ABSTRACT
AIM: To evaluate the effectiveness of motion-sensitised driven-equilibrium (MSDE)-prepared balanced magnetic resonance cholangiopancreatography (MRCP) in a gadolinium ethoxybenzyl diethylene triamine pentaacetic acid (Gd-EOB-DTPA)-enhanced study compared to conventional T2-weighted MRCP. MATERIALS AND METHODS: Fifteen patients (seven male and eight female patients) prospectively underwent conventional three-dimensional turbo spin-echo T2-weighted MRCP and MSDE-balanced MRCP using a 1.5 T MRI system after hepatobiliary phase image acquisition. For quantitative evaluation, the contrast-to-noise ratio (CNR) of the common hepatic duct to liver tissue was calculated. For qualitative analysis, two radiologists evaluated the depiction of the biliary system and main pancreatic duct (MPD) using a scoring system. Signal suppression of the portal vein (PV) and hepatic vein (HV) on MSDE-balanced MRCP was also scored. RESULTS: MSDE-balanced MRCP showed significantly higher CNR than T2-weighted MRCP. For all biliary structures, the mean depiction scores of MSDE-balanced MRCP were significantly higher than those of T2-weighted MRCP, whereas the mean depiction score of MPD with MSDE-balanced MRCP was significantly lower than that of T2-weighted MRCP. Signal suppression of the PV and HV was thought to be clinically sufficient. CONCLUSIONS: MSDE-balanced MRCP more clearly depicted biliary structures compared with T2-weighted MRCP in a Gd-EOB-DTPA-enhanced study. This sequence may be utilised for routine MRCP on Gd-EOB-DTPA-enhanced MRI.
Subject(s)
Biliary Tract/diagnostic imaging , Cholangiopancreatography, Magnetic Resonance/methods , Contrast Media , Gadolinium DTPA , Image Enhancement/methods , Adult , Aged , Aged, 80 and over , Feasibility Studies , Female , Humans , Image Interpretation, Computer-Assisted , Imaging, Three-Dimensional , Male , Middle Aged , Motion , Prospective Studies , Reproducibility of ResultsABSTRACT
AIM: To clarify whether the heterogeneity of non-cancerous liver parenchyma (NLP) in the hepatobiliary phase on gadoxetic acid enhanced magnetic resonance imaging (MRI) is correlated with hepatocellular carcinoma (HCC) development. MATERIALS AND METHODS: Institutional review board approval was obtained, and the requirements for informed consent were waived for this retrospective study. The imaging characteristics of 84 patients with chronic liver disease who underwent gadoxetic acid-enhanced 3T MRI between January 2013 and October 2014 were examined retrospectively. For the evaluation of the heterogeneity of the intensity in the hepatobiliary phase, the largest possible region of interest was placed on the NLP, and the skewness and kurtosis were calculated using ImageJ software. Skewness is the degree of asymmetry of a histogram, and kurtosis is a measure of the peak. Based on the median values of kurtosis and skewness, the patients were classified into four categories and the categories were compared between the 49 patients with HCC (HCC group) and the 35 patients without HCC (non-HCC group). RESULTS: Kurtosis was significantly higher in the HCC group compared to the non-HCC group (1.19±1.15 versus 0.43±0.83; p=0.0006). Skewness was significantly lower in the HCC group than in the non-HCC group (1.19±1.15 versus 0.43±0.83; p=0.0152). In a multivariate logistic analysis, the category showing lower-than-the-median (-0.1185) skewness and higher-than-the-median (0.547) kurtosis was significantly and independently associated with HCC development (p=0.0031). CONCLUSION: The heterogeneity of NLP in the hepatobiliary phase on gadoxetic acid enhanced MRI may reflect the development of HCC.
