bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of volume‐sensitive Cl− channels and ClC‐3 in acinar cells isolated from the rat lacrimal gland and submandibular salivary gland
Authors: Majid Aneela , Brown Peter D. , Best Len , Park Kyungpyo 
Published: August 2004

The expression of ClC‐3 was examined in rat lacrimal gland and submandibular salivary gland cells using RT‐PCR and Western analysis. Whole‐cell patch clamp methods were used to investigate the expression of volume‐sensitive anion channels in acinar cells isolated from these tissues.
Expression of mRNA encoding ClC‐3, and ClC‐3 protein, was found in rat submandibular gland by RT‐PCR and Western analysis. Rat lacrimal gland cells, however, did not appear to express mRNA encoding for ClC‐3, nor the ClC‐3 protein.
Volume‐sensitive anion conductances were observed in both rat lacrimal gland and submandibular salivary gland acinar cells. The conductance was of a similar size in the two cell types, but was much slower to activate in the lacrimal cells.
The properties of the conductances in lacrimal and submandibular cells were similar, e.g. halide selectivity sequence (PI > PCl > Paspartate) and inhibition by 4,4′‐diisothiocyanostilbene‐2,2′‐disulphonic acid, 5‐nitro‐2‐(3‐phenylpropylamino)‐benzoate and tamoxifen.
The data suggest that the expression of ClC‐3 is not an absolute requirement for the activity of volume‐sensitive anion channels in rat lacrimal gland acinar cells.
Synchronous oscillations of length and stiffness during loaded shortening of frog muscle fibres
Authors: Edman K. A. P. , Curtin N. A. 
Published: August 2004

A study was made of the damped oscillations in fibre length that are observed when isolated muscle fibres from the frog are released during the plateau of an isometric tetanus to shorten against a constant load (force clamp recording) near the isometric level (temperature, 1.0‐11.0 °C; initial sarcomere length, 2.25 μm).
The oscillatory length changes of the whole fibre were associated with similar length changes of marked consecutive segments along the fibre. The segmental length changes were initially in synchrony with the whole‐fibre movements but became gradually more disordered. At the same time the length oscillation of the whole fibre was progressively damped.
The fast length step that normally occurs at the outset of the load‐clamp manoeuvre was essential for initiating the oscillatory behaviour. Accordingly, no length oscillation occurred when the load clamp was arranged to start as soon as the selected tension level was reached during the rising phase of the tetanus.
The instantaneous stiffness was measured as the change in force that occurred in response to a high‐frequency (2‐4 kHz) length oscillation of the fibre. During the load‐clamp manoeuvre, when the tension was kept constant, the stiffness underwent periodic changes that correlated well in time with the damped oscillatory changes in fibre length. However, there was a phase shift between the stiffness oscillation and the oscillation of shortening velocity, the latter being in the lead of the stiffness response by 21.4 ± 0.8 ms (n = 19) at 1.8 ± 0.1 °C .
A mechanism is proposed to explain the oscillatory behaviour of the muscle fibre based on the idea that the quick length step at the outset of the load clamp leads to synchronous activity of the myosin cross‐bridges along the length of the fibre.
High Genetic Diversity of the Attachment (G) Protein of Human Metapneumovirus
Authors: Ishiguro Nobuhisa , Ebihara Takashi , Endo Rika , Ma Xiaoming , Kikuta Hideaki , Ishiko Hiroaki , Kobayashi Kunihiko 
Published: August 2004

ABSTRACT
Complete genes encoding the predicted nucleoprotein (N), phosphoprotein (P), matrix protein (M), fusion protein (F), M2-1protein, M2-2protein, small hydrophobic protein (SH), and attachmentprotein (G) of seven newly isolated human metapneumoviruses (hMPVs) were analyzed and compared with previously published data for hMPV genes. Phylogenetic analysis of the nucleotide sequences indicated that there were two genetic groups, tentatively named groups 1 and 2, similar to the grouping of human respiratory syncytial virus. Although the predicted amino acid sequences of N, P, M, F, and M2 were highly conserved between the two groups (amino acid identities, 96% for N, 85% for P, 97% for M, 94% for F, 95% for M2-1, and 90% for M2-2), the amino acid identities of the SH and G proteins were low (SH, 58%; G, 33%). Furthermore, each group could be subdivided into two subgroups by phylogenetic analysis, tentatively named subgroups 1A and 1B and subgroups 2A and 2B. The predicted amino acid sequences of G within members of each subgroup were highly conserved (amino acid identities, 88% for group 1A, 93% for group 1B, and 96% for group 2B). The G of hMPV is thought to be the major antigenic determinant and to play an important role in the production of neutralizing antibodies. Clarification of the antigenic diversity of G is important for epidemiological analysis and for establishment of strategies to prevent hMPV infection.
Evaluation of DNA Extraction and PCR Methods for Detection of
Enterocytozoon bienuesi
in Stool Specimens
Authors: Subrungruang Ittisak , Mungthin Mathirut , Chavalitshewinkoon-Petmitr Porntip , Rangsin Ram , Naaglor Tawee , Leelayoova Saovanee 
Published: August 2004

