bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Skewed TCR‐BV Repertoire Displayed at the Maternal–Fetal Interface of Women with Unexplained Pregnancy Loss
Authors: Wang Xipeng , Ma Zhengwen , Hong Yan , Zhao Aimin , Qiu Lihua , Lu Peihua , Lin Qide 
Published: August 2005

ProblemThe study was undertaken to investigate T‐cell receptor (TCR) variable β (BV)‐chain usage at maternal–fetal interface and explore the relationship between the skewed TCR‐BV usage and unexplained pregnancy losses.Method of studyA total of 57 patients with unexplained pregnancy loss including 39 cases with unexplained spontaneous abortion and 18 cases with unexplained recurrent spontaneous abortion (RSA) were chosen in Renji Hospital, Shanghai Second Medical University matched with 41 women with normal pregnancies in first trimester between September 2002 and November 2003. A high‐resolution spectratyping analysis of complementarity‐determining region 3 (CDR3) was used to detect and compare the degree and pattern of TCR‐BV repertoire usage at the maternal–fetal interface between patients with pregnancy loss and normal controls.ResultsThere were two comparisons of TCR usage performed between patients and controls, which included the degree (mean value of every TCR‐BVn expression) and the pattern (skewed TCR‐BVn frequency). The skewed TCR‐BVn is defined as an absolute BVn usage of >5% calculated from the formula or a double increased BVn usage compared with the mean value of normal BVn. According to the degree of TCR‐BV usage, BV2 (P = 0.046), BV10 (P = 0.016), and BV11 (P = 0.030) in spontaneous abortion group and BV19 (P = 0.038) in RSA group showed higher usage, while BV5.2 (P = 0.006 and P = 0.046) in both abortion groups showed significantly lower usage when compared with normal controls. About the pattern of skewed TCR‐BV distribution, we found that TCR‐BV2, ‐3, ‐6, and ‐7 were the four most common BV families in deciduas of patients with both types of abortion and normal controls. Women with spontaneous abortion demonstrated higher frequency of BV10 (P = 0.035) and lower frequencies of BV4 (P = 0.002) and BV5.2 (P = 0.003) in comparison with controls. In RSA, higher frequencies of BV15 (P = 0.018), BV19 (P = 0.049), and BV20 (P = 0.018), in the mean time, lower frequencies of BV4 (P = 0.026) and BV7 (P = 0.018) distributions were verified.ConclusionsOur results suggested that a significant skewed TCR‐BV repertoire occurred at the maternal–fetal interface with patients undergoing abortion. The specific skewed usages of TCR‐BV might be associated with the susceptibility to unexplained pregnancy loss.
Type 2 11β‐Hydroxysteroid Dehydrogenase Activity in Human Fallopian Tube and Correlation of Enzyme Levels with Endometrial Histopathology
Authors: Muneyyirci‐Delale Ozgul , Nacharaju Vijaya L. , Kessel Allan , Yang Lianfu , Jalou Sanaa , Dalloul Nezar , Rahman Monzila , Bowers Charles H. , Abulafia Ovadia 
Published: August 2005

ProblemThe inter‐conversion of hormonally active cortisol and inactive cortisone is catalyzed by 11β‐hydroxysteroid dehydrogenase (11β‐HSD). This conversion controls the level of active glucocorticoid concentration in tissues. As the fallopian tube plays a major role in the process of fertilization, we wanted to investigate whether 11β‐HSD is present in the human fallopian tube to control the glucocorticoid levels as in other tissues.Method of studyIsthmic, ampullary and fimbrial portions of the fallopian tube are obtained from patients undergoing hysterectomy and salpingo‐oopherectomy for symptomatic leomyomata uteri. 11β‐HSD activities were measured in the homogenates of the tube, cortisol as the steroid substrate. The enzyme activity was expressed as nanomolar cortisone formed per minute per gram of tissue (mean ± S.D.).ResultsA significant level of 11β‐HSD activity in oxidation direction was found in all three parts of the tube. There is no significant difference in the distribution of the enzyme activity throughout the tube. When tubal 11β‐HSD activity was compared with endometrial histology, the enzyme activity is significantly lower in proliferative endometrium when compared with secretory endometrium (P = 0.002). The enzyme activity in inactive endometrium is significantly higher than the active endometrium (P = 0.05).ConclusionThe presence of 11β‐HSD throughout the fallopian tube and its correlation to endometrial histology is indicative of its probable role in controlling the glucocorticoid levels in the tissue, which in turn may influence the fertilization process.
Altered Immunophenotypic Parameters in Infertile Women. Possible Role of Herpes Viremia
Authors: Thomas D , Michou V , Moustakarias T , Aleporou V , Matzavinos T , Mitsakos‐Barbagiannis K , Kalofoutis A , Tsilivakos V 
Published: August 2005