Subject(s)
Biomarkers , Carcinoma, Hepatocellular/diagnosis , Liver Neoplasms/diagnosis , Liver/pathology , Magnetic Resonance Imaging/methods , Adult , Aged , Aged, 80 and over , Chronic Disease , Female , Gadolinium DTPA , Humans , Logistic Models , Male , Middle Aged , Retrospective Studies , Young AdultABSTRACT
The pathogenicity of 331 blast isolates (Pyricularia oryzae Cavara) collected from different regions and ecosystems for rice cultivation in Bangladesh was evaluated by compatibility on 23 differential varieties (DV), each harboring a single blast resistance gene, and susceptible 'Lijiangxintuanheigu' (LTH). A wide variation in virulence was found among the isolates, and 267 races were classified using a new designation system. Virulence of blast isolates against DV carrying the resistance genes Pia, Pib, Pit, Pik-s, Piz-t, Pi12(t), Pi19(t), and Pi20(t), as well as avirulence against those carrying Pish, Pi9, Pita-2, and Pita, was distributed widely in Bangladesh. Cluster analysis of the compatibility data on the DV initially classified the isolates into groups I and II. The virulence spectra of the two groups differed mainly according to the reactions of the DV to Pii, Pi3, Pi5(t), Pik-m, Pi1, Pik-h, Pik, Pik-p, and Pi7(t). Group I isolates were distributed mainly in rainfed lowlands, whereas group II isolates were found mainly in irrigated lowlands; however, there were no critical differences in geographic distribution of the blast isolates. In total, 26 isolates, which could be used to identify the 23 resistance genes of the DV on the basis of their reaction patterns, were selected as a set of standard differential blast isolates. To our knowledge, this is the first clear demonstration of the diversity and differentiation of blast races in Bangladesh. This information will be used to develop a durable blast protection system in that country.
ABSTRACT
BACKGROUND AND PURPOSE: An accurate assessment of the hemodynamics of an intracranial dural AVF is necessary for treatment planning. We aimed to investigate the utility of 4D-MRA based on superselective pseudocontinuous arterial spin-labeling with CENTRA-keyhole and view-sharing (4D-S-PACK) for the vessel-selective visualization of intracranial dural AVFs. MATERIALS AND METHODS: We retrospectively analyzed the images of 21 patients (12 men and 9 women; mean age, 62.2 [SD,19.2] years) with intracranial dural AVFs, each of whom was imaged with DSA, 4D-S-PACK, and nonselective 4D-MRA based on pseudocontinuous arterial spin-labeling combined with CENTRA-keyhole and view-sharing (4D-PACK). The shunt location, venous drainage patterns, feeding artery identification, and Borden classification were evaluated by 2 observers using both MRA methods on separate occasions. Vessel selectivity was evaluated on 4D-S-PACK. RESULTS: Shunt locations were correctly evaluated in all 21 patients by both observers on both MRA methods. With 4D-S-PACK, observers 1 and 2 detected 76 (80.0%, P < .001) and 73 (76.8%, P < .001) feeding arteries of the 95 feeding arteries identified on DSA but only 39 (41.1%) and 46 (48.4%) feeding arteries with nonselective 4D-PACK, respectively. Both observers correctly identified 10 of the 11 patients with cortical venous reflux confirmed by DSA with both 4D-S-PACK and 4D-PACK (sensitivity = 90.9%, specificity = 90.9% for each method), and they made accurate Borden classifications in 20 of the 21 patients (95.2%) on both MRA methods. Of the 84 vessel territories examined, vessel selectivity was graded 3 or 4 in 73 (91.2%) and 66 (88.0%) territories by observers 1 and 2, respectively. CONCLUSIONS: 4D-S-PACK is useful for the identification of feeding arteries and accurate classifications of intracranial dural AVFs and can be a useful noninvasive clinical tool.
Subject(s)
Arteries , Magnetic Resonance Angiography , Angiography, Digital Subtraction/methods , Female , Humans , Magnetic Resonance Angiography/methods , Male , Middle Aged , Retrospective Studies , Spin LabelsABSTRACT
Cryosurgery is a recognized method for the treatment of mucoceles in the oral cavity. In this study, cryosurgery was used for mucoceles at the lip or buccal mucosa, and the effect and the indication were evaluated clinically. The subjects were patients with a clinical diagnosis of mucocele on the lip or buccal mucosa and who chose cryosurgery after procedures for both surgical excision and cryosurgery for the lesion were explained. Cryosurgery was performed with a freezing device using liquid nitrogen without local anesthesia. Twenty-four patients chose cryosurgery, including seven preschool children. There were no serious adverse events during and after cryosurgery. Healing progress after cryosurgery was not affected by patient age, lesion size, or how long the patients had the lesion. Two cases later underwent surgical excision because cryosurgery was not successful. Twenty-three patients chose surgical excision, one case had a recurrence. The number of younger patients who chose cryosurgery was significantly higher than that who chose surgical excision. This study suggests that cryosurgery is effective for mucoceles of the lip or buccal mucosa and is a simple and safe treatment method, especially for preschool children.
