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Dentin bonding is a dynamic process that involves the penetration of adhesive resin monomers into the extrafibrillar and intrafibrillar demineralized collagen matrix using a wet-bonding technique. However, adhesive resin monomers lack the capacity to infiltrate the intrafibrillar space, and the excess water that is introduced by the wet-bonding technique remains at the bonding interface. This imperfectly bonded interface is inclined to hydrolytic degradation, severely jeopardizing the longevity of bonded clinical restorations. The present study introduces a dentin bonding scheme based on a dry-bonding technique, combined with the use of extrafibrillar demineralization and a collagen-reactive monomer (CRM)-based adhesive (CBA). Selective extrafibrillar demineralization was achieved using 1-wt% high-molecular weight (MW) carboxymethyl chitosan (CMCS) within a clinically acceptable timeframe to create a less aggressive bonding substance for dentin bonding due to its selectively extrafibrillar demineralization capacity. CMCS demineralization decreased the activation of in situ collagenase, improved the shrinking resistance of demineralized collagen, and thus provided stronger and more durable bonding than traditional phosphoric acid etching. The new dentin bonding scheme that contained CMCS and CBA and used a dry-bonding technique achieved an encouraging dentin bonding strength and durability with low technical sensitivity. This bonding scheme can be used to improve the stability of the resin-dentin interface and foster the longevity of bonded clinical restorations., Competing Interests: The authors declare no competing interest., (© 2021 The Authors.)

Resin cements have been widely employed for bonding all-ceramic restorations in clinical practice, its color stability is directly related to long-term prosthetic effect of restorations. Discoloration of resin cements can be attributed to two causes: endogenous factors are generally related to material compositions and initiation mechanism of polymerization; exogenous factors are mainly related to stimulation of local oral environment. Color stability of resin cements has close relationship with esthetic effect of all-ceramic restorations. The aim of this literature review was to make a presentation and discussion systematically about color stability of resin cements commonly used clinically, its influence factors and influence on all-ceramic restorations, so as to provide a reference for the application of all-ceramic restorations.

An 8-week growth trial was conducted to investigate the effects of replacing dietary fishmeal with a plant protein blend on the growth performance, mucosal barrier integrity and the related regulation mechanism in Amur Sturgeon (Acipenser schrenckii) with initial weight of 87.48 g. Three isonitrogenous and isoenergetic diets were prepared. A basal diet containing 540 g/kg fishmeal (P0), whereas the other two diets were formulated by replacing 50% and 100% of FM with plant protein blend (soybean protein concentrate and cottonseed protein concentrate), and named as P50 and P100, respectively. Although essential amino acids, fatty acids, and available phosphorus had been balanced according to the nutrient requirement of sturgeon, compared with the fish of P0 and P50, the full plant protein diet (P100) significantly reduced growth performance and survival, and accompanied with serious spiral valve intestinal (SVI) damage. The increased tissue necrosis and failed responses in anti-oxidation, programming apoptosis, autophagy and cell proliferation system were regulated by inhibiting ERK1 phosphorylation, which indicated that SVI hypoimmunity and functional degradation were the main reasons for the high mortality and low utilization ability of plant protein in Amur sturgeon., (Copyright © 2019. Published by Elsevier Ltd.)

Through the high temperature solution method, three new phosphates, Cs 8 Pb 4 (P 2 O 7 ) 4 , CsLi 7 (P 2 O 7 ) 2 and LiCa(PO 3 ) 3 , were obtained. X-ray diffraction analyses suggest that they crystallize in the space group P1[combining macron] of the triclinic crystal system. However, the three-dimensional (3D) structures of Cs 8 Pb 4 (P 2 O 7 ) 4 and CsLi 7 (P 2 O 7 ) 2 consist of various isolated P 2 O 7 groups, while LiCa(PO 3 ) 3 is built up by the 1D (PO 3 ) ∞ chain. Detailed structural comparison suggests that the different sizes and coordination environments of Ca 2+ and Zn 2+ cations influence the whole structures of LiX(PO 3 ) 3 (X = Ca and Zn). IR spectroscopy and UV/Vis/NIR diffuse reflectance spectroscopy analyses have been performed to investigate their optical properties. First-principles theoretical studies were also carried out to aid the understanding of electronic structures and linear optical properties.

