bluuebunny/crossref_metadata_2025 huggingface dataset:
Suboptimal maternal nutrition during early fetal kidney development specifically promotes renal lipid accumulation following juvenile obesity in the offspring
Authors: Fainberg H. P. , Sharkey D. , Sebert S. , Wilson V. , Pope M. , Budge H. , Symonds M. E.
Published: July 2012

Reduced maternal food intake between early-to-mid gestation results in tissue-specific adaptations in the offspring following juvenile-onset obesity that are indicative of insulin resistance. The aim of the present study was to establish the extent to which renal ectopic lipid accumulation, as opposed to other markers of renal stress, such as iron deposition and apoptosis, is enhanced in obese offspring born to mothers nutrient restricted (NR) throughout early fetal kidney development. Pregnant sheep were fed either 100% (control) or NR (i.e. fed 50% of their total metabolisable energy requirement from 30–80 days gestation and 100% at all other times). At weaning, offspring were made obese and, at approximately 1 year, kidneys were sampled. Triglyceride content, HIF-1α gene expression and the protein abundance of the outer-membrane transporter voltage-dependent anion-selective channel protein (VDAC)-I on the kidney cortex were increased in obese offspring born to NR mothers compared with those born to controls, which exhibited increased iron accumulation within the tubular epithelial cells and increased gene expression of the death receptor Fas. In conclusion, suboptimal maternal nutrition coincident with early fetal kidney development results in enhanced renal lipid deposition following juvenile obesity and could accelerate the onset of the adverse metabolic, rather than cardiovascular, symptoms accompanying the metabolic syndrome.
The urgency of the development of CO
2
capture from ambient air
Authors: Lackner Klaus S. , Brennan Sarah , Matter Jürg M. , Park A.-H. Alissa , Wright Allen , van der Zwaan Bob
Published: July 2012

CO
2
capture and storage (CCS) has the potential to develop into an important tool to address climate change. Given society’s present reliance on fossil fuels, widespread adoption of CCS appears indispensable for meeting stringent climate targets. We argue that for conventional CCS to become a successful climate mitigation technology—which by necessity has to operate on a large scale—it may need to be complemented with air capture, removing CO
2
directly from the atmosphere. Air capture of CO
2
could act as insurance against CO
2
leaking from storage and furthermore may provide an option for dealing with emissions from mobile dispersed sources such as automobiles and airplanes.
Structural and functional analysis of phosphorylation-specific binders of the kinase ERK from designed ankyrin repeat protein libraries
Authors: Kummer Lutz , Parizek Petra , Rube Peter , Millgramm Bastian , Prinz Anke , Mittl Peer R. E. , Kaufholz Melanie , Zimmermann Bastian , Herberg Friedrich W. , Plückthun Andreas
Published: July 2012

We have selected designed ankyrin repeat proteins (DARPins) from a synthetic library by using ribosome display that selectively bind to the mitogen-activated protein kinase ERK2 (extracellular signal-regulated kinase 2) in either its nonphosphorylated (inactive) or doubly phosphorylated (active) form. They do not bind to other kinases tested. Crystal structures of complexes with two DARPins, each specific for one of the kinase forms, were obtained. The two DARPins bind to essentially the same region of the kinase, but recognize the conformational change within the activation loop and an adjacent area, which is the key structural difference that occurs upon activation. Whereas the rigid phosphorylated activation loop remains in the same form when bound by the DARPin, the more mobile unphosphorylated loop is pushed to a new position. The DARPins can be used to selectively precipitate the cognate form of the kinases from cell lysates. They can also specifically recognize the modification status of the kinase inside the cell. By fusing the kinase with
Renilla
luciferase and the DARPin to GFP, an energy transfer from luciferase to GFP can be observed in COS-7 cells upon intracellular complex formation. Phosphorylated ERK2 is seen to increase by incubation of the COS-7 cells with FBS and to decrease upon adding the ERK pathway inhibitor PD98509. Furthermore, the anti-ERK2 DARPin is seen to inhibit ERK phosphorylation as it blocks the target inside the cell. This strategy of creating activation-state–specific sensors and kinase-specific inhibitors may add to the repertoire to investigate intracellular signaling in real time.
Extreme hydrops fetalis and cardiovascular abnormalities in mice lacking a functional Adrenomedullin gene
Authors: Caron Kathleen M. , Smithies Oliver
Published: July 2012

Adrenomedullin, a recently identified potent vasodilator, is
expressed widely and has been suggested to have functions ranging from
reproduction to blood pressure regulation. To elucidate these functions
and define more precisely sites of Adm expression, we replaced the
coding region of the
Adm
gene in mice with a sequence
encoding enhanced green fluorescent protein while leaving the
Adm
promoter intact. We find that
Adm
−/−
embryos die at midgestation with
extreme hydrops fetalis and cardiovascular abnormalities, including
overdeveloped ventricular trabeculae and underdeveloped arterial walls.
These data suggest that genetically determined absence of
Adm
may be one cause of nonimmune hydrops fetalis in
humans.
Inhibition of melanoma tumor growth
in vivo
by survivin targeting
Authors: Grossman Douglas , Kim Paul J. , Schechner Jeffrey S. , Altieri Dario C.
Published: July 2012

A role of apoptosis (programmed cell death) in tumor
formation and growth was investigated by targeting the
apoptosis inhibitor survivin
in vivo
. Expression
of a phosphorylation-defective survivin mutant
(Thr
34
→Ala) triggered apoptosis in several human
melanoma cell lines and enhanced cell death induced by the
chemotherapeutic drug cisplatin
in vitro
. Conditional
expression of survivin Thr
34
→Ala in YUSAC2 melanoma cells
prevented tumor formation upon s.c. injection into CB.17 severe
combined immunodeficient-beige mice. When induced in established
melanoma tumors, survivin Thr
34
→Ala inhibited tumor
growth by 60–70% and caused increased apoptosis and reduced
proliferation of melanoma cells
in vivo
. Manipulation of
the antiapoptotic pathway maintained by survivin may be
beneficial for cancer therapy.
Analysis of the blind
Drosophila
mutant
ninaB
identifies the gene encoding the key enzyme for vitamin A formation
in vivo
Authors: von Lintig Johannes , Dreher Armin , Kiefer Cornelia , Wernet Mathias F. , Vogt Klaus
Published: July 2012