Subject(s)
Cryosurgery , Mouth Diseases , Mucocele , Child, Preschool , Cryosurgery/adverse effects , Humans , Mouth Diseases/surgery , Mucocele/diagnosis , Mucocele/surgery , Neoplasm Recurrence, LocalABSTRACT
BACKGROUND AND PURPOSE: Spiral MR imaging has several advantages compared with Cartesian MR imaging that can be leveraged for added clinical value. A multicenter multireader study was designed to compare spiral with standard-of-care Cartesian postcontrast structural brain MR imaging on the basis of relative performance in 10 metrics of image quality, artifact prevalence, and diagnostic benefit. MATERIALS AND METHODS: Seven clinical sites acquired 88 total subjects. For each subject, sites acquired 2 postcontrast MR imaging scans: a spiral 2D T1 spin-echo, and 1 of 4 routine Cartesian 2D T1 spin-echo/TSE scans (fully sampled spin-echo at 3T, 1.5T, partial Fourier, TSE). The spiral acquisition matched the Cartesian scan for scan time, geometry, and contrast. Nine neuroradiologists independently reviewed each subject, with the matching pair of spiral and Cartesian scans compared side-by-side, and scored on 10 image-quality metrics (5-point Likert scale) focused on intracranial assessment. The Wilcoxon signed rank test evaluated relative performance of spiral versus Cartesian, while the Kruskal-Wallis test assessed interprotocol differences. RESULTS: Spiral was superior to Cartesian in 7 of 10 metrics (flow artifact mitigation, SNR, GM/WM contrast, image sharpness, lesion conspicuity, preference for diagnosing abnormal enhancement, and overall intracranial image quality), comparable in 1 of 10 metrics (motion artifacts), and inferior in 2 of 10 metrics (susceptibility artifacts, overall extracranial image quality) related to magnetic susceptibility (P < .05). Interprotocol comparison confirmed relatively higher SNR and GM/WM contrast for partial Fourier and TSE protocol groups, respectively (P < .05). CONCLUSIONS: Spiral 2D T1 spin-echo for routine structural brain MR imaging is feasible in the clinic with conventional scanners and was preferred by neuroradiologists for overall postcontrast intracranial evaluation.
Subject(s)
Brain/diagnostic imaging , Magnetic Resonance Imaging/methods , Neuroimaging/methods , Adult , Aged , Artifacts , Female , Humans , Image Enhancement/methods , Male , Middle AgedABSTRACT
Extracellular matrix molecules are generally categorized as collagens, elastin, proteoglycans, or other noncollagenous structural/cell interaction proteins. Many of these extracellular proteins contain distinctive repetitive modules, which can sometimes be found in other proteins. We describe the complete primary structure of an alpha 1 chain of type XII collagen from chick embryonic fibroblasts. This large, structurally chimeric molecule identified by cDNA analysis combines previously unrelated molecular domains into a single large protein 3,124 residues long (approximately 340 kD). The deduced chicken type XII collagen sequence starts at the amino terminus with one unit of the type III motif of fibronectin, which is followed by one unit homologous to the von Willebrand factor A domain, then one more fibronectin type III module, a second A domain from von Willebrand factor, 6 units of type III motif and a third A domain, 10 consecutive units of type III motif and a fourth A domain, a domain homologous to the NC4 domain peptide of type IX collagen, and finally two short collagenous regions previously described as part of the partially sequenced collagen type XII molecule; an Arg-Gly-Asp potential cell adhesive recognition sequence is present in a hydrophilic region at the terminus of one collagenous domain. Antibodies raised to type XII collagen synthesized in a bacterial expression system recognized not only previously reported bands (220 kD et cetera) in tendons, but also bands with apparently different molecular sizes in fibroblasts and 4-d embryos. The antibodies stained a wide variety of extracellular matrices in embryos in patterns distinct from those of fibronectin or interstitial collagens. They prominently stained extracellular matrix associated with certain neuronal tissues, such as axons from dorsal root ganglia and neural tube. These studies identify a novel chimeric type of molecule that contains both adhesion molecule and collagen motifs in one protein. Its structure blurs current classification schemes for extracellular proteins and underscores the potentially large diversity possible in these molecules.
