bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of a
Bacillus
Phytase C Gene in
Pichia pastoris
and Properties of the Recombinant Enzyme
Authors: Guerrero-Olazarán Martha , Rodríguez-Blanco Lilí , Carreon-Treviño J. Gerardo , Gallegos-López Juan A. , Viader-Salvadó José M.
Published: July 2010

The cloning and expression of a native gene encoding a
Bacillus subtilis
phytase using
Pichia pastoris
as the host is described. In addition, the influence of N-glycosylation on the biochemical properties of the
B. subtilis
phytase, the influence of pH on the thermostability of the recombinant and native
B. subtilis
phytases, and the resistance of both phytases to shrimp digestive enzymes and porcine trypsin are also described. After 48 h of methanol induction in shake flasks, a selected recombinant strain produced and secreted 0.82 U/ml (71 mg/liter) recombinant phytase. This phytase was N-glycosylated, had a molecular mass of 39 kDa after N-deglycosylation, exhibited activity within a pH range of 2.5 to 9 and at temperatures of 25 to 70°C, had high residual activity (85% ± 2%) after 10 min of heat treatment at 80°C and pH 5.5 in the presence of 5 mM CaCl
2
, and was resistant to shrimp digestive enzymes and porcine trypsin. Although the recombinant
Bacillus
phytase had pH and temperature activity profiles that were similar to those of the corresponding nonglycosylated native phytase, the thermal stabilities of the recombinant and native phytases were different, although both were calcium concentration and pH dependent.
Isolation from
Ochrobactrum anthropi
of a Novel Class II 5-Enopyruvylshikimate-3-Phosphate Synthase with High Tolerance to Glyphosate
Authors: Tian Yong-Sheng , Xiong Ai-Sheng , Xu Jing , Zhao Wei , Gao Feng , Fu Xiao-Yan , Xu Hu , Zheng Jian-Li , Peng Ri-He , Yao Quan-Hong
Published: July 2010

Applying the genomic library construction process and colony screening, a novel
aro
A
gene encoding 5-enopyruvylshikimate-3-phosphate synthase from
Ochrobactrum anthropi
was identified, cloned, and overexpressed, and the enzyme was purified to homogeneity. Furthermore, site-directed mutagenesis was employed to assess the role of single amino acid residues in glyphosate resistance.
Regulation of the Expression of Genes Involved in NAD
De Novo
Biosynthesis in
Corynebacterium glutamicum
Authors: Teramoto Haruhiko , Suda Masako , Inui Masayuki , Yukawa Hideaki
Published: July 2010

Three genes,
nadA
,
nadB
, and
nadC
, involved in NAD
de novo
biosynthesis are broadly conserved in the genomes of numerous bacterial species. In the genome of
Corynebacterium glutamicum
,
nadA
and
nadC
but not
nadB
are annotated. The
nadA
and
nadC
genes are located in a gene cluster containing two other genes, designated
ndnR
and
nadS
herein.
ndnR
encodes a member of the Nudix-related transcriptional regulator (NrtR) family.
nadS
encodes a homologue of cysteine desulfurase involved in Fe-S cluster assembly. The gene cluster
ndnR
-
nadA
-
nadC
-
nadS
is genetically characterized herein. Mutant strains deficient in
nadA
,
nadC
, or
nadS
required exogenous nicotinate for growth, and the nicotinate auxotrophy was complemented by introduction of the corresponding gene in
trans
, indicating that each of these genes is essential for growth in the absence of an exogenous source of NAD biosynthesis. The results of reverse transcriptase PCR analyses and
ndnR
promoter-
lacZ
expression analyses revealed that the expression of
ndnR
,
nadA
,
nadC
, and
nadS
genes was markedly and coordinately repressed by nicotinate. The expression of these genes was enhanced by the disruption of
ndnR
, resulting in the loss of the nicotinate-responsive regulation of gene expression. These results suggest that NdnR acts as a transcriptional repressor of NAD
de novo
biosynthesis genes and plays an essential role in the regulation of the response to nicotinate.
The Abundance of Microbial Functional Genes in Grassy Woodlands Is Influenced More by Soil Nutrient Enrichment than by Recent Weed Invasion or Livestock Exclusion
Authors: Lindsay Elizabeth A. , Colloff Matthew J. , Gibb Nerida L. , Wakelin Steven A.
Published: July 2010

A diverse soil microbial community is involved in nitrogen cycling, and these microbes can be affected by land management practices and weed invasion. We surveyed 20 woodlands with a history of livestock grazing, with livestock recently excluded from 10 sites. We investigated whether soil nutrients were lower when grazing was excluded and higher when exotic grasses dominated the understory. Second, using quantitative real-time PCR, we investigated whether microbial nitrogen functional gene (NFG) abundance was altered with soil nutrient enrichment, livestock exclusion, and exotic grass invasion. The target genes were
chiA
(decomposition-ammonification),
nifH
(nitrogen fixation),
nirK
and
narG
(denitrification), and bacterial
amoA
(nitrification). Woodland soils were enriched in phosphorus and nitrogen compared to reference condition sites, but soil nutrients were not lower following livestock exclusion. Total nitrogen and
nifH
were negatively correlated in grazed woodlands, suggesting that aboveground herbivory reduces the capacity for belowground nitrogen fixation. Woodlands dominated by exotic grasses had higher levels of nitrate,
narG
, and
nirK
than those dominated by native grasses. We hypothesize that the increase in potential for denitrification was due to increases in soil nitrate, rather than changes in plant composition. Overall, soil physicochemistry explained more variation in NFG abundance than livestock presence or plant invasion, particularly for
chiA
and bacterial
amoA
, with significant relationships between the abundance of all five NFGs and total nitrogen or nitrate. All woodlands investigated had a history of anthropogenic disturbance and nutrification, and soil nutrient levels and the abundance of NFGs are likely to be related to long-term land management practices.
Next-Generation Sequencing Techniques for Eukaryotic Microorganisms: Sequencing-Based Solutions to Biological Problems
Authors: Nowrousian Minou
Published: July 2010