Visual pigments (rhodopsins) are composed of a chromophore (vitamin
A derivative) bound to a protein moiety embedded in the retinal
membranes. Animals cannot synthesize the visual chromophore
de
novo
but rely on the uptake of carotenoids, from which vitamin
A is formed enzymatically by oxidative cleavage. Despite its
importance, the enzyme catalyzing the key step in vitamin A formation
resisted molecular analyses until recently, when the successful cloning
of a cDNA encoding an enzyme with β,β-carotene-15,15′-dioxygenase
activity from
Drosophila
was reported. To prove its
identity with the key enzyme for vitamin A formation
in
vivo
, we analyzed the blind
Drosophila
mutant
ninaB
. In two independent
ninaB
alleles,
we found mutations in the gene encoding the
β,β-carotene-15,15′-dioxygenase. These mutations lead to a defect
in vitamin A formation and are responsible for blindness of these
flies.
Regulation of p53 stability and p53-dependent apoptosis by NADH quinone oxidoreductase 1
Authors: Asher Gad , Lotem Joseph , Cohen Batya , Sachs Leo , Shaul Yosef
Published: July 2012

The tumor suppressor gene wild-type
p53
encodes a
labile protein that accumulates in cells after different stress signals
and can cause either growth arrest or apoptosis. One of the p53
target genes, p53-inducible gene 3 (
PIG3
), encodes a
protein with significant homology to oxidoreductases, enzymes involved
in cellular responses to oxidative stress and irradiation. This fact
raised the possibility that cellular oxidation–reduction events
controlled by such enzymes also may regulate the level of p53. Here we
show that NADH quinone oxidoreductase 1 (NQO1) regulates p53 stability.
The NQO1 inhibitor dicoumarol caused a reduction in the level of
both endogenous and γ-irradiation-induced p53 in HCT116
human colon carcinoma cells. This reduction was prevented by the
proteasome inhibitors MG132 and lactacystin, suggesting enhanced p53
degradation in the presence of dicoumarol. Dicoumarol-induced
degradation of p53 also was prevented in the presence of simian virus
40 large T antigen, which is known to bind and to stabilize p53. Cells
overexpressing NQO1 were resistant to dicoumarol, and this finding
indicates the direct involvement of NQO1 in p53 stabilization. NQO1
inhibition induced p53 degradation and blocked wild-type p53-mediated
apoptosis in γ-irradiated normal thymocytes and in M1 myeloid
leukemic cells that overexpress wild-type
p53
.
Dicoumarol also reduced the level of p53 in its mutant form in M1
cells. The results indicate that NQO1 plays an important role in
regulating p53 functions by inhibiting its degradation.
Progesterone receptor and dopamine receptors are required in Δ
9
-tetrahydrocannabinol modulation of sexual receptivity in female rats
Authors: Mani Shailaja K. , Mitchell Andrea , O'Malley Bert W.
Published: July 2012

Ovarian steroids, estrogen and progesterone, influence the
sensitivity of certain neural processes to cannabinoid treatment by
modulation of brain dopaminergic activity. We examined the effects of
the active ingredient of cannabis,
Δ
9
-tetrahydrocannabinol (THC), on sexual behavior in
female rats and its influence on steroid hormone receptors and
neurotransmitters in the facilitation of sexual receptivity. Our
results revealed that the facilitatory effect of THC was inhibited by
antagonists to both progesterone and dopamine D
1
receptors.
To test further the idea that progesterone receptors (PR) and/or
dopamine receptors (D
1
R) in the hypothalamus are required
for THC-facilitated sexual behavior in rodents, antisense and sense
oligonucleotides to PR and D
1
R were administered
intracerebroventricularly (ICV) into the third cerebral ventricle of
ovariectomized, estradiol benzoate-primed rats. Progesterone- and
THC-facilitated sexual behavior was inhibited in animals treated with
antisense oligonucleotides to PR or to D
1
R. Antagonists to
cannabinoid receptor-1 subtype (CB
1
), but not to
cannabinoid receptor-2 subtype (CB
2
) inhibited
progesterone- and dopamine-facilitated sexual receptivity in female
rats. Our studies indicate that THC acts on the CB
1
cannabinoid receptor to initiate a signal transduction response that
requires both membrane dopamine and intracellular progesterone
receptors for effective induction of sexual behavior.
Subunit interactions influence the biochemical and biological properties of Hsp104
Authors: Schirmer Eric C. , Ware Danielle M. , Queitsch Christine , Kowal Anthony S. , Lindquist Susan L.
Published: July 2012