Subject(s)
Collagen/chemistry , Amino Acid Sequence , Animals , Base Sequence , Cell Adhesion , Cell Adhesion Molecules/chemistry , Cell Adhesion Molecules/ultrastructure , Chick Embryo , Cloning, Molecular , Collagen/genetics , Collagen/metabolism , Collagen/ultrastructure , DNA/genetics , Fibronectins/chemistry , Fibronectins/ultrastructure , Molecular Sequence Data , Oligonucleotides/chemistry , Oligopeptides , Polymerase Chain Reaction , Recombinant Proteins/immunology , Sequence Alignment , Tissue Distribution , von Willebrand Factor/chemistry , von Willebrand Factor/ultrastructureABSTRACT
Virulent Sabin-like poliovirus (VSLP) was isolated from river and sewage waters between October 1993 and September 1995 in Toyama Prefecture, Japan (Yoshida et al., Lancet 356, 1461-1463, 2000). In this study, to assess the possibility of an epidemic of poliomyelitis caused by a VSLP in Japan under the current vaccination policy of administration of live attenuated oral poliovirus vaccine (OPV), we determined titers of serum neutralizing antibodies to poliovirus 1 (PV-1) strains Sabin (vaccine strain), Mahoney (wild-type strain) and G4-12 (VSLP) in various groups of residents of Toyama Prefecture, Japan. The seropositivity and geometric mean neutralizing antibody titers against these strains in the individuals who obtained two doses of OPV were 99.1%, 94.5% and 95.5%, respectively, and 564, 186 and 194, respectively. Although the antibody titers to G4-12 were lower compared with those to Sabin, these results indicate that the OPV vaccination policy in Japan has been effective in preventing poliomyelitis caused by VSLPs. These results also suggest that (i) an epidemic of poliomyelitis caused by a VSLP has not occurred in Japan due to herd immunity, and (ii) the possibility of reemergence of VSLPs will be prevented if sufficient herd immunity is acquired immediately after completion of the OPV vaccination in accordance with the poliomyelitis eradication program.
Subject(s)
Antibodies, Viral/blood , Poliovirus Vaccine, Oral/immunology , Poliovirus/immunology , Adolescent , Adult , Aged , Child , Child, Preschool , Humans , Infant , Japan , Middle Aged , Neutralization Tests , Poliovirus Vaccine, Oral/administration & dosageABSTRACT
PURPOSE: Wall enhancement of saccular cerebral aneurysms has not been researched sufficiently. Our purpose of this study was to investigate the incidence of aneurysmal wall enhancement by the three-dimensional turbo spin-echo sequence with motion-sensitized driven equilibrium (MSDE-3D-TSE) imaging after gadolinium injection. METHODS: We retrospectively reviewed the pre- and postcontrast MSDE-3D-TSE images of 117 consecutive patients with intracranial aneurysms from September 2011 to July 2013. A total of 61 ruptured and 83 unruptured aneurysms of 61 patients with subarachnoid hemorrhage (SAH) and 56 non-SAH patients were enrolled in this study. We evaluated the wall enhancement of each aneurysm on postcontrast MSDE-3D-TSE images compared with precontrast images. We classified the aneurysmal wall enhancement into three groups as "Strong enhancement," "Faint enhancement," and "No enhancement." RESULTS: "Strong/Faint enhancement" of the aneurysm was detected in 73.8/24.6 % of the ruptured aneurysms and 4.8/13.3 % of the unruptured aneurysms. "No enhancement" was found in 1.6 % of the ruptured aneurysms and 81.9 % of the unruptured aneurysms. CONCLUSIONS: By magnetic resonance vessel wall imaging using the MSDE-3D-TSE sequence, wall enhancement was frequently observed on ruptured aneurysms. Therefore, aneurysmal wall enhancement may be an indicator of the ruptured condition, which is useful information for managing patients with SAH.