ProblemPurpose of this study was to reveal any alteration in peripheral blood lymphocytic concentrations of a large cohort of infertile women and to investigate the possible role of herpes viremia in the peripheral immunostimulation.Method of studyThe immunophenotypic characteristics and the presence of herpes viruses DNA in the peripheral blood of 168 infertile women were studied.ResultsPeripheral CD56+/CD16+ natural killer (NK) cell concentration, CD56+/CD16− NK cell concentration, white blood cell (WBC) concentration and lymphocyte concentration were statistically correlated to herpes viremia. Epstein–Barr virus (EBV) viremia is related with a limited reduction of CD56+/CD16− cell levels in the peripheral blood of infertile women with regard to the rest of herpes viruses. High T‐lymphocyte concentration, CD4+ T‐cell concentration and CD8+ T‐cell concentration was observed in women positive for three different kinds of herpes viruses (triple viremia) in the peripheral blood.ConclusionsAssuming that all women under study remained asymptomatic, these data suggest that subclinical herpesvirus viremia may be an important cause of peripheral immunostimulation in women with a history of infertility.
ASRI2005‐90  Vaginal formulations – how innocuous are they to the vaginal epithelial immune function?
Authors: Trifonova RT , Pasicznyk J‐M , Fichorova RN 
Published: August 2005

Problem:  The disruption of the epithelial barrier and activation of inflammatory pathways may lead to increased susceptibility to viral infection. Over‐the‐counter (OTC) vaginal products have received little attention in this regard, however the recent development of anti‐HIV‐1 microbicides has brought the importance of mucosal integrity to forefront. While vaginal microbicides represent a promising strategy for preventing HIV‐1 and other sexually transmitted infections, they can be efficacious only if they cause no epithelial toxicity or inflammation.Method:  We used a novel organotypic cervicovaginal model (VEC100, Mattek) to compare hygienic or spermicidal OTC to the anti‐HIV‐1 microbicide cellulose acetate 1,2‐benzenedicarboxylate (CAP).Results:  CAP film and gel formulations appeared non‐toxic in MTT and LDH viability assays and did not induce release of pro‐inflammatory mediators e.g. IL‐1, IL‐6 and IL‐8 as compared to controls. In contrast, a commercially available vaginal cleansing film containing Nonoxynol‐9 showed proinflammatory potential commeasurable to that of TNFα.Conclusions:  Our study demonstrates the usefulness of the VEC tissue model for immunoinflammatory characterization and safety assessment of vaginal products. It provides evidence that CAP in film and gel forms may be used as a safe anti‐HIV‐1 prevention agent or vehicle for other vaginal products. This study was supported by NICHD 1P01HD041761.
The Contemporary Presidency: Condoleezza Rice as NSC Advisor: A Case Study of the Honest Broker Role
Authors: BURKE JOHN P. 
Published: August 2005

“We need to plan as if things won’t go well.”

—George W. Bush, September 25, 2001 (Woodward 2002, 136)

“I want to know what the options are. A president cannot decide and make rational decisions unless I understand the feasibility of that which may have to happen.”