ABSTRACT

An evaluation of the sensitivities of three DNA extraction methods, i.e., FTA filter paper, a QIAamp stool mini kit, and a conventional phenol-chloroform method, by using specimens with known concentrations of
Enterocytozoon bieneusi
spores was performed. FTA filter paper and the QIAamp stool mini kit were the most sensitive methods, which could detect
E. bieneusi
in specimens with a concentration of 800 spores/ml. We also compared five previously described PCR methods that use five different primer pairs for the detection of
E. bieneusi
and showed that MSP3-MSP4B and EBIEF1-EBIER1 were the most sensitive primers. Although both sets of primers showed the same sensitivity, using the MSP3-MSP4B primers can directly provide genotypic information by sequencing. A blinded diagnostic test to compare PCR and light microscopy methods for the detection of
E. bieneusi
in stool specimens was also conducted. The use of FTA filter paper for DNA extraction together with the PCR method using the primer pair MSP3-MSP4B showed 100% sensitivity and 100% specificity for the detection of
E. bieneusi
in stool specimens, while the light microscopy method gave a sensitivity of 86.7% and a specificity of 100%.
Species of the Family
Helicobacteraceae
Detected in an Australian Sea Lion (
Neophoca cinerea
) with Chronic Gastritis
Authors: Oxley Andrew P. A. , Powell Mark , McKay David B. 
Published: August 2004

ABSTRACT

We describe the first case of gastritis in a male Australian sea lion (
Neophoca cinerea
) in which members of the family
Helicobacteraceae
, particularly the genus
Wolinella
, were detected. The sea lion exhibited clinical signs of gastrointestinal disease, including abdominal pain, lack of appetite, and lethargy. Examination of one ileal and five gastric biopsy specimens collected over a 10-year period revealed persistent fibrosis and/or superficial focal erosion and ulceration of the lamina propria. Spiral-shaped organisms 5 to 12 μm long were observed in two of the gut biopsy specimens. While
Helicobacter
species were detected by PCR in one of the gastric biopsy specimens,
Wolinella
species were detected in four of the five gastric specimens, including those in which spiral-shaped organisms were observed. Comparisons of biopsy specimen ribosomal DNA sequences with those obtained from the feces of this animal, the gastric tissue of a clinically healthy individual, and the feces of several other cohoused sea lions and fur seals revealed a separate and possibly novel gastric
Helicobacter
species. A possibly novel
Wolinella
species, along with
Wolinella succinogenes
, was also identified. These findings highlight the pathogenic potential of other members of this family in the etiopathogenesis of gastric disease in these animals.
Alanine-Threonine Polymorphism of
Helicobacter pylori
RpoB Is Correlated with Differential Induction of Interleukin-8 in MKN45 Cells
Authors: Lee Keun-Hwa , Cho Myung-Je , Yamaoka Yoshio , Graham David Y. , Yun Yeo-Jun , Woo So-Yon , Lim Chang-Young , Ko Kwan-Soo , Kim Bum-Joon , Jung Hyun-Chae , Lee Woo-Kon , Rhee Kwang-Ho , Kook Yoon-Hoh 
Published: August 2004

ABSTRACT

Geographical differences in the genetic diversity of
Helicobacter pylori
isolates were examined by analyzing
rpoB
sequences. An extremely high level of allelic diversity among
H. pylori
strains was found. The
rpoB
sequences of Asian and non-Asian (North and South American, European, and South African) strains were found to differ. An amino acid polymorphism (alanine and threonine RpoB types) was found at the 497th residue by deduced amino acid analysis. RpoB with a threonine residue (RpoB
Thr
) was uniquely present in East Asian countries, and two-thirds of the
H. pylori
isolate population in this region was RpoB
Thr
; however, this type was rare or absent in Western countries, where RpoB
Ala
predominated. RpoB
Thr
strains induced a much larger amount of interleukin-8, a chemokine that plays an important role in chronic inflammation, than RpoB
Ala
strains in cultured MKN45 cells.
Genetic Variability of the G Glycoprotein Gene of Human Metapneumovirus
Authors: Bastien Nathalie , Liu Li , Ward Diane , Taylor Tracy , Li Yan 
Published: August 2004