Subject(s)
Aneurysm, Ruptured/diagnostic imaging , Cerebral Arteries/diagnostic imaging , Image Enhancement/methods , Imaging, Three-Dimensional/methods , Intracranial Aneurysm/diagnostic imaging , Magnetic Resonance Angiography/methods , Adult , Aged , Aged, 80 and over , Algorithms , Aneurysm, Ruptured/pathology , Cerebral Angiography/methods , Cerebral Arteries/pathology , Contrast Media , Diagnosis, Differential , Female , Gadolinium , Humans , Image Interpretation, Computer-Assisted/methods , Intracranial Aneurysm/pathology , Male , Middle Aged , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
We have determined the nucleotide sequence of the cell binding domain region of the chicken fibronectin gene and analyzed it evolutionaly. We present here the complete nucleotide sequence of 4.3 kb HindIII/EcoRI segment from the clone lambda FC23 of the chicken fibronectin gene. There were five exons in this segment. When we lined up the amino acid of exons 28, 29 and 31, three alignments, known as the Type III repeat, appeared. Tetrapeptide, -RGDS-, called the cell binding domain, existed in the second repeat, coding exon 30. It was presumed that the Type III repeats were composed of two exons in the chicken gene, the same as in the rat and humans. We found repeatedly appearing amino-acid sequences such as -TIT- (three arrays in these Type III repeats) but also found one of the amino acids substituted in the tripeptide in these Type III repeats (seven arrays). We analyzed these repeats from the point of view of evolution. We used three of the nucleotide sequences (12-18 bp) coding such -TIT- repeats as a unit length for comparing the various homologies after dividing the coding region into 56 segments. The mutual homology of the divided segments to each one of three showed 53% on average. On the other hand, the mutual nucleotide homology of the Type III repeat was 44%. This suggested that the Type III repeat may have been developed by frequent duplication of small gene units.
Subject(s)
Fibronectins/genetics , Genes , Amino Acid Sequence , Animals , Base Sequence , Biological Evolution , Chickens , Cloning, Molecular , DNA/genetics , DNA/metabolism , DNA/ultrastructure , DNA Restriction Enzymes , Gene Amplification , Introns , Molecular Sequence DataABSTRACT
OBJECTIVE: To probe the utility of dynamic contrast-enhanced MRI (DCE-MRI) parameters in assessing the clinical characteristics of oral squamous cell carcinoma. METHODS: A total of 85 tumours were included. We applied the Tofts and Kermode model for the DCE-MRI data and obtained three dependent parameters: the influx forward volume transfer constant into the extravascular extracellular space (EES) from the plasma (K(trans)), the fractional volume of EES per unit volume of tissue (ve) and the fractional volume of plasma (vp). We evaluated the correlations between these parameters and the clinical stages. RESULTS: The T stage showed a negative correlation with the K(trans) (r = -0.2272; p = 0.0365), but it did not show a significant correlation with the other parameters. The N stage showed a negative correlation with K(trans) (r = -0.1948; p = 0.0404), and there were significant differences between N1 and N2+3 (0.119 ± 0.027 vs 0.096 ± 0.023 min(-1); p = 0.0198) and between N0 and N2+3 (0.114 ± 0.29 vs 0.096 ± 0.023 min(-1); p = 0.0288). CONCLUSION: A decrease in the K(trans) at the primary site was found in advanced N stage cases, which might indicate that the hypoxic status cause a high possibility of the metastasis. ADVANCES IN KNOWLEDGE: A decrease in the K(trans) at the primary site suggested the high possibility of an advanced N stage.