—George W. Bush, circa late November 2001 (Woodward 2004, 30)

“If we don’t have a case to make, I won’t send in the troops.”

—George W. Bush, January 6, 2003 (Woodward 2004, 256)
The role of honest broker has often been seen as a contributor to effective presidential decision making. This article explores the broker role of National Security Council Advisor Condoleezza Rice during three crucial decision‐making episodes in George W. Bush's first term: (1) deliberations in the aftermath of the September 11 attacks and the decision to go to war in Afghanistan; (2) deliberations that led to the war in Iraq; and (3) deliberations concerning Iraq's postwar stabilization and reconstruction. The article finds that the broker role declined over time. This decline does not appear to be linked to other roles of the NSC advisor such as policy advocacy, but it does appear to be affected by problems of organization and management as well as its fit with presidential expectations and support of the broker role.
IDENTIFICATION OF SALMONELLA‐INOCULATED BEEF USING A PORTABLE ELECTRONIC NOSE SYSTEM
Authors: BALASUBRAMANIAN S. , PANIGRAHI S. , LOGUE C.M. , MARCHELLO M. , SHERWOOD J.S. 
Published: August 2005

ABSTRACT A commercially available Cyranose‐320TM (Cyrano Sciences, Pasadena, CA) conducting polymer–based electronic nose system was used to analyze the volatile organic compounds emanating from vacuum‐packaged beef strip loins (Longissimus Lamborum) that were repackaged simulating retail store conditions and stored at 4 and 10°C after inoculating them with Salmonella Typhimurium. Two statistical techniques, i.e., linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA), were used to develop classification models from the collected sensor signals. The models were developed for the captured signals, which were scaled and not scaled. The performances of the developed models were validated by two different methods, leave‐one‐out cross validation and bootstrapping. The developed models classified the meat samples based on the Salmonella population into “No Salmonella” (microbial counts < 0.7 log10 cfu/g) and “Salmonella” (microbial counts ≥ 0.7 log10 cfu/g). Overall, scaling the signals improved the classification accuracies obtained. The quadratic discriminant‐based classification models validated by the bootstrapping technique performed better than the LDA‐based models. For the meat samples stored at 10°C, the highest classification accuracy obtained by the QDA method with bootstrapping validation was 80.5% when the signals were scaled. For the samples stored at 4°C, the QDA method provided the highest classification accuracy of 87.3% using bootstrapping validation. The results obtained prove that the electronic nose system could identify meat samples contaminated with S. Typhimurium at a population concentration level of 0.7–2.6 log10 cfu/g.
QUANTIFICATION OF ATTACHMENT STRENGTH OF SELECTED FOODBORNE PATHOGENS BY THE BLOT SUCCESSION METHOD
Authors: KIM T. , SILVA J.L. 
Published: August 2005

ABSTRACT Attachment strength of selected foodborne pathogens (Listeria monocytogenes, Listeria innocua, Listeria grayii, Salmonella Typhimurium , and Escherichia coli O157:H7) were quantified using the blot succession method. E. coli O157:H7 and S. Typhimurium had higher removal exponents than Listeria species. Regardless of initial cell number, L. monocytogenes had the lowest removal exponent (0.27), being the most difficult to remove from the glass surface. The other Listeria spp. had exponents of 0.38–0.52 while S. Typhimurium and E. coli O157:H7 had exponents of 0.9 or higher. This method can serve to quantify attachment or detachment strength on food contact surfaces or digestive tracts.
Integration of Signals through Crc and PtsN in Catabolite Repression of
Pseudomonas putida
TOL Plasmid pWW0
Authors: Aranda-Olmedo Isabel , Ramos Juan L. , Marqués Silvia 
Published: August 2005

