bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel Mutation of POU3F4 Causes Congenital Profound Sensorineural Hearing Loss in a Large Chinese Family
Authors: Wang Qiu‐Ju , Li Qing‐Zhong , Rao Shao‐Qi , Zhao Ya‐Li , Yuan Hu , Yang Wei‐Yan , Han Dong‐yi , Shen Yan 
Published: June 2006

AbstractObjectives/Hypothesis: It is known that approximately 5% of congenital profound hearing impaired cases are inherited in X‐linked inheritance. This study aimed at identifying its underlying molecular determinant(s) using a large, five‐generation Chinese family with multiple familial cases.Study Design: Model‐based linkage analysis and positional cloning.Methods: Model‐based genetic linkage analyses were performed with the use of microsatellite polymorphisms to determine disease locus. Mutation screening was performed within the family and unrelated population‐based controls to establish molecular evidence as to what caused the specific X‐linked inheritance pattern in the family.Results: Clinical investigations of the pedigree demonstrated the extremely high penetrance in the male members but no penetrance in the female members. Linkage analyses mapped the disease to the chromosomal region Xq13.I‐Xq23 (maximum X‐linkage logarithm of odds score = 3.27). Mutation screening of the candidate genes in the linkage region by direct sequencing revealed a de novo missense substitution (925T>C) in the well‐known deaf gene. POU3F4. Direct sequencing on 240 unrelated controls did not detect any mutation.Conclusions: Multiple analysis approaches demonstrated that these disorders in the family were caused by a founder mutation in the POU3F4 gene. Our findings provided confirmatory molecular evidence to support that development of congenital profound sensorineural hearing loss in the Chinese population results from a novel mutation in the same gene.
Effects of a Nerve‐Muscle Pedicle on the Denervated Rat Thyroarytenoid Muscle
Authors: Kumai Yoshihiko , Ito Takaaki , Udaka Naoko , Yumoto Eiji 
Published: June 2006

AbstractObjectives: To evaluate the effects of the nerve‐muscle pedicle (NMP) method on the rat thyroarytenoid (TA) muscle after transection of the recurrent laryngeal nerve (RLN).Study Design: Quantitative histologic assessment of the TA muscle after NMP implantation.Methods: Thirty‐six Wistar rats were divided into two groups: animals subjected to transection of the left RLN alone (DNV group) and animals subjected to transection of the left RLN followed by immediate transplantation of a NMP flap containing the sternohyoid (SH) muscle and ansa cervicalis nerve branch (NMP group). Animals were killed 2, 4, and 10 weeks after the treatments. The TA muscle stained with hematoxylin‐eosin was evaluated quantitatively. The pre‐ and postsynaptic structures of the neuromuscular junction (NMJ) in the TA muscle were analyzed histochemically. The myosin heavy chain (MyHC) isoforms were evaluated using immunohistochemistry and reverse‐transcriptase polymerase chain reaction (RT‐PCR).Results: In the NMP group, the TA muscle fiber recovered to almost normal at 10 weeks, and the ratio of the number of synaptophysin‐positive nerve terminals to that of α‐bungarotoxin‐positive acetylcholine receptors recovered to 79.8 ± 11.8% (P < .05, compared with the control). Immunohistochemistry and the RT‐PCR method after laser capture microdissection revealed the expression of MyHC isoforms type 2B and type 2A; the latter was detected in the SH muscle but not in the normal or denervated TA muscle.Conclusion: The NMP method was effective for recovering from the atrophic changes of the TA muscle after transection of the RLN. This was attributed to successful reinnervation by reconstruction of the NMJ, which might change MyHC isoform expression.
Darbepoetin‐alpha for the anaemia of myelofibrosis with myeloid metaplasia
Authors: Cervantes Francisco , Alvarez‐Larrán Alberto , Hernández‐Boluda Juan‐Carlos , Sureda Anna , Granell Miquel , Vallansot Rolando , Besses Carles , Montserrat Emili 
Published: June 2006

SummaryDarbepoetin‐α, a novel hyperglycosylated erythropoiesis‐stimulating protein, was administered to 20 patients with myelofibrosis with myeloid metaplasia and anaemia. The initial weekly dose, 150 μg, was increased to 300 μg when no response was observed after 4–8 weeks. Eight patients (40%) responded to treatment, including six complete and two partial responses, and five maintained their response at a median follow‐up of 12 months (range 4–22). Univariate analysis indicated that older age was the only factor associated with a favourable response to treatment (P = 0.006). None of the patients with appropriate serum erythropoietin levels responded. Treatment was usually well tolerated.
Suspects in the tale of lupus-associated thrombocytopenia
Authors: Ziakas P D , Routsias J G , Giannouli S , Tasidou A , Tzioufas A G , Voulgarelis M 
Published: June 2006