Point mutations in either of the two nucleotide-binding domains
(NBD) of Hsp104 (NBD1 and NBD2) eliminate its thermotolerance function
in vivo
.
In vitro
, NBD1 mutations
virtually eliminate ATP hydrolysis with little effect on
hexamerization; analogous NBD2 mutations reduce ATPase activity and
severely impair hexamerization. We report that high protein
concentrations overcome the assembly defects of NBD2 mutants and
increase ATP hydrolysis severalfold, changing
V
max
with little effect on
K
m
. In a complementary fashion, the
detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate
inhibits hexamerization of wild-type (WT) Hsp104, lowering
V
max
with little effect on
K
m
. ATP hydrolysis exhibits a Hill
coefficient between 1.5 and 2, indicating that it is influenced by
cooperative subunit interactions. To further analyze the effects of
subunit interactions on Hsp104, we assessed the effects of mutant
Hsp104 proteins on WT Hsp104 activities. An NBD1 mutant that
hexamerizes but does not hydrolyze ATP reduces the ATPase activity of
WT Hsp104
in vitro
.
In vivo
, this mutant
is not toxic but specifically inhibits the thermotolerance function of
WT Hsp104. Thus, interactions between subunits influence the ATPase
activity of Hsp104, play a vital role in its biological functions, and
provide a mechanism for conditionally inactivating Hsp104 function
in vivo
.
Visualization of a water-selective pore by electron crystallography in vitreous ice
Authors: Ren G. , Reddy V. S. , Cheng A. , Melnyk P. , Mitra A. K.
Published: July 2012

The water-selective pathway through the aquaporin-1 membrane
channel has been visualized by fitting an atomic model to a 3.7-Å
resolution three-dimensional density map. This map was determined by
analyzing images and electron diffraction patterns of
lipid-reconstituted two-dimensional crystals of aquaporin-1 preserved
in vitrified buffer in the absence of any additive. The aqueous pathway
is characterized by a size-selective pore that is ≈4.0 ± 0.5Å
in diameter, spans a length of ≈18Å, and bends by ≈25° as it
traverses the bilayer. This narrow pore is connected by wide,
funnel-shaped openings at the extracellular and cytoplasmic faces. The
size-selective pore is outlined mostly by hydrophobic residues,
resulting in a relatively inert pathway conducive to diffusion-limited
water flow. The apex of the curved pore is close to the locations of
the in-plane pseudo-2-fold symmetry axis that relates the N- and
C-terminal halves and the conserved, functionally important N76 and
N192 residues.
The crystal structure of the nitrogen regulation fragment of the yeast prion protein Ure2p
Authors: Umland Timothy C. , Taylor Kimberly L. , Rhee Sangkee , Wickner Reed B. , Davies David R.
Published: July 2012

The yeast nonchromosomal gene [URE3] is due to a prion form of
the nitrogen regulatory protein Ure2p. It is a negative regulator of
nitrogen catabolism and acts by inhibiting the transcription factor
Gln3p. Ure2p residues 1–80 are necessary for prion generation and
propagation. The C-terminal fragment retains nitrogen regulatory
activity, albeit somewhat less efficiently than the full-length
protein, and it also lowers the frequency of prion generation. The
crystal structure of this C-terminal fragment, Ure2p(97–354), at 2.3
Å resolution is described here. It adopts the same fold as the
glutathione
S
-transferase superfamily, consistent with
their sequence similarity. However, Ure2p(97–354) lacks a properly
positioned catalytic residue that is required for
S
-transferase activity. Residues within this regulatory
fragment that have been indicated by mutational studies to influence
prion generation have been mapped onto the three-dimensional structure,
and possible implications for prion activity are discussed.
The role of MHC class I glycoproteins in the regulation of induction of cell death in immunocytes by malignant melanoma cells
Authors: Fishman Daniel , Irena Brodyanski , Kellman-Pressman Sigal , Karas Michael , Segal Shraga
Published: July 2012

A deranged expression of MHC class I glycoproteins, characteristic
of a variety of malignancies, contributes to the ability of cancer to
avoid destruction by T cell-mediated immunity. An abrogation of the
metastatic capacity of B16 melanoma cells has been achieved by
transfecting an MHC class I-encoding vector into class I-deficient B16
melanoma clones [Gorelik, E., Kim, M., Duty, L. & Galili, U. (1993)
Clin. Exp. Metastasis
11, 439–452]. We report here
that the deranged expression of class I molecules by B16 melanoma cells
is more than a mere acquisition of the capacity to escape immune
recognition. Namely, cells of the B16 melanoma prompted splenic
lymphocytes to commit death after coculture. However, a class
I-expressing and nonmetastatic CL8-2 clone was found to be less potent
as an inducer of apoptosis than class I-deficient and
metastatic BL9 and BL12 clones. Both Thy1.2
+
and
Thy1.2
−
splenocytes underwent cell death when exposed to
the class I-deficient BL9 clone. A proportion of CD4
+
and
CD8
+
cells among splenocytes exposed to the BL9 clone was
lower than that observed in a coculture with cells of the CL8-2 clone.
Consistently, none of the melanoma clones studied produced a ligand to
the FAS receptor (FAS-L). Thus, our results provide evidence that
(
i
) the production of FAS-L may not be the sole
mechanism by which malignant cells induce apoptosis in
immunocytes, and (
ii
) absence of MHC class I
glycoproteins plays an important role in preventing the elimination of
potential effector immunocytes by tumor cells.
Genetic and pharmacological disruption of neurokinin 1 receptor function decreases anxiety-related behaviors and increases serotonergic function
Authors: Santarelli Luca , Gobbi Gabriella , Debs Pierre C. , Sibille Etienne L. , Blier Pierre , Hen René , Heath Mark J. S.
Published: July 2012

Alterations in serotonin (5-hydroxytriptamine, 5-HT),
norepinephrine, and γ-aminobutyric acid have been linked to the
pathophysiology of anxiety and depression, and medications that
modulate these neurotransmitters are widely used to treat mood
disorders. Recently, the neuropeptide substance P (SP) and its
receptor, the neurokinin 1 receptor (NK1R), have been proposed as
possible targets for new antidepressant and anxiolytic therapies.
However, animal and human studies have so far failed to provide a clear
consensus on the role of SP in the modulation of emotional states. Here
we show that both genetic disruption and acute pharmacological blockade
of the NK1R in mice result in a marked reduction of anxiety and
stress-related responses. These behavioral changes are paralleled by an
increase in the firing rate of 5-HT neurons in the dorsal raphe
nucleus, a major source of serotonergic input to the forebrain. NK1R
disruption also results in a selective desensitization of 5-HT1A
inhibitory autoreceptors, which resembles the effect of sustained
antidepressant treatment. Together these results indicate that the SP
system powerfully modulates anxiety and suggest that this effect is at
least in part mediated by changes in the 5-HT system.
Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice
Authors: Fruebis Joachim , Tsao Tsu-Shuen , Javorschi Sandrine , Ebbets-Reed Dana , Erickson Mary Ruth S. , Yen Frances T. , Bihain Bernard E. , Lodish Harvey F.
Published: July 2012

