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In this work, a biodegradable and injectable in situ gel-forming controlled drug delivery system based on thermosensitive poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) hydrogel was studied. The prepared PCEC hydrogel undergoes temperature-dependent sol-gel-sol transition, which is a flowing sol at ambient temperature and turns into a non-flowing gel at around physiological body temperature. Furthermore, the sol-gel phase transition mechanism was investigated using (13)C-nuclear magnetic resonance imaging and a laser diffraction particle size analyzer. The in vitro release behaviors of several model drugs, including a hydrophilic small-molecule drug, a hydrophobic small-molecule drug and a macromolecular protein drug, from PCEC hydrogel were also investigated in detail. The results showed that the model drugs could be released from the PCEC hydrogel system over a sustained period. In addition, an anaesthesia assay was conducted using the tail flick latency (TFL) test to evaluate the in vivo controlled drug delivery effect of the PCEC hydrogel system. In the TFL assay, a lidocaine-loaded PCEC hydrogel produced significantly longer-lasting local anaesthetic effects compared with lidocaine aqueous solution at the same dose. Therefore, PCEC hydrogel is promising for use as an injectable local drug delivery system.

Rapid local transdermal anaesthetic is desirable in clinic. In this paper, lidocaine loaded poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCL-PEG-PCL) nanoparticles were prepared, and a novel transdermal lidocaine formulation: lidocaine loaded PCL-PEG-PCL nanoparticles in F127 hydrogel (Nano-Lido Gel), was demonstrated. These lidocaine loaded PCL-PEG-PCL nanoparticles with mean particle size of ca. 200 nm had drug loading of about 40%. The efficiency of transdermal anaesthesia of four treatments: EMLA cream (E), Nano-Lido Gel (N), EMLA cream with brief focal ultrasound pretreatment (EU), and Nano-Lidocaine Gel with brief focal ultrasound pretreatment (NU), was evaluated by tail-flick latency test assay in rats. Results indicated that the topical anaesthesia onset time in NU was 5 times and 2.5 times shorter than that in E and EU. The efficiency of anaesthesia in NU, expressed as maximum possible effects (MPE) value, was significantly higher than that in other treatments. It provided a novel path to develop rapid transdermal anaesthesia by combination of ultrasound pretreatment and lidocaine nano-formulation based on polymeric nanoparticles.

Poly(epsilon-caprolactone)-poly(ethylene glycol) (PCL-PEG) copolymers are important synthetic biomedical materials with amphiphilicity, controlled biodegradability, and great biocompatibility. They have great potential application in the fields of nanotechnology, tissue engineering, pharmaceutics, and medicinal chemistry. This review introduced several aspects of PCL-PEG copolymers, including synthetic chemistry, PCL-PEG micro/nanoparticles, PCL-PEG hydrogels, and physicochemical and toxicological properties.

A new azo dyes-decolorizing fungi strain QQ was isolated from activated sludge. It was identified as Penicillium sp. based on 26S rRNA gene sequence analysis. The study indicated that strain QQ could aerobically decolorize Reactive Brilliant Red X-3B by the way of bioadsorption, and nutrient-poor medium was more beneficial for adsorption. Decolorization rate was inversely proportional to the size of mycelial pellets. The optimum pH was observed at 4 or 5 for X-3B decolorization. There was still 70% color removal when salinity increased to 6%. By contrast with aerobic decolorization, the degradation of azo dyes occurred under anaerobic conditions, and some azo dyes could be absolutely decolorized. Furthermore, the decolorization of azo dyes by fungal-bacterial cocultures was investigated. The results demonstrated that strain QQ and Sphingomonas xenophaga QYY cocultures performed better than any single strain did. Weak acidity conditions and the presence of small amount of surfactant could enhance the ability of consortium to decolorize azo dyes.

