Asthma and atopic dermatitis as risk factors for rheumatoid arthritis: a bidirectional mendelian randomization study.

BACKGROUND: Previous observational studies have shown an association between asthma, atopic dermatitis (AD) and rheumatoid arthritis (RA). However, the bidirectional cause-effect chain between asthma and AD and RA has not been proven yet. METHODS: We performed bidirectional two-sample Mendelian randomization (TSMR) and selected single nucleotide polymorphisms (SNPs) associated with asthma, AD, and RA as instrumental variables. All of the SNPs were obtained from the latest genome-wide association study in Europeans. Inverse variance weighted (IVW) was the main method used in MR analysis. MR-Egger, weighted model, simple model, and weighted median were used for quality control. The robustness of the results was tested by sensitivity analysis. RESULTS: Asthma was found to be the largest effect size for RA susceptibility using the IVW method (OR, 1.35;95%CI, 1.13-1.60; P, 0.001), followed by AD (OR, 1.10;95%CI, 1.02-1.19; P, 0.019). In contrast, there was no causal relationship between RA and asthma (IVW: P = 0.673) or AD (IVW: P = 0.342). No pleiotropy or heterogeneity was found in the sensitivity analysis. CONCLUSION: Findings from this study showed a causal relationship between genetic susceptibility to asthma or AD and increased risk of RA, but do not support a causal relationship between genetic susceptibility to RA and asthma or AD.

FAM172A supervises ER (endoplasmic reticulum) stress-triggered autophagy in the epidural fibrosis process.

Backgrounds: Lumbar laminectomy is usually utilized for lumbar disc herniation (LDH), but also causes epidural fibrosis (EF) process associated with abnormal proliferation of fibroblasts. FAM172A is associated with ER stress and cell proliferation, but its mechanism was unclear, especially in the process of EF. Methods: Therefore, the regulation of FAM172A on the calcium flux and autophagy in fibroblasts were investigated by inducing ER stress with tunicamycin and upexpression or downexpression of FAM172A. The calcium flux was determined using Fluo-3, and autophagy was examined with immunofluorescence or western blot for LC3, Beclin-1, ATG-5, and p62. Moreover, the apoptotic protein of Bax and Bcl-2 was detected, too. Furthermore, the laminectomy model was constructed and then dealt with overexpression of FAM172A. Results: Tunicamycin-induced endoplasmic reticulum (ER) stress and autophagy process in fibroblasts were associated with the calcium flux regulated by FAM172A, especially in EF cells. Besides, tunicamycin induced autophagy and suppressed cell apoptosis of fibroblasts. Furthermore, FAM72A repressed the proliferation of fibroblasts and the process of EF in the laminectomy model through the mediation of the autophagy process. Conclusions: Tunicamycin-induced endoplasmic reticulum (ER) stress in fibroblasts was associated with calcium flux mediated by FAM172A. FAM72A participated in the autophagy regulation of fibroblasts and maybe the key interaction regulator of apoptosis and autophagy in fibroblasts, especially for epidural scar cells.

Associations of Dietary and Circulating Vitamin E Level With Metabolic Syndrome. A Meta-Analysis of Observational Studies.

Objective: The associations of dietary and circulating vitamin E level with metabolic syndrome (MetS) remains conflicting. This meta-analysis of observational study was therefore employed to investigate the issue above. Methods: The PubMed, Web of Science and Embase database were searched up to April 2021. The observational studies on the associations of dietary and circulating vitamin E level with MetS were specified. The pooled relative risk (RR) of MetS for the highest vs. lowest dietary and circulating vitamin E level, and the standard mean difference (SMD) of dietary and circulating vitamin E level for MetS vs. control subjects, were calculated. Results: A total of 25 observational studies with 51,276 participants, were included in this meta-analysis. The overall multi-variable adjusted RR demonstrated that the dietary vitamin E level was inversely associated with MetS (RR = 0.92, 95%CI: 0.85-1.00; P = 0.044). In addition, the dietary vitamin E level in MetS was also lower than that in control subjects according to the overall combined SMD (SMD = -0.08, 95%CI: -0.14 to -0.02; P = 0.024). On the other hand, the overall multi-variable adjusted RR showed no significant relationship between the circulating vitamin E level and MetS (RR = 1.46, 95%CI: 0.85-2.48; P = 0.17). However, the circulating vitamin E level in MetS was lower than that in control subjects according to the overall combined SMD (SMD = -0.58, 95%CI: -1.04 to -0.13; P = 0.013). Conclusions: The results of this meta-analysis suggest that the dietary vitamin E level is inversely associated with MetS. On the other hand, current evidence is still insufficient to conclude a relationship between the circulating vitamin E level and MetS. More well-designed prospective cohort studies are needed to address the issues further.