ABSTRACT
Human metapneumovirus (hMPV) has been associated with respiratory illnesses like those caused by human respiratory syncytial virus (HRSV) infection. Similar to other pneumoviruses, genetic diversity has been reported for hMPV. Little information is currently available on the genetic variability of the G glycoprotein (G), which is the most variable gene in RSV and avian pneumovirus. The complete nucleotide sequences of the G open reading frame (ORF) of 24 Canadian hMPV isolates were determined. Phylogenetic analysis showed the existence of two major groups or clusters (1 and 2). All but one of the hMPV isolates that we examined belonged to cluster 1. Additional genetic variability was observed in cluster 1, which separated into two genetic subclusters. Within cluster 1 the nucleotide sequence identity for the G ORF was 74.2 to 100%, and the identity for the predicted amino acid sequence was 61.4 to 100%. The G genes of cluster 1 isolates were more divergent from the cluster 2 isolates, with 45.6 to 50.5% and 34.2 to 37.2% identity levels for the nucleotide and amino acid sequences, respectively. Sequence analysis also revealed changes in stop codon usage, resulting in G proteins of different lengths (217, 219, 228, and 236 residues). Western blot analysis with the use of hMPV-specific polyclonal antisera to each hMPV cluster showed significant antigenic divergence between the G proteins of clusters 1 and 2. These results suggest that the G protein of hMPV is continuously evolving and that the genetic diversity observed for the hMPV genes is reflected in the antigenic variability, similar to HRSV.
Use of
tuf
Sequences for Genus-Specific PCR Detection and Phylogenetic Analysis of 28 Streptococcal Species
Authors: Picard François J. , Ke Danbing , Boudreau Dominique K. , Boissinot Maurice , Huletsky Ann , Richard Dave , Ouellette Marc , Roy Paul H. , Bergeron Michel G. 
Published: August 2004

ABSTRACT

A 761-bp portion of the
tuf
gene (encoding the elongation factor Tu) from 28 clinically relevant streptococcal species was obtained by sequencing amplicons generated using broad-range PCR primers. These
tuf
sequences were used to select
Streptococcus
-specific PCR primers and to perform phylogenetic analysis. The specificity of the PCR assay was verified using 102 different bacterial species, including the 28 streptococcal species. Genomic DNA purified from all streptococcal species was efficiently detected, whereas there was no amplification with DNA from 72 of the 74 nonstreptococcal bacterial species tested. There was cross-amplification with DNAs from
Enterococcus durans
and
Lactococcus lactis
. However, the 15 to 31% nucleotide sequence divergence in the 761-bp
tuf
portion of these two species compared to any streptococcal
tuf
sequence provides ample sequence divergence to allow the development of internal probes specific to streptococci. The
Streptococcus
-specific assay was highly sensitive for all 28 streptococcal species tested (i.e., detection limit of 1 to 10 genome copies per PCR). The
tuf
sequence data was also used to perform extensive phylogenetic analysis, which was generally in agreement with phylogeny determined on the basis of 16S rRNA gene data. However, the
tuf
gene provided a better discrimination at the streptococcal species level that should be particularly useful for the identification of very closely related species. In conclusion,
tuf
appears more suitable than the 16S ribosomal RNA gene for the development of diagnostic assays for the detection and identification of streptococcal species because of its higher level of species-specific genetic divergence.
High-Throughput Detection of Pathogenic Yeasts of the Genus
Trichosporon
Authors: Diaz Mara R. , Fell Jack W. 
Published: August 2004

ABSTRACT

The need for a rapid and accurate method for the detection of fungal pathogens has become imperative as the incidence of fungal infections has increased dramatically. Herein, we tested the Luminex 100, a novel flow cytometer, for the detection of the medically important genus
Trichosporon
. This genus was selected as our proof-of-concept model due to the close phylogenetic relationship between the species. The method, which is based on a nucleotide hybridization assay, consists of a combination of different sets of fluorescent beads covalently bound to species-specific capture probes. Upon hybridization, the beads bearing the target amplicons are classified by their spectral addresses with a 635-nm laser. Quantitation of the hybridized biotinylated amplicon is based on fluorescence detection with a 532-nm laser. We tested in various multiplex formats 48 species-specific and group-specific capture probes designed in the D1/D2 region of ribosomal DNA, internal transcribed spacer regions, and intergenic spacer region. Species-specific biotinylated amplicons were generated with three sets of primers to yield fragments from the three regions. The assay was specific and fast, as it discriminated species differing by 1 nucleotide and required less than 50 min following amplification to process a 96-well plate. The sensitivity of the assay allowed the detection of 10
2
genome molecules in PCRs and 10
7
to 10
8
molecules of biotinylated amplification product. This technology provided a rapid means of detection of
Trichosporon
species with the flexibility to identify species in a multiplex format by combining different sets of beads.
Molecular Epidemiology of
Enterobacteriaceae
Isolates Producing Extended-Spectrum β-Lactamases in a French Hospital
Authors: Lavigne Jean-Philippe , Bouziges Nicole , Chanal Catherine , Mahamat Aba , Michaux-Charachon Sylvie , Sotto Albert 
Published: August 2004