Subject(s)
Carcinoma, Squamous Cell/pathology , Magnetic Resonance Imaging/methods , Mouth Neoplasms/pathology , Aged , Contrast Media , Female , Gadolinium DTPA , Humans , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Staging , Prospective StudiesABSTRACT
Both the epithelium and the mesenchyme of the larval small intestine of anurans undergoes metamorphic conversion into the adult counterparts. The conversion of the mesenchyme has been poorly understood especially at the molecular level, whereas the changes of the epithelium have been extensively studied. The present study investigated the metamorphic changes of the mesenchyme of tadpoles of bullfrog, Rana catesbeiana, focusing on the expression of genes of type I collagen. By using the cDNA clones coding for a 1(I) and a 2(I) collagen as probes, expression of each collagen gene was examined. These genes were drastically up-regulated at the climax period of spontaneous metamorphosis, which was precociously mimicked by treating tadpoles with thyroid hormone. The increased expression of these genes at the climax stage was well correlated with the conversion of the thin larval mesenchyme to more thick and dense adult connective tissues of the intestine. In situ hybridization identified the fibroblasts that were actively expressing the collagen genes and, therefore, were thought to be responsible for the remodeling. These results strongly suggest that the expression of type I collagen genes is regulated during the intestinal remodeling in a cell-type specific and thyroid hormone-dependent manner.
Subject(s)
Collagen/genetics , Gene Expression Regulation, Developmental , Intestine, Small/metabolism , Metamorphosis, Biological/physiology , Thyroid Hormones/metabolism , Amino Acid Sequence , Animals , Base Sequence , Blotting, Southern , Collagen/classification , DNA, Complementary , Humans , Intestine, Small/embryology , Molecular Sequence Data , RNA, Messenger , Rana catesbeiana , Sequence Homology, Amino AcidABSTRACT
The present study determined nucleotide sequences of the full-length cDNA of alpha2 chain of bullfrog type I collagen. Hybridization of a bullfrog cDNA library with human alpha1 type I collagen cDNA yielded a clone named 6A-1 which was 3449 bp long and lacked a 5' region of the gene. A 5' region containing the translation initiation site was amplified by the reverse transcription polymerase chain reaction using poly(A)+RNA from tadpole tail tissues as template, and oligonucleotides encoding the translation initiation region of mammalian fibrillar collagens and the Gly-X-Y repeat region of clone 6A-1 as primers. As a result we obtained a 1518 bp long clone Y31. A 355 bp long clone Y31-9 was produced by extending clone Y31 from its ATG codon to a 127 bp upstream region. Combining these three clones, the complete nucleotide sequence of the full-length cDNA was determined which contained 4692 bp as a whole and 4065 bp in the open reading frame. The comparison of its structure with known collagen cDNAs of various vertebrates showed that the cDNA obtained codes for alpha2(I) chain of bullfrog. Its deduced amino acid sequence revealed the complete conservation of seven cysteine residues in the C-propeptide and three lysine residues in the N-telopeptide through the helical domain. Northern blot analysis revealed that the thyroid hormone regulated the expression of alpha2(I) collagen in an organ-dependent manner: intense up-regulation in the back skin and intestine, weak and transient up-regulation in the liver, and initial down-regulation, but later up-regulation in the tail. Prolactin increased its expression in both the back skin and tail. These results suggested that the expression of bullfrog alpha2(I) collagen is cooperatively regulated by these two metamorphosis-regulating hormones.
Subject(s)
Collagen/genetics , Amino Acid Sequence , Animals , Chickens , Cloning, Molecular , DNA, Complementary , Gene Expression Regulation , Humans , Invertebrates , Mammals , Metamorphosis, Biological , Molecular Sequence Data , Prolactin/physiology , Rana catesbeiana , Sequence Homology, Amino Acid , Thyroid Hormones/physiologyABSTRACT
The skin develops and differentiates during embryogenesis, which is concertedly regulated by a variety of genes. The present study isolated from the rat embryonic skin a novel differentiation-associated gene named Kdap (keratinocyte differentiation-associated protein) by suppression subtractive hybridization between the skin of 14day postcoitus (dpc) embryo (the prehair-germ stage) and that of 17dpc embryo (the hair-germ stage). Its mRNA contained four spliced forms in these tissues. The gene encoded a protein of total 98 amino acids with a calculated molecular mass of 11kDa and an isoelectric point of 6.1 as an unspliced form. The two splicing zones were well conserved among rat, mouse, and human. This protein had a high hydrophobic N-terminal region, a possible signal sequence, and contained two putative N-myristoylation sites and two casein kinase II phosphorylation sites. In situ hybridization experiments detected Kdap transcripts exclusively in the suprabasal cell layers of the embryonic epidermis. Intense expression was also seen in suprabasal cells in regions of infundibulum of the hair follicle. These results indicated that Kdap provides a new insight into the mechanism of differentiation and the maintenance of stratified epithelia.