Relationship Between Nut Consumption and Metabolic Syndrome: A Meta-Analysis of Observational Studies.

Objective: Our aim was to examine the relationship between nut consumption and metabolic syndrome (MetS). Methods: The electronic databases of PubMed, Web of Science, and Embase were searched up to November 2018 for observational studies on the relationship between nut consumption and MetS. The pooled relative risk (RR) of MetS for the highest versus lowest category of nut consumption, as well as their corresponding 95% confidence intervals (CIs) were calculated. Results: A total of 11 observational studies (6 cross-sectional and 5 prospective cohort studies), which involved a total of 89,224 participants, were identified for this meta-analysis. The overall multivariable adjusted RR showed that nut consumption was negatively associated with MetS (RR = 0.84; 95% CI, 0.76-0.92; p < 0.001). Of interest, subgroup analysis confirmed that such findings existed in tree nuts (RR = 0.97; 95% CI, 0.94-1.00; p = 0.04), but not in peanuts (RR = 1.01; 95% CI, 0.96-1.06; p = 0.68). Conclusions: The existing evidence suggested that nut consumption was negatively associated with MetS. However, such an inverse relationship only existed in tree nuts, not in peanuts. More well-designed studies with detailed specifications of nut varieties are needed to further elaborate the issues examined in this study.

Red meat, poultry, and egg consumption with the risk of hypertension: a meta-analysis of prospective cohort studies.

The objective of this study was to examine the associations of red meat, poultry, and egg consumption with the risk of hypertension (HTN). The electronic databases of PubMed, Web of Science, and Embase were searched up to August 2017, for prospective cohort studies on the associations between red meat, poultry, or egg consumption with the risk of HTN. The pooled relative risk (RR) of HTN for the highest vs. lowest category of red meat, poultry, and egg consumption as well as their corresponding 95% confidence interval (CI) were calculated. A total of eight articles made up of 10 prospective cohort studies, which involved 351,819 participants and 5000 HTN cases, were included in this meta-analysis. Specifically, nine studies were related to red meat consumption, and the overall multi-variable adjusted RR showed a positive association between red meat consumption and the risk of HTN (RR = 1.22, 95% CI: 1.11-1.35; P < 0.001). In the subgroup analysis that consisted of five studies, both processed (RR = 1.12, 95% CI: 1.02-1.23; P = 0.02) and unprocessed (RR = 1.19, 95% CI: 1.04-1.36; P = 0.01) red meat were associated with a higher risk of HTN. In addition, in the six studies related to poultry consumption, the overall multi-variable adjusted RR demonstrated that poultry consumption was also associated with a higher risk of HTN (RR = 1.15, 95% CI: 1.03-1.28; P = 0.015). Moreover, three of the studies that were included were related to egg consumption, and the overall multi-variable-adjusted RR showed that egg consumption was associated with a lower risk of HTN (RR = 0.79, 95% CI: 0.68-0.91; P = 0.001). The existing evidence suggested that red meat (both processed and unprocessed) and poultry consumption were associated with a higher risk of HTN, while egg consumption was associated with a lower risk of HTN. Owing to the limited number of studies, more well-designed prospective cohort studies are needed to further elaborate the issues examined in this study.

Relationship Between Serum Zinc Level and Metabolic Syndrome: A Meta-Analysis of Observational Studies.