Adipocyte complement-related protein (30 kDa) (Acrp30), a secreted
protein of unknown function, is exclusively expressed in differentiated
adipocytes; its mRNA is decreased in obese humans and mice. Here we
describe novel pharmacological properties of the protease-generated
globular head domain of Acrp30 (gAcrp30). Acute treatment of mice with
gAcrp30 significantly decreased the elevated levels of plasma free
fatty acids caused either by administration of a high fat test meal or
by i.v. injection of Intralipid. This effect of gAcrp30 was caused, at
least in part, by an acute increase in fatty acid oxidation by muscle.
As a result, daily administration of a very low dose of gAcrp30 to mice
consuming a high-fat/sucrose diet caused profound and sustainable
weight reduction without affecting food intake. Thus, gAcrp30 is a
novel pharmacological compound that controls energy homeostasis and
exerts its effect primarily at the peripheral level.
CIN85 phosphorylation is essential for EGFR ubiquitination and sorting into multivesicular bodies
Authors: Schroeder Barbara , Srivatsan Subhashini , Shaw Andrey , Billadeau Daniel , McNiven Mark A.
Published: July 2012

Ubiquitination of the epidermal growth factor receptor (EGFR) by cbl and its cognate adaptor cbl-interacting protein of 85 kDa (CIN85) is known to play an essential role in directing this receptor to the lysosome for degradation. The mechanisms by which this ubiquitin modification is regulated are not fully defined, nor is it clear where this process occurs. In this study we show that EGFR activation leads to a pronounced src-mediated tyrosine phosphorylation of CIN85 that subsequently influences EGFR ubiquitination. Of importance, phospho-CIN85 interacts with the Rab5-positive endosome, where it mediates the sequestration of the ubiquitinated receptor into multivesicular bodies (MVBs) for subsequent degradation. These findings provide novel insights into how src- kinase–based regulation of a cbl adaptor regulates the fate of the EGFR.
NESdb: a database of NES-containing CRM1 cargoes
Authors: Xu Darui , Grishin Nick V. , Chook Yuh Min
Published: July 2012

The leucine-rich nuclear export signal (NES) is the only known class of targeting signal that directs macromolecules out of the cell nucleus. NESs are short stretches of 8–15 amino acids with regularly spaced hydrophobic residues that bind the export karyopherin CRM1. NES-containing proteins are involved in numerous cellular and disease processes. We compiled a database named NESdb that contains 221 NES-containing CRM1 cargoes that were manually curated from the published literature. Each NESdb entry is annotated with information about sequence and structure of both the NES and the cargo protein, as well as information about experimental evidence of NES-mapping and CRM1-mediated nuclear export. NESdb will be updated regularly and will serve as an important resource for nuclear export signals. NESdb is freely available to nonprofit organizations at http://prodata.swmed.edu/LRNes .
Caloramator quimbayensis sp. nov., an anaerobic, moderately thermophilic bacterium isolated from a terrestrial hot spring
Authors: Rubiano-Labrador Carolina , Baena Sandra , Díaz-Cárdenas Carolina , Patel Bharat K. C.
Published: July 2012

An anaerobic, moderately thermophilic, terminal-spore-forming bacterium, designated strain USBA AT, was isolated from a terrestrial hot spring located at an altitude of 2683 m in the Andean region of Colombia (04° 50′ 14.0″ N 75° 32′ 53.4″ W). Cells of strain USBA AT were Gram-stain-positive, straight to slightly curved rods (0.9×2.5 µm), that were arranged singly or in pairs, and were motile by means of flagella. Growth occurred at 37–55 °C and pH 6.0–8.0, with a doubling time of 2 h under the optimal conditions (50 °C and pH 7.0). Glucose fermentation in strain USBA AT required yeast extract or peptone (each at 0.2 %, w/v). The novel strain fermented sugars, amino acids, Casamino acids, propanol, propionate, starch and dextrin, but no growth was observed on galactose, lactose, xylose, histidine, serine, threonine, benzoate, butyrate, lactate, pyruvate, succinate, methanol, ethanol, glycerol, casein, gelatin or xylan. The end products of glucose fermentation were formate, acetate, ethanol and lactate. Strain USBA AT did not grow autotrophically (with CO2 as carbon source and H2 as electron donor) and did not reduce thiosulfate, sulfate, elemental sulfur, sulfite, vanadium (V) or Fe (III) citrate. Growth of strain USBA AT was inhibited by ampicillin, chloramphenicol, kanamycin, penicillin and streptomycin (each at 10 µg ml−1). The predominant fatty acids were iso-C15 : 0, C16 : 0 and iso-C17 : 0 and the genomic DNA G+C content was 32.6 mol%. 16S rRNA gene sequence analysis indicated that strain USBA AT belonged in the phylum

Firmicutes

and that its closest relative was

Caloramator viterbiensis

JW/MS-VS5T (95.0 % sequence similarity). A DNA–DNA relatedness value of only 30 % was recorded in hybridization experiments between strain USBA AT and

Caloramator viterbiensis

DSM 13723T. Based on the phenotypic, chemotaxonomic and phylogenetic evidence and the results of the DNA–DNA hybridization experiments, strain USBA AT represents a novel species of the genus