Biodegradable poly(epsilon-caprolactone)/poly(ethylene glycol) (PCL/PEG) copolymer nanoparticles showed potential application in drug delivery systems. In this article, monodisperse poly(epsilon-caprolactone)/poly(ethylene glycol)/poly(epsilon-caprolactone) (PCL/PEG/PCL, PCEC) nanoparticles, approximately 40 nm, were prepared by solvent extraction method using acetone as the organic solvent. These PCL/PEG/PCL nanoparticles did not induce hemolysis in vitro and did not show toxicity in vitro or in vivo. The prepared PCL/PEG/PCL nanoparticles were employed to load doxorubicin by a pH-induced self-assembly method. In vitro release study indicated that doxorubicin release from nanoparticles at pH 5.5 was faster than that at pH 7.0. The encapsulation of doxorubicin in PCL/PEG/PCL nanoparticles enhanced the cytotoxicity of doxorubicin on a C-26 cell line in vitro. Meanwhile, compared with free doxorubicin, doxorubicin in nanoparticles could more efficiently treat mice bearing subcutaneous C-26 tumors. The doxorubicin-loaded PCL/PEG/PCL nanoparticles might be a novel doxorubicin formulation for cancer therapy.

Objective: To establish a gas chromatography method for simultaneous determination of organochlorine and pyrethroid pesticide residues in Viscum coloratum by cloud-point extraction (CPE)., Method: Pesticides were extracted with the non-ionic surfactant Triton X-100. The apparatus was gas chromatography with electron capture detector and the separation was performed on an Hp-5 column. The pesticide residues were calculated by external standard method., Result: Good linear relation was obtained over the range of 5-500 microg L(-1) for organochlorine and 10-1,000 microg L(-1) for pyrethroid. The limits of detection was 1.5-7.5 microg kg(-1). The average recoveries of organochlorine and pyrethroid were 74.15% -111.6% with corresponding RSD of 4.0% -9.1%., Conclusion: The sample and rapid method was applied to pesticide residues determination.

Background: Infiltration of a long-lasting anaesthetic is helpful during the post-operative period. The recently developed local drug delivery system, biodegradable nanoparticles in a thermo-sensitive hydrogel (nanogel system), may possibly provide an extended duration of drugs. Therefore, we evaluated whether prolonged infiltration anaesthesia could be achieved by loading lidocaine into this delivery system., Methods: Thirty male rats were randomized into five groups of six rats each: saline; 2% hydrochloride lidocaine solution; lidocaine-loaded nanogel system and its compositing formulations, namely lido-nano gel; lido-nano; and lidogel. Durations of local anaesthesia with subcutaneously injected agents were measured by tail flick latency tests in a randomized, blind fashion., Results: Lido-nano gel produced effective anaesthesia for 360+/-113 min, compared with 150+/-33 min by lidogel, 180+/-37 min by lido-nano, and 110+/-45 min by lidocaine solution (P<0.001, means+/-SD), and elicited complete sensory blockade for 300+/-114 min, compared with 75+/-37 min by lidogel, 105+/-53 min by lido-nano, and 60+/-33 min by lidocaine solution (P<0.001, means+/-SD) without severe skin/systemic toxicity., Conclusion: Lidocaine-loaded biodegradable nanoparticles in hydrogel produced prolonged infiltration anaesthesia in rats without severe toxicity, indicating a possible way to develop long-lasting local anaesthetics.

Purpose: Honokiol showed potential application in cancer treatment, but its poor water solubility restricts its clinical application greatly. So, we designed a self-assembled monomethoxy poly(ethylene glycol)-poly(epsilon-caprolactone) (MPEG-PCL) micelle to load honokiol to overcome its poor water solubility., Methods: We synthesized MPEG-PCL diblock copolymer that could self-assemble into monodisperse micelles at the particle size of ca.18 nm in water. Honokiol was loaded into MPEG-PCL micelle by direct dissolution method assisted by ultrasound, without any surfactants, organic solvents, and vigorous stirring., Results: The blank MPEG-PCL micelles (100 mg/mL) did not induce any hemolysis in vitro and showed very low toxicity ex vivo and in vivo. Honokiol could be molecularly incorporated into MPEG-PCL micelles at the drug loading of about 20% by direct dissolution method assisted by ultrasound. After loaded into MPEG-PCL micelles, honokiol maintained its molecular structure and anticancer activity in vitro. Honokiol could be sustained released from MPEG-PCL micelles in vitro. The honokiol loaded MPEG-PCL micelles could be lyophilized without any adjuvant., Conclusion: The prepared honokiol formulation based on self-assembled MPEG-PCL micelle was stable, safe, effective, easy to produce and scale up, and showed potential clinical application.

