Epidemiology and clinical profiles of cutaneous graft versus host disease in allogeneic peripheral blood stem cell transplantation.

INTRODUCTION: The epidemiology of cutaneous graft versus host disease (GVHD) in allogeneic peripheral blood stem cell transplantation (PBSCT) in Malaysia has not been described. MATERIALS AND METHODS: We retrospectively analysed 691 allogeneic PBSCT patients between 2010-2017 in two centers. RESULTS: The prevalence of cutaneous GVHD was 31.4% (217/691). No associations were detected with race, age or gender of donor and recipients. Cutaneous GVHD was associated with host cytomegalovirus (CMV) seropositivity (p<0.01), conditioning (p<0.01), GVHD prophylaxis (p=0.046) and survival (p<0.01). Majority developed the acute form (58.1%;126/217). Biopsies in 20.7% (45/217) showed 55.6% positivity for GVHD. Overall, involvement was non-severe. A majority demonstrated complete response (CR) to first-line corticosteroids (70.0%;152/217). Secondline therapies (extracorporeal phototherapy (ECP), psolaren ultraviolet A (PUVA), mycophenolate, tumour necrosis factor (TNF) inhibitors, interleukins inhibitors, or CD20 monoclonal antibodies) were required in 65/217, with 38.5% CR. Second-line therapy was associated with gender (p=0.042), extra-cutaneous GVHD (p=0.021), treatment outcomes (p=0.026) and survival (p=0.048). Mortality in cutaneous GVHD was 24.0% with severe sepsis being the leading cause at Day 100 (7.8%) and 5-years (7.8%), and relapsed disease at 2-years (32.7%). In steroid refractoriness, severe GVHD caused 30.8% mortality. In cutaneous GVHD, survival at Day 100 was 95.4%; 80.2% at 2-years and 73.1% at 5-years. The median survival in cutaneous GVHD was significantly shorter at 55 months, compared to those without GVHD at 69 months (p=0.001). CONCLUSION: Cutaneous involvement is the commonest clinical manifestation of GVHD. A larger national study is warranted to further analyse severity and outcome of multiorgan GVHD, and factors associated with steroid refractoriness.

iNKT cells prevent obesity-induced hepatic steatosis in mice in a C-C chemokine receptor 7-dependent manner.

Non-alcoholic fatty liver disease and non-alcoholic steatohepatitis are characterized by an increase in hepatic triglyceride content with infiltration of immune cells, which can cause steatohepatitis and hepatic insulin resistance. C-C chemokine receptor 7 (CCR7) is primarily expressed in immune cells, and CCR7 deficiency leads to the development of multi-organ autoimmunity, chronic renal disease and autoimmune diabetes. Here, we investigated the effect of CCR7 on hepatic steatosis in a mouse model and its underlying mechanism. Our results demonstrated that body and liver weights were higher in the CCR7-/- mice than in the wild-type (WT) mice when they were fed a high-fat diet. Further, glucose tolerance and insulin sensitivity were markedly diminished in CCR7-/- mice. The number of invariant natural killer T (iNKT) cells was reduced in the livers of the CCR7-/- mice. Moreover, liver inflammation was detected in obese CCR7-/- mice, which was ameliorated by the adoptive transfer of hepatic mononuclear cells from WT mice, but not through the transfer of hepatic mononuclear cells from CD1d-/- or interleukin-10-deficient (IL-10-/-) mice. Overall, these results suggest that CCR7+ mononuclear cells in the liver could regulate obesity-induced hepatic steatosis via induction of IL-10-expressing iNKT cells.

Annual prevalence and economic burden of genital warts in Korea: Health Insurance Review and Assessment (HIRA) service data from 2007 to 2015.