Caloramator

, for which the name Caloramator quimbayensis sp. nov. is proposed. The type strain is USBA AT ( = CMPUJ U833T  = DSM 22093T).
Erythrobacter jejuensis sp. nov., isolated from seawater
Authors: Yoon Byoung-Jun , Lee Dong-Heon , Oh Duck-Chul
Published: July 2012

A Gram-staining-negative, yellow-pigmented, non-motile, strictly aerobic, rod-shaped bacterium, designated strain CNU001T, was isolated from seawater collected on the coast of Jeju Island, South Korea, and subjected to a polyphasic taxonomic study. The temperature, pH and NaCl ranges for growth were 10–30 °C, pH 6.0–10.0 and 2.0–5.0 %, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain CNU001T belonged to the genus

Erythrobacter

in the family

Erythrobacteraceae

, with

Erythrobacter longus

DSM 6997T (96.6 % sequence similarity),

Erythrobacter gaetbuli

SW-161T (96.3 %),

Erythrobacter vulgaris

022 2-10T (96.2 %),

Erythrobacter nanhaisediminis

T30T (96.1 %) and other members of the genus

Erythrobacter

(<96.0 %) identified as the novel strain’s closest relatives. The major cellular fatty acids were C18 : 1ω7c and C17 : 1ω6c. The polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, one sphingoglycolipid, one unidentified aminolipid and six other unidentified lipids. The major respiratory quinone was ubiquinone-10 (UQ-10) and the genomic DNA G+C content of the novel strain was 58.9 mol%. On the basis of phenotypic, phylogenetic and genotypic data, strain CNU001T represents a novel species within the genus

Erythrobacter

, for which the name Erythrobacter jejuensis sp. nov. is proposed. The type strain is CNU001T ( = KCTC 23090T  = JCM 16677T).
Stomatobaculum longum gen. nov., sp. nov., an obligately anaerobic bacterium from the human oral cavity
Authors: Sizova Maria V. , Muller Paul , Panikov Nicolai , Mandalakis Manolis , Hohmann Tine , Hazen Amanda , Fowle William , Prozorov Tanya , Bazylinski Dennis A. , Epstein Slava S.
Published: July 2012

A strictly anaerobic Gram-stain-variable but positive by structure, non-spore-forming bacterium designated

Lachnospiraceae

bacterium ACC2 strain DSM 24645T was isolated from human subgingival dental plaque. Bacterial cells were 4–40 µm long non-motile rods, often swollen and forming curved filaments up to 200 µm. Cells contained intracellular, poorly crystalline, nanometre-sized iron- and sulfur-rich particles. The micro-organism was able to grow on yeast extract, trypticase peptone, milk, some sugars and organic acids. The major metabolic end-products of glucose fermentation were butyrate, lactate, isovalerate and acetate. The growth temperature and pH ranges were 30–42 °C and 4.9–7.5, respectively. Major fatty acids were C14 : 0, C14 : 0 DMA (dimethyl aldehyde), C16 : 0, C16 : 1ω7c DMA. The whole-cell hydrolysate contained meso-diaminopimelic acid, indicating peptidoglycan type A1γ. The DNA G+C content was calculated to be 55.05 mol% from the whole-genome sequence and 55.3 mol% as determined by HPLC. There were no predicted genes responsible for biosynthesis of respiratory lipoquinones, mycolic acids and lipopolysaccharides. Genes associated with synthesis of teichoic and lipoteichoic acids, diaminopimelic acid, polar lipids and polyamines were present. According to the 16S rRNA gene sequence phylogeny, strain DSM 24645T formed, together with several uncultured oral clones, a separate branch within the family

Lachnospiraceae

, with the highest sequence similarity to the type strain of

Moryella indoligenes

at 94.2 %. Based on distinct phenotypic and genotypic characteristics, we suggest that strain DSM 24645T represents a novel species in a new genus, for which the name Stomatobaculum longum gen. nov., sp. nov. is proposed. The type strain of Stomatobaculum longum is DSM 24645T ( = HM-480T; deposited in BEI Resources, an NIH collection managed by the ATCC).
Galbibacter marinus sp. nov., isolated from deep-sea sediment
Authors: Li Chongping , Lai Qiliang , Fu Yuanyuan , Chen Shuangxi , Shao Zongze
Published: July 2012

A taxonomic study was carried out on a novel bacterium, designated strain ck-I2-15T, which was isolated from deep-sea sediment collected from the South-west Indian Ocean Ridge. Cells of strain ck-I2-15T were Gram-reaction-negative, rod-shaped, non-motile, moderately halophilic and capable of denitrification. Growth was observed with 0–9 % (w/v) NaCl and at temperatures of 10–37 °C. The novel strain was unable to degrade gelatin. The dominant cellular fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH and summed feature 9 (iso-C17 : 1ω9c and/or 10-methyl C16 : 0). The major respiratory quinone was MK6 and the polar lipid profile comprised phosphatidylethanolamine, one unidentified phospholipid, two unidentified aminolipids, one unidentified glycolipid and four other unidentified lipids. The G+C content of the genomic DNA was 38 mol%. 16S rRNA gene sequence comparison indicated that strain ck-I2-15T was most closely related to

Galbibacter mesophilus

Mok-17T (92.9 % sequence similarity), followed by ‘Joostella atrarenae’ M1-2 (92.8 %),

Joostella marina

En5T (92.7 %) and

Zhouia amylolytica

HN-171T (91.6 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain ck-I2-15T formed a clade with the genus

Galbibacter

, within the family

Flavobacteriaceae

. Several phenotypic properties allowed strain ck-I2-15T to be distinguished from its closest phylogenetic relatives. On the basis of the phenotypic and phylogenetic data, strain ck-I2-15T represents a novel species of the genus

Galbibacter

, for which the name Galbibacter marinus is proposed. The type strain is ck-I2-15T ( = CCTCC AB 209062T = LMG 25228T = MCCC 1A03044T).
SOME EFFECTS OF KÖ 1313 IN ANIMALS AND MAN
Authors: EKUE J.M. KOFI , SHANKS R.G.
Published: July 2012

Kö 1313 competitively inhibited the increases in heart rate produced by isoprenaline in anaesthetized dogs.