Objective: This research sought to summarize the evidence regarding the relationship between serum zinc level and metabolic syndrome (MetS).Methods: The electronic databases of PubMed, Web of Science, and Embase were searched up to October 2017 for observational studies on the association between serum zinc level and MetS. The standard mean difference (SMD) and its corresponding 95% confidence interval (CI) of the serum zinc level for MetS versus control participants were calculated. In addition, the pooled odds ratio (OR) and relative risk (RR) of MetS for the highest versus lowest category of serum zinc level, as well as their corresponding 95% CI, were also calculated.Results: A total of 11 observational studies (8 cross-sectional, 1 case-control, and 2 cohort studies) were included in this meta-analysis. The combined SMD demonstrated that the serum zinc level in MetS was higher than that in control participants (SMD = 0.11; 95% CI, 0.03-0.19; p = 0.009). Moreover, the overall multivariable-adjusted RR showed that the increased serum zinc level was associated with a higher risk of MetS (RR = 1.82; 95% CI, 1.33-2.50; p < 0.001). On the contrary, the overall multivariable-adjusted OR showed that there was no significant relationship between serum zinc level and MetS (OR = 1.00; 95% CI, 0.99-1.01; p = 0.841).Conclusions: Although the serum zinc level in participants with MetS was significantly higher than that in control ones, the existing evidence was still insufficient to conclude a definite relationship between serum zinc level and MetS. More well-designed prospective cohort studies are needed to elaborate the concerned issues further.

Circulating parathyroid hormone and risk of hypertension: A meta-analysis.

OBJECTIVES: To examine the relationship between circulating parathyroid hormone (PTH) level and risk of hypertension (HTN). METHODS: The electronic databases of PubMed, Web of Science and Embase were searched up to December 2017, for prospective cohort studies on the relationship between circulating PTH level and risk of HTN. The pooled relative risk (RR) of HTN for the highest versus lowest category of circulating PTH level as well as their corresponding 95% confidence interval (CI) were calculated. RESULTS: A total of six prospective cohort studies, which involved 18,994 participants and 5040 HTN cases, were included in this meta-analysis. The overall multi-variable adjusted RR showed a positive relationship between circulating PTH level and risk of HTN (RR = 1.35, 95%CI: 1.09 to 1.67; P = 0.006). A substantial level of heterogeneity was observed among the studies (P < 0.001, I2 = 77.6%). No evidence of publication bias was observed among the studies according to Begg's rank-correlation test (P = 0.452). CONCLUSIONS: The existing evidence suggests that an increase in circulating PTH level may be associated with a higher risk of HTN. However, due to the limited number of included studies, more well-designed prospective cohort studies are needed to further elaborate the issues examined in this study.

Associations of vegetable and fruit consumption with metabolic syndrome. A meta-analysis of observational studies.

OBJECTIVE: To examine the associations of vegetable and/or fruit consumption with metabolic syndrome (MetS). DESIGN: Meta-analysis of observational studies. SETTING: The electronic databases of PubMed, Web of Science and EMBASE were searched up to September 2017 for observational studies concerning the associations of vegetable and/or fruit consumption with MetS. The pooled relative risk (RR) of MetS for the highest v. the lowest category of vegetable and/or fruit consumption, as well as their corresponding 95 % CI, were calculated. RESULTS: A total of twenty-six observational studies (twenty cross-sectional, one case-control and five cohort studies) were included in the meta-analysis. Specifically, sixteen studies were related to vegetable consumption and the overall multivariable-adjusted RR evidenced a negative association between vegetable consumption and MetS (RR=0·89, 95 % CI 0·85, 0·93; P<0·001). For fruit consumption, sixteen studies were included and the overall multivariable-adjusted RR demonstrated that fruit consumption was inversely associated with MetS (RR=0·81, 95 % CI 0·75, 0·88; P<0·001). For vegetable and fruit consumption, eight studies were included; the overall multivariable-adjusted RR showed that vegetable and fruit consumption was also negatively associated with MetS (RR=0·75, 95 % CI 0·63, 0·90; P=0·002). CONCLUSIONS: The existing evidence suggests that vegetable and/or fruit consumption is negatively associated with MetS. More well-designed prospective cohort studies are needed to elaborate the concerned issues further.

Associations of coffee consumption with circulating level of adiponectin and leptin. A meta-analysis of observational studies.

