frontier
Authors: Magnus David 
Published: June 2010

There are many kinds of clinical trials. The regulatory framework within which most drug development takes place appears to be the one that is to be applied to the development of novel stem cell-based clinical trials. In the standard drug development model, appropriate pre-clinical research is conducted (largely based upon non-human animal models), and investigators or research sponsors (typically a company) submit an investigational new drug (IND) application to the Food and Drug Administration (FDA).If approved, typical clinical trials start with Phase I, which is usually a trial (with a small number of participants) to determine the maximum tolerable dose of a drug. Phase I trials are often referred to as “safety studies” because the primary goal of Phase I research is to determine if an intervention is safe (not whether it works).
Damage Control: Unintended Pregnancy in the United States Military
Authors: Ponder Kathryn L. , Nothnagle Melissa 
Published: June 2010

Military reproductive health policies affect large numbers of women. In 2006 servicewomen numbered nearly 350,000 and comprised 14.5% of active-duty forces and 17.4% of the reserve force. In addition, approximately 165,000 female dependents of active duty military personnel and 157,000 female dependents of reserve duty personnel are between the ages of 12 and 22 and are eligible for military health care services. Dependents of military personnel are eligible for military health care coverage until age 21, or up to the age of 23 if considered a full-time student.About 10% of active duty female servicemembers become pregnant each year. Pregnant servicewomen are considered non-deployable and are usually not assigned to overseas commands. Pregnant women in the Army, the service branch with the greatest number of women on active duty, can choose to remain on active duty, request temporary leave from service, or request separation from service. Separation options include either discharge, honorable or uncharacterized depending on rank, or transfer to individual ready reserve.
Teaching Health Law
Authors: Apel Susan B. 
Published: June 2010

Interdisciplinary teaching can be a hard sell to the legal academic community. Over almost three decades, I have spoken at conferences on a variety of subjects. When I have presented on this particular topic, however, I have drawn my most meager crowds. Is it because we think interdisciplinary pedagogy is a bad idea, that we are ill-equipped, or that it is generally too difficult to do successfully? After a dozen years of creating and teaching an interdisciplinary course in law and medicine, I confess that it is now my other, law-only courses that I feel I have to justify. Why is an interdisciplinary approach to teaching law the exception rather than the rule?My path to creating an interdisciplinary course in law and medicine (MedLaw) began indirectly and with impatience at my own teaching of family law. I had been a professor of family law for decades, and I had developed a sophisticated set of simulations that asked students to negotiate a settlement in a marital dissolution case.
Prevalence of Cysticercus bovis in Australian cattle
Authors: Pearse BHG , Traub RJ , Davis A , Cobbold R , Vanderlinde PB 
Published: June 2010

Objective  The first national abattoir survey of Cysticercus bovis (‘beef measles’) in cattle was conducted in February 2008.Methods  During the data collection period, 493,316 cattle were subjected to standard postmortem procedures, including incision of the masseter and heart muscles. On‐site veterinarians were asked to record the location of any C. bovis cysts, as well as the National Livestock Identification System ear tag numbers of infected animals. Veterinarians were asked to submit samples for laboratory confirmation by histology and polymerase chain reaction testing.Results  Of the 23 samples submitted, none was positive for C. bovis by either diagnostic method.Conclusions  Occasional, isolated diagnoses of beef measles are still made in most states of Australia, but since the last regional surveys were conducted 30 years ago, when the estimated prevalence was 50 to 200 per 100,000 cattle slaughtered, the parasite has become extremely rare.
Culturing marine bacteria – an essential prerequisite for biodiscovery
Authors: Joint Ian , Mühling Martin , Querellou Joël 
Published: June 2010