In this article, poly(epsilon-caprolactone)-pluronic-poly(epsilon-caprolactone) (PCFC) copolymer was synthesized by ring-opening polymerization and characterized by 1H-NMR and GPC. Cationic PCFC nanoparticles were prepared at one-step by modified emulsion solvent evaporation method using cetyltrimethylammonium bromide as the stabilizer. The cytotoxicity of PCFC nanoparticles was studied here with and without serum. The obtained cationic PCFC nanoparticles were employed to condense and adsorb DNA onto its surface. And it could protect plasmid GFP (pGFP) from enzymatic degradation and acidic degradation in a certain period. Release behavior of pGFP from the pGFP/PCFC nanoparticles complex was also studied in vitro. The obtained cationic PCEC nanoparticles might have great potential application in DNA delivery.

In this article, a novel composite drug delivery system, honokiol nanoparticles in biodegradable hydrogels based on chitosan (CS) and beta-glycerophosphate (beta-GP), was prepared. CS/beta-GP solution was liquid at room temperature and turned into gel as temperature increased. With increase in beta-GP concentration, the sol-gel transition temperature decreased accordingly. Honokiol nanoparticles with diameter of about 30 nm were prepared by emulsion solvent evaporation method. The sol-gel transition temperature of CS/beta-GP system decreased as F-127 presented in honokiol nanoparticles. In vitro release profiles were studied, the results showed that honokiol could be slowly released from CS/beta-GP gel over at least 2 weeks and the release rate was greatly influenced by initial drug loading. The described injectable hydrogels based on chitosan (CS) and beta-glycerophosphate (beta-GP) might have potential application as local drug delivery for honokiol.

In this article, poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCL-PEG-PCL, PCEC) nanoparticles were successfully prepared for honokiol delivery in vitro. Blank or honokiol loaded PCL-PEG-PCL nanoparticles were prepared in moderate condition by solvent diffusion method without using any surfactants. The prepared blank PCL-PEG-PCL nanoparticles are mono-dispersed and smaller than 200 nm. The particle size increased with increase in polymer concentration and oil-water (O/W) ratio. The prepared PCL-PEG-PCL nanoparticles (40 mg/mL, ca. 106 nm) did not induce hemolysis in vitro. And the 50% inhibiting concentration (IC50) of it (48 h) on HEK293 cells was higher than 5 mg/mL. Honokiol could be efficiently loaded into PCL-PEG-PCL nanoparticles and released from these nanoparticles in an extended period in vitro. After honokiol (HK) was entrapped into PCL-PEG-PCL nanoparticles, the particle size increased with the increase in HK/PCEC mass ratio in feed, and the encapsulated honokiol retained potent anticancer activity in vitro. The PCL-PEG-PCL nanoparticle was suitable for honokiol delivery, and such honokiol loaded PCL-PEG-PCL nanoparticle was a novel honokiol formulation.

A novel CBL-interacting protein kinase (CIPK) gene, ZmCIPK16, was isolated from maize (Zea mays), which has been certified to have two copies in the genome. The ZmCIPK16 is strongly induced in maize seedlings by PEG, NaCl, ABA, dehydration, heat and drought, but not by cold. A yeast two-hybrid assay demonstrated that ZmCIPK16 interacted with ZmCBL3, ZmCBL4, ZmCBL5, and ZmCBL8. Bimolecular fluorescence complementation (BiFC) assays prove that ZmCIPK16 can interact with ZmCBL3, ZmCBL4, ZmCBL5, and ZmCBL8 in vivo. Subcellular localization showed that ZmCIPK16 is distributed in the nucleus, plasma membrane and cytoplasm; this is different from the specific localization of ZmCBL3, ZmCBL4, and ZmCBL5, which are found in the plasma membrane. The results also showed that overexpression of ZmCIPK16 in the Arabidopsis sos2 mutant induced the expression of the SOS1 gene and enhanced salt tolerance. These findings indicate that ZmCIPK16 may be involved in the CBL-CIPK signaling network in maize responses to salt stress.