The association between coffee consumption and the circulating level of adiponectin and leptin has been evaluated in several epidemiological studies with conflicting results. Therefore, the aim of this study was to examine the associations of coffee consumption with the circulating level of adiponectin and leptin. A comprehensive literature search up to January 2018, using PUBMED, EMBASE and Web of Science databases, was conducted to identify the relevant observational studies that examined the associations of coffee consumption with the circulating level of adiponectin and leptin. A total of twelve cross-sectional studies were included in this meta-analysis. According to the combined standard mean difference (SMD) between the highest and the lowest coffee intake category, coffee consumption was associated with a higher circulating adiponectin level (SMD = 0.11, 95%CI: 0.06-0.17; P < .001). Subgroup analysis confirmed such findings in females (SMD = 0.11, 95%CI: 0.02-0.20; P = .01), but not in males (SMD = 0.03, 95%CI: -0.08 to 0.14; P = .59). In addition, the combined SMD showed that coffee consumption was negatively associated with the circulating level of leptin (SMD = -0.19, 95%CI: -0.28 to -0.10; P < .001). The results of this meta-analysis suggested that coffee consumption was associated with a higher circulating level of adiponectin. Additionally, we showed that coffee consumption was inversely associated with the circulating level of leptin. More well-designed prospective cohort studies and randomised controlled trials are needed to further elaborate the concerned issues.

Is coffee consumption associated with a lower level of serum C-reactive protein? A meta-analysis of observational studies.

The association between coffee consumption and the level of C-reactive protein (CRP) has been evaluated in several epidemiological studies with conflicting results. This study aims to examine the relationship between coffee consumption and the serum CRP level. A comprehensive literature search up to August 2017, using PUBMED, EMBASE and Web of Science databases, was conducted to identify the relevant observational studies that examined the association between coffee consumption and the serum CRP level. A total of nine cross-sectional studies were included in this meta-analysis. According to the combined standard mean difference (SMD) between the highest and the lowest coffee intake category, coffee consumption was associated with a lower level of serum CRP level (SMD = -0.34, 95%CI: -0.62 to -0.06; p = .016). Subgroup analysis for CRP marker showed that coffee consumption was associated with a lower level of serum high-sensitivity CRP (hsCRP) (SMD = -0.51, 95%CI: -0.88 to -0.14; p = .007), but not standard CRP (SMD = 0.02, 95%CI: -0.28 to 0.32; p = .913). The existing evidence suggested that coffee consumption was associated with a lower level of serum CRP. More well-designed prospective cohort studies are needed to elaborate the concerned issues further.

ATF3 promotes migration and M1/M2 polarization of macrophages by activating tenascin­C via Wnt/ß­catenin pathway.

There are different polarization states of macrophages, including the classically activated M1 phenotype and the alternatively activated M2 phenotype. These have different functions in the inflammation process. Activating transcription factor 3 (ATF3) is a key transcriptional regulator that inhibits the inflammatory response. However, the effects of ATF3 on migration and anti­inflammatory control mechanisms of macrophages have not been thoroughly investigated. The present study investigated the effect of ATF3 on macrophage migration and M1/M2 polarization. Results revealed that overexpression of ATF3 promoted macrophage migration and the expression of the M2 phenotype markers [cluster of differentiation (CD) 163, mannose receptor C type 1, arginase 1 and peroxisome proliferator­activated receptor γ] and inhibited expression of the M1 phenotype markers (monocyte chemoattractant protein­1, inducible nitric oxide synthase, CD16 and tumor necrosis factor­α), whereas knockdown of ATF3 resulted in a contrary effect. In addition, the wingless­type MMTV integration site family member (Wnt)/ß­catenin signaling pathway was activated and the expression level of tenascin (TNC) was significantly upregulated by overexpression of ATF3. Additionally, inhibition of Wnt/ß­catenin signaling significantly attenuated the upregulatory effect of ATF3 on TNC. Finally, the effect of ATF3 on macrophage migration and markers of the M1 or M2 state was investigated using TNC­specific siRNA. In conclusion, the results of the present study suggested that ATF3 promotes macrophage migration and reverses M1­polarized macrophages to the M2 phenotype by upregulation of TNC via the Wnt/ß­catenin signaling pathway.

mTOR signal transduction pathways contribute to TN-C FNIII A1 overexpression by mechanical stress in osteosarcoma cells.