Objective: To evaluate the efficacy, safety and remission rates of pruritus of bezafibrate and UDCA combination therapy in the treatment of refractory PBC. Methods: PubMed, Embase, The Cochrane Library databases, Science direct, Web of Science, CBM, WangFang Data, CNKI, VIP databases were searched to collect randomized controlled trials, crossover trials and self-control clinical trials of combination therapy of UDCA and bezafibrate with UDCA monotherapy for PBC up to June, 2018. RevMan 5.3 software was used for meta-analysis. Two evaluators independently screened the literature, extracted the data, and evaluated the risk of bias of relevant study. Results: Eleven studies, including 465 patients were included. Ursodeoxycholic acid combined with bezafibrate had greatly improved liver biochemical indicators ( P < 0.01) and pruritus scores in patients with refractory primary biliary cholangitis ( MD = -2.97, 95% CI : -4.34~ -1.60, P < 0.01). However, there was no statistically significant differences in adverse events ( RR = 1.28, 95% CI : 0.96 to 1.70, P = 0.09), and mortality rate ( RR = 2.58, 95% CI : 0.57 to 11.73, P = 0.22) between the two groups. Conclusion: Ursodeoxycholic acid combined with bezafibrate may improve the biochemical response and pruritus score of refractory PBC, but has no significant effect on adverse events and mortality rate.

A new barium borate halide, Ba3B10O17Br2, has been obtained. A detailed structural comparison with other barium borate halides suggested that Ba3B10O17Br2 is the first barium borate halide with a B-O layered structure. First-principles theoretical studies were conducted to aid understanding of the electronic structure and optical properties.

Objective: To explore the effects of oleic acid and palmitic acid on lipid deposition and mTOR/S6K1/SREBP-1c pathways in HepG2 cells. Methods: The model of steatosis was established with induction of oleic acid and palmitic acid and was intervened by rapamycin. The changes in lipid droplets were observed after staining the cells with oil Red O. Intracellular triglyceride (TG) contents in cells were measured by TG kit. mTOR, S6K1, and SREBP-1c mRNA expression levels were detected using QRT-PCR. Western blot was used to determine protein expression levels of mTOR, S6K1 and SREBP-1c. Results: Both fatty acids increased lipid droplets in HepG2 cells. Fatty degeneration with elevated TG occurred with significant changes in oleic acid group lipids. Rapamycin alleviated lipid deposition caused by oleic acid and palmitic acid and inhibited their induction of increased expression of mTOR, S6K1, and SREBP-1c. QRT-PCR and Western blot results showed that mRNA and protein expressions of mTOR, S6K1, and SREBP-1c in oleic acid and palmitic acid group were significantly higher than the control group ( P < 0.05). The increase was more pronounced in the palmitic acid group ( P < 0.05); however, after rapamycin intervention, the expression of mRNA and protein in the three groups were significantly lower ( P < 0.05), and the change in palmitic acid group was more pronounced ( P < 0.05). Conclusion: Oleic acid and palmitic acid can induce lipid deposition in HepG2 cells and increase expression of every component of mTOR/S6K1/SREBP-1c pathway; however, Oleic acid-induced lipid deposition is more pronounced, and the mTOR, S6K1, and SREBP-1c pathway change is more obvious in palmitic acid. Rapamycin has high potent inhibitory effect on palmitic acid-induced lipid deposition. These results specify that lipid synthesis involved in the mTOR/S6K1/SREBP-1c pathways are mainly associated to palmitic acid in HepG2 cells, whereas other signaling pathway may mediate oleic acid-induced lipid synthesis.

Injection of triamcinolone acetonide is a non-operative treatment for early-stage Dupuytren disease in Caucasians, but its effectiveness in non-Caucasians is unclear. We report averaged 5-year follow-up results of 37 patients (49 affected hands) with early-stage Dupuytren disease for patients in Taiwan (non-Caucasian) who received a single dose of 5 mg triamcinolone acetonide injection into nodules monthly for 3 months. Using ultrasound, we recorded no progression of sizes of the modules following injection after 6 months. After an average 5-year follow-up, two patients with three hands (6%) experienced reactivation of the treated nodules. None required surgical intervention. Ultrasound examination showed that sizes of the treated Dupuytren nodules decreased significantly by 40% 6 months after injection and 56% at the final follow-up. We conclude that in these Chinese patients in Taiwan with early Dupuytren nodules, triamcinolone acetonide injection was effective in reducing the size of the Dupuytren nodules and maintaining long-term durable control of the nodular growth., Level of Evidence: III.