ABSTRACT

In 2002, 80 isolates of
Enterobacteriaceae
producing extended-spectrum β-lactamases (ESBLs) were collected from infected patients in our hospital.
Enterobacter aerogenes
was the most common bacterium isolated from all specimens (36.5%). The ESBLs were predominantly (90%) TEM derivatives (TEM-24, TEM-3). Pulsed-field gel electrophoresis highlighted that
E. aerogenes
,
Klebsiella pneumoniae
, and
Citrobacter koseri
had a clonal propagation.
Cryptococcus
Infection in Tropical Australia
Authors: Jenney Adam , Pandithage Kishan , Fisher Dale A. , Currie Bart J. 
Published: August 2004

ABSTRACT

Eighteen cases of disease caused by the saprophytic fungi
Cryptococcus neoformans
and
Cryptococcus bacillisporus
are described from the Northern Territory of Australia. The majority of infections were with
Cryptococcus bacillisporus
and in the rural Aboriginal population, often causing pulmonary mass lesions.
Evaluation of a New Automated, Standardized Generic Nucleic Acid Extraction Method (Total Nucleic Acid Isolation Kit) Used in Combination with Cytomegalovirus DNA Quantification by COBAS AMPLICOR CMV MONITOR
Authors: Gärtner Barbara Christine , Fischinger Johannes Michael , Litwicki Anke , Roemer Klaus , Mueller-Lantzsch Nikolaus 
Published: August 2004

ABSTRACT
The new generic Total Nucleic Acid Isolation kit improved the detection limit of a cytomegalovirus (CMV) PCR from 400 to 200 copies/ml. Sensitivity and specificity in 30 CMV-positive and 100 negative samples were 100%. Analytical performance was excellent. The kit provides a reliable, standardized, and time-saving tool for DNA extraction.
Infection of a Japanese Patient by Genotype 4 Hepatitis E Virus While Traveling in Vietnam
Authors: Koizumi Yuko , Isoda Norio , Sato Yukihiro , Iwaki Takaaki , Ono Kazunori , Ido Kenichi , Sugano Kentaro , Takahashi Masaharu , Nishizawa Tsutomu , Okamoto Hiroaki 
Published: August 2004

ABSTRACT
Cases of imported hepatitis E in industrialized countries infected with a genotype 1 hepatitis E virus (HEV) have been identified. We report a 56-year-old Japanese man who acquired infection with a genotype 4 HEV with 98.8% identity to a Vietnamese isolate after ingestion of uncooked shellfish while traveling in Vietnam.
Mycobacterium lentiflavum
as an Emerging Causative Agent of Cervical Lymphadenitis
Authors: Piersimoni Claudio , Goteri Gaia , Nista Domenico , Mariottini Alessandro , Mazzarelli Gianna , Bornigia Stefano 
Published: August 2004

ABSTRACT

A lymph node excision was performed on a 45-year-old woman with left cervical swelling. The disorder which developed after the patient had undergone oral surgery for a severe periodontal disease failed to respond to antimicrobial chemotherapy. A mycobacterial strain subsequently identified by high-performance liquid chromatography analysis of cell wall mycolic acids as
Mycobacterium lentiflavum
grew from the excised specimen. This case and previously published reports highlight the relevance of
M. lentiflavum
as an emerging causative agent of mycobacterial cervical lymphadenitis.
Preliminary inventory and classification of indigenous afromontane forests on the Blyde River Canyon Nature Reserve, Mpumalanga, South Africa
Authors: Lötter Mervyn C , Beck Hans T 
Published: August 2004

Abstract
Background
Mixed evergreen forests form the smallest, most widely distributed and fragmented biome in southern Africa. Within South Africa, 44% of this vegetation type has been transformed. Afromontane forest only covers 0.56 % of South Africa, yet it contains 5.35% of South Africa's plant species. Prior to this investigation of the indigenous forests on the Blyde River Canyon Nature Reserve (BRCNR), very little was known about the size, floristic composition and conservation status of the forest biome conserved within the reserve. We report here an inventory of the forest size, fragmentation, species composition and the basic floristic communities along environmental gradients.

