bluuebunny/crossref_metadata_2025_split huggingface dataset:
Long‐term Protection in Hamsters against Human Parainfluenza Virus Type 3 Following Mucosal or Combinations of Mucosal and Systemic Immunizations with Chimeric Alphavirus‐based Replicon Particles
Authors: Greer C. E. , Zhou F. , Goodsell A. , Legg H. S. , Tang Z. , Zur Megede J. , Uematsu Y. , Polo J. M. , Vajdy M. 
Published: October 2007

AbstractNo licensed vaccines are available to protect against parainfluenza virus type 3 (PIV3), a significant health risk for infants. In search of a safe vaccine, we used an alphavirus‐based chimeric vector, consisting of Sindbis virus (SIN) structural proteins and Venezuelan equine encephalitis virus (VEE) replicon RNA, expressing the PIV3 hemagglutinin‐neuraminidase (HN) glycoprotein (VEE/SIN‐HN). We compared different routes of intramuscular (IM), intranasal (IN), or combined IN and IM immunizations with VEE/SIN‐HN in hamsters. Six months after the final immunization, all hamsters were protected against live PIV3 IN challenge in nasal turbinates and lungs. This protection appeared to correlate with antibodies in serum, nasal turbinates and lungs. This is the first report demonstrating mucosal protection against PIV3 for an extended time following immunizations with an RNA replicon delivery system.
Comparison of five xylan synthesis mutants reveals new insight into the mechanisms of xylan synthesis
Authors: Brown David M. , Goubet Florence , Wong Vicky W. , Goodacre Royston , Stephens Elaine , Dupree Paul , Turner Simon R. 
Published: October 2007

SummaryPrevious studies using co‐expression analysis have identified a large number of genes likely to be involved in secondary cell‐wall formation. However, the function of very few of these genes is known. We have studied the cell‐wall phenotype of irx7, irx8 and irx9, three previously described irregular xylem (irx) mutants, and irx14 and parvus‐3, which we now show also to be secondary cell‐wall mutants. All five mutants, which have mutations in genes encoding putative glycosyltransferases, exhibited large decreases in xylan. In addition, all five mutants were found to have the same specific defect in xylan structure, retaining MeGlcUA but lacking GlcUA side branches. Polysaccharide analysis by carbohydrate gel electrophoresis (PACE) was used to determine the xylan structure in Arabidopsis, and revealed that side branches are added to approximately one in every eight xylose residues. Interestingly, this ratio is constant in all the lines analysed despite the wide variation in xylan content and the absence of GlcUA branches. Xylanase digestion of xylan from wild‐type plants released a short oligosaccharide sequence at the reducing end of the xylan chain. MALDI‐TOF MS analysis indicated that this sequence of sugars was absent in xylan from irx7, irx8 and parvus‐3 mutants, but was present in irx9 and irx14. This is consistent with previous NMR analysis of xylan from irx7, irx8 and irx9, and suggests that PARVUS may be involved in the synthesis of a xylan primer whereas IRX14 may be required to synthesize the xylan backbone. This hypothesis is supported by assays showing that irx9 and irx14 are both defective in incorporation of radiolabel from UDP 14C‐xylose. This study has important implications for both our understanding of xylan biosynthesis and the functional analysis of cell‐wall biosynthesis genes.
Effect of an inhaled adenosine A2A agonist on the allergen‐induced late asthmatic response
Authors: Luijk B. , Van Den Berge M. , Kerstjens H. A. M. , Postma D. S. , Cass L. , Sabin A. , Lammers J.‐W. J. 
Published: October 2007

Background:  Adenosine receptor activation is suggested to play a role in asthmatic airway inflammation. Inhibition of adenosine receptors may have an effect on the late asthmatic response (LAR) after allergen inhalation and this mechanism could offer a potential new treatment in asthma.Methods:  We evaluated the effect of an inhaled adenosine‐2A (A2A)‐receptor agonist (GW328267X), 25 μg, in 15 nonsmoking atopic asthmatics who underwent an inhaled allergen challenge following twice daily treatment for 1 week in a double‐blind, placebo‐ and fluticasone propionate (250 μg) controlled study.Results:  In contrast to fluticasone, treatment with the A2A‐receptor agonist neither significantly protect against the allergen‐induced early and late asthmatic reaction, nor the accompanying inflammatory response as measured by sputum total cell counts, number of EG2+ cells, and the concentration of interleukin‐8 and eosinophil cationic protein.Conclusion:  The inhaled A2A‐receptor agonist, GW328267X, 25 μg does not affect the allergen‐induced LAR or the associated inflammatory response in asthma.
Galectin‐10 mRNA is overexpressed in peripheral blood of aspirin‐induced asthma
Authors: Devouassoux G. , Pachot A. , Laforest L. , Diasparra J. , Freymond N. , Van Ganse E. , Mougin B. , Pacheco Y. 
Published: October 2007