ABSTRACT

Toluene degradation in
Pseudomonas putida
KT2440 pWW0 plasmid is subjected to catabolite repression. Pu and P
S1
promoters of the pWW0 TOL plasmid are down-regulated in vivo during exponential growth in rich medium. In cells growing on minimal medium, yeast extract (YE) addition mimics exponential-phase rich medium repression of these promoters. We have constructed and tested mutants in a series of global regulators described in
Pseudomonas
. We describe that a mutant in
crc
(
c
atabolite
r
epression
c
ontrol) partially relieves YE repression. Macroarray experiments show that
crc
transcription is strongly increased in the presence of YE, inversely correlated with TOL pathway expression. On the other hand, we have found that induced levels of expression from Pu and P
S
in the presence of YE are partially derepressed in a
ptsN
mutant of
P. putida
. PtsN but not Crc seems to directly interfere with XylR activation at target promoters. The effect of the double mutation in
ptsN
and
crc
is not the sum of the effects of each independent mutation and suggests that both regulators are elements of a common regulatory pathway. Basal expression levels from these promoters in the absence of inducer are still XylR dependent and are also repressed in the presence of yeast extract. Neither
crc
nor
ptsN
could relieve this repression.
Characterization of an Unusual Cold-Active β-Glucosidase Belonging to Family 3 of the Glycoside Hydrolases from the Psychrophilic Isolate
Paenibacillus
sp. Strain C7
Authors: Shipkowski Stephanie , Brenchley Jean E. 
Published: August 2005

ABSTRACT

We selected for spore-forming psychrophilic bacteria able to use lactose as a carbon source and one isolate, designated
Paenibacillus
sp. strain C7, that was phylogenetically related to, but distinct from both
Paenibacillus macquariensis
and
Paenibacillus antarcticus
. Some
Escherichia coli
transformants obtained with genomic DNA from this isolate hydrolyzed X-Gal (5-bromo-4-chloro-3-indoyl-β-
d
-galactopyranoside) only below 30°C, an indication of cold-active β-galactosidase activity. Sequencing of the cloned insert revealed an open reading frame encoding a 756-amino acid protein that, rather than belonging to a family typically known for β-galactosidase activity, belonged to glycoside hydrolase family 3, a family of β-glucosidases. Because of this unusual placement, the recombinant enzyme (BglY) was purified and characterized. Consistent with its classification, the enzyme had seven times greater activity with the glucoside substrate ONPGlu (
o
-nitrophenyl-β-
d
-glucopyranoside) than with the galactoside substrate ONPGal (
o
-nitrophenyl-β-
d
-galactopyranoside). In addition, the enzyme had, with ONPGlu, a thermal optimum around 30 to 35°C, activity over a broad pH range (5.5 to 10.9), and an especially low

K
m

(<0.003 mM). Further examination of substrate preference showed that the BglY enzyme also hydrolyzed other aryl-β-glucosides such as helicin, MUG (4-methylumbelliferyl-β-
d
-glucopyranoside), esculin, indoxyl-β-
d
-glucoside (a natural indigo precursor), and salicin, but had no activity with glucosidic disaccharides or lactose. These characteristics and substrate preferences make the BglY enzyme unique among the family 3 β-glucosidases. The hydrolysis of a variety of aryl-β-glucosides suggests that the enzyme may allow the organism to use these substrates in the environment and that its low

K
m

on indoxyl-β-
d
-glucoside may make it useful for producing indigo.
Naturally Occurring DNA Transfer System Associated with Membrane Vesicles in Cellulolytic
Ruminococcus
spp. of Ruminal Origin
Authors: Klieve Athol V. , Yokoyama Melvin T. , Forster Robert J. , Ouwerkerk Diane , Bain Peter A. , Mawhinney Erin L. 
Published: August 2005