Results
A total of 2111 ha of forest occurs on Blyde River Canyon Nature Reserve. The forest is fragmented, with a total of 60 forest patches recorded, varying from 0.21 ha to 567 ha in size. On average, patch size was 23 ha. Two forest communities – high altitude moist afromontane forest and low altitude dry afromontane forest – are identified. Sub-communities are recognized based on canopy development and slope, respectively. An altitudinal gradient accounts for most of the variation within the forest communities.

Conclusion
BRCNR has a fragmented network of small forest patches that together make up 7.3% of the reserve's surface area. These forest patches host a variety of forest-dependent trees, including some species considered rare, insufficiently known, or listed under the Red Data List of South African Plants. The fragmented nature of the relatively small forest patches accentuates the need for careful fire management and stringent alien plant control.
Effect of haemoperfusion on plasma paraquat concentration in vitro and in vivo
Authors: Hong Sae-Yong , Yang Jong-Oh , Lee Eun-Young , Kim Soo-Hyun 
Published: August 2004

This study was to observe the paraquat (PQ) reduction rate after haemoperfusion (HP) on the groups of a relatively large number: 50 survivors out of 105 patients with acute PQ poisoning. We started off by measuring the clearance of haemodialysis (HD) and HP for the PQ in vitro. At the blood flow of 250 mL/min, the PQ clearance was greater in HP than in HD during the first 90 minutes: 215 versus 175 mL/min at 30 minutes, 213 versus 201 mL/min at 60 minutes, and 199 versus 179 mL/min at 90 minutes. The clearance in HP decreased rapidly after two hours. By the end of the dialysis, however, the final concentration in container decreased to 5.7 mg/mL in HD and 1.5 mg/mL in HP, which implied that although HD was cleared more effectively during the later stages, the overall elimination was greater in HP. Following this preliminary investigation, we performed HP on all the patients in order to assess the extracorporeal elimination. One hundred and five patients who had swallowed one to three mouthfuls of PQ (24.5% w/v) solution were subjected to the in vivo study. The reduction rate of PQ was checked out by measuring the PQ concentration in plasma before and after four hours of HP. Seeing the reduction rate was significantly higher in the survivors group than in the nonsurvivors group (80.39 ± 19.9 versus 67.29 ± 19.2%, P <0.01), we concluded that adequate HP appears to be an indispensable treatment for patients with acute PQ poisoning.
Interpretation of infrared spectra of dioctahedral smectites in the region of OH-stretching vibrations
Authors: Zviagina Bella B. , McCarty Douglas K. , Środoń Jan , Drits Victor A. 
Published: August 2004

AbstractDioctahedral smectite samples of a wide range of compositions (beidellites, montmorillonites, nontronites, Fe-rich montmorillonites and Al-rich nontronites) were studied by infrared (IR) spectroscopy. A special sample-preparation technique was used to eliminate the contribution of molecular water. The OH-stretching regions of the spectra were decomposed and curve-fitted, and the individual OH-stretching bands were assigned to all the possible types of OH-bonded cation pairs that involve Al, Mg and Fe. The integrated optical densities of the OH bands were assumed to be proportional to the contents of the specific types of OH-linked cation pairs with the absorption coefficients being the same for all individual OH bands. Good agreement between the samples’ octahedral cation compositions calculated from the IR data and those given by crystal-chemical formulae was obtained for a representative collection of samples in terms of a unique set of individual OH-band positions that vary within narrow wavenumber intervals. This has allowed us to minimize the ambiguity in spectra decomposition caused by the poor resolution of smectite spectra and confirmed the validity of the resulting band identification.The bands associated with specific OH-bonded cation pairs in the spectra of smectites are, on the whole, shifted to greater wavenumbers with respect to the corresponding bands in micas. In addition to OH bands that refer to the smectite structure, AlOHAl and AlOHFe bands of the pyrophyllite structural fragments were identified. The band-position variation ranges overlap in a few cases (AlOHFe and MgOHMg; AlOHAl of smectite and AlOHFe of pyrophyllite-like component).Unambiguous interpretation of the OH-stretching vibrations was found to be possible only for smectite samples with known chemical compositions, so that IR data cannot be used for quantitative determination of octahedral cation composition of mixtures of dioctahedral 2:1 phyllosilicates. In the case of the studied monomineral smectites with known chemical compositions, IR data provided information on the short-range order/disorder in the distribution of octahedral cations along cation-OH-cation directions. This information can be employed, in conjunction with the data of other spectroscopic and diffraction techniques, in the analysis of short-range octahedral cation distribution.