This study evaluated the annual prevalence of anogenital warts (AGW) caused by human papillomavirus (HPV) and analysed the trend in annual per cent changes (APC) by using national claims data from the Health Insurance Review and Assessment of Korea, 2007-2015. We also estimated the socio-economic burden and co-morbidities of AGW. All analyses were performed based on data for primary A63.0, the specific diagnosis code for AGW. The socio-economic cost of AGW was calculated based on the direct medical cost, direct non-medical cost and indirect cost. The overall AGW prevalence and socio-economic burden has increased during the last 9 years. However, the prevalence of AGW differed significantly by sex. The female prevalence increased until 2012, and decreased thereafter (APC + 3·6%). It would fall after the introduction of routine HPV vaccination, principally for females, in Korea. The male prevalence increased continuously over time (APC + 11·6%), especially in those aged 20-49 years. Referring to the increasing AGW prevalence and its disease burden, active HPV infection control surveillance and prevention in males are worth consideration.

Effects of ionised or chelated water-soluble mineral mixture supplementation on growth performance, nutrient digestibility, blood characteristics, meat quality and intestinal microbiota in broilers.

An experiment was conducted to study the effects of dietary supplementation of water-soluble ionised or chelated mineral mixture on growth performance, nutrient digestibility, blood characteristics, relative organ weight, meat quality and excreta microflora in broilers. A total of 408 Arbor Acres broilers (17 birds in 8 replicate pens) were randomly allocated into one of the following three treatments: (1) Control/basal diet (CON), (2) T1 (basal diet + 0.5% ionised mineral mixture solution, pH 3.0) and (3) T2 (basal diet + 0.5% chelated mineral mixture solution, pH 3.0). The body weight gain was greater and feed conversion ratio was lower in broilers supplemented with ionised or chelated mineral liquid complex compared to CON during the grower and overall phase of the experiment. No significant effect in the concentration of Ca and P in the blood was observed in birds supplemented with ionised or chelated mineral mixture solution. No adverse effects were observed in organ weight and meat quality with ionised or chelated mineral mixture supplementation. Regarding intestinal microbiota counts there was a reduction of Escherichia coli counts in the small intestine in ionised mineral supplemented birds. In the large intestine, E. coli as well as Salmonella populations were reduced in ionised mineral supplemented birds. In conclusion, ionised or chelated minerals have partial positive effects in improving growth performance and reducing pathogenic bacteria load in the gastro-intestinal tract.

Color moiré simulations in contact-type 3-D displays.

A new method of color moiré fringe simulation in the contact-type 3-D displays is introduced. The method allows simulating color moirés appearing in the displays, which cannot be approximated by conventional cosine approximation of a line grating. The color moirés are mainly introduced by the line width of the boundary lines between the elemental optics in and plate thickness of viewing zone forming optics. This is because the lines are hiding some parts of pixels under the viewing zone forming optics, and the plate thickness induces a virtual contraction of the pixels. The simulated color moiré fringes are closely matched with those appearing at the displays.

Deep neuromuscular block improves the surgical conditions for laryngeal microsurgery.

BACKGROUND: Adequate neuromuscular block is required throughout laryngeal microsurgery. We hypothesized that the surgical conditions would improve under a deeper level of rocuronium-induced neuromuscular block. METHODS: Seventy-two patients undergoing laryngeal microsurgery were randomly allocated to either the 'post-tetanic counts 1-2' (PTC1-2) group or the 'train-of-four counts 1-2' (TOFcount1-2) group according to the level of neuromuscular block used. Two different doses of rocuronium (1.2 or 0.5 mg kg(-1)) were used after anaesthetic induction, and two respective targets of neuromuscular block (post-tetanic counts ≤2 or train-of-four count of 1 or 2) were used. Surgical conditions were assessed by the surgeon using a five-point rating scale (extremely poor/poor/acceptable/good/optimal), and clinically acceptable surgical conditions were defined as those which were rated acceptable, good, or optimal. The occurrence of vocal cord movement and postoperative adverse events was assessed. RESULTS: The surgical conditions were significantly different between the PTC1-2 and TOFcount1-2 groups (extremely poor/poor/acceptable/good/optimal: 0/2/1/7/26 and 3/10/2/14/7, respectively, P<0.001). The incidence of clinically acceptable surgical conditions was significantly higher in the PTC1-2 group than in the TOFcount1-2 group (94 vs 64%, P=0.003). The percentage of patients who exhibited vocal cord movement was significantly lower in the PTC1-2 group than in the TOFcount1-2 group (3 vs 39%, P<0.001). The incidence of postoperative adverse events was not significantly different except for the less frequent occurrence of mouth dryness in the PTC1-2 group (P=0.035). CONCLUSIONS: Deep neuromuscular block (post-tetanic count of 1-2) surgical conditions in patients undergoing laryngeal microsurgery improves. CLINICAL TRIAL REGISTRATION: NCT01980069.