Objective: To evaluate the reliability and validity of the Chinese version on comprehensive scores for financial toxicity (COST), based on the patient-reported outcome measures. Methods: A total of 118 cancer patients were face-to-face interviewed by well-trained investigators. Cronbach's α and Pearson correlation coefficient were used to evaluate reliability. Content validity index (CVI) and exploratory factor analysis (EFA) were used to evaluate the content validity and construct validity, respectively. Results: The Cronbach's α coefficient appeared as 0.889 for the whole questionnaire, with the results of test-retest were between 0.77 and 0.98. Scale-content validity index (S-CVI) appeared as 0.82, with item-content validity index (I-CVI) between 0.83 and 1.00. Two components were extracted from the Exploratory factor analysis, with cumulative rate as 68.04% and loading>0.60 on every item. Conclusion: The Chinese version of COST scale showed high reliability and good validity, thus can be applied to assess the financial situation in cancer patients.

Objective: To assess the accuracy of computed tomographic colonography (CTC) for detection of colorectal neoplasm. Methods: Publications prior to January 2016 from the Medline, Embase, CNKI, WANFANG, and VIP literature databases were systematically reviewed. A QUADAS checklist was used to assess the quality of the studies. According to the sizes of tumor (≥6 mm or ≥10 mm), diagnostic test accuracy indexes (area under the curve, sensitivity and specific) were pooled and stratified. Spearman correlation and curve of summary receiver operating characteristic (SROC) were applied to comprehensively assess the threshold effect. A bi-variate mixed-effects model was used for testing the overall merging value and heterogeneity. Results: A total of 19 articles ( n =11 540) were included in the analysis. A total of 18 studies were from Europe and Unite States, with 1 from Asia. Numbers of studies regarding information on general population, high-risk groups and clinical symptoms were 9, 5 and 5, respectively. In 19 articles, data on the accuracy of CTC diagnosis (≥6 mm group) was recorded, with another 17 on the group ≥10 mm. Area under the SROC curve in the ≥6 mm group was 0.92 (95 % CI : 0.92-0.94). It was estimated that the pooled sensitivity and specificity were 0.80 (95 % CI : 0.73-0.86) and 0.89 (95 % CI : 0.86-0.92) respectively. In area under the SROC curve, the pooled sensitivity and specificity in ≥10 mm group were 0.87(95 %CI :0.80-0.92), 0.97(95 %CI :0.95-0.98), respectively. Results from the subgroup showed that the sensitivity of with or without the fecal tagging groups were 0.84 and 0.67 in the ≥6 mm group, 0.92 and 0.76 in the ≥10 mm group. It was also found that in the experienced or less experienced readers, rates of sensitivity were 0.83 and 0.75 in the ≥6 mm group, 0.91 and 0.79 in the ≥10 mm group. Conclusions: CTC had high diagnostic efficiency for colorectal neoplasm, especially in the ≥10 mm group. Fecal tagging and experience of the reasers can improve the diagnostic sensitivity.

Objective: To evaluate the effects of epigallocatechin-3-gallate (EGCG) modification on the bonding stability of an etch-and-rinse adhesive to intraradicular dentin, and to find a new strategy to improve the stability of bonding interface. Methods: EGCG was incorporated into Single Bond 2 (SB2) with the concentration of 200 mg/L and 400 mg/L respectively to fabricated experimental adhesives group A and group B, while Single Bond 2 without EGCG was used as control group. Laser scanning confocal microscope (LSCM) and scanning electron microscope (SEM) were used to observe the bacterial biofilm adherent to the surface of the cured adhesive. Micro-Raman spectrum was used to test the degree of conversion (DC) of adhesives. The push-out bond strength of instant testing and aging with thermocycling for 5 000 times were also tested. Results: Group A and group B showed inhibiting effect on the biofilm formation of Enterococcus faecalis and performed better with higher concentration. No significant differences were detected in DC among group A ([69.73±0.68]%), group B ([69.03±1.65]%) and control group ([70.06±1.62]%) ( P> 0.05), and the immediate push-out bond strength of control group ([10.45±2.00] MPa) was not compromised compared to group A ([10.02±2.03] MPa) and group B ([9.95±3.03] MPa) ( P> 0.05). After thermocycling for 5 000 times, group A ([7.01±1.39] MPa) and group B ([7.62±1.88] MPa) showed significantly higher push-out bond strength than control group did ([5.08±1.56] MPa) ( P< 0.05). Conclusions: EGCG modified etch-and-rinse adhesives showed anti-bacterial effect and enhanced bonding stability of intraradicular dentin-adhesive interfaces.