Kö 1313 was 35 times more active than propranolol in blocking an isoprenaline induced tachycardia in anaesthetized dogs but only three times more active in blocking the increase in femoral blood flow produced by the injection of isoprenaline into the femoral artery. These observations suggest that Kö 1313 may have some cardio‐selective activity.

When a series of doses of Kö 1313 was injected intravenously into anaesthetized dogs, the lower doses decreased heart rate while the larger doses increased heart rate. The same doses of propranolol reduced heart rate.

In lightly anaesthetized dogs, a series of doses of Kö 1313 progressively increased heart rate while propranolol produced little change. These observations indicate that Kö 1313 has slight intrinsic sympathomimetic activity.

The oral administration of Kö 1313 and propranolol had no effect on resting heart rate in man but reduced the increases produced by head‐up tilt, standing and exercise.
THIOPURINOL: COMPARATIVE ENZYME INHIBITION AND PROTEIN BINDING STUDIES WITH ALLOPURINOL, OXIPURINOL AND 6‐MERCAPTOPURINE
Authors: DEAN B.M. , PERRETT D. , SIMMONDS H.A. , GRAHAME R.
Published: July 2012

The metabolites of thiopurinol have been investigated in intact human and pig erythrocytes using [6‐ 14C] ‐thiopurinol, high voltage electrophoresis and automated cation exchange chromatography.

Pre‐incubation of human erythrocytes in vitro with thiopurinol increased the formation of hypoxanthine from [8‐14C]‐inosine and simultaneously reduced IMP synthesis.

Reduced inosine formation, following one week of thiopurinol therapy, has also been observed in vitro using the intact red cells from three patients suffering from gout.

Binding of thiopurinol, and to lesser extents of 6‐mercaptopurine, uric acid, oxipurinol and allopurinol, has been demonstrated electrophoretically to human and pig serum proteins.

Thiopurinol is shown to have a greater binding capacity for purified human serum albumin than uric acid or 6‐mercaptopurine at varying substrate and albumin concentrations covering the physiological range. Under these conditions the binding of uric acid is reduced in the presence of thiopurinol.

A comparison of the distribution of thiopurinol, 6‐mercaptopurine, allopurinol and oxipurinol in whole blood has revealed that allopurinol and oxipurinol are not irreversibly bound to cellular proteins; whereas 30% and 13% of the total cellular uptake was irreversibly bound in the case of thiopurinol and 6‐mercaptopurine respectively at substrate levels of 1 nmol per μl blood.
ACUTE EFFECT OF ANTIPYRETIC ANALGESICS, ALONE OR IN COMBINATION WITH ALCOHOL, ON HUMAN PSYCHOMOTOR SKILLS RELATED TO DRIVING
Authors: LINNOILA M. , SEPPÄLÄ T. , MATTILA M.J.
Published: July 2012

The effect of acetylsalicylic acid (1 g), indomethacin (50 mg), and phenylbutazone (200 mg) on psychomotor skills was examined double blind on 180 volunteer students. Ninety students received ethyl alcohol (0.5 g/kg) and 90 subjects an equal volume of placebo drink in combination with the drugs.

Psychomotor skills were measured with a choice reaction test, two co‐ordination tests, and a divided attention test, having correlation with traffic behaviour. The subjects assessed their feelings of performance by means of a rating scale. The tests were done 30, 90 and 150 min after the administration of the agents.

Acetylsalicylic acid proved inactive whereas both indomethacin and phenylbutazone impaired eye‐hand co‐ordination and divided attention. Acetylsalicylic acid did not interact with alcohol to a measurable extent whereas indomethacin in combination with alcohol proved less harmful than without it. The deleterious effects of phenylbutazone and alcohol were additive.

An impairment of psychomotor skills related to driving by indomethacin and phenylbutazone should be considered when prescribing these drugs to active out‐patients.
α‐ADRENOCEPTOR BLOCKING DRUGS, PRESSOR RESPONSES TO NORADRENALINE, AND THE ANKLE JERK IN MAN
Authors: DE WHITE C. B. , RICHENS A.
Published: July 2012

Changes in pressor responses to infused noradrenaline and in triceps surae jerk amplitude have been measured after the i.v. administration of: chlorpromazine, thioridazine, prochlorperazine, dimethothiazine, indoramin, benzoctamine and clonidine.

Each of the drugs blocked pressor responses to noradrenaline and reduced triceps surae jerk amplitude, but little correlation was found between their potencies in the two systems.

The effects of the drugs on the triceps surae jerk are not incompatible with an action at a spinal α‐adrenoceptor site, but other pharmacological factors may be involved, such as penetration of the drugs into the central nervous system, and differences in their uptake1 blocking activity.
Assessment of drugs in schizophrenia. Diagnosis and patient selection.
Authors: Forrest AD
Published: July 2012

It would seem that some set of operational definitions are required for the spectrum of psychotic patients often embraced by the rubric schizophrenia. The problems of acute (first‐admission) and chronic patients have been described. At the present time ‘relapse re‐ admissions’ would seem to constitute the best population for drug evaluations. The importance of factors such as age, sex, ethnics, geography and length of history has been emphasized. Finally the importance of carefully excluding patients who would be at risk from pre‐existing disease or hypersensitivity has been emphasized.
Assessment of drugs in schizophrenia. Basic trial design.
Authors: Lader M
Published: July 2012

Clinical trials with major tranquilizers must take into account the clinical features of patients with schizophrenia and pharmacokinetic and pharmacodynamic properties of the drug. The objectives of the trial must be carefully defined so that appropriate selection criteria for patients, rating instruments and dosage schedules can be selected. It is useful to monitor physiological and biochemical actions of major tranquilizers as well as the clinical effects.
ABSORPTION OF DRUGS BY THE LUNG
Authors: SHENFIELD GILLIAN M. , EVANS MARION E. , PATERSON J.W.
Published: July 2012

Ten patients undergoing diagnostic bronchoscopy had radioisotopically labelled drugs put directly into their bronchi; two received sodium cromoglycate, two salbutamol, three salmefamol and three rimiterol.