Subject(s)
Epithelium/metabolism , Phosphoproteins/genetics , Amino Acid Sequence , Animals , Animals, Newborn , Base Sequence , Cell Differentiation , DNA, Complementary/chemistry , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Embryo, Mammalian/metabolism , Epithelium/embryology , Female , Gene Expression Regulation, Developmental , In Situ Hybridization , Keratinocytes/cytology , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Inbred F344 , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid , Skin/embryology , Skin/growth & development , Skin/metabolism , Tissue DistributionABSTRACT
The ability of cells to organize collagen fibrils is fundamental to a variety of processes found in embryogenesis, wound healing, fibrosis, and scar formation. We previously isolated a monoclonal antibody (mAb A3A5) which inhibits human fibroblast-mediated collagen gel contraction, an in vitro model producing the process of collagen morphogenesis. Human fibronectin (FN) has been shown to be the antigen of A3A5. The present study aimed at identifying the A3A5 epitope to reveal the mode of binding between collagen, FN, and fibroblasts in the process of gel contraction. The epitope was sought in FN fragments obtained by pepsin digestion and in recombinant FN fragments expressed in Escherichia coli by determining their immunological reactivity with A3A5, and was identified as a short segment consisting of the fourth through the amino half of the fifth FN type III. We propose a new functional domain of FN which plays a crucial role in the binding of fibroblasts to collagen fibrils and is involved in collagen morphogenesis.
Subject(s)
Antibodies, Monoclonal/immunology , Collagen/metabolism , Epitope Mapping , Fibroblasts/metabolism , Fibronectins/chemistry , Amino Acid Sequence , Antibodies, Monoclonal/pharmacology , Binding Sites , Blotting, Western , Collagen/chemistry , Epitopes/chemistry , Fibronectins/immunology , Fibronectins/metabolism , Gels , Gene Deletion , Humans , Molecular Sequence Data , Mutagenesis, Site-Directed , Pepsin A/metabolism , Peptide Fragments/chemistry , Peptide Fragments/immunology , Polymerase Chain Reaction , Recombinant ProteinsABSTRACT
Two cDNA subfragments containing the cell-attachment site of human fibronectin (FN) were expressed as beta-galactosidase fusion proteins in E. coli. The products were purified to homogeneity by monoclonal antibody affinity chromatography and assayed for activity in a standard cell-adhesion assay. A fusion protein containing an 80 kDa fragment of human FN appeared functionally equivalent to intact FN purified from human plasma, whereas a truncated fusion protein of 33 kDa still containing a previously postulated cell-attachment site was approx. 50-fold less active. Our study establishes a system for analyzing adhesive protein function by DNA manipulation, rules out any major role for eukaryotic post-translational modifications in FN adhesive function, and localizes additional functional activity to a 1.3 kb region.
Subject(s)
Escherichia coli/genetics , Fibronectins/genetics , Amino Acid Sequence , Animals , Binding Sites , Biological Assay , Cell Adhesion , Cell Line , Cricetinae , Fibronectins/physiology , Humans , Kidney , Peptide Fragments/genetics , Peptide Fragments/physiology , Protein Processing, Post-Translational , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/pharmacology , beta-GalactosidaseABSTRACT
This work was designed to evaluate the toxicity of inhalable particles [less than/equal to] 10 microm in aerodynamic diameter (PM(10)) collected from the urban air in São Paulo, Brazil, to the mucociliary apparatus using the frog palate preparation. Seven groups of frog palates were immersed in different concentrations of PM(10) diluted in Ringer's solution during 120 min: 0 (control, n = 31); 50 (n = 10); 100 (n = 9); 500 (n = 28); 1,000 (n = 10); 5,000 (n = 11); and 10,000 microg/m(3) (n = 10). Mucociliary transport and transepithelial potential difference were determined at 0, 30, 60, and 120 min exposure. Additional groups (control and 500 microg/m(3)) were studied by means of morphometric analyses (quantification of the amount of intraepithelial and surface mucins), measurement of cilia beat frequency, and quantification of total glutathione. Mucociliary transport and transepithelial potential difference were significantly decreased at higher concentrations of PM(10) (p = 0.03 and p = 0.02, respectively). Exposure to PM(10) also elicited a significant decrease of total glutathione (p = 0. 003) and depletion of neutral intraepithelial mucins (p = 0.0461). These results show that PM(10) can promote significant alterations in ciliated epithelium in vitro.