Gene pathways and cell cycle-related genes in cultured avian primordial germ cells.

Primordial germ cells (PGC) from early embryos are applicable to various kinds of research, including the production of transgenic animals. Primordial germ cells eventually migrate and differentiate into germ cells in the gonads, where they settle and rapidly proliferate. However, the proliferation rate of PGC is low in early embryos, and there are many significant pathways that mediate PGC activity. Therefore, in vitro culture of PGC from early embryos with efficient growth factors has been necessary. Recently, we cultured chicken PGC from embryonic d 2.5 with basic fibroblast growth factor and characterized the PGC through analysis of cell morphology, survival, proliferation, and apoptosis. However, large-scale analyses of genes expressed in cultured PGC and the genes involved in associated pathways are limited. The objective of the present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between PGC and their somatic counterpart, chicken embryonic fibroblasts (CEF). We identified 795 genes that were expressed more predominantly in PGC and 824 genes that were expressed more predominantly in CEF. Among the predominant genes in PGC, 201 were differentially identified in 106 pathways. Among the predominant genes in CEF, 242 were differentially identified in 99 pathways. To further validate the genes involved in at least one candidate pathway, those involved in the cell cycle (12 predominant genes in PGC and 8 predominant genes in CEF) were examined by real-time PCR. To the best of our knowledge, this study is the first to investigate signaling and metabolic pathways in cultured PGC.

Apolipoprotein B-48 is the product of a messenger RNA with an organ-specific in-frame stop codon.

The primary structure of human apolipoprotein (apo) B-48 has been deduced and shown by a combination of DNA excess hybridization, sequencing of tryptic peptides, cloned complementary DNAs, and intestinal messenger RNAs (mRNAs) to be the product of an intestinal mRNA with an in-frame UAA stop codon resulting from a C to U change in the codon CAA encoding Gln2153 in apoB-100 mRNA. The carboxyl-terminal Ile2152 of apoB-48 purified from chylous ascites fluid has apparently been cleaved from the initial translation product, leaving Met2151 as the new carboxyl-terminus. These data indicate that approximately 85% of the intestinal mRNAs terminate within approximately 0.1 to 1.0 kilobase downstream from the stop codon. The other approximately 15% have lengths similar to hepatic apoB-100 mRNA even though they have the same in-frame stop codon. The organ-specific introduction of a stop codon to a mRNA appears unprecedented and might have implications for cryptic polyadenylation signal recognition and RNA processing.

The first step in sugar transport: crystal structure of the amino terminal domain of enzyme I of the E. coli PEP: sugar phosphotransferase system and a model of the phosphotransfer complex with HPr.