Over the past 5 years, large-scale sequencing has been revolutionized by the development of several so-called next-generation sequencing (NGS) technologies. These have drastically increased the number of bases obtained per sequencing run while at the same time decreasing the costs per base. Compared to Sanger sequencing, NGS technologies yield shorter read lengths; however, despite this drawback, they have greatly facilitated genome sequencing, first for prokaryotic genomes and within the last year also for eukaryotic ones. This advance was possible due to a concomitant development of software that allows the
de novo
assembly of draft genomes from large numbers of short reads. In addition, NGS can be used for metagenomics studies as well as for the detection of sequence variations within individual genomes, e.g., single-nucleotide polymorphisms (SNPs), insertions/deletions (indels), or structural variants. Furthermore, NGS technologies have quickly been adopted for other high-throughput studies that were previously performed mostly by hybridization-based methods like microarrays. This includes the use of NGS for transcriptomics (RNA-seq) or the genome-wide analysis of DNA/protein interactions (ChIP-seq). This review provides an overview of NGS technologies that are currently available and the bioinformatics analyses that are necessary to obtain information from the flood of sequencing data as well as applications of NGS to address biological questions in eukaryotic microorganisms.
A Systems Biology Approach To Modeling
Vibrio cholerae
Gene Expression under Virulence-Inducing Conditions
Authors: Kanjilal Sanjat , Citorik Robert , LaRocque Regina C. , Ramoni Marco F. , Calderwood Stephen B.
Published: July 2010

Vibrio cholerae
is a Gram-negative bacillus that is the causative agent of cholera. Pathogenesis
in vivo
occurs through a series of spatiotemporally controlled events under the control of a gene cascade termed the ToxR regulon. Major genes in the ToxR regulon include the master regulators
toxRS
and
tcpPH
, the downstream regulator
toxT
, and virulence factors, the
ctxAB
and
tcpA
operons. Our current understanding of the dynamics of virulence gene expression is limited to microarray analyses of expression at selected time points. To better understand this process, we utilized a systems biology approach to examine the temporal regulation of gene expression in El Tor
V. cholerae
grown under virulence-inducing conditions
in vitro
(AKI medium), using high-resolution time series genomic profiling. Results showed that overall gene expression in AKI medium mimics that of
in vivo
studies but with less clear temporal separation between upstream regulators and downstream targets. Expression of
toxRS
was unaffected by growth under virulence-inducing conditions, but expression of
toxT
was activated shortly after switching from stationary to aerating conditions. The
tcpA
operon was also activated early during mid-exponential-phase growth, while the
ctxAB
operon was turned on later, after the rise in
toxT
expression. Expression of
ctxAB
continued to rise despite an eventual decrease in
toxT
. Cluster analysis of gene expression highlighted 15 hypothetical genes and six genes related to environmental information processing that represent potential new members of the ToxR regulon. This study applies systems biology tools to analysis of gene expression of
V. cholerae
in vitro
and provides an important comparator for future studies done
in vivo
.
Structural Study of the
Serratia entomophila
Antifeeding Prophage: Three-Dimensional Structure of the Helical Sheath
Authors: Sen Anindito , Rybakova Daria , Hurst Mark R. H. , Mitra Alok K.
Published: July 2010

The sheath of the
Serratia entomophila
antifeeding prophage, which is pathogenic to the New Zealand grass grub
Costelytra zealandica
, is a 3-fold helix formed by a 4-fold symmetric repeating motif disposed around a helical inner tube. This structure, determined by electron microscopy and image processing, is distinct from that of the other known morphologically similar bacteriophage sheaths.
A Monoacylglycerol Lipase from
Mycobacterium smegmatis
Involved in Bacterial Cell Interaction
Authors: Dhouib Rabeb , Laval Françoise , Carrière Frédéric , Daffé Mamadou , Canaan Stéphane
Published: July 2010

MSMEG
_
0220
from
Mycobacterium smegmatis
, the ortholog of the
Rv0183
gene from
M. tuberculosis
, recently identified and characterized as encoding a monoacylglycerol lipase, was cloned and expressed in
Escherichia coli
. The recombinant protein (rMSMEG_0220), which exhibits 68% amino acid sequence identity with Rv0183, showed the same substrate specificity and similar patterns of pH-dependent activity and stability as the
M. tuberculosis
enzyme. rMSMEG_0220 was found to hydrolyze long-chain monoacylglycerol with a specific activity of 143 ± 6 U mg
−1
. Like Rv0183 in
M. tuberculosis
, MSMEG_0220 was found to be located in the cell wall. To assess the
in vivo
role of the homologous proteins, an
MSMEG
_
0220
disrupted mutant of
M. smegmatis
(MsΔ0220) was produced. An intriguing change in the colony morphology and in the cell interaction, which were partly restored in the complemented mutant containing either an active (ComMsΔ0220) or an inactive (ComMsΔ0220S111A) enzyme, was observed. Growth studies performed in media supplemented with monoolein showed that the ability of both MsΔ0220 and ComMsΔ0220S111A to grow in the presence of this lipid was impaired. Moreover, studies of the antimicrobial susceptibility of the MsΔ0220 strain showed that this mutant is more sensitive to rifampin and more resistant to isoniazid than the wild-type strain, pointing to a critical structural role of this enzyme in mycobacterial physiology, in addition to its function in the hydrolysis of exogenous lipids.
Expression of Tropodithietic Acid Biosynthesis Is Controlled by a Novel Autoinducer
Authors: Geng Haifeng , Belas Robert
Published: July 2010