Background: Bone metastases occur frequently in patients with advanced cancer and are a serious complication of cancer. The decision to treat is often individualized, based on each patient's clinical presentation, life expectancy, and quality of life., Methods: We reviewed the current literature pertaining to management of metastatic disease to bone, and the medical, radiotherapeutic, and surgical treatment options for management of bone metastasis are discussed., Results: Current management of skeletal metastasis includes analgesia, systemic therapy, radiation therapy, and surgery. We propose treatment algorithms for management of vertebral and nonvertebral bone metastases and suggest individualized interventions based on clinical presentation., Conclusions: Management of bone metastases is complex and requires a multidisciplinary approach. The goal of treatment is often palliative, and intervention and treatment regimens should be individualized based on the specific clinical presentation of each patient.

Background: Radiation therapy is a common and effective treatment modality in the management of skeletal metastases. Recent advances in technology permitting delivery of an ablative radiation dose with an image-guided stereotactic approach improve the therapeutic threshold., Methods: The authors reviewed the literature on conventional external-beam radiation therapy and summarized the emerging data about image-guided stereotactic body radiation therapy (SBRT) for vertebral oligometastasis., Results: Pain control can be achieved effectively with conventional external-beam radiation therapy and may be further improved with image-guided spinal SBRT. Image-guided SBRT allows delivery of an ablative radiation dose with minimal toxicity, may potentially improve local tumor control, and may enhance clinical outcomes for histologies that are considered radioresistant. However, further understanding of long-term normal tissue toxicity is lacking., Conclusions: Radiotherapy options are expanding for patients with skeletal metastases. Image-guided spinal SBRT can deliver a safe ablative radiation dose to improve pain control and potentially local tumor control. Randomized clinical trials are ongoing to assess clinical benefits and outcome with spinal SBRT.

EphB2 is a receptor tyrosine kinase of the Eph family and ephrin-B1 is one of its transmembrane ligands. In the embryo, EphB2 and ephrin-B1 participate in neuronal axon guidance, neural crest cell migration, the formation of blood vessels, and the development of facial structures and the inner ear. Interestingly, EphB2 and ephrin-B1 can both signal through their cytoplasmic domains and become tyrosine-phosphorylated when bound to each other. Tyrosine phosphorylation regulates EphB2 signaling and likely also ephrin-B1 signaling. Embryonic retina is a tissue that highly expresses both ephrin-B1 and EphB2. Although the expression patterns of EphB2 and ephrin-B1 in the retina are different, they partially overlap, and both proteins are substantially tyrosine-phosphorylated. To understand the role of ephrin-B1 phosphorylation, we have identified three tyrosines of ephrin-B1 as in vivo phosphorylation sites in transfected 293 cells stimulated with soluble EphB2 by using mass spectrometry and site-directed mutagenesis. These tyrosines are also physiologically phosphorylated in the embryonic retina, although the extent of phosphorylation at each site may differ. Furthermore, many of the tyrosines of EphB2 previously identified as phosphorylation sites in 293 cells (Kalo, M. S., and Pasquale, E. B. (1999) Biochemistry 38, 14396-14408) are also phosphorylated in retinal tissue. Our data underline the complexity of ephrin-Eph bidirectional signaling by implicating many tyrosine phosphorylation sites of the ligand-receptor complex.