Osteosarcoma is the most common primary malignant bone tumor with a very poor prognosis. Treating osteosarcoma remains a challenge due to its high transitivity. Tenascin-C, with large molecular weight variants including different combinations of its alternative spliced FNIII repeats, is specifically over expressed in tumor tissues. This study examined the expression of Tenascin-C FNIIIA1 in osteosarcoma tissues, and estimated the effect of mechanical stimulation on A1 expression in MG-63 cells. Through immunohistochemical analysis, we found that the A1 protein was expressed at a higher level in osteosarcoma tissues than in adjacent normal tissues. By cell migration assay, we observed that there was a significant correlation between A1 expression and MG-63 cell migra-tion. The relation is that Tenascin-C FNIIIA1 can promote MG-63 cell migration. According to our further study into the effect of mechanical stimulation on A1 expression in MG-63 cells, the mRNA and protein levels of A1 were significantly up-regulated under mechanical stress with the mTOR molecule proving indispensable. Meanwhile, 4E-BP1 and S6K1 (downstream molecule of mTOR) are necessary for A1 normal expression in MG-63 cells whether or not mechanical stress has been encountered. We found that Tenascin-C FNIIIA1 is over-expressed in osteosar-coma tissues and can promote MG-63 cell migration. Furthermore, mechanical stress can facilitate MG-63 cell migration though facilitating A1 overexpression with the necessary molecules (mTOR, 4E-BP1 and S6K1). In con-clusion, high expression of A1 may promote the meta-stasis of osteosarcoma by facilitating MG-63 cell migration. Tenascin-C FNIIIA1 could be used as an indicator in metastatic osteosarcoma patients.

Tumoricidal effects of a selenium (Se)-polysaccharide from Ziyang green tea on human osteosarcoma U-2 OS cells.

Selenium(Se)-enriched green tea consumption in human diets is well-known to reduce the risk of a variety of diseases. Here, we isolated a Se-polysaccharide (Se-ZYTP) from Se-enriched Ziyang green tea and investigated its antitumor activity on human osteosarcoma U-2 OS cells in vitro and in vivo. Se-ZYTP contained 94.5% of carbohydrate and 2.1% of uronic acid, as well as 2.14 µg/g Se, revealing that Se-ZYTP was an acidic Se-conjugated polysaccharide. Monosaccharide composition analysis indicated that Se-ZYTP consisted of mannose, rhamnose and fucose in molar ratios of 2.4:1.5:1.2:0.2:0.1:0.3:0.3. In vitro, both MTT and LTH assays proved that Se-ZYTP (25, 50, 100 and 200 µg/ml) could significantly inhibit the proliferation of human osteosarcoma U-2 OS cells in a concentration-dependent fashion (P<0.05 or P<0.01). In U-2 OS cancer xenograft model in BALB/c athymic mice, Se-ZYTP oral administration at three doses of 100, 200 and 400mg/kg body weight (B.W.) daily for 28 days resulted in obvious tumor regression as compared to model control (P<0.05 or P<0.01). In addition, body weights of mice in control or Se-ZYTP treated groups did not differ significantly and no mice died during experiment, suggesting the safety of Se-ZYTP. Therefore, we postulate that Se-ZYTP might have cancer-preventive and cancer-therapeutic benefit for human osteosarcoma.

Wavelet analysis for electrocardiogram variation of arrhythmia patients / 生物医学工程学杂志

To explore the energy proportion of the QRS complexes in the electrocardiogram (ECG), a new method, wavelet analysis, has been presented. Using wavelet transform of ECG, we can calculate the energy proportion of QRS complexes in multiple scales. The electrocardiograms (ECGs) were used as the experimental data, which were collected from a young (21-34 yr) group and an elderly (68-81 yr) group of healthy subjects, as well as from a group of arrhythmia patients (66-81 yr). the data analysis was performed with the energy proportion of the QRS complexes in the ECG using Mexican-Hat as a mother wavelet in multiple scales. Results showed that the energy proportion of the QRS complexes had no changes with ages increasing (P > 0.44), but in the same age group, the arrhythmia patients' energy proportion of the QRS complexes near 17Hz are obviously less than that in the healthy group (P < 0.01), so the energy proportion of the QRS complexes calculated by wavelet analysis can be used as a feature index to judge whether a person is a sinus arrhythmia patient or not.

[Establishment and primary application of sandwich ELISA method to detect tenascin-C].