Summary
Immunologically mediated thrombocytopenia is a frequent clinical manifestation in patients with systemic lupus erythematosus (SLE). Autoantibodies targeting platelet membrane glucoproteins have a central role in peripheral platelet destruction. Autoantibodies against thrombopoietin are also present in about one-third of patients, but their pathogenetic role is obscure. Thirty-eight serum samples from SLE patients were tested for anti-platelet antibodies, anti-thrombopoietin antibodies and levels of circulating thrombopoietin. Bone marrow histology was also assessed. Thirty-nine per cent of sera displayed anti-thrombopoietin antibodies and 29% had circulating anti-platelet antibodies. Anti-thrombopoietin antibodies were associated with lower thrombopoietin concentrations, and lower mean platelet values in long-term follow-up. Anti-platelet antibodies were present in about 40% of thrombocytopenic and non-thrombocytopenic individuals but were absent in patients who had recovered from thrombocytopenia, supporting their pathogenetic role. Both autoantibodies were absent in control sera from patients with rheumatoid arthritis and primary Sjögren’s syndrome. Decreased bone marrow cellularity, normal or low number of hypolobulated, pyknotic megakaryocytes and stromal alterations were prominent findings in thrombocytopenic SLE patients, suggesting a defect in megakaryopoiesis. These findings were not evident in specimens from patients with idiopathic thrombocytopenic purpura who had increased megakaryocytes, normal cellularity and absence of stromal alterations. In conclusion, peripheral destruction due to platelet autoantibodies, anti-thrombopoetin antibodies, lower effective circulating thrombopoetin and impaired compensatory response due to bone marrow damage interact in SLE and thrombocytopenia ensues.
Signalling network with a bistable hysteretic switch controls developmental activation of the σF transcription factor in Bacillus subtilis
Authors: Igoshin Oleg A. , Price Chester W. , Savageau Michael A. 
Published: June 2006

SummaryThe sporulation process of the bacterium Bacillus subtilis unfolds by means of separate but co‐ordinated programmes of gene expression within two unequal cell compartments, the mother cell and the smaller forespore. σF is the first compartment‐specific transcription factor activated during this process, and it is controlled at the post‐translational level by a partner‐switching mechanism that restricts σF activity to the forespore. The crux of this mechanism lies in the ability of the anti‐sigma factor SpoIIAB (AB) to form alternative complexes either with σF, holding it in an inactive form, or with the anti‐anti‐sigma factor SpoIIAA (AA) and a nucleotide, either ATP or ADP. In the complex with AB and ATP, AA is phosphorylated on a serine residue and released, making AB available to capture σF in an inactive complex. Subsequent activation of σF requires the intervention of the SpoIIE serine phosphatase to dephosphorylate AA, which can then attack the AB–σF complex to induce the release of σF. By incorporating biochemical, biophysical and genetic data from the literature we have constructed an integrative mathematical model of this partner‐switching network. The model predicts that the self‐enhancing formation of a long‐lived complex of AA, AB and ADP transforms the network into an essentially irreversible hysteretic switch, thereby explaining the sharp, robust and irreversible activation of σF in the forespore compartment. The model also clarifies the contributions of the partly redundant mechanisms that ensure correct spatial and temporal activation of σF, reproduces the behaviour of various mutants and makes strong, testable predictions.
Multi‐micronutrient Sprinkles including a low dose of iron provided as microencapsulated ferrous fumarate improves haematologic indices in anaemic children: a randomized clinical trial
Authors: Christofides Anna , Asante Kwaku Poku , Schauer Claudia , Sharieff Waseem , Owusu‐Agyei Seth , Zlotkin Stanley 
Published: June 2006