Purpose: The purpose of the study was to determine the incidence of GERD associated with prescription NSAID consumption., Methods: All Georgia Medicaid patients > 25 years of age and continuously eligible for 1996, 1997, and 1998 were included in the study. Patients were excluded if they received a GERD diagnoses during 1996 and 1997. Patients were observed in 1998 and classified into GERD and control cohorts. Comorbidities, demographics, and NSAID prescription consumption were retained and modeled with logistic regression., Results: The absolute risk of developing GERD without previous NSAID consumption was 0.38. The absolute risk of developing GERD for those patients who consumed one or more NSAID prescriptions during 1996 and 1997 was 0.80. Thus, the relative risk of GERD for NSAID patients was 2.11. GERD was significantly associated with one or more NSAID prescriptions (OR = 1.82), age (OR = 1.05 for 5 year range), gender (OR = 1.31 for females), asthma (OR = 3.24), obesity (OR = 2.77), hiatal hernia (OR = 4.17), tobacco use (OR = 2.56), and alcohol (OR = 1.83). The initial NSAID prescription was responsible for the greatest marginal increase in GERD., Conclusions: Our study suggests that NSAIDs are associated with GERD especially for females, alcohol and tobacco users, and patients with asthma, hiatal hernia, or obesity.

The Eph family of receptor tyrosine kinases and the Abl family of non-receptor tyrosine kinases have both been implicated in tissue morphogenesis. They regulate the organization of the actin cytoskeleton in the developing nervous system and participate in signaling pathways involved in axon growth. Both Eph receptors and Abl are localized in the neuronal growth cone, suggesting that they play a role in axon pathfinding. Two-hybrid screens identified regions of Abl and Arg that bind to the EphB2 and EphA4 receptors, suggesting a novel signaling connection involving the two kinase families. The association of full-length Abl and Arg with EphB2 was confirmed by co-immunoprecipitation and found to involve several distinct protein interactions. The SH2 domains of Abl and Arg bind to tyrosine-phosphorylated motifs in the juxtamembrane region of EphB2. A second, phosphorylation-independent interaction with EphB2 involves non-conserved sequences in the C-terminal tails of Abl and Arg. A third interaction between Abl and EphB2 is probably mediated by an intermediary protein because it requires tyrosine phosphorylation of EphB2, but not the binding sites for the Abl SH2 domain. The connection between EphB2 and Abl/Arg appears to be reciprocal. Activated EphB2 causes tyrosine phosphorylation of Abl and Arg, and vice versa. Interestingly, treatment of COS cells and B35 neuronal-like cells with ephrin-B1 to activate endogenous EphB2 decreased the kinase activity of endogenous Abl. These data are consistent with the opposite effects that Eph receptors and Abl have on neurite ougrowth and suggest that Eph receptors and Abl family kinases have shared signaling activities.

Intact soil-core microcosms were used to compare persistence of Pseudomonas chlororaphis 3732RN-L11 in fallow soil and on wheat roots with field releases at diverse sites. Parallel field and microcosm releases at four sites in 1996 were repeated with addition of one site in 1997. Microcosms were obtained fresh and maintained at 60% soil water holding capacity in a growth chamber at 70% relative humidity, a 12-hour photoperiod, and constant temperature. Persistence of 3732RN-L11 was measured at each site in field plots and microcosms at 7-21 day intervals, and in duplicate microcosms sampled at an independent laboratory. Linear regression slopes of field plot and microcosm persistence were compared for each site, and between identical microcosms sampled at different sites, using log10 transformed plate counts. Microcosm persistence closely matched field plots for wheat roots, but persistence in fallow soil differed significantly in several instances where persistence in field plots was lower than in microcosms. Analysis of weather variations at each site indicated that rainfall events of 30-40 mm caused decreased persistence in fallow soil. Cooler temperatures enhanced persistence in field plots at later time points. Inter-laboratory comparison of regression slopes showed good agreement for data generated at different sites, though in two instances, longer sampling periods at one site caused significant differences between the sites. Soil characteristics were compared and it was found that fertility, namely the carbon to nitrogen ratio, and the presence of expanding clays, were related to persistence. These microcosm protocols produced reliable data at low cost, and were useable for pre-release risk analyses for microorganisms.