The interactions between marine prokaryotic and eukaryotic microorganisms are crucial to many biological and biogeochemical processes in the oceans. Often the interactions are mutualistic, as in the symbiosis between phytoplankton, e.g., the dinoflagellate
Pfiesteria piscicida
and
Silicibacter
sp. TM1040, a member of the
Roseobacter
taxonomic lineage. It is hypothesized that an important component of this symbiosis is bacterial production of tropodithietic acid (TDA), a biologically active tropolone compound whose synthesis requires the expression of
tda
ABCDEF
(
tdaA-F
), as well as six additional genes (
cysI
,
malY
,
paaIJK
, and
tdaH
). The factors controlling
tda
gene expression are not known, although growth in laboratory standing liquid cultures drastically increases TDA levels. In this report, we measured the transcription of
tda
genes to gain a greater understanding of the factors controlling their expression. While the expression of
tdaAB
was constitutive,
tdaCDE
and
tdaF
mRNA increased significantly (3.7- and 17.4-fold, respectively) when cells were grown in standing liquid broth compared to their levels with shaking liquid culturing. No transcription of
tdaC
was detected when a
tdaC
p
::
lacZ
transcriptional fusion was placed in 11 of the 12 Tda
−
mutant backgrounds, with
cysI
being the sole exception. The expression of
tdaC
could be restored to 9 of the remaining 11 Tda
−
mutants—
tdaA
and
tdaH
failed to respond—by placing wild-type (Tda
+
) strains in close proximity or by supplying exogenous TDA to the mutant, suggesting that TDA induces
tda
gene expression. These results indicate that TDA acts as an autoinducer of its own synthesis and suggest that roseobacters may use TDA as a quorum signal.
DNA Uptake Sequence-Mediated Enhancement of Transformation in
Neisseria gonorrhoeae
Is Strain Dependent
Authors: Duffin Paul M. , Seifert H. Steven
Published: July 2010

Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen
Neisseria gonorrhoeae. Neisseria
spp. have been shown to preferentially take up and transform their own DNA by recognizing the nonpalindromic 10- or 12-nucleotide sequence 5′-

AT

GCCGTCTGAA-3′ (additional semiconserved nucleotides are underlined), termed the DNA uptake sequence (DUS10 or DUS12). Here we investigated the effects of the DUS on transformation and DNA uptake for several laboratory strains of
N. gonorrhoeae
. We found that all strains showed efficient transformation of DUS containing DNA (DUS10 and DUS12) but that the level of transformation with DNA lacking a DUS (DUS0) was variable in different strains. The DUS-enhanced transformation was 20-fold in two strains, FA1090 and FA19, but was approximately 150-fold in strains MS11 and 1291. All strains tested provide some level of DUS0 transformation, and DUS0 transformation was type IV pilus dependent. Competition with plasmid DNA revealed that transformation of MS11 was enhanced by the addition of excess plasmid DNA containing a DUS while FA1090 transformation was competitively inhibited. Although FA1090 was able to mediate much more efficient transformation of DNA lacking a DUS than was MS11, DNA uptake experiments showed similar levels of uptake of DNA containing and lacking a DUS in FA1090 and MS11. Finally, DNA uptake was competitively inhibited in both FA1090 and MS11. Taken together, our data indicate that the role of the DUS during DNA transformation is variable between strains of
N. gonorrhoeae
and may influence multiple steps during transformation.
Complete Genome Sequence of
Halalkalicoccus jeotgali
B3
T
, an Extremely Halophilic Archaeon
Authors: Roh Seong Woon , Nam Young-Do , Nam Seong-Hyeuk , Choi Sang-Haeng , Park Hong-Seog , Bae Jin-Woo
Published: July 2010

Halalkalicoccus jeotgali
B3
T
, isolated from salt-fermented seafood from South Korea, is an extremely halophilic archaeon belonging to the family
Halobacteriaceae
. Here, we present the complete genome sequence of the type strain
H. jeotgali
B3
T
(3,698,650 bp, with a G+C content of 62.5%), which consists of one chromosome and six plasmids. This is the first complete genome sequence of the
Halalkalicoccus
species.
Tachycardia, reduced vagal capacity, and age-dependent ventricular dysfunction arising from diminished expression of the presynaptic choline transporter
Authors: English Brett A. , Appalsamy Martin , Diedrich Andre , Ruggiero Alicia M. , Lund David , Wright Jane , Keller Nancy R. , Louderback Katherine M. , Robertson David , Blakely Randy D.
Published: July 2010