AIM: To establish a sandwich method to detect tenascin-c on the basis of preparation of monoclonal antibodies (mAbs) against tenascin-C (TN-C). METHODS: The ascites of three stains of mAbs (No. 1A8, 3H7 and 4D6) were prepared and purified. The mAbs were conjugated with HRP and paired, respectively. The recombinant TN-C was taken as standard to analyze the optimal combination between mAbs. The sera TN-C concentrations of patients with osteosarocoma and the normal persons were evaluated with the sandwich ELISA method. RESULTS: Among these mAbs, the sensitivity was obtained when combined the coated 1A8 with HRP-4D6. The sera TN-C significantly higher than the normal controls. CONCLUSION: The sandwich ELISA method to detect TN-C was established successfully. The sera TN-C concentrations of patients with osteosarcoma and the normal persons were found distinct with the sandwich method.

Clinical value of signal transducers and activators of transcription 3 (STAT3) gene expression in human osteosarcoma.

The dysregulation of signal transducers and activators of transcription 3 (STAT3) has been reported to be associated with tumor progression, angiogenesis and metastasis. The purpose of this study was to analyze the clinical value of STAT3 expression in human osteosarcoma. First, semi-quantitative RT-PCR was performed to detect the expression of STAT3 mRNA in normal bone tissues, chondroma tissues and osteosarcoma tissues. Then, immunohistochemistry was performed to detect the expression of STAT3 protein in 76 osteosarcoma tissues and the relationship of STAT3 protein expression with clinicopathologic factors or prognosis of osteosarcoma patients. RNA interference (RNAi) technology was employed to inhibit STAT3 expression. MTT and flow cytometric assays were performed to analyze the effect of STAT3 inhibition on proliferation and apoptosis of osteosarcoma cells. Finally, the expression of STAT3-related target genes were also determined. Results showed that osteosarcoma tissues showed significantly higher expression levels of STAT3 mRNA than normal bone or chondroma tissues (P<0.05). Immunohistochemistry showed that the staining of STAT3 protein was mainly located in cytoplasm of osteosarcoma cells in osteosarcoma tissue samples. The high level of STAT3 protein was associated with poor tumor differentiation and presentation of metastasis (P=0.039 and 0.022). Moreover, the 5-year overall and relapse-free survival rates for osteosarcoma patients with high STAT3 expression were lower than those for patients with low STAT3 expression. In addition, the status of STAT3 protein expression was an independent prognostic factor for both disease-free survival (P=0.0235) and overall survival (P=0.0032). RNAi-mediated STAT3 inhibition could induce proliferation inhibition and apoptosis enhancement in osteosarcoma cells, which might be associated with inhibition of some anti-apoptosis genes. Overall, STAT3 plays crucial roles in osteosarcoma development and might become a potential molecular target for gene therapy of human osteosarcomas.

Dual sources of vitronectin in the human lower urinary tract: synthesis by urothelium vs. extravasation from the bloodstream.

Vitronectin (VN), secreted into the bloodstream by liver hepatocytes, is known to anchor epithelial cells to basement membranes through interactions with cell surface integrin receptors. We report here that VN is also synthesized by urothelial cells of urothelium in vivo and in vitro. In situ hybridization, dideoxy sequencing, immunohistochemistry, and ELISA of urothelial cell mRNA, cDNA, tissue, and protein extracts demonstrated that the VN gene is active in vivo and in vitro. The expression of VN by urothelium is hypothesized to constitute one of several pathways that anchor basal cells to an underlying substratum and explains why urothelial cells adhere to glass and propagate under serum-free conditions. Therefore, two sources of VN in the human urinary bladder are recognized: 1) localized synthesis by urothelial cells and 2) extravasation of liver VN through fenestrated capillaries. When human plasma was fractionated by denaturing heparin affinity chromatography, VN was isolated in a biologically active form that supported rapid spreading of urothelial cells in vitro under serum-free conditions. This activity was inhibited by the matricellular protein SPARC via direct binding of VN to SPARC through a Ca(+2)-dependent mechanism. A novel form of VN, isolated from the same heparin affinity chromatography column and designated as the VN(c) chromatomer, also supported cell spreading but failed to interact with SPARC. Therefore, the steady-state balance among urothelial cells, their extracellular milieu, and matricellular proteins constitutes a principal mechanism by which urothelia are anchored to an underlying substrata in the face of constant bladder cycling.