Semaphorins comprise a large family of phylogenetically conserved secreted and transmembrane glycoproteins, many of which have been implicated in repulsive axon guidance events. The transmembrane semaphorin Sema-1a in Drosophila is expressed on motor axons and is required for the generation of neuromuscular connectivity. Sema-1a can function as an axonal repellent and mediates motor axon defasciculation. Here, by manipulating the levels of Sema-1a and the cell adhesion molecules fasciclin II (Fas II) and connectin (Conn) on motor axons, we provide further evidence that Sema-1a mediates axonal defasciculation events by acting as an axonally localized repellent and that correct motor axon guidance results from a balance between attractive and repulsive guidance cues expressed on motor neurons.

The enteric peptides, guanylin and uroguanylin, are local regulators of intestinal secretion by activation of receptor-guanylate cyclase (R-GC) signaling molecules that produce cyclic GMP (cGMP) and stimulate the cystic fibrosis transmembrane conductance regulator-dependent secretion of Cl- and HCO3-. Our experiments demonstrate that mRNA transcripts for guanylin and uroguanylin are markedly reduced in colon polyps and adenocarcinomas. In contrast, a specific uroguanylin-R-GC, R-GCC, is expressed in polyps and adenocarcinomas at levels comparable with normal colon mucosa. Activation of R-GCC by uroguanylin in vitro inhibits the proliferation of T84 colon cells and elicits profound apoptosis in human colon cancer cells, T84. Therefore, down-regulation of gene expression and loss of the peptides may interfere with renewal and/or removal of the epithelial cells resulting in the formation of polyps, which can progress to malignant cancers of the colon and rectum. Oral replacement therapy with human uroguanylin was used to evaluate its effects on the formation of intestinal polyps in the Min/+ mouse model for colorectal cancer. Uroguanylin significantly reduces the number of polyps found in the intestine of Min/+ mice by approximately 50% of control. Our findings suggest that uroguanylin and guanylin regulate the turnover of epithelial cells within the intestinal mucosa via activation of a cGMP signaling mechanism that elicits apoptosis of target enterocytes. The intestinal R-GC signaling molecules for guanylin regulatory peptides are promising targets for prevention and/or therapeutic treatment of intestinal polyps and cancers by oral administration of human uroguanylin.

Background: Hyperhomocysteinemia has been associated with coronary atherosclerosis in many, but not all, prospective and retrospective studies. Some on these inconsistencies may be attributed to methodological variabilities., Methods: In the present study, three newly commercially available assays and one in-house HPLC assay for total homocysteine (tHcy) were utilized in 99 subjects with angiographically documented atherosclerosis and in 91 community controls matched by age, gender, and smoking history. The in-house assay, a modified Fortin and Genest HPLC method, was compared with the Bio-Rad HPLC assay, the Abbott IMx((R)) fluorescence polarization immunoassay, and a Bio-Rad enzyme-linked immunoassay (EIA) microtiter method., Results: Correlation coefficient values between the in-house HPLC assay and the Bio-Rad HPLC, the Abbott IMx, and the Bio-Rad EIA assays were 0.95, 0.96 and 0.90, respectively. Although tHcy concentrations were higher in cases compared with controls by all four methods, the difference reached statistical significance only with the in-house HPLC procedure (median, 13.5 +/- 6.7 micromol/L in cases vs 10.9 +/- 4.8 micromol/L in controls; P <0. 01, adjusting for covariates), where it was an independent predictor of case or control status, along with hypertension, total cholesterol, and triglycerides. The tHcy distributions in cases and controls demonstrated significant overlap. The number of atherosclerotic major coronary vessels was associated with significantly higher tHcy (P <0.01 for trend) in all four methods., Conclusions: The three commercial assays for tHcy differed in analytical and clinical performance. Analytically, the Abbott IMx method showed the best comparability with the in-house assay, but clinically, the three commercial methods were similar and did not distinguish cases from controls.