Healthy cardiovascular function relies on a balanced and responsive integration of noradrenergic and cholinergic innervation of the heart. High-affinity choline uptake by cholinergic terminals is pivotal for efficient ACh production and release. To date, the cardiovascular impact of diminished choline transporter (CHT) expression has not been directly examined, largely due to the transporter's inaccessibility in vivo. Here, we describe findings from cardiovascular experiments using transgenic mice that bear a CHT genetic deficiency. Whereas CHT knockout (CHT−/−) mice exhibit early postnatal lethality, CHT heterozygous (CHT+/−) mice survive, grow, and reproduce normally and exhibit normal spontaneous behaviors. However, the CHT+/−mouse heart displays significantly reduced levels of high-affinity choline uptake accompanied by significantly reduced levels of ACh. Telemeterized recordings of cardiovascular function in these mice revealed tachycardia and hypertension at rest. After treadmill exercise, CHT+/−mice exhibited slower heart rate recovery, consistent with a diminished cholinergic reserve, a contention validated through direct vagal nerve stimulation. Echocardiographic and histological experiments revealed an age-dependent decrease in fractional shortening, increased left ventricular dimensions, and increased ventricular fibrosis, consistent with ventricular dysfunction. These cardiovascular phenotypes of CHT+/−mice encourage an evaluation of humans bearing reduced CHT expression for their resiliency in maintaining proper heart function as well as risk for cardiovascular disease.
ANG II inhibits insulin-mediated production of PI 3,4,5-trisphosphates via a Ca2+-dependent but PKC-independent pathway in the cardiomyocytes
Authors: Ikushima Masashi , Ishii Masaru , Ohishi Mitsuru , Yamamoto Koichi , Ogihara Toshio , Rakugi Hiromi , Kurachi Yoshihisa
Published: July 2010

Insulin resistance (IR) is a condition where different organs are refractory to insulin stimulation of glucose uptake. ANG II has been suggested to be involved in the development of IR in the heart. The precise mechanism by which this occurs is still unknown. Here we have used dynamic fluorescent imaging techniques to show that ANG II inhibits insulin production of phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3] in cardiac myocytes. Fluorophore (Venus)-conjugated cAMP-dependent protein kinase-pleckstrin homology domain, which specifically binds to PI(3,4,5)P3, was transfected in neonatal rat cardiac myocytes. Insulin induced a robust increase in the fluorescence intensity at the cell surface, which was diminished by application of ANG II. The inhibitory action of ANG II was antagonized by RNH-6270 (an angiotensin type 1 receptor antagonist) but not by PD-122370 (an angiotensin type 2 receptor antagonist). BAPTA-AM (Ca2+chelator) largely attenuated the ANG II effect, whereas K-252b (PKC inhibitor) did not. Furthermore, an elevation of intracellular Ca2+induced by ionomycin mimicked the ANG II effect. Therefore, it is suggested that ANG II antagonizes insulin-mediated production of PI(3,4,5)P3via a Ca2+-dependent but PKC-independent pathway in cardiac myocytes.
Tachybradycardia in the isolated canine right atrium induced by chronic sympathetic stimulation and pacemaker current inhibition
Authors: Joung Boyoung , Shinohara Tetsuji , Zhang Hong , Kim Daehyeok , Choi Eue-Keun , On Yong-Keun , Piccirillo Gianfranco , Chen Peng-Sheng , Lin Shien-Fong
Published: July 2010

The mechanisms of sinoatrial node (SAN) dysfunction in patients with chronically elevated sympathetic tone and reduced pacemaker current ( If; such as heart failure) are poorly understood. We simultaneously mapped membrane potential and intracellular Ca2+ in the Langendorff-perfused canine right atrium (RA). Blockade of either If (ZD-7288) or sarcoplasmic reticulum Ca2+ release (ryanodine) alone decreased heart rate by 8% ( n = 3) and 16% ( n = 3), respectively. Combined treatment of ZD-7288 and ryanodine consistently resulted in prolonged (≥3 s) sinus pauses (PSPs) ( n = 4). However, the middle SAN remained as the leading pacemaking site after these treatments. Prolonged exposure with isoproterenol (0.01 μmol/l) followed by ZD-7288 completely suppressed SAN but triggered recurrent ectopic atrial tachycardia. Cessation of tachycardia was followed by PSPs in five of eight RAs. Isoproterenol initially increased heart rate by 75% from baseline with late diastolic intracellular Ca2+ elevation (LDCAE) from the superior SAN. However, after a prolonged isoproterenol infusion, LDCAE disappeared in the superior SAN, the leading pacemaker shifted to the inferior SAN, and the rate reduced to 52% above baseline. Caffeine (2 ml, 20 mmol/l) injection after a prolonged isoproterenol infusion produced LDCAE in the SAN and accelerated the SAN rate, ruling out sarcoplasmic reticulum Ca2+ depletion as a cause of Ca2+ clock malfunction. We conclude that in an isolated canine RA preparation, chronically elevated sympathetic tone results in abnormal pacemaking hierarchy in the RA, including suppression of the superior SAN and enhanced pacemaking from ectopic sites. Combined malfunction of both membrane and Ca2+ clocks underlies the mechanisms of PSPs.
U.S. Food and Drug Administration Approval: Ofatumumab for the Treatment of Patients with Chronic Lymphocytic Leukemia Refractory to Fludarabine and Alemtuzumab
Authors: Lemery Steven J. , Zhang Jenny , Rothmann Mark D. , Yang Jun , Earp Justin , Zhao Hong , McDougal Andrew , Pilaro Anne , Chiang Raymond , Gootenberg Joseph E. , Keegan Patricia , Pazdur Richard
Published: July 2010

Purpose: To describe the data and analyses that led to the U.S. Food and Drug Administration (FDA) approval of ofatumumab (Arzerra, GlaxoSmithKline) for the treatment of patients with chronic lymphocytic leukemia (CLL) refractory to fludarabine and alemtuzumab.