ABSTRACT

A genetic transformation system with similarities to those reported for gram-negative bacteria was found to be associated with membrane vesicles of the ruminal cellulolytic genus
Ruminococcus.
Double-stranded DNA was recovered from the subcellular particulate fraction of all the cellulolytic ruminococci examined. Electron microscopy revealed that the only particles present resembled membrane vesicles. The likelihood that the DNA was associated with membrane vesicles (also known to contain cellulosomes) was further supported by the adherence of the particles associated with the subcellular DNA to cellulose powder added to culture filtrates. The particle-associated DNA comprised a population of linear molecules ranging in size from <20 kb to 49 kb (
Ruminococcus
sp. strain YE73) and from 23 kb to 90 kb (
Ruminococcus albus
AR67). Particle-associated DNA from
R. albus
AR67 represented DNA derived from genomic DNA of the host bacterium having an almost identical HindIII digestion pattern and an identical 16S rRNA gene. Paradoxically, particle-associated DNA was refractory to digestion with EcoRI, while the genomic DNA was susceptible to extensive digestion, suggesting that there is differential restriction modification of genomic DNA and DNA exported from the cell. Transformation using the vesicle-containing fraction of culture supernatant of
Ruminococcus
sp. strain YE71 was able to restore the ability to degrade crystalline cellulose to two mutants that were otherwise unable to do so. The ability was heritable and transferred to subsequent generations. It appears that membrane-associated transformation plays a role in lateral gene transfer in complex microbial ecosystems, such as the rumen.
Diel Infection of a Cyanobacterium by a Contractile Bacteriophage
Authors: Kao C. Cheng , Green Susan , Stein Barry , Golden Susan S. 
Published: August 2005

ABSTRACT

Light was found to strongly influence the infection of a freshwater cyanobacterium (
Synechococcus elongatus
PCC 7942) by a contractile DNA phage named AS-1. Phage progeny production was correlated with the amount of light in the laboratory and occurred in a diel pattern under natural light. At least one effect of light on AS-1 infection is at the level of adsorption.
Expression of 3-Ketoacyl-Acyl Carrier Protein Reductase (
fabG
) Genes Enhances Production of Polyhydroxyalkanoate Copolymer from Glucose in Recombinant
Escherichia coli
JM109
Authors: Nomura Christopher T. , Taguchi Kazunori , Gan Zhihua , Kuwabara Kazuhiro , Tanaka Tomoyo , Takase Kazuma , Doi Yoshiharu 
Published: August 2005

ABSTRACT

Polyhydroxyalkanoates (PHAs) are biologically produced polyesters that have potential application as biodegradable plastics. Especially important are the short-chain-length-medium-chain-length (SCL-MCL) PHA copolymers, which have properties ranging from thermoplastic to elastomeric, depending on the ratio of SCL to MCL monomers incorporated into the copolymer. Because of the potential wide range of applications for SCL-MCL PHA copolymers, it is important to develop and characterize metabolic pathways for SCL-MCL PHA production. In previous studies, coexpression of PHA synthase genes and the 3-ketoacyl-acyl carrier protein reductase gene (
fabG
) in recombinant
Escherichia coli
has been shown to enhance PHA production from related carbon sources such as fatty acids. In this study, a new
fabG
gene from
Pseudomonas
sp. 61-3 was cloned and its gene product characterized. Results indicate that the
Pseudomonas
sp. 61-3 and
E. coli
FabG proteins have different substrate specificities in vitro. The current study also presents the first evidence that coexpression of
fabG
genes from either
E. coli
or
Pseudomonas
sp. 61-3 with
fabH
(F87T) and PHA synthase genes can enhance the production of SCL-MCL PHA copolymers from nonrelated carbon sources. Differences in the substrate specificities of the FabG proteins were reflected in the monomer composition of the polymers produced by recombinant
E. coli
. SCL-MCL PHA copolymer isolated from a recombinant
E. coli
strain had improved physical properties compared to the SCL homopolymer poly-3-hydroxybutyrate. This study defines a pathway to produce SCL-MCL PHA copolymer from the fatty acid biosynthesis that may impact on PHA production in recombinant organisms.
Enhanced Reactivity of
Rhizopus oryzae
Lipase Displayed on Yeast Cell Surfaces in Organic Solvents: Potential as a Whole-Cell Biocatalyst in Organic Solvents
Authors: Shiraga Seizaburo , Kawakami Masayuki , Ishiguro Masaji , Ueda Mitsuyoshi 
Published: August 2005