Self-assembled polymeric micelles are widely applied in drug delivery system (DDS). In this study, honokiol (HK) loaded micelles were prepared from biodegradable poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) copolymers. Micelles were prepared by self-assembly of triblock copolymer PCEC in distilled water triggered by its amphiphilic character without any organic solvent. Drug loading and encapsulation efficiency were determined by adjusting the weight ratio of HK and PCEC. The particle size and zeta potential distribution of obtained micelles were determined using Malvern laser particle sizer, and spherical geometry were observed on atomic force microscope (AFM). Otherwise, the thermo-sensitivity of honokiol-loaded micelles was monitored. And the cytotoxicity results of drug loaded micelles showed that the encapsulated honokiol remained potent antitumor effect. Moreover, in vitro release profile demonstrated a significant difference between rapid release of free honokiol and much slower and sustained release of HK-loaded micelles. These results suggested that we have successfully prepared honokiol-loaded micelles in an improved method which is safer and more efficient. The prepared micelles might be potential carriers for honokiol delivery in cancer chemotherapy.

In this work, a series of biodegradable triblock poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) copolymers were successfully synthesized by ring-opening copolymerization, and were characterized by (1)H NMR, FT-IR, GPC, and DSC. Aqueous solutions of PECE copolymers underwent thermosensitive sol-gel-sol transition as temperature increases when the concentration was above corresponding critical gel concentration (CGC). Sol-gel-sol phase transition diagrams were recorded using test tube inverting method, which depended on hydrophilic/hydrophobic balance in macromolecular structure, as well as some other factors, including topology of triblock copolymers and solution composition of the hydrogel. As a result, the sol-gel-sol transition temperature range could be varied, which might be very useful for its application as injectable drug delivery systems. The in vivo gel formation and degradation behavior was conducted by injecting aqueous PECE solution into KunMing mice subcutaneously. In vitro degradation behavior, in vitro drug release behavior, and cytotoxicity were also investigated in this paper. Therefore, owing to great thermosensitivity and biodegradability of these copolymers, PECE hydrogel is believed to be promising for in situ gel-forming controlled drug delivery system.

A novel biphenyl-degrading bacterial strain LA-4 was isolated from activated sludge. It was identified as Dyella ginsengisoli according to phylogenetic similarity of 16S rRNA gene sequence. This isolate could utilize biphenyl as sole source of carbon and energy, which degraded over 95 mg/L biphenyl within 36 h. The major metabolites formed from biphenyl, such as 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA) and benzoic acid, were identified by LC-MS. The crude cell extract of strain LA-4 exhibited the activity of 2,3-dihydroxybiphenyl 1,2-dioxygenase (2,3-DHBD) and the kinetic parameters K(m) was 26.48 micromol/L and V(max) was 8.12 U/mg protein. A conserved region of the biphenyl dioxygenase gene bphA1 of strain LA-4 was amplified by PCR and confirmed by DNA sequencing.

The performances and microbial population changes in laboratory-scale membrane bioreactor (MBR) augmented with Sphingomonas xenophaga QYY were investigated in the present study. It was demonstrated that after 30 days acclimation, the non-augmented MBR system were able to degrade bromoamine acid (BAA) well. However, the efficiency of the system decreased with BAA concentration increasing. While the augmented MBR showed higher capability, in which the color and COD removal were more than 90% and 50%, respectively. By ribosomal intergenic spacer analysis (RISA), it was found that BAA-utilizing populations gradually increased to become the dominant species in the non-augmented MBR. However, the augmented MBR possessed relatively stable treatment abilities, in which the introduced strain QYY could be persistent and co-exist well with the indigenous populations.

Biodegradable polyetherester copolymer (PCL/PEG/PCL, PCEC) was synthesized by ring-opening polymerization of epsilon-caprolactone initiated by poly(ethylene glycol) (PEG). The PCEC nanoparticles were prepared by solvent diffusion method or w/o/w double emulsion method. The obtained particles' morphology was observed on scanning electron microscopy, and the particle size distribution was determined using Malvern laser particle sizer. Bovine serum albumin was used as the model water-soluble protein drug, which was successfully encapsulated in PCEC nanoparticles, the drug release behavior was studied in detail. The hydrolytic degradation behavior of the PCEC nanoparticles was also studied.