Subject(s)
Air Pollutants/toxicity , Mucociliary Clearance/drug effects , Palate/drug effects , Aerosols , Animals , Palate/pathology , Palate/physiology , Rana catesbeianaABSTRACT
We mobilized peripheral blood stem cells (PBSC) following CHOP plus rituximab (CHOP-R) therapy, and compared with the findings following CHOP therapy without rituximab. All patients were given G-CSF starting from day 11 after CHOP therapy. Patients in the CHOP-R group (n=8) were given rituximab on day 12. Target CD34(+) cells number was collected in a single leukapheresis on day 14, from all the eight patients in the CHOP-R group. PBSC mobilization kinetics, CD34(+) cells yield and colony-forming ability in the graft collection, toxicity during mobilization, and engraftment after transplantation of CHOP-R group were not significantly different from those in the CHOP group (n=8). In all patients given CHOP-R therapy, CD20(+) cells and immunoglobulin heavy chain (IgH) rearrangement in the graft collection were undetectable by flow-cytometric analysis and Southern blot analysis, respectively, but with PCR analysis two of eight grafts were positive for IgH rearrangement. While further studies are needed to evaluate the efficacy of purging and the outcome of patients undergoing autologous transplantation, CHOP-R therapy can be safely and effectively used in the mobilization phase of PBSC collection, without excess clinical toxicity or deleterious effect on PBSC mobilization kinetics or engraftment time.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Hematopoietic Stem Cell Mobilization/methods , Lymphoma, B-Cell/therapy , Peripheral Blood Stem Cell Transplantation/methods , Transplantation Conditioning/methods , Adult , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal, Murine-Derived , Antineoplastic Combined Chemotherapy Protocols/toxicity , Cyclophosphamide/administration & dosage , Doxorubicin/administration & dosage , Female , Gene Rearrangement , Graft Survival , Granulocyte Colony-Stimulating Factor/administration & dosage , Humans , Immunoglobulin Heavy Chains/genetics , Kinetics , Leukapheresis/methods , Male , Middle Aged , Prednisone/administration & dosage , Rituximab , Vincristine/administration & dosageABSTRACT
The aim of this study was to analyze how functional activation in the supplementary motor area (SMA) and sensorimotor cortex (SMC) is related to bimanual coordination using functional magnetic resonance imaging. Subjects included 24 healthy volunteers, 15 of whom were right-handed and 9 left-handed. Three kinds of activation tasks, all of which required the repetitive closing and opening of a fist, were performed: unimanual movement of the nonpreferred hand (task A); simultaneous, agonistic movement of both hands (task B); simultaneous, antagonistic movement of both hands (task C). The SMA activation during task C was more pronounced than that during the other two tasks for right and left handers. The results suggested that the activation of the SMA, at least during a simple motion used in the present study, was little influenced by whether the motion was unimanual or bimanual but instead how the bimanual motion was composed of the motion element of a single hand. The SMC activation during task C was significantly larger than that during task B, whereas hemispheric differences in the activation were not found. This indicated that the complexity of the bimanual movement also affected the SMC activation.
Subject(s)
Motor Cortex/physiology , Psychomotor Performance/physiology , Somatosensory Cortex/physiology , Adult , Functional Laterality/physiology , Humans , Magnetic Resonance Imaging , MaleABSTRACT
This study analyzed the diagnostic procedures of 391 patients with a pulmonary nodule due to lung cancer encountered in the practice from 1987 to 1997. Transbronchial biopsy and cytological procedures under fiberoptic bronchoscopy made a diagnosis in 329 (84.1%) of the patients. In 266 patients with endoscopically invisible tumor, transbronchial biopsy gave a positive result in 74.1% and the rest were diagnosed by cytological procedures. Transbronchial biopsy with cytological procedures should be considered as the initial diagnostic procedure. The most appropriate method and combining methods can improve the diagnostic yield in patients with pulmonary nodules.