BACKGROUND: The bacterial phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS) transports exogenous hexose sugars through the membrane and tightly couples transport with phosphoryl transfer from PEP to the sugar via several phosphoprotein intermediates. The phosphate group is first transferred to enzyme I, second to the histidine-containing phosphocarrier protein HPr, and then to one of a number of sugar-specific enzymes II. The structures of several HPrs and enzymes IIA are known. Here we report the structure of the N-terminal half of enzyme I from Escherichia coli (EIN). RESULTS: The crystal structure of EIN (MW approximately 30 kDa) has been determined and refined at 2.5 A resolution. It has two distinct structural subdomains; one contains four alpha helices arranged as two hairpins in a claw-like conformation. The other consists of a beta sandwich containing a three-stranded antiparallel beta sheet and a four-stranded parallel beta sheet, together with three short alpha helices. Plausible models of complexes between EIN and HPr can be made without assuming major structural changes in either protein. CONCLUSIONS: The alpha/beta subdomain of EIN is topologically similar to the phosphohistidine domain of the enzyme pyruvate phosphate dikinase, which is phosphorylated by PEP on a histidyl residue but does not interact with HPr. It is therefore likely that features of this subdomain are important in the autophosphorylation of enzyme I. The helical subdomain of EIN is not found in pyruvate phosphate dikinase; this subdomain is therefore more likely to be involved in phosphoryl transfer to HPr.

Fundus autofluorescence imaging patterns in central serous chorioretinopathy according to chronicity.

PurposeTo investigate the time-period characteristics associated with morphologic changes in central serous chorioretinopathy (CSC) using fundus autofluorescence (FAF).Patients and methodsRetrospective, cross-sectional observational case series. Patients were classified into three groups: acute and chronic according to the onset of subjective symptoms of 6 weeks and sequelae patients who have history and symptoms but no serous retinal detachment (SRD). We compared FAF images to obtain characteristic findings according to the chronicity.ResultsA total of 52 eyes were included in this study. Acute CSC eyes were characterized by decreased FAF intensity at the leakage point in 13/22 eyes (56.5%) and staining patterns with various levels of fluorescence signal (hyperautofluorescent (10 eyes, 43.5%), hypoautofluorescent (1 eye, 4.3%), and minimal changes (12 eyes, 52.2%)) in the area of SRD. In chronic CSC eyes, hyperautofluorescent (14 eyes, 63.6%) or minimal changes (8 eyes, 36.4%) were observed in the area of SRD. Discrete dots with increased FAF intensity were observed in chronic CSC eyes (P<0.001). Eyes with sequelae of CSC had mixed FAF patterns over areas of retinal pigment epithelium (RPE) atrophy in seven eyes (100%, P<0.001)) and descending tracts which showed various FAF intensities according to the RPE and photoreceptor status (P<0.001).ConclusionFAF imaging patterns in CSC eyes differ according to the course of the disease, reflecting RPE and outer retinal changes. Detailed investigation using FAF could help to estimate the duration of CSC and determine the proper treatment modality.

Evaluation of parameters obtained from the Sysmex XN-2000 for predicting the recovery of the absolute neutrophil count and platelets after hematopoietic stem cell transplantation.

INTRODUCTION: We analyzed abilities of parameters from Sysmex XN-2000 (Sysmex, Kobe, Japan) to predict absolute neutrophil count (ANC) and platelet recovery after hematopoietic stem cell transplantation (HSCT) in patients with hematologic malignancies. METHODS: We prospectively analyzed 911 follow-up peripheral blood samples from 44 HSCT-performed patients and evaluated the performances of the following parameters: WBC, immature granulocyte (IG), hematopoietic stem and progenitor cells (HPC), immature reticulocyte fraction (IRF), immature platelet fraction (IPF), platelet distribution width (PDW), mean platelet volume (MPV), and platelet larger cell ratio (P-LCR). RESULTS: When compared to four other parameters, the identification of initiation in IG (%)/HPC (%) increase enabled earlier prediction of ANC recovery to >500/µL and >1000/µL with more time benefit of 3.5-6.5 days/2.0-5.0 days and 3.0-6.0 days/2.0-5.0 days, respectively. When compared to IPF (%), the identification of initiation in PDW, MPV, and P-LCR (%) increase enabled earlier prediction of platelet recovery to >20 000/µL and >50 000/µL with more time benefit of 2.5-3.5 days and 2.0-3.0 days, respectively. However, the standard deviation of time benefit obtained from IG (%)/HPC (%)/PDW/MPV/P-LCR (%) was consistently large (3.0-4.3 days). CONCLUSIONS: There is a systematic pattern where a rise in most of the studied parameters can be observed in most patients before ANC/platelet recovery. However, the interindividual variation between the time of rise of these parameters and ANC/platelet recovery is large, and therefore, using these parameters to predict recovery in the individual patient is probably not meaningful in the clinical setting.