Experimental Design: The FDA reviewed the results of a planned interim analysis of a single-arm trial, enrolling 154 patients with CLL refractory to fludarabine, and a supportive dose-finding, activity-estimating trial in 33 patients with CLL. Patients in the primary efficacy study received ofatumumab weekly for eight doses, then every 4 weeks for an additional four doses; patients in the supportive trial received four weekly doses. In the primary efficacy study, endpoints were objective response rate and response duration.

Results: For regulatory purposes, the primary efficacy population consisted of 59 patients with CLL refractory to fludarabine and alemtuzumab. In this subgroup, the investigator-determined objective response rate was 42% [99% confidence interval (CI), 26–60], with a median duration of response of 6.5 months (95% CI, 5.8–8.3); all were partial responses. The most common adverse reactions in the primary efficacy study were neutropenia, pneumonia, pyrexia, cough, diarrhea, anemia, fatigue, dyspnea, rash, nausea, bronchitis, and upper respiratory tract infections. Infusion reactions occurred in 44% of patients with the first infusion (300 mg) and 29% with the second infusion (2,000 mg). The most common serious adverse reactions were infections, neutropenia, and pyrexia.

Conclusions: On October 26, 2009, the FDA granted accelerated approval to ofatumumab for the treatment of patients with CLL refractory to fludarabine and alemtuzumab, on the basis of demonstration of durable tumor shrinkage. Clin Cancer Res; 16(17); 4331–8. ©2010 AACR.
Influence of Oct1/Oct2-Deficiency on Cisplatin-Induced Changes in UrinaryN-Acetyl-β-D-Glucosaminidase
Authors: Franke Ryan M. , Kosloske Ashley M. , Lancaster Cynthia S. , Filipski Kelly K. , Hu Chaoxin , Zolk Oliver , Mathijssen Ron H. , Sparreboom Alex
Published: July 2010

Purpose: This study aimed to test the influence of functional renal organic cation transporters (OCT2 in humans, Oct1 and Oct2 in mice) on biomarkers of cisplatin nephrotoxicity, such as urinary activity of N-acetyl-β-D-glucosaminidase (NAG).

Experimental Design: Temporal cisplatin-induced nephrotoxicity was assessed by histopathology and biomarkers. Cisplatin-mediated NAG changes and survival were determined in wild-type and Oct1/2(-/-) mice. Identification of OCT2 inhibitors was done in transfected 293Flp-In cells, and the NCI60 cell line panel was used to assess contribution of OCT2 to cisplatin uptake in cancer cells.

Results: Classical biomarkers such as blood urea nitrogen and serum creatinine were not elevated until 72 hours after cisplatin administration and substantial kidney damage had occurred. Oct1/2(-/-) mice had 2.9-fold lower NAG by 4 hours (P < 0.0001) and 2.3-fold increased survival (P = 0.0097). Among 16 agents, cimetidine strongly inhibited uptake of tetraethylammonium bromide (P = 0.0006) and cisplatin (P < 0.0001), but did not have an influence on cisplatin uptake in SK-OV-3 cells, the cancer line with the highest OCT2 mRNA levels. In wild-type mice, cimetidine inhibited cisplatin-induced NAG changes (P = 0.016 versus cisplatin alone) to a degree similar to that seen in Oct1/2(-/-) mice receiving cisplatin (P = 0.91). Cumulative NAG activity of >0.4 absorbance units (AU) was associated with 21-fold increased odds for severe nephrotoxicity (P = 0.0017), which was linked with overall survival (hazard ratio, 8.1; 95% confidence interval, 2.1-31; P = 0.0078).

Conclusions: Cimetidine is able to inhibit OCT2-mediated uptake of cisplatin in the kidney, and subsequently ameliorate nephrotoxicity likely with minimal effect on uptake in tumor cells. Clin Cancer Res; 16(16); 4198–206. ©2010 AACR.
Transcriptional Profiling of Polycythemia Vera Identifies Gene Expression Patterns Both Dependent and Independent from the Action of JAK2V617F
Authors: Berkofsky-Fessler Windy , Buzzai Monica , Kim Marianne K-H. , Fruchtman Steven , Najfeld Vesna , Min Dong-Joon , Costa Fabricio F. , Bischof Jared M. , Soares Marcelo B. , McConnell Melanie Jane , Zhang Weijia , Levine Ross , Gilliland D. Gary , Calogero Raffaele , Licht Jonathan D.
Published: July 2010

Purpose: To understand the changes in gene expression in polycythemia vera (PV) progenitor cells and their relationship to JAK2V617F.

Experimental Design: Messenger RNA isolated from CD34+ cells from nine PV patients and normal controls was profiled using Affymetrix arrays. Gene expression change mediated by JAK2V617F was determined by profiling CD34+ cells transduced with the kinase and by analysis of leukemia cell lines harboring JAK2V617F, treated with an inhibitor.

Results: A PV expression signature was enriched for genes involved in hematopoietic development, inflammatory responses, and cell proliferation. By quantitative reverse transcription-PCR, 23 genes were consistently deregulated in all patient samples. Several of these genes such as WT1 and KLF4 were regulated by JAK2, whereas others such as NFIB and EVI1 seemed to be deregulated in PV by a JAK2-independent mechanism. Using cell line models and comparing gene expression profiles of cell lines and PV CD34+ PV specimens, we have identified panels of 14 JAK2-dependent genes and 12 JAK2-independent genes. These two 14- and 12-gene sets could separate not only PV from normal CD34+ specimens, but also other MPN such as essential thrombocytosis and primary myelofibrosis from their normal counterparts.