All four drugs were rapidly absorbed but higher peak plasma levels per unit dose were seen with sodium cromoglycate and salbutamol than with the other two drugs.

It is suggested that the lung metabolizes salmefamol and rimiterol but does not metabolize salbutamol or sodium cromoglycate.
Experience with the use of a corticosteroid aerosol.
Authors: Chatterjee SS
Published: July 2012

1. In 48 patients with chronic asthma who had been receiving treatment with beclomethasone dipropionate aerosol (BDA) at a daily dose of 400 microgram for several years, observations were made on the effect of increasing the dose to 800 microgram daily for 6 months. 2. There was subjective improvement in 37 of the 48 patients, but the only objective evidence of improvement was a slight reduction in airways resistance. 3. The higher dose of BDA did not produce impairment of pituitary‐ adrenal function or an increase in the incidence of oropharyngeal candidiasis.
Long‐term results with beclomethasone dipropionate aerosol in children with bronchial asthma: why does it sometimes fail?
Authors: Gwynn CM , Smith JM
Published: July 2012

1. Beclomethasone dipropionate aerosol has been shown to be a highly effective treatment for asthma in childhood, with virtual absence of side effects at this age. 2. When treatment is unsuccessful, this is usually due to failure to take it correctly and regularly. 3. A good response is usually associated with an improvement in ventilatory function and a marked increase in growth velocity.
Beclomethasone dipropionate aerosol in treatment of perennial and seasonal rhinitis: a review of five years' experience.
Authors: Brown HM , Storey G , Jackson FA
Published: July 2012

1. Five years' experience in the use of beclomethasone dipropionate aerosol (BDA) in the treatment of 315 patients with upper respiratory tract allergy is reviewed. 2. A total of 223 patients with perennial rhinitis was treated. In 23, where the nasal allergy had recurred after oral corticosteroid therapy for asthma was withdrawn, BDA was effective in 69% of cases. A similar success rate (68%) was recorded in 169 patients suffering from perennial allergic rhinitis alone, but a satisfactory response was observed in only 45% of 31 patients with nasal polypi. 3. In 92 patients with seasonal allergic rhinitis freedom from symptoms was achieved in 80%. 4. A total of approximately 534 patient‐years of treatment has been recorded without any evidence of side‐effects either clinically or on nasal biopsy.
Beclomethasone dipropionate aerosol and nasal mucosa.
Authors: Poynter D
Published: July 2012

1. Biopsies taken from the inferior turbinates of patients undergoing treatment with beclomethasone dipropionate aerosol for allergic rhinitis showed no changes attributable to this treatment. 2. A reduction in oedema and eosinophilia was noted but there was no effect on the thickening of the basement membrane.
A simple gas chromatographic method for the determination of nomifensine in plasma and a comparison of the method with other available techniques.
Authors: Chamberlain J. , Hill HM
Published: July 2012

1. A rapid simple gas chromatographic technique for determining nomifensine in plasma is described. 2. Nomifensine is rapidly absorbed after oral administration to man. 3. Following a 100‐mg dose, peak levels of an acid‐labile conjugate of 2–3 microgram/ml are reached about 1.5 h after administration. 4. The conjugate is cleared with a half‐life of between 1 and 2 hours.
Dynamic interaction of nomifensine with alcohol.
Authors: Taeuber K.
Published: July 2012

1. Nine healthy volunteers participated in a comparative study of the effects of nomifensine, nomifensine plus alcohol, and placebo plus alcohol, on aspects of psychomotor performance. 2. The study was carried out according to a Latin square design and each treatment was separated from the preceding treatment by 7 days. 3. Placebo plus alcohol impaired performance, increased pulse rate and blood pressure, and increased feelings of activity and euphoria. 4. Nomifensine plus alcohol produced the same subjective and objective changes as placebo plus alcohol, but in no instance were changes any greater. 5. Nomifensine alone produced none of these changes. 6. It was concluded that single doses of nomifensine did not potentiate the effect of alcohol.
Effects of nomifensine in vitro on uptake of 5‐hydroxytryptamine and dopamine into human platelets.
Authors: Ehsanullah RS , Turner P.
Published: July 2012

1. The effect of nomifensine on the uptake of 5‐hydroxytryptamine (5‐ HT) and dopamine (DA) into human platelet‐rich plasma has been studied. 2. A significant inhibition of DA uptake was observed at nomifensine 10(‐6) M. 3. 5‐HT uptake was significantly inhibited only at nomifensine 10(‐4) M or more. 4. These results are consistent with those from previous animal experiments and suggest that the human platelet may be used as a model for studying DA uptake as well as that of 5‐HT.
Toxicology of clobazam.
Authors: Schutz E
Published: July 2012

1 Extensive toxicological investigations of clobazam are summarized. 2 LD50 values after oral administration ranged from 100 mg/kg in the dog to 6000 mg/kg in the rat. 3 Long‐term repeated dose studies have shown withdrawal effects in the dog and monkey (notably convulsions leading to death), similar to those known to occur with other benzodiazepines. 4 It is concluded that the overall safety and tolerability of clobazam have been demonstrated in the preclinical animal studies.
Clobazam: pharmacological and therapeutic profile.
Authors: Hanks GW
Published: July 2012