AbstractHome‐fortification of complementary foods with micronutrients (including iron) as Sprinkles is a new strategy to control iron deficiency and anaemia in developing countries. However, the most effective dose and form of iron is not known. The purpose of this study was to compare the efficacy of various doses (12.5, 20 or 30 mg) and treatment methods (multi‐micronutrient Sprinkles vs. ferrous sulphate drops) on haemoglobin (Hb) concentration after 8 weeks of treatment in anaemic children. In total, 133 anaemic Ghanaian children (Hb 70–99 g L−1) aged 6–18 months were randomly assigned to one of five daily interventions for 8 weeks. Out of the five interventions, four used Sprinkles, and one used iron drops. Of the four Sprinkles groups, three included 12.5, 20 or 30 mg of iron as ferrous fumarate, and one included 20 mg of iron as ferric pyrophosphate. The iron drops group included 12.5 mg of iron as liquid ferrous sulphate. Hb concentrations were measured at baseline, week 3 and week 8. The primary outcome measure was Hb concentration at 8 weeks after treatment. We compared differences in Hb and ferritin concentrations and prevalence of iron deficiency anaemia (Hb < 100 g L−1 and soluble transferrin receptor concentrations >8.5 mg L−1) from baseline to 8 weeks within and between groups. Adherence and reporting of side effects (staining of the teeth, ease of use, diarrhoea and darkening of stools) were compared between groups. Mean change in Hb was 1.4 g L−1 (SD = 1.8) (P = 0.0001). Change in Hb concentrations from baseline to 8 weeks was significant in all groups (P = 0.0001–0.0007), with no differences across groups. Geometric means of serum ferritin varied from 18.6 to 44.0 µg L−1 at baseline. At week 8, these means were in the interval of 48.0–78.3 µg L−1, with no group differences. Prevalence of iron deficiency anaemia decreased significantly from baseline to 8 weeks in all groups with the exception of the iron drops group, with no group differences. Adherence was lower in the drops group (64%) as compared with Sprinkles groups (84%). Greater staining of the teeth and less ease of use were reported in the drops group as compared with Sprinkles groups. A dose as low as 12.5 mg of iron as ferrous fumarate when provided as Sprinkles may be effective in anaemic children.
Depth-Related Gradients of Viral Activity in Lake Pavin
Authors: Colombet J. , Sime-Ngando T. , Cauchie H. M. , Fonty G. , Hoffmann L. , Demeure G. 
Published: June 2006

ABSTRACT
High-resolution vertical sampling and determination of viral and prokaryotic parameters in a deep volcanic lake shows that in the absence of thermal stratification but within light, oxygen, and chlorophyll gradients, host availability empirically is prevalent over the physical and chemical environments and favors lytic over lysogenic “viral life cycles.”
Cadmium Ion Biosorption by the Thermophilic Bacteria
Geobacillus stearothermophilus
and
G. thermocatenulatus
Authors: Hetzer Adrian , Daughney Christopher J. , Morgan Hugh W. 
Published: June 2006

ABSTRACT

This study reports surface complexation models (SCMs) for quantifying metal ion adsorption by thermophilic microorganisms. In initial cadmium ion toxicity tests, members of the genus
Geobacillus
displayed the highest tolerance to CdCl
2
(as high as 400 to 3,200 μM). The thermophilic, gram-positive bacteria
Geobacillus stearothermophilus
and
G. thermocatenulatus
were selected for further electrophoretic mobility, potentiometric titration, and Cd
2+
adsorption experiments to characterize Cd
2+
complexation by functional groups within and on the cell wall. Distinct one-site SCMs described the extent of cadmium ion adsorption by both studied
Geobacillus
sp. strains over a range of pH values and metal/bacteria concentration ratios. The results indicate that a functional group with a deprotonation constant pK value of approximately 3.8 accounts for 66% and 80% of all titratable sites for
G. thermocatenulatus
and
G. stearothermophilus
, respectively, and is dominant in Cd
2+
adsorption reactions. The results suggest a different type of functional group may be involved in cadmium biosorption for both thermophilic strains investigated here, compared to previous reports for mesophilic bacteria.
Evaluation of the Duopath
Legionella
Lateral Flow Assay for Identification of
Legionella pneumophila
and
Legionella
Species Culture Isolates
Authors: Helbig Jürgen Herbert , Lück Paul Christian , Kunz Britta , Bubert Andreas 
Published: June 2006

ABSTRACT

Duopath
Legionella
(Merck KGaA, Darmstadt, Germany) is a new immunochromatographic assay for the simultaneous identification of cultured
L. pneumophila
and
Legionella
species other than
L. pneumophila
. In tests of 89
L. pneumophila
strains and 87
Legionella
strains other than
L. pneumophila
representing 41 different species, Duopath and a widely used latex agglutination assay detected
L. pneumophila
with 100% and 98% accuracy, respectively, whereas the percentages differed significantly for other
Legionella
spp. (93% versus 37% [
P
< 0.001]). Since many countries’ regulations require the identification of
Legionella
spp. in water and environmental samples, the use of Duopath
Legionella
to comply with those regulations could contribute to significantly fewer false-negative results.
Prevalence and Identity of
Cryptosporidium
spp. in Pig Slurry
Authors: Xiao Lihua , Moore John E. , Ukoh Ukeme , Gatei Wangeci , Lowery Colm J. , Murphy Thomas M. , Dooley James S. G. , Millar B. Cherie , Rooney Paul J. , Rao Juluri R. 
Published: June 2006