Fast and sensitive liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS) method for the specific determination of viaminate in 14 kinds of rat tissues pre-treated with simple procedure was developed and validated. Biological samples were prepared by direct precipitation to skin, stomach, intestine, and liver and extracted by liquid-liquid extraction to lung, kidney, muscle, spleen, brain, fat, testes (male and female), eye, and heart. After addition of menaquinon as internal standard to tissue homogenate, the supernatant was injected into the isocratic chromatographic system using a Waters Symmetry C8 column and methanol-water-formic acid (93:7:0.1) as the eluent. The eluate was completely led into an APCI interface with selected ion monitoring mode and the analytes were quantified using triple quadrupole MS. The assays were successfully validated in the ranges 0.02-20 ng/mL for lung, 0.02-10 ng/mL for kidney, spleen, muscle, brain, fat, eye, and heart, 0.05-10 ng/mL for testicle, 0.4-100 ng/mL for liver, skin and intestine, and 1.0-200 ng/mL for stomach. The accurate and precise studies showed good reproducibility with coefficients of variation below 8.5% and the recoveries range from 90 to 109%. The analytes were chemically stable under all relevant conditions and the assays were applied in tissue distribution study. The results showed that the viaminate concentration was high in skin, low in kidney, and almost undetectable in eye and brain.

In this article, a novel local hydrophobic drug delivery system: nanoparticles in thermo-sensitive hydrogel, was demonstrated. First, honokiol, as a model hydrophobic drug, loaded poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) nanoparticles were prepared by emulsion solvent evaporation method, and then were incorporated into thermo-sensitive F127 hydrous matrix. The obtained injectable hydrophobic drug delivery system can act as a depot for sustained release of honokiol in situ. The lower critical solution temperature (LCST) of the composite matrix increases with increase in the mass of incorporated nanoparticles, or with decrease in the amount of residual organic solvent in the system. Honokiol release profile in vitro was studied, and the results showed that honokiol could be sustained released from the system. The described injectable drug delivery system might have great potential application for local delivery of hydrophobic drugs such as honokiol.

In this article, blank anionic poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) and anionic mannan modified PCEC (MPCEC) nanoparticles with nearly the same particle size and zeta potential were prepared by emulsion solvent evaporation method. Human basic fibroblast growth factor (bFGF) was absorbed onto anionic nanoparticles surface due to electrostatic interaction. The obtained bFGF-nanoparticles complexes were injected subcutaneously into C57BL/6 mice at 20 microg of bFGF/dose on week 0, 1, 2 and 3. The mice serum was collected on week 4, and bFGF-specific autoantibody total IgG, IgG1 and IgG2a titer in serum was determined by ELISA. The results indicated that the autoantibody IgG, IgG1 and IgG2a titer of the mice immunized by bFGF-MPCEC complexes were higher than that immunized by either bFGF-PCEC or bFGF-Alum. This phenomenon might be due to that mannan functionalized MPCEC nanoparticles could be targeted to dendritic cells (DCs) to improve humoral immunity. The prepared anionic mannan modified PCEC nanoparticles (MPCEC) might have great potential application in vaccine delivery systems.

In this article, human basic fibroblast growth factor (bFGF) loaded PCL-PEG-PCL (PCEC) nanoparticles were prepared by modified W1/O/W2 double emulsion solvent evaporation method. The bFGF encapsulated in PCEC nanoparticles could be released in sustained release behavior in vitro. After subcutaneous single-dose injection of bFGF loaded PCEC nanoparticles at 20 microg of bFGF per dose in mice, the anti-bFGF special autoantibody IgG continued to grow until 8 weeks after the immunization and was still kept at high level at week 11. At the same time, HK293 cell viability assay in vitro indicated that the cytotoxicity of blank PCEC nanoparticles was low but dose dependent. For some success in controlling tumor growth had been met by neutralizing bFGF reported previously, the bFGF loaded PCEC nanoparticles prepared in this paper might have potential application as anti-tumor vaccine.

A bacterial strain that degrades phenanthrene was isolated from activated sludge of wastewater treatment plant from a chemical corporation. The strain can utilize phenanthrene as sole source of carbon and energy. It was identified as Burkholderia sp. according to morphological and 16S rRNA gene sequence analysis. Phenanthrene (100 mg x L(-1)) was removed 74.1% within 84 h by this strain. It was suggested that the strain possessed the salicylic acid pathway by LC-MS and UV methods. And the strain contains an inductive catechol 2,3-dioxygenase (C230). The kinetic constants Km and Vmax were 10.93 micromol x L(-1) and 106.38 micromol x min(-1), receptively. It indicates that the affinity of the endoenzyme to catechol is great and catechol is a better substrate.