SummaryThe potential for using marine microbes for biodiscovery is severely limited by the lack of laboratory cultures. It is a long‐standing observation that standard microbiological techniques only isolate a very small proportion of the wide diversity of microbes that are known in natural environments from DNA sequences. A number of explanations are reviewed. The process of establishing laboratory cultures may destroy any cell‐to‐cell communication that occurs between organisms in the natural environment and that are vital for growth. Bacteria probably grow as consortia in the sea and reliance on other bacteria for essential nutrients and substrates is not possible with standard microbiological approaches. Such interactions should be considered when designing programmes for the isolation of marine microbes. The benefits of novel technologies for manipulating cells are reviewed, including single cell encapsulation in gel micro‐droplets. Although novel technologies offer benefits for bringing previously uncultured microbes into laboratory culture, many useful bacteria can still be isolated using variations of plating techniques. Results are summarized for a study to culture bacteria from a long‐term observatory station in the English Channel. Bacterial biodiversity in this assemblage has recently been characterized using high‐throughput sequencing techniques. Although Alphaproteobacteria dominated the natural bacterial assemblage throughout the year, Gammaproteobacteria were the most frequent group isolated by plating techniques. The use of different gelling agents and the addition of ammonium to seawater‐based agar did lead to the isolation of a higher proportion of Alphaproteobacteria. Variation in medium composition was also able to increase the recovery of other groups of particular interest for biodiscovery, such as Actinobacteria.
A comparison of canopy evapotranspiration for maize and two perennial grasses identified as potential bioenergy crops
Authors: HICKMAN GEORGE C. , VANLOOCKE ANDY , DOHLEMAN FRANK G. , BERNACCHI CARL J. 
Published: June 2010

AbstractIn the Midwestern US, perennial rhizomatous grasses (PRGs) are considered one of the most promising vegetation types to be used as a cellulosic feedstock for renewable energy production. The potential widespread use of biomass crops for renewable energy production has sparked numerous environmental concerns, including the impacts of land‐use change on the hydrologic cycle. We predicted that total seasonal evapotranspiration (ET) would be higher for PRGs relative to maize resulting from higher leaf area and a prolonged growing season. We further predicted that, compared with maize, higher aboveground biomass associated with PRGs would offset the higher ET and increase water‐use efficiency (WUE) in the context of biomass harvests for liquid biofuel production. To test these predictions, ET was estimated during the 2007 growing season for replicated plots of Miscanthus×giganteus (miscanthus), Panicum virgatum (switchgrass), and Zea mays (maize) using a residual energy balance approach. The combination of a 25% higher mean latent heat flux (λET) and a longer growing season resulted in miscanthus having ca. 55% higher cumulative ET over the growing season compared with maize. Cumulative ET for switchgrass was also higher than maize despite similar seasonal‐mean λET. Based on total harvested aboveground biomass, WUE was ca. 50% higher for maize relative to miscanthus; however, when WUE calculated from only maize grain biomass was compared with WUE calculated from miscanthus harvested aboveground biomass, this difference disappeared. Although WUE between maize and miscanthus differed postsenescence, there were no differences in incremental WUE throughout the growing season. Despite initial predictions, aboveground biomass for switchgrass was less than maize; thus WUE was substantially lower for switchgrass than for either maize scenario. These results indicate that changes in ET due to large‐scale implementation of PRGs in the Midwestern US would likely influence local and regional hydrologic cycles differently than traditional row crops.
Fasting tests of insulin secretion and sensitivity predict future prediabetes in Japanese with normal glucose tolerance
Authors: Onishi Yukiko , Hayashi Tomoshige , Sato Kyoko Kogawa , Ogihara Takehide , Kuzuya Nobuaki , Anai Motonobu , Tsukuda Katsunori , Boyko Edward J , Fujimoto Wilfred Y , Kikuchi Masatoshi 
Published: June 2010