Conclusions: A subset of the aberrant gene expression in PV progenitor cells can be attributed to the action of the mutant kinase, but there remain a significant number of genes characteristic of the disease but deregulated by as yet unknown mechanisms. Genes deregulated in PV as a result of the action of JAK2V617F or independent of the kinase may represent other targets for therapy. Clin Cancer Res; 16(17); 4339–52. ©2010 AACR.
Children's Welfare in Soviet Russia: Society and the State, 1917-1930s
Authors: Smirnova Tatiana
Published: July 2010

The Bolsheviks did not alienate citizens from helping find solutions to the problems afflicting children. Many social actions deemed as "useful" by the Soviet authorities were met with support by the regime. These included the "Week of the Homeless Child", school self-taxation, local societies of the "Friend of the Children", and others. Establishing its control over "useful" public ventures, the Government eventually absorbed them. On the surface, the proliferation of public ventures in the area of children's welfare, such as patronage by industrial enterprises, labor unions and other groups and the growth of various advisory boards and children's inspections, appeared to be a result of growing social initiative. In reality the government's support of public work led to de facto state and party control. In order to carry out successful public initiatives, the population had to adapt to the particulars of Bolshevik rule.
The Constitutional Bases of Society and the Historical and Cultural Heritage of the Soviet Past in Modern Russia
Authors: Sokolov Andrey
Published: July 2010

In modern Russia problems of civil society's genesis discussed very widely. The author consecutively examines the social-cultural practices, which were generated by more then 70 years of Soviet history, and were reflected in low, constitutions and other legislative acts. He illustrated how these practices influenced the interaction between power and society, especially in labor and everyday Soviet life. The general attention was devoted to analysis of action of these practices after fall of Soviet system – how its may restrain or stimulate formation of civil society.
Molecular phylogeny and historical biogeography of the bee genus Colletes Latreille, 1802 (Hymenoptera: Apiformes: Colletidae), based on mitochondrial COI and nuclear 28S sequence data
Authors: Laurenne Nina , Almeida Eduardo , Quicke Donald , Kuhlmann Michael
Published: July 2010

A phylogenetic analysis of the intrageneric relationships within the bee genus Colletes using a combined data set of the mitochondrial cytochrome oxidase 1 (COI) and 28S sequences is presented and its implications for historical biogeography are investigated. We analyzed 91 out of 469 described Colletes species (19.4%) from the entire range of distribution and 25 outgroup taxa representing all colletid subfamilies closely related to Colletinae. Eight different combinations of alignment parameters were used for the 28S data, and our combined data set comprised between 1801 and 1845 aligned nucleotides with COI contributing 339 informative bases and 28S between 318 and 360 informative sites. Our results corroborate the monophyly of about half of the Old World subgenera and show the need for a taxonomic re-delimitation of other groups currently accepted. The phylogenetic results confirm the South American origin of Colletes and multiple faunal exchange events between North America and Eurasia. The last continent colonized was Africa south of the Sahara, which experienced at least two independent invasions.
Salticidae (Arachnida: Araneae) from Oriental, Australian and Pacific Regions: Astilodes and Urogelides, new genera from Australia
Authors: Żabka Marek
Published: July 2010

Astilodes mariae gen. et sp.n. and Urogelides daviesae gen. et sp.n. are diagnosed and described from eastern and south-eastern Queensland (Australia). Information on distribution and biota requirements for both generic type species are given. On the basis of morphological characters the genus Astilodes is temporarily classified within the Astioida clade, while Urogelides is placed in the Heliophaninae subfamily.
The Tibiotarsal Articulation and Intertibiotarsal Leg Sclerite in Dictyoptera (Insecta)
Authors: Schmidt Christian , Barton Ariane , Klass Klaus-Dieter , Eulitz Ute
Published: July 2010

The skeletal morphology of the transition area between the tibia and the tarsus is described for Dictyoptera, with a focus on Blaberus craniifer (Blattodea) and Sphodromantis viridis (Mantodea), but with 8 mantodean and 39 blattodean species (including 3 termite species) examined altogether. We emphasise the occurrence in most dictyopterans of a small sclerite located on the anteroventral side of the leg, between the margins of the tibia and the tarsus facing each other, the intertibiotarsal sclerite (abbreviated as itts: tibio-tarsal band/ligament sensu D. Neff and co-workers). The itts region is shown on SEM micrographs for a representative subset of our taxon sample. Comparison with selected outgroup species and data from the literature indicates that sclerite itts is autapomorphic for Dictyoptera and further confirms the monophyly of the group. Details of sclerite itts yield characters for phylogenetic work in Dictyoptera, such as a dorsal extension of the sclerite limited to a subset of our mantodean sample. In Cryptocercidae cockroaches and in termites sclerite itts is vestigial or absent, respectively. This could be interpreted as supporting termites to be the sister group of all remaining Dictyoptera, followed by Cryptocercidae. In the light of recent phylogenetic work, however, this rather appears to be a reductive apomorphy, which then further supports Cryptocercidae to be the sister group of termites. Functional aspects of the tibiotarsal articulation in Dictyoptera are also addressed, with a focus on the roles of and interdependence among the dorsal tibiotarsal articulation, the itts sclerite, and the tendons originating from the proximal rim of the tarsus (bearing the tarsal muscles); on this issue a functional model is presented. Most importantly, the effects of the tarsal muscles are being modulated with the status of itts (expanded or bent underneath).
A new tribe, genus and species of Nemonychidae from Baltic amber (Coleoptera: Curculionoidea: Nemonychidae: Cimberidinae)
Authors: Riedel Alexander
Published: July 2010