1 Clobazam is a 1,5‐benzodiazepine the pharmacological profile of which differs from that of the 1,4 benzodiazepines in that it displays a wide separation of psychosedative or 'tranquillizing' properties from impairment of motor coordination. This is associated with a relative lack of muscle relaxant activity. 2 In man it seems to be an effective anxiolytic agent when given in daily doses of 20‐30 mg but produces minimal effects on psychomotor performance at these dose levels. 3 Clobazam has a long elimination half‐life of about 18 h; the main metabolite is N‐desmethylclobazam. Clobazam seems to share the good safety and tolerability characteristics of the 1,4 benzodiazepines.
Studies on sleep and performance with a triazolo‐1, 4‐thienodiazepine (brotizolam).
Authors: Nicholson AN , Stone BM , Pascoe PA
Published: July 2012

1 Brotizolam, a triazolo‐1,4‐thienodiazepine, was studied in healthy young adults. Electroencephalographic sleep variables and subjective effects, and performance on a visuo‐motor coordination task were measured. 2 In the sleep studies six males each ingested 0.2, 0.4 and 0.6 mg brotizolam overnight. All doses increased total sleep time, improved the sleep efficiency index, and reduced drowsy sleep and number of awakenings. Brotizolam 0.4 and 0.6 mg also reduced awake activity and increased stage 2 sleep. There was some evidence of a delay to the first REM period, but only 0.6 mg reduced the total duration of REM sleep. There were no changes in slow wave sleep. 3. In the performance studies six females each ingested 0.4 mg in the morning and 0.2, 0.4 and 0.6 mg brotizolam at night. After morning ingestion of 0.4 mg there was impaired performance from 0.5 to 5.5 h. There were no residual effects after 0.2 mg brotizolam, but with 0.4 mg there was a residual effect at 9.5 h, and 0.6 mg led to impairments up to 15.0 h after ingestion. 4 Brotizolam is a short‐acting hypnotic. In doses around 0.2 mg it has useful hypnotic activity free of adverse effects on sleep and residual effects on performance. With 0.4 mg the hypnotic effect is enhanced with only minimal residual effects.
The effect of beta‐adrenoceptor agonists and antagonists on gastric emptying in man.
Authors: Rees MR , Clark RA , Holdsworth CD , Barber DC , Howlett PJ
Published: July 2012

1. The effect of beta‐adrenoceptor agonists and antagonists on gastric emptying of a solid meal labelled with Indium113m was measured by a gamma camera in healthy volunteers or patients with hypertension. Each subject acted as his own control. 2 In ten subjects given isoprenaline (10 or 20 mg) sublingually 30 min before the meal, gastric emptying half‐times were significantly prolonged (P less than 0.05). No effect was observed when isoprenaline was given immediately before the meal. 3 Salbutamol, 4 mg four times daily for 1 week also significantly prolonged gastric emptying in four subjects (P less than 0.05). 4 The effect of isoprenaline on gastric emptying was blocked by propranolol 40 mg four times daily for 1 week. 5 In ten subjects propranolol 40 mg four times daily alone for 1 week was found to significantly speed up gastric emptying (P less than 0.005), suggesting that emptying of the normal stomach is subject to some degree of adrenergic inhibition.
Pharmacological screening of new chemical entities in human subjects and methods of data handling.
Authors: Sullivan TJ , Croft AR , Hanka R , Roberts SJ
Published: July 2012

1 A system is described for screening new chemical entities in normal human subjects, which is designed to search for novel drug actions at an early stage of drug development. 2 In single (subject) blind experiments, doses are increased incrementally from one subject to next, with immediate feed‐back of results from each dose. 3 As many physiological variables as possible are recorded, for which a purpose‐ designed apparatus system was constructed, and connected to a computer. 4 To overcome problems of inter‐subject variability, the need for sensitivity, and to identify drug‐related effects from the mass of data generated, a novel data management system was developed, and physiological profiles of volunteer subjects were built up to provide control data. 5 Results from a validation experiment in which isoprenaline was infused intravenously at different rates are presented and discussed.
Pharmacokinetics of a sustained‐release theophylline formulation.
Authors: Trembath PW , Boobois SW
Published: July 2012

1 Plasma theophylline concentrations following administration of sustained‐release (SR) theophylline tablets were determined in ten healthy volunteers using a dose of 190 mg or 380 mg 12 hourly. 2 The plasma theophylline levels during the first 12 h period confirmed the sustained‐release formulation characteristics, with the plasma drug concentrations reaching a plateau for the last 6 hours. 3 During the fifth 12 h dosing period the mean maximum and minimum plasma theophylline concentrations were 7.25 and 4.30 microgram/ml after 190 mg SR theophylline 12 hourly (n = 6) and 12.96 and 7.36 microgram/ml after 380 mg 12 hourly (n = 5), although there was marked between‐ subject variation in plasma theophylline concentrations. 4 One subject withdrew from the study due to side effects, which were more common when the higher dose of SR theophylline was given.
Antipyrene clearance in Indian villagers.
Authors: Desai NK , Sheth UK , Mucklow JC , Fraser HS , Bulpitt CJ , Jones SW , Dollery CT
Published: July 2012

1 Antipyrine clearance has been measured using saliva samples in 50 Maharashtrans from a village to the north of Bombay. 2 All subjects were very lean and were also anaemic probably as a result of hookworm infestation. 3 Antipyrine clearance was 36% faster than in White Londoners studied previously and more than twice as fast as in Asian immigrants living in London. 4 Clearance was 25% faster in men than in women but volume of distribution was also greater in men and mean half‐ lives did not differ significantly between the sexes. 5 There was a negative correlation between antipyrine clearance and haemoglobin concentration. 6 The study has identified geographic differences in antipyrine metabolism which could be clinically important. The role of anaemia merits further study.