AbstractAims/Introduction:  Reduced insulin sensitivity and secretion are important in the pathogenesis of type 2 diabetes. Their relationships to prediabetes, impaired glucose tolerance (IGT) and impaired fasting glucose (IFG) have been previously studied with the oral glucose tolerance test (OGTT). We investigated whether or not baseline measures of insulin secretion and sensitivity obtained from fasting blood specimens were related to the development of prediabetes and how these measures compared with those based on the OGTT.Materials and Methods:  In 152 Japanese subjects with normal glucose tolerance, we measured baseline plasma glucose and insulin after an overnight fast and during a 75 g OGTT, insulin resistance index (homeostasis model assessment [HOMA‐IR]), and insulin secretion (insulinogenic index [30 min insulin − fasting insulin] ÷ [30 min glucose − fasting glucose] or HOMA‐β).Results:  At a 5–6 year (mean 5.7 years) follow‐up examination, we confirmed 36 cases of prediabetes. After adjusting for age, sex, family history of diabetes, body mass index, and 2‐h plasma glucose, the odds ratio comparing the lowest tertile (≤0.82) of insulinogenic index with the highest tertile (≥1.43) was 6.98 (95% confidence interval, 1.96–24.85) and was 10.72 (2.08–55.3) comparing the lowest tertile (≤76.3) of HOMA‐β with the highest tertile (≥122.1), whereas the respective odds ratios of HOMA‐IR were 3.74 (1.03–13.57) and 10.89 (1.93–61.41) comparing the highest tertile (≥1.95) with the lowest tertile (≤1.25).Conclusions:  Lower insulin secretion and sensitivity are independent risk factors for prediabetes. Clinically practical identification of those at risk for prediabetes is obtainable from HOMA‐β and HOMA‐IR, both of which are measured in fasting state. (J Diabetes Invest, doi: 10.1111.j.2040‐1124.2010.00041.x, 2010)
Rapid Quantification of Viable
Campylobacter
Bacteria on Chicken Carcasses, Using Real-Time PCR and Propidium Monoazide Treatment, as a Tool for Quantitative Risk Assessment
Authors: Josefsen M. H. , Löfström C. , Hansen T. B. , Christensen L. S. , Olsen J. E. , Hoorfar J. 
Published: June 2010

ABSTRACT

A number of intervention strategies against
Campylobacter
-contaminated poultry focus on postslaughter reduction of the number of cells, emphasizing the need for rapid and reliable quantitative detection of only viable
Campylobacter
bacteria. We present a new and rapid quantitative approach to the enumeration of food-borne
Campylobacter
bacteria that combines real-time quantitative PCR (Q-PCR) with simple propidium monoazide (PMA) sample treatment. In less than 3 h, this method generates a signal from only viable and viable but nonculturable (VBNC)
Campylobacter
bacteria with an intact membrane. The method's performance was evaluated by assessing the contributions to variability by individual chicken carcass rinse matrices, species of
Campylobacter
, and differences in efficiency of DNA extraction with differing cell inputs. The method was compared with culture-based enumeration on 50 naturally infected chickens. The cell contents correlated with cycle threshold (

C
T

) values (
R
2
= 0.993), with a quantification range of 1 × 10
2
to 1 × 10
7
CFU/ml. The correlation between the
Campylobacter
counts obtained by PMA-PCR and culture on naturally contaminated chickens was high (
R
2
= 0.844). The amplification efficiency of the Q-PCR method was not affected by the chicken rinse matrix or by the species of
Campylobacter
. No Q-PCR signals were obtained from artificially inoculated chicken rinse when PMA sample treatment was applied. In conclusion, this study presents a rapid tool for producing reliable quantitative data on viable
Campylobacter
bacteria in chicken carcass rinse. The proposed method does not detect DNA from dead
Campylobacter
bacteria but recognizes the infectious potential of the VBNC state and is thereby able to assess the effect of control strategies and provide trustworthy data for risk assessment.
Novel, Versatile, and Tightly Regulated Expression System for
E
scherichia coli
Strains
Authors: Choi Young J. , Morel Lyne , Le François Teffanie , Bourque Denis , Bourget Lucie , Groleau Denis , Massie Bernard , Míguez Carlos B. 
Published: June 2010

ABSTRACT

A novel tightly regulated gene expression system was developed for
Escherichia coli
by applying the regulatory elements of the
Pseudomonas putida
F1
cym
and
cmt
operons to control target gene expression at the transcriptional level by using
p
-isopropylbenzoate (cumate) as an inducer. This novel expression system, referred to as the cumate gene switch, includes a specific expression vector, pNEW, that contains a partial T
5
phage promoter combined with the
Pseudomonas
-based synthetic operator and the
cymR
repressor protein-encoding gene designed to express constitutively in the host strain. The induction of transcription relies on the addition of the exogenous inducer (cumate), which is nontoxic to the culture, water soluble, and inexpensive. The characteristics and potential of the expression system were determined. Using flow cytometry and fed-batch fermentations, we have shown that, with the newly developed cumate-regulated system, (i) higher recombinant product yields can be obtained than with the pET (isopropyl-β-
d
-thiogalactopyranoside [IPTG])-induced expression system, (ii) expression is tightly regulated, (iii) addition of cumate quickly results in a fully induced and homogenous protein-expressing population in contrast to the bimodal expression profile of an IPTG-induced population, (iv) expression can be modulated by varying the cumate concentration, and (v) the cumate-induced population remains induced and fully expressing even at 8 h following induction, resulting in high yields of the target protein Furthermore, the cumate gene switch described in this article is applicable to a wide range of
E. coli
strains.
Transcriptomic Insights into the Physiology ofAspergillus nigerApproaching a Specific Growth Rate of Zero
Authors: Jørgensen Thomas R. , Nitsche Benjamin M. , Lamers Gerda E. , Arentshorst Mark , van den Hondel Cees A. , Ram Arthur F. 
Published: June 2010