Plasma lipid transport in the preruminant calf, Bos spp: primary structure of bovine apolipoprotein A-I.

The preruminant calf (Bos spp.) is a model of considerable interest with regard to hepatic and intestinal lipoprotein metabolism (Bauchart et al., J. Lipid Res. (1989) 30, 1499-1514 and Laplaud et al., J. Lipid Res. (1990) 31, 1781-1792). As a preliminary step towards future experiments dealing with HDL metabolism in the calf, we have purified apoA-I from this animal and determined its complete amino acid sequence. Thus, approx. 10% of calf apoA-I was shown to contain a propeptide, with the sequence Arg-His-Phe-Trp-Gln-Gln. Enzymatic cleavage of apoA-I resulted in 10 proteolytic peptides. The complete apoA-I sequence was obtained after alignment of peptides on the basis of their homologies with those from rabbit apoA-I. Thus calf apoA-I consists of 241 amino acid residues, and exhibits high sequence homology with all mammalian apoA-I's studied to date. The bovine protein contained 10 hydrophobic amphipathic helical regions, occurring between residues 43-64, 65-86, 87-97, 98-119, 120-141, 142-163, 164-184, 185-206, 207-217 and 218-241. A computer-constructed phylogenetic tree showed that bovine apoA-I was more closely related to its dog counterpart, including the presence of a single methionine, than to the corresponding macaque and human proteins. Comparative predictions of the respective antigenic structures of human and bovine apoA-I's using the Hopp-Woods algorithm indicated similar positions for all 13 detectable antigenic sites, among which 7 were of identical, or closely related, amino acid composition. This finding was confirmed by demonstration of partial immunological identity between the two proteins upon immunodiffusion analysis, a result obtained using a monospecific rabbit antiserum against bovine apoA-I. Finally, comparison of sequence homology between bovine apoA-I and the lecithin:cholesterol acyl transferase (LCAT) activating region of human apoC-I suggests that several LCAT activating domains may be present in calf apoA-I.

The primary structure of human apolipoprotein A-IV.

Human apolipoprotein (apo) A-IV was purified from chylous ascites fluid. Proteolytic peptides produced by trypsin and Staphylococcus aureus V8 proteinase digestions were purified by high-performance liquid chromatography and sequenced. Human apoA-IV contains 376 amino acid residues. The peptide-derived sequence generally matches two previously reported DNA-derived amino acid sequences except for discrepancies in five positions. In order to examine these discrepancies further, one complete apoA-IV cDNA clone and another partial clone were sequenced. Comparison of all the available information indicates that the peptide-derived sequence reported here is accurate. Sequencing errors probably account for some of the discrepancies between the two primary sequences predicted by earlier nucleotide analyses. In certain positions, however, bona fide sequence heterogeneity or cloning artifact cannot be excluded.

Nuclear morphometry and prognosis in favorable histology Wilms' tumor: A prospective reevaluation.