In this article, a novel biodegradable and pH-sensitive hydrogel based on polycaprolactone, poly(ethylene glycol) and methylacrylic acid (MAA), was prepared by UV-initiated free radical polymerization. The obtained hydrogel was characterized by (1)H NMR and FTIR. The acute toxicity tests and histopathological study were performed in BALB/c mice. In acute oral toxicity test, mice were orally administered with a total 15 g/kg body weight (b.w.) of P(CL-MAA-EG) hydrogels, and were observed continuously for 14 days. For histopathologic study, samples including heart, liver, lung, kidneys, spleen, stomach, and intestine, were histochemically prepared and stained with hematoxylin-eosin for histopathologic examination. No mortality or significant signs of acute toxicity was observed during the whole observation period, and no macroscopic alteration was found in the organs. Histopathological analysis of various organs also did not show any significant pathological changes. Thus, the maximal tolerance dose of P(CL-MAA-EG) hydrogels was calculated to be higher than 15 g/kg b.w. in BALB/c mice. It was suggested that the studied P(CL-MAA-EG) hydrogel in this article were nontoxic after acute oral administration and it might be a promising candidate as a novel oral drug carrier., ((c) 2007 Wiley Periodicals, Inc.)

In this paper, a new kind of biodegradable aliphatic polyetheresteramide copolymers (PEEA) based on epsilon-caprolactone, 6-aminocaproic acid, and poly(ethylene glycol) (PEG) were synthesized by melt polymerization method. The obtained copolymers were characterized by 1H-NMR. The thermal properties of PEEA copolymers were studied by DSC and TGA/DTA under nitrogen atmosphere. The water absorption and hydrolytic degradation behavior was also studied in detail. With the increase in PEG content or the decrease in caprolactone content, the water absorption of the copolymers increased accordingly. For the hydrolytic degradation behavior, with the increase in PEG content or caprolactone content, the degradation rate increased then.

Hydrolytic degradable PBT/PEG copolymer was synthesized by macromolecular transesterification method from PBT and PEG macromonomers. The resultant copolymers were characterized by (1)H-NMR and GPC. The non-isothermal crystallization behavior of these copolymers was studied by differential scanning calorimetry (DSC). The water absorption and hydrolytic degradation behavior of PBT/PEG copolymers were also studied in detail.

Purpose: We investigated aberrant methylation patterns in esophageal adenocarcinoma and correlated the findings to patient survival and tumor recurrence., Experimental Design: Gene promoter methylation was performed in 82 samples from 41 esophagectomy patients consisting of 41 adenocarcinoma samples, each with its adjacent nonmalignant tissue, which included one sample with Barretts metaplasia. The methylation status of seven genes was determined. Epigenetic silencing was confirmed using immunohistochemical staining. Kaplan-Meier plots were constructed using disease-specific survival as the primary end point and the interval from surgery to tumor recurrence as the secondary end point. The association of clinicopathological and biomolecular risk factors to survival and recurrence was performed using the Log-rank test and Cox proportional hazards model for multivariate analysis., Results: Methylation frequencies of the genes analyzed were APC, 68%; E-cadherin, 66%; O(6)-methylguanine DNA methyltransferase, 56%; ER, 51%; p16, 39%; DAP-kinase, 19%; and TIMP3, 19%. DNA methylation of some genes individually showed only trends toward diminished survival, whereas patients whose tumors had >50% of their gene profile methylated had both significantly poorer survival (P = 0.04) and earlier tumor recurrence (P = 0.05) than those without positive methylation. By multivariate analysis, the hazard ratios (HRs) with positive methylation status were more powerful predictors of survival [HR 2.7 (1.14-6.45; 95% confidence interval)] and tumor recurrence [HR 2.5 (1.11-5.6)] than age (HR 2.03 and 1.96, respectively) or stage (HR 1.48 and 1.67, respectively)., Conclusions: Our data suggest that positive methylation status for multiple genes in esophageal adenocarcinoma is a predictor of poor prognosis.