ABSTRACTThe physiology of filamentous fungi at growth rates approaching zero has been subject to limited study and exploitation. With the aim of uncoupling product formation from growth, we have revisited and improved the retentostat cultivation method forAspergillus niger. A new retention device was designed allowing reliable and nearly complete cell retention even at high flow rates. Transcriptomic analysis was used to explore the potential for product formation at very low specific growth rates. The carbon- and energy-limited retentostat cultures were highly reproducible. While the specific growth rate approached zero (<0.005 h−1), the growth yield stabilized at a minimum (0.20 g of dry weight per g of maltose). The severe limitation led to asexual differentiation, and the supplied substrate was used for spore formation and secondary metabolism. Three physiologically distinct phases of the retentostat cultures were subjected to genome-wide transcriptomic analysis. The severe substrate limitation and sporulation were clearly reflected in the transcriptome. The transition from vegetative to reproductive growth was characterized by downregulation of genes encoding secreted substrate hydrolases and cell cycle genes and upregulation of many genes encoding secreted small cysteine-rich proteins and secondary metabolism genes. Transcription of known secretory pathway genes suggests thatA. nigerbecomes adapted to secretion of small cysteine-rich proteins. The perspective is thatA. nigercultures as they approach a zero growth rate can be used as a cell factory for production of secondary metabolites and cysteine-rich proteins. We propose that the improved retentostat method can be used in fundamental studies of differentiation and is applicable to filamentous fungi in general.
Production of a New Thiopeptide Antibiotic, TP-1161, by a Marine
Nocardiopsis
Species
Authors: Engelhardt Kerstin , Degnes Kristin F. , Kemmler Michael , Bredholt Harald , Fjærvik Espen , Klinkenberg Geir , Sletta Håvard , Ellingsen Trond E. , Zotchev Sergey B. 
Published: June 2010

ABSTRACT

Twenty-seven marine sediment- and sponge-derived actinomycetes with a preference for or dependence on seawater for growth were classified at the genus level using molecular taxonomy. Their potential to produce bioactive secondary metabolites was analyzed by PCR screening for genes involved in polyketide and nonribosomal peptide antibiotic synthesis. Using microwell cultures, conditions for the production of antibacterial and antifungal compounds were identified for 15 of the 27 isolates subjected to this screening. Nine of the 15 active extracts were also active against multiresistant Gram-positive bacterial and/or fungal indicator organisms, including vancomycin-resistant
Enterococcus faecium
and multidrug-resistant
Candida albicans
. Activity-guided fractionation of fermentation extracts of isolate TFS65-07, showing strong antibacterial activity and classified as a
Nocardiopsis
species, allowed the identification and purification of the active compound. Structure elucidation revealed this compound to be a new thiopeptide antibiotic with a rare aminoacetone moiety. The
in vitro
antibacterial activity of this thiopeptide, designated TP-1161, against a panel of bacterial strains was determined.
Deletion of a
fur
-Like Gene Affects Iron Homeostasis and Magnetosome Formation in
Magnetospirillum gryphiswaldense
Authors: Uebe René , Voigt Birgit , Schweder Thomas , Albrecht Dirk , Katzmann Emanuel , Lang Claus , Böttger Lars , Matzanke Berthold , Schüler Dirk 
Published: June 2010