Fibroblast growth factor-10 signals development of von Brunn's nests in the exstrophic bladder.

von Brunn's nests have long been recognized as precursors of benign lesions of the urinary bladder mucosa. We report here that von Brunn's nests are especially prevalent in the exstrophic bladder, a birth defect that predisposes the patient to formation of bladder cancer. Cells of von Brunn's nest were found to coalesce into a stratified, polarized epithelium which surrounds itself with a capsule-like structure rich in types I, III, and IV collagen. Histocytochemical analysis and keratin profiling demonstrated that nested cells exhibited a phenotype similar, but not identical, to that of urothelial cells of transitional epithelium. Immunostaining and in situ hybridization analysis of exstrophic tissue demonstrated that the FGF-10 receptor is synthesized and retained by cells of von Brunn's nest. In contrast, FGF-10 is synthesized and secreted by mesenchymal fibroblasts via a paracrine pathway that targets basal epithelial cells of von Brunn's nests. Small clusters of 10pRp cells, positive for both FGF-10 and its receptor, were observed both proximal to and inside blood vessels in the lamina propria. The collective evidence points to a mechanism where von Brunn's nests develop under the control of the FGF-10 signal transduction system and suggests that 10pRp cells may be the original source of nested cells.

Insights into lignin primary structure and deconstruction from Arabidopsis thaliana COMT (caffeic acid O-methyl transferase) mutant Atomt1.

The Arabidopsis mutant Atomt1 lignin differs from native lignin in wild type plants, in terms of sinapyl (S) alcohol-derived substructures in fiber cell walls being substituted by 5-hydroxyconiferyl alcohol (5OHG)-derived moieties. During programmed lignin assembly, these engender formation of benzodioxane substructures due to intramolecular cyclization of their quinone methides that are transiently formed following 8-O-4' radical-radical coupling. Thioacidolytic cleavage of the 8-O-4' inter-unit linkages in the Atomt1 mutant, relative to the wild type, indicated that cleavable sinapyl (S) and coniferyl (G) alcohol-derived monomeric moieties were stoichiometrically reduced by a circa 2 : 1 ratio. Additionally, lignin degradative analysis resulted in release of a 5OHG-5OHG-G trimer from the Atomt1 mutant, which then underwent further cleavage. Significantly, the trimeric moiety released provides new insight into lignin primary structure: during polymer assembly, the first 5OHG moiety is linked via a C8-O-X inter-unit linkage, whereas subsequent addition of monomers apparently involves sequential addition of 5OHG and G moieties to the growing chain in a 2 : 1 overall stoichiometry. This quantification data thus provides further insight into how inter-unit linkage frequencies in native lignins are apparently conserved (or near conserved) during assembly in both instances, as well as providing additional impetus to resolve how the overall question of lignin macromolecular assembly is controlled in terms of both type of monomer addition and primary sequence.

FGF-10 and its receptor exhibit bidirectional paracrine targeting to urothelial and smooth muscle cells in the lower urinary tract.

Control of the regenerative properties of urothelial tissue would greatly aid the clinician in the management of urinary tract disease and disorders. Fibroblast growth factor 10 (FGF-10) is a mitogen which is particularly promising as a protein therapy for urothelial injury. The spatial synthesis, transport, targeting, and mechanistic pathway of FGF-10 and its receptor were studied in a human urothelial cell culture model and in fixed sections of lower urinary tract tissue. Synthesis of FGF-10 was restricted to mesenchymal fibroblasts, and secreted FGF-10 exhibited paracrine transport to two proximal sites, transitional epithelium and smooth muscle cell bundles, both of which were also the exclusive sites of FGF-10 receptor synthesis. The addition of recombinant FGF-10 to quiescent urothelial cells in vitro was sufficient to stimulate DNA synthesis. This stimulation was through a pathway independent of the epidermal growth factor receptor pathway. Deconvolution, light and transmission electron microscopic studies captured FGF-10 and its receptor in association with the urothelial cell surface, in cytoplasm, and within nuclei, observations that describe the mechanism that transduces the mitogenic signal in these tissues. Localization of the FGF-10 receptor to the superficial urothelial layer is clinically significant because intravesical administration of FGF-10 may provide the clinician a means to control the turnover of transitional epithelium in bladder disorders such as interstitial cystitis.