A new tribe of Nemonychidae is proposed based on inclusions in Baltic amber: Kuschelomacerini trib.n., for Kuschelomacer kerneggeri gen. & sp.n. It can be attributed to the subfamily Cimberidinae but lacks some of the derived characters of the two extant tribes of this subfamily. The excellent preservation allows providing the most detailed description of a fossil Nemonychid weevil to date. Presumably, the species developed on Pinaceae common in the amber forests. Thus, the association of extant Cimberidinae with Pinaceae dates back at least until the Eocene.
The Bali Roadmap for Global Climate Policy—New Horizons and Old Pitfalls
Authors: Sterk Wolfgang , Watanabe Rie , Arens Christof , Mersmann Florian , Ott Hermann
Published: July 2010

The international climate negotiations have seen endless struggles between countries from South and North for almost 17 years, ever since the initiation of negotiations by the International Negotiation Committee (INC) for the United Nations Framework Convention on Climate Change (UNFCCC). The 13th meeting of the Conference of the Parties to the UNFCCC and the 3rd meeting of the Conference of the Parties serving as the meeting of the Parties to the Kyoto Protocol (COP 13 / CMP 3) held in Bali in December 2007 (the Bali conference) could mark the beginning of a rapprochement. Parties agreed on initiating a new “Ad-hoc working group on Long-Term Cooperative Action under the Convention” (AWG-LCA) that aims to negotiate a post-2012 agreement with participation of all parties, including the US and developing countries, by the end of 2009 at COP 15 / CMP 5 in Copenhagen. This article examines the outcomes of the Bali conference, focussing on the negotiations regarding post-2012, flexible mechanisms, financial mechanisms, technology transfer and deforestation. Finally, the article concludes that the Bali Conference saw a significant shift in the battle lines, a rearrangement of positions and alliances that might well announce a decisive new era in global climate policy and provides a real chance to agree on an effective and workable post-2012 agreement in Copenhagen.
The European Commission's proposed Directive on the geological storage of carbon dioxide
Authors: Tomescu Mihai , Brockett Scott , Doppelhammer Martina
Published: July 2010

The 2007 Spring European Council called for the European Union (EU) to be at the forefront of efforts to combat climate change and endorsed ambitious targets aimed at setting the European energy system on a sustainable path. To ensure that these targets are met, the European Commission put forward a set of proposed policy instruments, the Climate and Energy Package, in January 2008. A key element of the adopted Package is the proposed Directive on the geological storage of carbon dioxide, a technology which can help tackle emissions from energy generation from fossil fuel sources which will otherwise compromise EU climate objectives. This paper lays out policy options to incentivise the deployment of carbon dioxide capture and storage (CCS) in the EU and presents the proposed Directive.
Environmental judgments by the Court of Justice and their duration
Authors: Krämer Ludwig
Published: July 2010

This article discusses some data concerning the environmental jurisdiction of the EC Court of Justice during 2006 and 2007, and compares them with previous years. During this time, the Court—including the Court of First Instance—issued about one decision per week. The majority of cases concerned nature conservation issues, followed by waste and air-climate change matters. About two thirds of all cases were submitted to the Court by the European Commission. Preliminary cases (Article 234 EC Treaty) took, on average, a time of 19 months, cases under Article 230 21 months, and cases under Article 226 EC Treaty 18 months. However, if one includes in the cases under Article 226 the compulsory pre-judicial period, the average duration of procedures was 47 months, thus almost four years. A closer analysis of the data reveals that this long duration is mainly attributable to the EC Commission.
Perspectives for European Water Management Law Report of a conference in Brussels, 3 and 4 April 2008
Authors: Bach Simone
Published: July 2010

On 3rd and 4th April 2008 a conference containing the “Perspectives of European Water Management Law” took place in Brussels with participation of about 80 representatives of the European Commission and universities, authorities, enterprises and associations of the individual Member States of the European Community. The focus of the conference was on the Water Framework Directive 2000/60/EC and its implementation into national law, whereby the “polluter pays principle” arouse special interest. Another important and controversial topic was whether privatisation of the water sector would be advantageous or disadvantageous in regard to efficient allocation and sustainable environmental protection. Beyond the conference dealt with the public procurement in the water sector in relation to the directive 2004/17/EC, with the reorganisation of European protection against floods in the course of the EU flood directive 2007/60/EC and the effects of climate change on hydrologic balance.
Legal and design issues arising in linking the EU ETS with existing and emerging emissions trading schemes
Authors: Anderson Jason ,   Mace
Published: July 2010

Since the EU Emissions Trading Scheme (EU ETS) came into being in 2003, both the opportunity and incentive to link the EU ETS to other cap-and-trade schemes that aim to reduce greenhouse gases has increased dramatically. This paper provides an overview of the key design features of existing and emerging emissions trading schemes around the world. It then considers the political and technical significance of these design features for a decision on whether or not to link the EU ETS to these schemes and explores the possible legal nature of an agreement that could link various potential linking partners at the national and sub-national levels.
Cross-border Development Plans—An Outline of a Useful Planning Instrument for European Border Regions
Authors: Kment Martin
Published: July 2010