At cardiac catheterization, analog images obtained using cinefilm are translated into digital images, and images appearing on the CRT are filtered by a sharpen filter. We investigated the effect of the sharpening filter on vessel diameter as measured by quantitative coronary arteriography. We acquired images of a vessel phantom filled with contrast material using an X-ray image intensifier. Vessel diameters measured by quantitative coronary arteriography were 1 mm, 1.5 mm, 2 mm, 3 mm, 4 mm, and 6 mm. Results showed that vessel diameters were decreased when the sharpening filter was used and that more intense filtering decreased the measured diameter further. When the diameter of the vessel phantom was less than 4 mm, the diameter was smaller and the ratio of decrease was larger. Vessel diameters of 2 mm, 3 mm, 4 mm, and 6 mm measured a maximum of 2.9 smaller, while those of 1 mm and 1.5 mm measured a maximum of 9.2 smaller.

Although dynamic cardiomyoplasty (DCMP) is currently being evaluated as an alternative to end-stage congestive heart failure, the overall results of DCMP are variable and inconclusive. We evaluated the effect of classic DCMP on systolic and diastolic cardiac function in normal heart using reliable indicators which minimize the influences of load conditions. Six experimental dogs were evaluated with the acute nonpreconditioning model. The slope of the linear preload recruitable stroke work relationship (Mw) showed a significant increase with latissimus dorsi muscle (LDM) stimulation (postwrap non-stimulation 59.1+/-6.3, postwrap stimulation 98.6+/-9.7 erg cm(-3) x 10(3); P < 0.01), and the x-intercept (V0) was unchanged; these were utilized as the indicators of left ventricular systolic function. The constant of pressure decay (tau) increased after LDM wrap (prewrap 45.8+/-6.0, postwrap nonstimulation 69.3+/-10.3, postwrap stimulation 72.3+/-13.9 ms; P < 0.05), and the peak filling rate was unchanged after LDM wrap, which were utilized as the indicators of diastolic function. We concluded that classic dynamic cardiomyoplasty is effective in assisting systolic cardiac function, but may to some degree have a detrimental effect on the diastolic cardiac function.

In their experimental approach to dynamic cardiomyoplasty (DCMP), the authors hypothesized that a wrapping method using the proximal part of a free latissimus dorsi muscle (LDM) graft might augment ventricular contractility more than the classic Carpentier's wrapping method. The authors divided 12 mongrel dogs into a pedicled LDM graft group (Group 1, n = 6) and a free LDM graft group (Group 2, n = 6) to evaluate the properties of the different wrapping methods. To evaluate the effect of DCMP on left ventricular systolic and diastolic function, the authors used precise indicators that minimize the influences of load conditions. The slope of the linear preload recruitable stroke work relationship (Mw) and an X-intercept (Vo) were utilized as the indicator of left ventricular systolic function. The constants of pressure decay (tau) and peak filling rate (PFR) were measured to determine diastolic function. All experimental animals were evaluated with the acute, non preconditioning model. Mw was significantly increased with LDM stimulation in both groups (postwrap non stimulation 59.1 +/- 6.3; postwrap stimulation 98.6 +/- 9.7 erg.cm-3. 10(3); p < 0.01 in Group 1, postwrap non stimulation 66 +/- 6.7; postwrap stimulation 155 +/- 15.7 erg.cm-3.10(3), p < 0.001 in Group 2). Stimulated free LDM grafts significantly increased the Mw in comparison to pedicled grafts (p = 0.011). Vo was unchanged in both groups and there was no significant difference between the two groups. Tau increased after LDM wrap in both groups (p < 0.05), but there was no difference between the two groups (tau; prewrap 45.8 +/- 6.0; postwrap non stimulation 69.3 +/- 10.3; postwrap stimulation 72.3 +/- 13.9 msec in Group 1, prewrap 50.0 +/- 6.0; post wrap non stimulation 61.8 +/- 5.0; post wrap stimulation in 64.3 +/- 4.7 msec in Group 2). Peak filling rate was unchanged after LDM wrap in both groups. Free LDM grafts significantly increased left ventricular systolic function compared to the pedicled LDM. Although myocardial relaxation was impaired after LDM wrap in both groups, there was no difference between the two groups. The authors' results emphasize the explicit benefit of utilization of a free LDM graft in respect to left ventricular systolic function; the free LDM grafts have no gross detrimental effect on diastolic function when compared with a pedicle graft in the acute model.