ABSTRACT

Magnetotactic bacteria synthesize specific organelles, the magnetosomes, which are membrane-enveloped crystals of the magnetic mineral magnetite (Fe
3
O
4
). The biomineralization of magnetite involves the uptake and intracellular accumulation of large amounts of iron. However, it is not clear how iron uptake and biomineralization are regulated and balanced with the biochemical iron requirement and intracellular homeostasis. In this study, we identified and analyzed a homologue of the
f
erric
u
ptake
r
egulator Fur in
Magnetospirillum gryphiswaldense
, which was able to complement a
fur
mutant of
Escherichia coli
. A
fur
deletion mutant of
M. gryphiswaldense
biomineralized fewer and slightly smaller magnetite crystals than did the wild type. Although the total cellular iron accumulation of the mutant was decreased due to reduced magnetite biomineralization, it exhibited an increased level of free intracellular iron, which was bound mostly to a ferritin-like metabolite that was found significantly increased in Mössbauer spectra of the mutant. Compared to that of the wild type, growth of the
fur
mutant was impaired in the presence of paraquat and under aerobic conditions. Using a Fur titration assay and proteomic analysis, we identified constituents of the Fur regulon. Whereas the expression of most known magnetosome genes was unaffected in the
fur
mutant, we identified 14 proteins whose expression was altered between the mutant and the wild type, including five proteins whose genes constitute putative iron uptake systems. Our data demonstrate that Fur is a regulator involved in global iron homeostasis, which also affects magnetite biomineralization, probably by balancing the competing demands for biochemical iron supply and magnetite biomineralization.
Structure and Function of CinD (YtjD) ofLactococcus lactis, a Copper-Induced Nitroreductase Involved in Defense against Oxidative Stress
Authors: Mermod Mélanie , Mourlane Frédéric , Waltersperger Sandro , Oberholzer Anselm E. , Baumann Ulrich , Solioz Marc 
Published: June 2010

ABSTRACTInLactococcus lactisIL1403, 14 genes are under the control of the copper-inducible CopR repressor. This so-called CopR regulon encompasses the CopR regulator, two putative CPx-type copper ATPases, a copper chaperone, and 10 additional genes of unknown function. We addressed here the function of one of these genes,ytjD, which we renamedcinD(copper-inducednitroreductase). Copper, cadmium, and silver inducedcinD in vivo, as shown by real-time quantitative PCR. A knockout mutant ofcinDwas more sensitive to oxidative stress exerted by 4-nitroquinoline-N-oxide and copper. Purified CinD is a flavoprotein and reduced 2,6-dichlorophenolindophenol and 4-nitroquinoline-N-oxide withkcatvalues of 27 and 11 s−1, respectively, using NADH as a reductant. CinD also exhibited significant catalase activityin vitro. The X-ray structure of CinD was resolved at 1.35 Å and resembles those of other nitroreductases. CinD is thus a nitroreductase which can protectL. lactisagainst oxidative stress that could be exerted by nitroaromatic compounds and copper.
Xer1-Mediated Site-Specific DNA Inversions and Excisions in
Mycoplasma agalactiae
Authors: Czurda Stefan , Jechlinger Wolfgang , Rosengarten Renate , Chopra-Dewasthaly Rohini 
Published: June 2010

ABSTRACT

Surface antigen variation in
Mycoplasma agalactiae
, the etiologic agent of contagious agalactia in sheep and goats, is governed by site-specific recombination within the
vpma
multigene locus encoding the Vpma family of variable surface lipoproteins. This high-frequency Vpma phase switching was previously shown to be mediated by a Xer1 recombinase encoded adjacent to the
vpma
locus. In this study, it was demonstrated in
Escherichia coli
that the Xer1 recombinase is responsible for catalyzing
vpma
gene inversions between recombination sites (RS) located in the 5′-untranslated region (UTR) in all six
vpma
genes, causing cleavage and strand exchange within a 21-bp conserved region that serves as a recognition sequence. It was further shown that the outcome of the site-specific recombination event depends on the orientation of the two
vpma
RS, as direct or inverted repeats. While recombination between inverted
vpma
RS led to inversions, recombination between direct repeat
vpma
RS led to excisions. Using a newly developed excision assay based on the
lacZ
reporter system, we were able to successfully demonstrate under native conditions that such Xer1-mediated excisions can indeed also occur in the
M. agalactiae
type strain PG2, whereas they were not observed in the control
xer1
-disrupted VpmaY phase-locked mutant (PLMY), which lacks Xer1 recombinase. Unless there are specific regulatory mechanisms preventing such excisions, this might be the cost that the pathogen has to render at the population level for maintaining this high-frequency phase variation machinery.