Background:  In sensitive patients, aspirin is associated with nasal and bronchial inflammation, eliciting local symptoms. Although the disease is clinically well characterized, its physiopathology is incompletely understood and noninvasive procedures, allowing an effective distinction between aspirin‐induced asthma (AIA) and aspirin‐tolerant asthma (ATA) are missing.Objectives:  The aims of the study were to compare AIA and ATA cohorts for clinical characteristics and to screen peripheral blood for differential mRNA expression.Methods:  Patients experiencing symptoms following aspirin ingestion were considered as aspirin sensitive. Peripheral blood was collected to quantify mRNA expression, using microarray technology and quantitative RT‐PCR.Results:  Data indicated that AIA and ATA share large number of similarities for clinical phenotype. Screening of mRNA expression using microarray showed an overexpression of galectin‐10 mRNA in AIA (AIA/ATA ratio = 1.9, P < 0.05). Results were confirmed using qRT‐PCR. A positive correlation was established between microarray and qRT‐PCR results for galectin‐10 mRNA expression (r = 0.92, P < 0.0001). Finally, qRT‐PCR results were validated on a subset of asthmatics and controls, showing an increased expression of galectin‐10 mRNA in AIA vs ATA (P < 0.001) and vs controls (P < 0.01).Conclusions:  Our results demonstrate that AIA and ATA remain difficult to distinguish using clinical criteria. Employing two molecular biological methods, we demonstrate that galectin‐10 mRNA is overexpressed in AIA, suggesting a novel candidate gene and a potentially innovative pathway for mucosal inflammation in aspirin intolerance.
Rapid detection of the cariogenic pathogens Streptococcus mutans and Streptococcus sobrinus using loop‐mediated isothermal amplification
Authors: Nagashima S. , Yoshida A. , Ansai T. , Watari H. , Notomi T. , Maki K. , Takehara T. 
Published: October 2007

Introduction:  Streptococcus mutans and Streptococcus sobrinus are associated with the development of dental caries in humans. In this study, we developed a rapid, sensitive method for detecting these major cariogenic pathogens using loop‐mediated isothermal amplification (LAMP). The assay procedure is quite simple: the amplification is carried out in a single tube under isothermal conditions at 63°C, and the result can be obtained in less than 1 h.Methods:  Initially, a set of six primers was designed by targeting S. mutans‐specific and S. sobrinus‐specific regions, identified using the genomic subtractive hybridization technique. We evaluated the specificities and sensitivities of these assays. Furthermore, we detected and quantified these bacteria in saliva and carious dentin from eight children.Results:  The sensitivities of the S. mutans‐specific and S. sobrinus‐specific LAMP methods, examined using agarose gel electrophoresis, were each one cell for a 30‐min reaction. The detection limits using real‐time turbidimetry analysis were 1 to 107 cells (3.28 × 101 to 3.28 × 108 fg S. mutans template DNA) per reaction tube and 1 to 105 cells (2.72 × 103 to 2.72 × 108 fg S. sobrinus template DNA) per reaction tube. Using these assays, we detected and quantified these cariogenic bacteria for evaluation of the LAMP assay for clinical diagnosis.Conclusions:  Our results suggest that the LAMP‐based assay in combination with subtractive hybridization is valuable for preparing species‐specific primers for closely related species. Furthermore, the LAMP‐based assay will be a useful tool for the rapid and sensitive prediction of dental caries.
Nitrite reductase in Streptoccocus mutans plays a critical role in the survival of this pathogen in oral cavity
Authors: Choudhury T. , Sato E. F. , Inoue M. 
Published: October 2007