ABSTRACT

Cryptosporidium
spp. were detected in 25 of 56 pig slurry samples from 33 Irish farms by PCR and DNA sequencing. The organisms detected included
C. suis
,
Cryptosporidium
pig genotype II, and
C. muris.
We concluded that
Cryptosporidium
oocysts can persist in treated slurry and potentially contaminate surface water through improper discharge or uncontrolled runoff.
Structure of Trichamide, a Cyclic Peptide from the Bloom-Forming Cyanobacterium
Trichodesmium erythraeum
, Predicted from the Genome Sequence
Authors: Sudek Sebastian , Haygood Margo G. , Youssef Diaa T. A. , Schmidt Eric W. 
Published: June 2006

ABSTRACT

A gene cluster for the biosynthesis of a new small cyclic peptide, dubbed trichamide, was discovered in the genome of the global, bloom-forming marine cyanobacterium
Trichodesmium erythraeum
ISM101 because of striking similarities to the previously characterized patellamide biosynthesis cluster. The
tri
cluster consists of a precursor peptide gene containing the amino acid sequence for mature trichamide, a putative heterocyclization gene, an oxidase, two proteases, and hypothetical genes. Based upon detailed sequence analysis, a structure was predicted for trichamide and confirmed by Fourier transform mass spectrometry. Trichamide consists of 11 amino acids, including two cysteine-derived thiazole groups, and is cyclized by an N—C terminal amide bond. As the first natural product reported from
T. erythraeum
, trichamide shows the power of genome mining in the prediction and discovery of new natural products.
Cysteine Metabolism in
Legionella pneumophila
: Characterization of an
l
-Cystine-Utilizing Mutant
Authors: Ewann Fanny , Hoffman Paul S. 
Published: June 2006

ABSTRACT

Growth of
Legionella pneumophila
on buffered charcoal-yeast extract (BCYE) medium is dependent on
l
-cysteine (but not
l
-cystine), which is added in excess over what is required for nutrition. We investigated the biochemical and genetic bases for this unusual requirement and determined that much of the
l
-cysteine in BCYE medium is rapidly oxidized to
l
-cystine and is unavailable to the bacteria. Analysis of cysteine consumption during bacterial growth indicated that of the 11% consumed, 3.85% (∼0.1 mM) was incorporated into biomass. The activities of two key cysteine biosynthetic enzymes (serine acetyltransferase and cysteine synthase) were not detected in cell extracts of
L. pneumophila
, and the respective genes were not present in the genome sequences, confirming cysteine auxotrophy. Kinetic studies identified two energy-dependent cysteine transporters, one with high affinity (apparent

K
m

, 3.29 μM) and the other with low affinity (apparent

K
m

, 93 μM), each of which was inhibited by the uncoupling agent carbonyl cyanide
m
-chlorophenylhydrazone. Cystine was not transported by
L. pneumophila
; however, a mutant strain capable of growth on
l
-cystine (CYS1 mutant) transported
l
-cystine with similar kinetics (

K
m

, 4.4 μM and 90 μM). Based on the bipartite kinetics, requirement for proton motive force, and inhibitor studies, we suggest that a high-affinity periplasmic binding protein and a major facilitator/symporter (low affinity) mediate uptake. The latter most likely is functional at high cysteine concentrations and most likely displays altered substrate specificity in the CYS-1 mutant. Our studies provide biochemical evidence to support a general view that
L. pneumophila
is restricted to an intracellular lifestyle in natural environments by an inability to utilize cystine, which most likely ensures that the dormant cyst-like transmissible forms do not germinate outside suitable protozoan hosts.
Development of
Bacteroides
16S rRNA Gene TaqMan-Based Real-Time PCR Assays for Estimation of Total, Human, and Bovine Fecal Pollution in Water
Authors: Layton Alice , McKay Larry , Williams Dan , Garrett Victoria , Gentry Randall , Sayler Gary 
Published: June 2006