Exercise is associated with changes in lipids that may protect against coronary heart disease (CHD). In this study of 28 triathletes, we analyzed acute changes in serum lipid and lipoprotein concentrations after completion of the 1995 World Championship Hawaii Ironman Triathlon. With standard laboratory assays, we demonstrate significant decreases in total cholesterol, VLDL cholesterol, ApoB100, and Lp(a). Total HDL cholesterol increased significantly immediately after the race. With a novel proton NMR spectroscopy assay, we demonstrate that smaller diameter LDL particles, corresponding to small, dense LDL, declined by 62%. Moreover, larger HDL subclasses, whose levels are inversely associated with CHD, increased significantly by 11%. Smaller HDL subclasses, which have been directly associated with CHD in some studies, acutely decreased by 16%. Therefore, exercise not only acutely induces changes in lipoprotein concentrations among the standard species in a manner that favorably affects CHD risk, but also induces favorable changes in specific lipoprotein subclass size distribution that also may alter CHD risk independently of the total lipoprotein serum concentration.

A soluble sulfotransferase from porcine serum which catalyzes the transfer of sulfate from adenosine 3'-phosphate 5'-phosphosulphate (PAPS) to 2'-fucosyllactose (2'-FL) was purified 36,333-fold using a combination of conventional and affinity chromatographic steps. The purified enzyme preparation after non-denaturing discontinuous-PAGE exhibited a molecular mass of about 80 kDa by reducing SDS-PAGE. However, when a partially purified enzyme preparation was subjected to gel filtration on Sephacryl S-300, the enzyme activity eluted in the void volume, which indicated that the native enzyme existed as an oligomer. The purified enzyme showed Km values of 9.15 microM for PAPS and 15.38 mM for 2'-FL at the optimum pH value of 7.4. The substrate specificity of the purified enzyme was evaluated with various sugars that are structurally similar to sialyl LewisX (sLeX). Results indicated that 3'-sialyllactose and lactose were efficient acceptors of sulfation, whereas 6'-sialyllactose and 6'-sialyllactosamine were poor substrates for this sulfotransferase. Further, the reaction product analysis revealed that the sulfate substitution, when using 2'-FL as the substrate, was at the C-6 position of the galactose residue. Coincidentally, a similar enzyme activity was also found in porcine lymphoid tissues such as, lymph nodes (peripheral and mesenteric) and spleen. Collectively, these findings suggest that this enzyme might be involved in the synthesis of the ligand for L-selectin.

A microsomal galactose-6-O-sulfotransferase (Gal-6-O-Stase) from porcine lymph nodes, able to transfer the sulfate group from adenosine 3'-phosphate 5'-phosphosulphate (PAPS) onto 2'-fucosyllactose (2'-FL) and other sialyl LewisX (sLex)-related sugars, has been purified and characterized. The enzyme was purified to about 35,000-fold by a combination of conventional and affinity chromatographic steps. The purified enzyme preparation exhibited two protein bands at around 80-90 and 170 kDa on 7.5% SDS-PAGE under reducing conditions. Both of these protein bands always comigrated in the gel when peak fractions containing Gal-6-O-Stase activity from the 3',5'-ADP-agarose column were subjected to 6% SDS-PAGE under reducing conditions. These protein bands also showed similar binding patterns to WGA (wheat germ agglutinin), Con A (concanvalin A), and EBA (elderberry agglutinin). Similarly, when the enzyme preparation after the hydroxylapatite step was photolabeled with 8-azido-[32P]-PAPS, both 80-90 and 170 kDa protein bands were labeled in a specific manner. These results suggest a possible association of these two protein bands with the enzyme activity. The carbohydrate substrate specificity of this enzyme suggests that it is well suited to catalyze the sulphonation at the C-6 position of the galactose residues of oligosaccharides that are structurally similar to sLex. Furthermore, a survey of several porcine organs revealed that this enzyme was selectively expressed in lymphoid tissues such as lymph nodes (peripheral and mesenteric) and spleen. These findings suggest that this enzyme may be involved in the assembly of 3'-sialyl-6'-sulfo Lewisx, the major capping group of HEV-ligands for L-selectin., (Copyright 1999 Academic Press.)

The semaphorins comprise a large family of conserved glycoproteins, several members of which have been shown to function in repulsive neuronal growth cone guidance. We show here that Drosophila Semaphorin I (Sema I), a transmembrane semaphorin expressed on embryonic motor and CNS axons, is required for correct guidance of motor axons and for the formation of CNS pathways. In mutant embryos lacking Sema I, motor axons stall and fail to defasciculate at specific choice points where normally they would project to their muscle targets. In addition, a specific CNS fascicle fails to form correctly in these embryos. Rescue and ectopic expression experiments show that Sema I is required in neurons to mediate axon guidance decisions. These studies further suggest that like secreted semaphorins, transmembrane semaphorins can function as repulsive guidance cues for specific axon guidance events during neurodevelopment.