Background/aims:  The mechanisms of nitric oxide (NO) production by bacteria in the oral cavity are still not clearly defined but salivary streptococci have been reported to generate NO. The aim of this study was to clarify the mechanism of nitrite metabolism and generation of NO by Streptococcus mutans, a major pathogen of dental caries.Methods:  We searched the genomic database of oral pathogens for nitrite reductase and used a polymerase chain reaction (PCR) to clone the nirJ gene from S. mutans GS5. His‐tagged recombinant NirJ protein was expressed in Escherichia coli BL21 and characterized. We constructed a nirJ gene‐disrupted mutant strain of S. mutans (ΔnirJ) to analyze the physiological significance of nirJ.Results:  S. mutans generates NO from nitrite, probably as a result of the possession of nitrite reductase. We cloned the nirJ gene from S. mutans GS5 by PCR. The recombinant NirJ protein catalyzed the reduction of nitrite with a Km value of 3.37 μm and a specific activity of 2.5 μmol/min/mg of protein at 37°C. Biochemical analysis revealed that the nitrite‐reducing activity of the mutant (ΔnirJ) strain was significantly lower than that of the wild‐type strain. The growth of the mutant strain, but not of the wild‐type strain, was strongly suppressed by the presence of physiological levels of nitrite (∼0.2 mm) in saliva.Conclusion:  These observations suggest that the elimination of nitrite and/or the generation of NO are important for the survival of S. mutans in the oral cavity.
Quantitative messenger RNA expression of Toll‐like receptors and interferon‐α1 in gingivitis and periodontitis
Authors: Kajita K. , Honda T. , Amanuma R. , Domon H. , Okui T. , Ito H. , Yoshie H. , Tabeta K. , Nakajima T. , Yamazaki K. 
Published: October 2007

Introduction:  In addition to bacteria, viruses have been reportedly implicated in periodontitis. However, the available data are confined to Toll‐like receptor 2 (TLR2) and TLR4, which recognize bacterial products in periodontitis. In the present study, we investigated the expression levels of TLR5, ‐7, and ‐9 messenger RNAs (mRNAs) in addition to those of TLR2 and ‐4, and compared gingivitis and periodontitis. Interferon‐α1 (IFN‐α1), which is important for the antiviral response, was also compared.Methods:  Gene expression was analyzed using quantitative real‐time polymerase chain reaction for 59 periodontitis and 27 gingivitis tissue samples together with viral serology in some patients. The presence of plasmacytoid dendritic cells (pDCs), a robust producer of IFN‐α, was immunohistochemically analyzed in an additional seven periodontitis and two gingivitis specimens.Results:  The expression levels of TLR2, ‐4, ‐7, and ‐9 were significantly higher in periodontitis lesions than gingivitis lesions. The expression level of TLR5 was comparable to levels of TLR2 and ‐4; however, no significant difference was found between gingivitis and periodontitis. Although the expression of IFN‐α1 mRNA was higher in periodontitis lesions compared with gingivitis lesions, the level was quite low. Only a few pDCs were found in some periodontitis specimens. No difference was found for antibody‐positivity between gingivitis and periodontitis.Conclusion:  This is the first study to show that a variety of TLRs are up‐regulated in periodontitis lesions compared with gingivitis lesions, suggesting that diverse microbial and possibly viral antigens are involved in the pathogenic mechanisms for periodontal diseases. However, the ligands recognized by the various TLRs in periodontal lesions remain to be determined.
Bacterial examination of endodontic infections by clonal analysis in concert with denaturing high‐performance liquid chromatography
Authors: Jacinto R. C. , Gomes B. P. F. A. , Desai M. , Rajendram D. , Shah H. N. 
Published: October 2007

Background/aims:  The aim of this study was to examine the diversity of bacterial species in the infected root canals of teeth associated with endodontic abscesses by cloning and sequencing techniques in concert with denaturing high‐performance liquid chromatography.Methods:  Samples collected from five infected root canals were subjected to polymerase chain reaction (PCR) with universal 16S ribosomal DNA primers. Products of these PCRs were cloned and sequenced. Denaturing high‐performance liquid chromatography (DHPLC) was used as a screening method to reduce the number of clones necessary for DNA sequencing.Results:  All samples were positive for the presence of bacteria and a range of 7–13 different bacteria were found per root canal sample. In total, 48 different oral clones were detected among the five root canal samples. Olsenella profusa was the only species present in all samples. Porphyromonas gingivalis, Dialister pneumosintes, Dialister invisus, Lachnospiraceae oral clone, Staphylococcus aureus, Pseudoramibacter alactolyticus, Peptostreptococcus micros and Enterococcus faecalis were found in two of the five samples. The majority of the taxa were present in only one sample, for example Tannerella forsythia, Shuttleworthia satelles and Filifactor alocis. Some facultative anaerobes that are frequently isolated from endodontic infections such as E. faecalis, Streptococcus anginosus and Lactobacillus spp. were also found in this study.Conclusion:  Clonal analysis of the microflora associated with endodontic infections revealed a wide diversity of oral species.
N. Femoralis, N. Obturatorius and N. Ischiadicus: Deviation in Creation in the Dogs
Authors: Mihelić D. , Mihelić D. , Slavica A. , Deždek D. , Trbojević‐Vukičević T. , Džaja P. , Majić‐Balić I. 
Published: October 2007