ABSTRACT

Bacteroides
species are promising indicators for differentiating livestock and human fecal contamination in water because of their high concentration in feces and potential host specificity. In this study, a real-time PCR assay was designed to target
Bacteroides
species (AllBac) present in human, cattle, and equine feces. Direct PCR amplification (without DNA extraction) using the AllBac assay was tested on feces diluted in water. Fecal concentrations and threshold cycle were linearly correlated, indicating that the AllBac assay can be used to estimate the total amount of fecal contamination in water. Real-time PCR assays were also designed for bovine-associated (BoBac) and human-associated (HuBac)
Bacteroides
16S rRNA genes. Assay specificities were tested using human, bovine, swine, canine, and equine fecal samples. The BoBac assay was specific for bovine fecal samples (100% true-positive identification; 0% false-positive identification). The HuBac assay had a 100% true-positive identification, but it also had a 32% false-positive rate with potential for cross-amplification with swine feces. The assays were tested using creek water samples from three different watersheds. Creek water did not inhibit PCR, and results from the AllBac assay were correlated with those from
Escherichia coli
concentrations (
r
2
= 0.85). The percentage of feces attributable to bovine and human sources was determined for each sample by comparing the values obtained from the BoBac and HuBac assays with that from the AllBac assay. These results suggest that real-time PCR assays without DNA extraction can be used to quantify fecal concentrations and provide preliminary fecal source identification in watersheds.
Genotyping of Human and Porcine
Yersinia enterocolitica
,
Yersinia intermedia
, and
Yersinia bercovieri
Strains from Switzerland by Amplified Fragment Length Polymorphism Analysis
Authors: Boghenbor Kathrin Kuehni , On Stephen L. W. , Kokotovic Branko , Baumgartner Andreas , Wassenaar Trudy M. , Wittwer Matthias , Bissig-Choisat Beatrice , Frey Joachim 
Published: June 2006

ABSTRACT

In this study, 231 strains of
Yersinia enterocolitica
, 25 strains of
Y. intermedia
, and 10 strains of
Y. bercovieri
from human and porcine sources (including reference strains) were analyzed using amplified fragment length polymorphism (AFLP), a whole-genome fingerprinting method for subtyping bacterial isolates. AFLP typing distinguished the different
Yersinia
species examined. Representatives of
Y. enterocolitica
biotypes 1A, 1B, 2, 3, and 4 belonged to biotype-related AFLP clusters and were clearly distinguished from each other.
Y. enterocolitica
biotypes 2, 3, and 4 appeared to be more closely related to each other (83% similarity) than to biotypes 1A (11%) and 1B (47%). Biotype 1A strains exhibited the greatest genetic heterogeneity of the biotypes studied. The biotype 1A genotypes were distributed among four major clusters, each containing strains from both human and porcine sources, confirming the zoonotic potential of this organism. The AFLP technique is a valuable genotypic method for identification and typing of
Y. enterocolitica
and other
Yersinia
spp.
Differential
inlA
and
inlB
Expression and Interaction with Human Intestinal and Liver Cells by
Listeria monocytogenes
Strains of Different Origins
Authors: Werbrouck Hadewig , Grijspeerdt Koen , Botteldoorn Nadine , Van Pamel Els , Rijpens Nancy , Van Damme Jo , Uyttendaele Mieke , Herman Lieve , Van Coillie Els 
Published: June 2006

ABSTRACT

In this study, a number of
Listeria monocytogenes
strains of different origins were evaluated for in vitro invasion capacity for various human cell types (monocytic THP-1, enterocytic Caco-2, and hepatocytic HepG2 cells) and for expression levels of specific virulence genes. For THP-1 cells, no differences between clinical and nonclinical
L. monocytogenes
strains in invasion capacity or in production of the proinflammatory cytokine interleukin-8 (IL-8) were observed, whereas for the Caco-2 and HepG2 cells, significant differences in invasion capacity were noticed. On average, the clinical strains showed a significantly lower invasion capacity than the nonclinical
L. monocytogenes
strains. Furthermore, it was shown that the clinical strains induce lower IL-8 levels in HepG2 cells than do the nonclinical strains. This observation led us to study the mRNA expression levels of
inlA
,
inlB
, and
ami
, important virulence genes mediating adhesion and invasion of eukaryotic cells, by real-time reverse transcription-PCR for 27 clinical and 37 nonclinical
L. monocytogenes
strains. Significant differences in
inlA
and
inlB
expression were observed, with clinical strains showing a lower expression level than nonclinical strains. These observations were in accordance with in vitro invasion of Caco-2 and HepG2 cells, respectively. The results of this study indicate that differential expression levels of
inlA
and
inlB
possibly play a role in the virulence capacities of
L. monocytogenes
strains. The lower capacity of clinical strains to invade HepG2 cells and to induce IL-8 is possibly a mechanism of immune evasion used by specific
L. monocytogenes
strains.