The effectiveness of dynamic cardiomyoplasty (DCMP) remains controversial. We hypothesized that effectiveness of DCMP using the latissimus dorsi muscle graft (LDMG) depends on the wrapping method. We analyzed pressure-volume relations (PVR), the left ventricular stroke work (LVSW), and the left ventricular end diastolic volume (LVEDV) changes during nonstimulation and stimulation of the LDMG to evaluate the effect of a new wrapping method of DCMP on the LVSW and the LVEDV changes. The new wrapping technique was evaluated in an acute animal experimental model. In 12 mongrel dogs, we performed continuous measurement of the dimensional and pressure dates of the left ventricle (LV) after the DCMP. The measurement was performed 15 min after wrapping during 5 periods. The duration of one measurement period was 15 s. The animals were divided into 2 groups according to the wrapping method. The heart was wrapped with the LDMG using 2 different methods. For Method 1, Carpentier's method, the heart was wrapped primarily with the distal part of the LDMG, the lateral segment. The vasculoneural pedicle of the latissimus dorsi muscle (LDM) was preserved. For Method 2, the LDM was separated, and the vasculoneural pedicle was cut. The medical sternotomy was performed. The thoracodorsal artery of LDMG was anastomosed to the right internal mammary artery, and the thoracodorsal vein was anastomosed to the right atrial appendage. The heart was wrapped primarily with the proximal part of the "free LDMG," the transverse segment. Based on the PVR loops, the changes of the LVSW and the LVEDV in both experimental groups were analyzed. The paired t-test was used for statistical analysis. Using Method 1, the LVSW and the LVEDV showed no significant changes during stimulation (stim) of the LDMG, compared with nonstimulation (nonstim) (LVSW: nonstim, 970 +/- 168 erg x 10(3); stim, 1,181 +/- 203 erg x 10(3); p = 0.126 and LVEDV: nonstim, 36.6 +/- 6.7 ml; stim, 37.2 +/- 6.8 ml; p = 0.36). Using Method 2, the LVSW was increased, and the LVEDV was decreased during stimulation of the free LDMG, compared with nonstimulation (LVSW: nonstim, 694 +/- 117 erg x 10(3); stim, 846 +/- 104 erg x 10(3); p < 0.001 and LVEDV: nonstim, 47.7 +/- 2.8 ml; stim, 46.8 +/- 2.7 ml; p < 0.001). The stimulated free LDMG wrapping of the heart seems to be a more effective wrapping method for DCMP, and it results in an increase of the LVSW and a decrease of the LVEDV, compared with the original Carpentier's method.

We previously demonstrated that the MHC class II molecule on murine sperm and CD4-like molecule on the egg vitelline membrane are involved in fertilization. In this study, the RNA transcripts in murine eggs corresponding to parts of the CD4 gene and lck gene in thymocytes were demonstrated by means of the reverse transcriptase (RT)/polymerase chain reaction (PCR) followed by the sequencing of the PCR products. The deduced sequences potentially encode the amino acid sequence of the transmembrane to the cytoplasmic domain of CD4 and that of the N-terminal domain of p56lck respectively. These findings indicate that a signal transducing complex similar to that in immune T cells is expressed at the transcriptional level in murine eggs.

Erythropoietin (EPO) has been used widely for correcting anemia in hemodialyzed (HD) patients. Enhancement of phagocytic function during EPO treatment of HD patients has been studied, but no data have been available on the effect of EPO on neutrophil chemiluminescence (CL) after challenge with phorbol myristate acetate (PMA). CL was measured in prehemodialysis whole blood samples from 15 stabilized patients and 15 normal healthy control subjects (C) after challenge with PMA. Before EPO treatment, CL was noted to be significantly higher in HD patients than in C, which changed significantly after 5 weeks of treatment (Rx) and continued for 13 weeks of Rx. There was a significant increase in hematocrit in these HD patients after 5 weeks that persisted until the 13th week. It was concluded that there is a significant decrease in whole blood CL in response to challenge with PMA during correction of anemia in HD patients treated with EPO. This study demonstrated that EPO could decrease enhanced PMA-activated reactive oxygen metabolite production and suggested that this decrease may protect against tissue damage, including red blood cell hemolysis in the uremic milieu.