SummaryTwo hundred and twelve dog cadavers belonging to different breeds were examined, to investigate the formation of the femoral, obturator and the sciatic nerve. Besides the commonly described formation patterns of the mentioned nerves, some variations were also found. These variations were not gender‐related, but on the other hand we discovered a certain correlation between the variations appearing in the formation of the femoral, obturator and the sciatic nerve. In 74.05% of cases, the femoral nerve was formed from ventral branches of the 4th, 5th and 6th lumbar nerve, and 16.98% of the dogs had the nerve formed from ventral branches of the 3rd, 4th and 5th lumbar nerve. Many dogs (i.e. 2.30%) had the femoral nerve formed from the ventral branches of the 5th, 6th and 7th, the 3rd, 4th, 5th and 6th or the 4th, 5th, 6th and 7th lumbar nerve, respectively. In 1.88% of dogs in particular, the femoral nerve was formed from ventral rami of the 4th and 5th lumbar nerve. In 66.98% of the examined dogs, the obturator nerve was formed from the ventral branches of the 4th, 5th and 6th lumbar nerve, followed by 16.59% of the dogs with the obturator nerve formed from the ventral rami of the 4th and 5th lumbar nerve and 9.43% of dogs in which the nerve was formed from the ventral branches of the 5th and 6th lumbar nerve. In 4.71% of dogs, the obturator nerve was formed from the ventral rami of the 4th, 5th, 6th and 7th lumbar nerve, while only 2.30% of the examined dogs had the same nerve formed from the ventral branches of the 5th, 6th and 7th lumbar nerve. The sciatic nerve was formed from ventral branches of the 6th and 7th lumbar nerve and the 1st sacral nerve in 86.79% of the dogs. In 5.18% of cases, the same nerve was formed from a junction of the ventral branches of the 7th lumbar and the 1st and 2nd sacral nerve, and, in the same percentage of cases, it was formed from a junction of ventral branches of the 6th and 7th lumbar nerve and the 1st and 2nd sacral nerve. In 2.83% of the dogs, it was formed from a junction of the ventral branches of the 6th and 7th lumbar and the 1st sacral nerve. The correlation of variations established in the formation of the femoral, obturator and the sciatic nerve was not statistically significant.
Arterial Supply of the Penis in Agoutis (Dasyprocta prymnolpha, Wagler, 1831)
Authors: De Carvalho M. A. M. , Machado Junior A. A. N. , Bezerra e Silva R. A. , Menezes D. J. A. , Conde Júnior A. M. , Righi D. A. 
Published: October 2007

SummaryThe arterial vascularization of agoutis’ penis (Dasyprocta prymnolopha) were analysed using ten male adults from ‘Núcleo de Estudos e Preservação de Animais Silvestres da Universidade Federal do Piauí’ (FUFPI/IBAMA n° 02/99). Among the total number of specimens, six animals had natural death and were members of the research collection of the Laboratory of Anatomy, and four were killed after anaesthesia. Stained bi‐centrifugated‐Cis‐I‐4 latex was injected in arterial vessels responsible for penis vascularization throughout the abdominal portion of aorta. The samples were fixed in 10% formaldehyde solution and arteries were dissected. The penile artery is originated as a branch of internal pudendal artery. At the level of ischiatic arch, the penile artery project two branches, the penile dorsal and the deep arteries; those arteries irrigates the penile dorsal surface and the corpus cavernosum penis. The penile dorsal arteries have an independent course up to the glans penis. Based on the conditions of this work a remarkable similarity regarding the distribution of vessels destined to the agouti penis when compared to other domestic, wild and lagomorph rodents as rabbits.
Oocyte Morphometry in Female Dogs (Canis familiaris, Linnaeus, 1758)
Authors: Diagone K. V. , Vicente W. R. R. , Pacheco M. R. , Mateus O. 
Published: October 2007

SummaryIn the present study, the ovaries of 15 healthy bitches divided into three groups according to age were analysed by histology and morphometry: group 1 (1–3 years), group 2 (>3–5 years) and group 3 (>5–7 years). After ovariosalpingohysterectomy, the ovaries were fixed, routinely processed for embedding in paraffin and stained with haematoxylin–eosin. The following morphometric parameters were analysed: maximum and minimum diameter (μm), perimeter (μm), area (μm2) and roundness of the cytoplasm and nucleus of oocytes from different types of follicles. Significant differences in the cytoplasmic and nuclear parameters of follicular oocytes between the experimental groups were determined by anova and the Tukey test (5%). For the biotechnology of reproduction, the present results showed that in vitro maturation yielded the best performance for oocytes from primordial follicles in group 3 females and for oocytes embedded in secondary and tertiary follicles in group 2 females. In addition, the present findings will help in the diagnosis and prognosis of diseases affecting bitches, and will especially contribute to a better understanding of these cells by researchers in the field of histology and canine reproduction.
Effects of Castration on the Expression of TrkB and p75NTR in the Vas Deferens and Male Accessory Genital Glands of the Rat
Authors: Mirabella N. , Squillacioti C. , De Luca A. , Paino G. 
Published: October 2007