ABSTRACT

Immobilization of enzymes on some solid supports has been used to stabilize enzymes in organic solvents. In this study, we evaluated applications of genetically immobilized
Rhizopus oryzae
lipase displayed on the cell surface of
Saccharomyces cerevisiae
in organic solvents and measured the catalytic activity of the displayed enzyme as a fusion protein with α-agglutinin. Compared to the activity of a commercial preparation of this lipase, the activity of the new preparation was 4.4 × 10
4
-fold higher in a hydrolysis reaction using
p
-nitrophenyl palmitate and 3.8 × 10
4
-fold higher in an esterification reaction with palmitic acid and
n
-pentanol (0.2% H
2
O). Increased enzyme activity may occur because the lipase displayed on the yeast cell surface is stabilized by the cell wall. We used a combination of error-prone PCR and cell surface display to increase lipase activity. Of 7,000 colonies in a library of mutated lipases, 13 formed a clear halo on plates containing 0.2% methyl palmitate. In organic solvents, the catalytic activity of 5/13 mutants was three- to sixfold higher than that of the original construct. Thus, yeast cells displaying the lipase can be used in organic solvents, and the lipase activity may be increased by a combination of protein engineering and display techniques. Thus, this immobilized lipase, which is more easily prepared and has higher activity than commercially available free and immobilized lipases, may be a practical alternative for the production of esters derived from fatty acids.
First Archaeal Inorganic Polyphosphate/ATP-Dependent NAD Kinase, from Hyperthermophilic Archaeon
Pyrococcus horikoshii
: Cloning, Expression, and Characterization
Authors: Sakuraba Haruhiko , Kawakami Ryushi , Ohshima Toshihisa 
Published: August 2005

ABSTRACT

The gene (PH1074) encoding the NAD kinase of the hyperthermophilic archaeon
Pyrococcus horikoshii
was identified in the genome database, cloned, and functionally expressed in
Escherichia coli
. The recombinant enzyme was purified to homogeneity by heat treatment at 90°C for 20 min and one successive HiTrap affinity chromatography step. The purified enzyme was easily precipitated by dialysis against phosphate buffer without NaCl and imidazole and was usually stored in buffer containing 0.5 M NaCl and 0.5 M imidazole to avoid precipitation. The molecular mass of the active enzyme was determined to be 145 kDa by a gel filtration method, and the enzyme was composed of a tetramer of 37-kDa subunits. The archaeal enzyme utilized several nucleoside triphosphates, such as GTP, CTP, UTP, and ITP, as well as ATP and inorganic polyphosphates [poly(P)] as phosphoryl donors for NAD phosphorylation. The enzyme utilized poly(P)
27
(the average length of the phosphoryl chain was 27) as the most active inorganic polyphosphate for NAD phosphorylation. Thus, this enzyme is categorized as an inorganic polyphosphate/ATP-dependent NAD kinase. The enzyme was the most thermostable NAD kinase found to date: its activity was not lost by incubation at 95°C for 10 min. The enzyme showed classical Michaelis-Menten-type kinetics for NAD and ATP, but not for poly(P)
27
. The

K
m

values for NAD were determined to be 0.30 and 0.40 mM when poly(P)
27
and ATP, respectively, were used as the phosphoryl donors. The

K
m

value for ATP was 0.29 mM, and the concentration of poly(P)
27
which gave half of the maximum enzyme activity was 0.59 mM. The enzyme required several metal cations, such as Mg
2+
, Mn
2+
, or Ni
2+
, for its activity. The deduced amino acid sequence showed a low level of identity to those of
E. coli
ATP-dependent NAD kinase (31%) and the inorganic polyphosphate/ATP-dependent NAD kinase of
Mycobacterium tuberculosis
(29%). This is the first description of the characteristics of a poly(P)/ATP-dependent NAD kinase from a hyperthermophilic archaeon.