This article addresses the legal framework, the benefits and the difficulties of cross-border development plans. They will be presented by examining the advantages and disadvantages of two possible models with respect to a range of relevant criteria. The most important criteria are elements of procedural law, especially public participation; the preparation of planning regulations; the structure of the planning entity; and the implementation of templates into national legal systems. This analysis will point out, that the greatest obstacle to be overcome is assimilating the legal systems of the states involved—irrespective of which model is taken into account.
No wheels, no driver, can't travel. The EU sustainable consumption and production action plan, a framework of policies towards eco-design of products for material efficiency?
Authors: Shinn Melissa
Published: July 2010

The EU has a clear need to reduce its resource and material consumption to stay within biological and equitable limits. The EU Sustainable Consumption and Production and Sustainable Industry Policy Action Plan (SCP/SIP AP) published in 2008 is the new platform for development of policies on product eco-design and consumption. This article investigates the policies that could or do constitute drivers for product eco-design for material efficiency through various possible existing or foreseen policies. The scope includes the initiatives of the SCP/SIP AP itself and the various strategies and pieces of legislation that could be seen as part of the SCP legislative tool box, including the resources strategy, products and waste legislation. The underlying question is whether there is a comprehensive and effective sustainable consumption and production policy within the EU, and how far it goes.
Applying voluntary tools to achieve Sustainable Consumption and Production: collaboration between the European Commission and the retail sector
Authors: Aichinger Herbert , Schnabl Gernot , Parenti Alberto
Published: July 2010

This article analyses recent activities undertaken by the European Commission in collaboration with retailers and other stakeholders to help achieve sustainable consumption and production (SCP) as proposed in its SCP/SIP Action Plan. The focus is on the setting up and functioning of a Retail Forum. These activities are seen in the wider context of the use of “soft law” and, in particular, of voluntary environmental agreements as an effective alternative to regulatory measures in promoting sustainable development both along the supply chain and as regards consumers. The article suggests that the voluntary approach can be extremely useful to promote sustainable consumption and production provided that some essential conditions are met: voluntary commitments must be verifiable, they need the right level of ambition and they must involve a wide range of actors. In particular, the article explores the feasibility of developing a code of conduct for the retail sector.
'Gold plating' of European Environmental Measures?
Authors:   Squintani,   Aragão,   Macrory,   Jans,   Wegener
Published: July 2010

Minimum harmonization in European environmental law essentially means that the Member States have the power to lay down more stringent standards in this area of regulation than those laid down by European legislation. However, there were indications which seemed to suggest that Member States make very little use of their powers to lay down or maintain more stringent national standards. The general question to be dealt with in this paper is whether Member States actually use their power to lay down or maintain more stringent environmental standards after European harmonization. Based upon the papers and discussion at the meeting of the Avosetta Group of European Environmental Lawyers in Monção (Portugal), 6 and 7th February 2009, this paper aims to paint an impressionist picture of the contemporary practice of gold plating (or not) in the Member States.
Enforcing Non-Discrimination in Transboundary Environmental Impact Assessment: Advantages for EU Citizens from the Transposition of the Espoo and Aarhus Conventions?
Authors: Marsden Simon
Published: July 2010

This article examines the relationship between international and European law with respect to transboundary environmental impact assessment (TEIA), which under the UNECE Convention on Environmental Impact Assessment in a Transboundary Context (Espoo) applies requirements for EIA to the relationship between states known as 'Parties of origin' and 'affected Parties'. Information is shared and participation in the Party of origin procedure by the public in affected as well as origin states is required (non-discrimination); these provisions are enhanced under the related Convention on Access to Information, Public Participation in Decision-Making and Access to Justice in Environmental Matters (Aarhus), which also contains provisions enabling enforcement. The purpose is to analyse whether EU citizens have greater opportunities to enforce these rights than citizens of state Parties to the two treaties that are not members of the EU. Procedure and practice under the transposing directives on EIA and public participation is examined, and conclusions are drawn that although to a large extent EU membership is advantageous to EU citizens involved with TEIA, certain constraints concerning public participation and access to justice remain.
Climate change and EU legal initiatives regarding water availability
Authors: Krämer Ludwig
Published: July 2010

Discussions on climate change legislation concentrate on measures relating to the emission of greenhouse gases. The present contribution tries to have a look at the impact of water availability within the European Union and to examine the need for EU legislation in this area, starting from the fact that the EU has, until now, not considered water availability to be a topic which deserves specific attention by the Union. The articles points at the serious impact which water scarcity is likely to have in particular, though not exclusively, in Southern European countries and passes in review a number of possible options for EU wide legislation.
The environmental complaint in EU law
Authors: Krämer Ludwig
Published: July 2010

The European Commission which has a large discretion to decide which infringements of EC environmental law it will pursue, has recently published communications which announce that it will concentrate on the transposition of EC legislation into the law of the Member States. This omits to structurally address the practical application of environmental legislation. Strong environmental groups which could contribute to the effective application of EC environmental law, lack; this feature makes the environmental sector different from most other economic or social sectors, where strong interests—professional bodies, trade unions, competitors, trade and industry associations, farmers' associations—promote the defense of their respective interests. All the more is the environment dependent on the commitment of the civil society to preserve and protect it. The contribution examines the instrument of the environmental complaint in EU law and criticizes the Commission's attempt to stop using the complaint system as a tool to assess practical application of EC environmental law in the Member States, though it is almost the only one existing.