PURPOSE: This study was designed to evaluate the ability of a previously published nuclear morphometry discriminant function to predict disease-free survival in patients with Wilms' tumor. PATIENTS AND METHODS: We identified 218 patients with stage I-IV Wilms' tumor of favorable histology who were entered onto the National Wilms' Tumor Study (NWTS) between January 1, 1990 and April 15, 1994. The nuclear morphometry score was calculated for each patient as follows: MV(f) = (0.02 x AGE) + (1.17 x SNRF) + (90.6 x LEFD) - 94, with AGE denoting age at diagnosis in months, SNRF the skewness of the nuclear roundness factor, and LEFD the lowest value of nuclear ellipticity as measured by the feret diameter method. Relative risks of relapse were estimated for the total score and for each of its components. Sensitivity and specificity were determined for the criterion of "MV(f) is greater than -0.35" as a predictor of relapse. RESULTS: By contrast with previously published results, neither the SNRF nor the LEFD made any contribution to the prediction of disease-free survival. Sensitivity and specificity of the criterion of "MV(f) is greater than -0.35" were 71% and 56%, respectively. CONCLUSION: Re-evaluation of a published nuclear morphometry score showed that it did not predict disease-free survival in patients with Wilms' tumor. The earlier study very likely overestimated the predictive power of nuclear morphometry by using the same data set both to develop the score and to evaluate its properties. Because of the huge number of combinations of nuclear morphometry measurements that may enter into the multivariate discriminant function, use of appropriate statistical methods is essential to estimate accurately the sensitivity and specificity.

Laparoscopic bowel injury in an animal model: monocyte migration and apoptosis.

BACKGROUND: Unrecognized laparoscopic bowel injury has a delayed and covert presentation. Differences in monocyte migration and apoptosis between laparoscopic and open bowel injury were determined. METHODS: For this study, 24 rabbits were divided into laparoscopic (n = 9) and open surgical (n = 9) bowel injury groups and a control group (n = 6) without bowel injury. Bowel injury was created using monopolar electrocautery. The animals were killed 1 day, 1 week, and 2 weeks after surgery. Monocyte migration assay was performed across a modified Boyden chamber. Apoptosis was assessed by DNA fluorescent stain H-33342. RESULTS: In laparoscopy, monocyte apoptosis was decreased (p < 0.001), and migration was increased (p < 0.05), as compared with the open group. Apoptosis increased over time in both study groups, and was higher than in the control group (p < 0.001). Migration was decreased in both study groups as compared with the control group (p < 0.05) CONCLUSIONS: These results suggest decreased immune system priming with laparoscopic bowel injury, which may contribute to the masking of relevant signs and symptoms of peritonitis.

Sepsis affects most routine and cell population data (CPD) obtained using the Sysmex XN-2000 blood cell analyzer: neutrophil-related CPD NE-SFL and NE-WY provide useful information for detecting sepsis.

INTRODUCTION: The Sysmex XN-2000 analyzer can assess 36 routine and 57 cell population data (CPD) items. In this study, we evaluated these items as sepsis biomarkers. METHODS: We enrolled 280 normal control (NC) and 130 sepsis patients. The sepsis patients were classified as uncomplicated or complicated sepsis. Routine and CPD items were determined, and the results were compared at between the NC and sepsis groups, uncomplicated and complicated sepsis groups, and survivors and nonsurvivors. RESULTS: For the detection of sepsis, CPD items NE-SFL [defined as the fluorescent light intensity of the neutrophil area on the WDF (white blood cell differential) scattergram] and NE-WY (defined as the fluorescent light distribution width of the neutrophil area on the WDF scattergram) showed comparative or higher AUC of 0.909 and 0.905, respectively, when compared with routine items such as hematocrit, hemoglobin, RBC, RDW, immature granulocytes count, lymphocytes count, and neutrophils count. For the discrimination of sepsis severity, only platelet-related items showed higher AUC (0.723 - 0.748) than lactic acid (0.695). For the prediction of 28-day mortality, only CV and SD of RDW showed higher AUC (0.766 and 0.732 each) than lactic acid (0.712). CONCLUSIONS: Sepsis patients demonstrated significant changes in routine and CPD items related to RBC, neutrophils, lymphocytes, and platelets when compared to NCs. Increase in CPD items NE-SFL and NE-WY, which may indicate neutrophil immaturity or activation, could be useful for the detection of sepsis patients, in conjunction with currently used surrogate sepsis biomarkers. However, these items did not efficiently contribute to the discrimination of sepsis severity or predict mortality.