The complete nucleotide sequence of the triple gene block one (TGB 1) of cymbidium mosaic potexvirus (CymMV) was compared to those from other potex-, carla-, furo- and hordeiviruses. Seven conserved motifs in the TGB 1, including the ATP-GTP binding domain (P-Loop) consensus GXXGKTSTS, were found in all four virus genera. We propose that all TGBV can be classified into phylogenetic clusters based on their TGB 1 homolog genes. These clusters can be further delineated to form subgroups. The first cluster comprises the potexviruses which are further subdivided into three subgroups; BaMV, FMV, PlaMV and PapMV (subgroup Ia); CymMV, PAMV, NMV, SMYEaV and WC1MV (subgroup Ib) and PVX (subgroup Ic). The second cluster comprises carlaviruses with a dual subgrouping; CVB, LSV, PVM, PMV and ASPV (subgroup IIa) and LVX (subgroup IIb). The third cluster carries the most diverse of TGBV comprising furoviruses PCV, PMTV and BSBV (subgroup IIIa) and hordeiviruses PSLV, BSMV and LRSV (subgroup IIIb). The phylogenetic relationships of triple gene block viruses (TGBV) based on the TGB 1 homolog gene indicates a convergent evolution.

Rapid recovery is an essential component in the anesthesia of OPD surgery. In this study midazolam, the short-duration benzodiazepine, was used as an intravenous anesthetic in OPD surgery. At the end of the operation flumazenil, the specific antagonist of benzodiazepines, was given to reverse the effect of midazolam. Recovery of these patients was compared with those in the control group who received saline instead of flumazenil. In a randomized, placebo-controlled clinical study the antagonistic effect of flumazenil on midazolam was investigated in fifty ASA class I-II gynecologic outpatients subjected to D & C procedure under anesthesia induced with midazolam (0.3 mg/kg). They were divided into group A (flumazenil group) and group B (placebo group) with 25 patients in each. At the end of operation patients in group A were given 0.2 mg flumazenil intravenously to antagonize the residual effect of midazolam while patients in group B were given 2 ml normal saline intravenously as control. Our results showed that patients in group A exhibited a rapid and steady return of consciousness scaled by alertness, orientation of time and place and activity collaboration at 5, 30 and 60 min intervals following administration of flumazenil (p less than 0.005). There were no significant changes found in the hemodynamic or respiratory aspect between groups (p greater than 0.05). All the patients tolerated midazolam and flumazenil well. It is concluded that the use of flumazenil to antagonize the residual effect of midazolam is safe and effective as far as outpatient procedure is concerned.

One hundred and sixty-one cases of chyluria have been studied. The possible aetiological factors, the approach to investigation and the methods and results of treatment have been discussed. In this series, the diagnosis was established by the presence of chyle in the urine. Lymphography was carried out in 90 patients for the localisation of the site of lymphatic reflux while cystoscopic examination after fatty meal was useful in determining the side with the heavier leakage. One hundred and nineteen cases responded to conservative treatment and only 42 cases required operative treatment. Stripping of the renal pedicle via the retroperitoneal route gave the best results.

Our experience of 76 pelvic exenterations for advanced pelvic malignancies is presented, with emphasis on the results and complications. The overall operative mortality rate of 14% is acceptable, and a five-year survival rate of 20% has been achieved. The procedure has a definite role to play in the management of advanced but otherwise localized pelvic malignancies.

Sixty-five cases of hemospermia seen over a ten-year period were reviewed. No specific etiology was disclosed, although renal tuberculosis was seen in 11 per cent of cases, indicating that hemospermia is a self-limiting and benign condition which does not require full urologic investigation.

We treated 50 patients with chyluria by stripping the renal pedicle. Until further studies reveal the etiology of chyluria ligation and stripping of the pedicle remain the treatment of choice and this fact has been borne out by our long-term results.