SummaryBrain‐derived neurotrophic factor (BDNF) is a growth factor belonging to the family of neurotrophins. Neurotrophins exert their effects by binding to corresponding receptors, which are formed by the tyrosine receptor kinases TrkA, TrkB and TrkC, and the low‐affinity p75NTR receptor. The role of neurotrophins in the biology of male genital organs is far from clear. In particular, little is known about the influence of sex hormones on the expression of neurotrophin receptors. In the present study, using immunohistochemistry, we investigated the distribution of TrkB and p75NTR in the vas deferens and accessory male genital glands in normal and castrated rats. In normal rats, both TrkB and p75NTR immunoreactivities were localized in the epithelial layer of the vas deferens and prostate gland. The nerves were immunoreactive in all the examined organs. Castration induced the loss of both TrkB and p75NTR immunoreactivities in the epithelial layer. On the contrary, both receptors persisted or, as in the case of p75NTR, increased in the nerves after castration. These results show that receptors that bind BDNF are expressed in the internal male genital organs of the rat and that their expression is differently regulated by androgen hormones. It can be hypothesized that BDNF, via interacting with p75NTR, has a role in the castration‐induced regression of the sympathetic nerves.
Effects of Betamethasone Treatment on Central Myelination in Fetal Sheep: An Electron Microscopical Study
Authors: Raschke C. , Schmidt S. , Schwab M. , Jirikowski G. 
Published: October 2007

SummaryThe long‐term effect of betamethasone on the myelination of commissural and associational fibres was investigated in fetal sheep. We studied the corpus callosum and subcortical white matter by electron microscopy. Axons were subdivided into classes according to their axonal diameter: class I: ≤0.65 μm; class II: 0.66–0.84 μm; class III: ≥0.85 μm. Under control conditions, the different functions of the white matter tracts examined were reflected by three morphological criteria: (1) there was a diverse percentage of axonal classes in the investigated areas. In corpus callosum the axons of class II predominate (47.1%). In the subcortical white matter, class I axons with small diameter are in majority (40.8%). (2) In the subcortical white matter more axons are present, with especially large diameter and hence of axonal class III. (3) The axons of subcortical white matter have thicker myelin sheaths than those of the corpus callosum. Betamethasone administration caused a significant decrease of class II axons in the corpus callosum (36.9%). In corpus callosum, axons of all classes present thicker myelin sheaths. Betamethasone administration resulted in a change in the formation of the myelin sheath in the commissural fibres of the corpus callosum but not in the associational fibres of the subcortical white matter. This could be the morphological correlate to behavioral and cognitive changes known to occur in humans after prenatal glucocorticoid treatment.
Changes in the Myosin Heavy Chain 2a and 2b Isoforms of the Anterior Belly of the Digastric Muscle Before and After Weaning in Mice
Authors: Yoshii M. , Sakiyama K. , Abe S. , Agematsu H. , Mitarashi S. , Tamatsu Y. , Ide Y. 
Published: October 2007

SummaryDuring the process of growth and development, the digastric muscle is subjected to marked functional changes, including the change from suckling to mastication. In particular, because the anterior belly of the digastric muscle, which is one of the suprahyoid muscles, plays an important role in mastication. Therefore, this muscle seems to undergo a marked functional change before and after weaning. However, the details remain unknown. Here, to clarify the changes in the muscle fibre characteristics of the anterior belly of the digastric muscle before and after weaning, we examined myosin heavy chain isoforms at the protein (immunohistochemistry) and mRNA (transcription) levels. As a control, the changes in the muscle fibre characteristics of the sternohyoid muscle, which is anatomically aligned in the same direction as the anterior belly of the digastric muscle, were analyzed. The results showed that, in the anterior belly of the digastric muscle that is involved in mandibular movements in mice, the ratio of a fast‐contraction isoform with strong contractile force increased after weaning. We believe that this occurred in response to a functional change from suckling to mastication. On the other hand, there was little change in the composition of sternohyoid muscle.