NF-kappa B mediates the adaptation of human U937 cells to hydrogen peroxide.

Low doses of oxidative stress can induce cellular resistance to subsequent higher doses of the same stress. By using human U937 leukemia cells, we previously demonstrated that H(2)O(2) can induce such an adaptive response without elevating the cellular capacity to degrade H(2)O(2), and were able to confer the cells a cross-resistance to an H(2)O(2)-independent lethal stimulus, C(2)-ceramide. In this study, it was found that the adaptation is accompanied by the translocation of cytoplasmic NF-kappa B to the nuclei. This event was promoted or abolished when either IKK alpha or a dominant negative mutant of I kappa B, respectively, was overexpressed. The overexpression of IKK alpha also resulted in the suppression of H(2)O(2)-induced cell death and DNA fragmentation, whereas these events were accelerated by the expression of the I kappa B mutant. The protective effect of IKK alpha was accompanied neither by an elevation of protein levels of various antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase, nor by an increase in the cellular capacity to consume H(2)O(2). Moreover, the overexpression of IKK alpha resulted in an enhancement of H(2)O(2)-induced resistance to C(2)-ceramide. The overall data suggest that NF-kappa B mediates the H(2)O(2) adaptation induced in a manner independent of H(2)O(2)-degrading activity.

Molecular cloning and functional expression of bovine brain GABA transaminase.

We isolated a cDNA that encodes the bovine brain gamma-aminobutyrate transaminase (GABA-T; EC 2.6.1.19) from the lambda gt 11 cDNA library, which showed a high degree of sequence similarity to the corresponding enzymes from various sources. Northern blot analysis revealed two differentially expressed GABA-T transcripts of approximately 2.0 and 6.0 kb in the bovine tissues. Southern blot analysis indicates that the two GABA-T transcripts are encoded in a greater-than 10-kb, single-copy gene. Bovine GABA-T cDNA was expressed in E. coli using the pGEX bacterial- expression vector system. The overexpressed GABA-T was enzymatically active after purification, and it had very similar kinetic parameters when compared with those of other mammalian GABA-Ts.

A quantitative histologic analysis of collagen subtypes: the primary obstructed and refluxing megaureter of childhood.

OBJECTIVES: To analyze and identify collagen subtypes in the primary obstructed and refluxing megaureter of childhood. METHODS: Anticollagen monoclonal antibodies to collagen types I, III, and IV were used in control ureters (n = 4), obstructed (n = 7), and refluxing (n = 13) megaureters. Additionally, all were stained with Masson's trichrome to further define the extracellular matrix. After staining and serial sectioning, representative ureteral sections, focusing on the muscularis and lamina propria regions, were digitized and analyzed with a color image analysis system. RESULTS: Immunohistochemical studies demonstrated increases in collagen types I and III for both obstructed and refluxing megaureters compared with controls (P <0.05). Collagen type IV was not detected in statistically significant amounts in any ureter. In control ureters most was type I (83% +/- 9%) collagen. Obstructed megaureters produced similar results with virtually all collagen being type I (84% +/- 26%) with very little type III collagen present, 5.3% +/- 3%. Refluxing megaureters contained only 55% +/- 15% type I collagen. However, there was an increase in type III collagen (16% +/- 4%) versus (4.5% +/- 2%) in controls (P <0.05). CONCLUSIONS: These data suggest that the greater contribution of type III collagen may play a role in the pathophysiology of refluxing megaureters. Because type III collagen is a less distensible fiber, it may cause an intrinsically stiffer ureter and play a role in the lower surgical success in the reimplantation of refluxing megaureters.