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PubMed1995

Modified ultrafiltration improves cerebral metabolic recovery after circulatory arrest.

Modified ultrafiltration uses hemofiltration of the patient and bypass circuit after separation from cardiopulmonary bypass to reverse hemodilution and edema. This study investigated the effect of modified ultrafiltration on cerebral metabolic recovery after deep hypothermic circulatory arrest. Twenty-six 1-week-old piglets (2 to 3 kg) were supported by cardiopulmonary bypass (37 degrees C) at 100 ml.kg-1.min-1 and cooled to 18 degrees C. Animals underwent 90 minutes of circulatory arrest followed by rewarming to 37 degrees C. After being weaned from cardiopulmonary bypass, animals were divided into three groups: controls (n = 10); modified ultrafiltration for 20 minutes (n = 9); transfusion of hemoconcentrated blood for 20 minutes (n = 7). Global cerebral blood flow was measured by xenon 133 clearance methods: stage I--before cardiopulmonary bypass; stage II--5 minutes after cardiopulmonary bypass; and stage III--25 minutes after cardiopulmonary bypass. Cerebral metabolic rate of oxygen consumption, cerebral oxygen delivery, and hematocrit value were calculated for each time point. At point III, the hematocrit value (percent) was elevated above baseline in the ultrafiltration and transfusion groups (44 +/- 1.8, 42 +/- 1.8 versus 28 +/- 1.7, 30 +/- 0.7, respectively, p < 0.05). Cerebral oxygen delivery (ml.100 gm-1.min-1) increased significantly above baseline at point III after ultrafiltration (4.98 +/- 0.32 versus 3.85 +/- 0.16, p < 0.05) or transfusion (4.59 +/- 0.17 versus 3.89 +/- 0.06, p < 0.05) and decreased below baseline in the control group (2.77 +/- 0.19 versus 3.81 +/- 0.16, p < 0.05). Ninety minutes of deep hypothermic circulatory arrest resulted in impaired cerebral metabolic oxygen consumption (ml.100 gm-1.min-1) at point III in the control group (1.95 +/- 0.15 versus 2.47 +/- 0.07, p < 0.05) and transfusion group (1.72 +/- 0.10 versus 2.39 +/- 0.15, p < 0.05). After modified ultrafiltration, however, cerebral metabolic oxygen consumption at point III had increased significantly from baseline (3.12 +/- 0.24 versus 2.48 +/- 0.13, p < 0.05), indicating that the decrease in cerebral metabolism immediately after deep hypothermic circulatory arrest is reversible and may not represent permanent cerebral injury. Use of modified ultrafiltration after cardiopulmonary bypass may reduce brain injury associated with deep hypothermic circulatory arrest.

Animals↗
PubMed1995

Modified Pereyra bladder neck suspension: 10-year mean followup using outcomes analysis in 125 patients.

PURPOSE: We attempt to determine the long-term success rate of the modified Pereyra bladder neck suspension, identify preoperative factors predictive of long-term outcome, and investigate the influence of method and length of followup on reported continence status after modified Pereyra bladder neck suspension. MATERIALS AND METHODS: The charts of 177 patients who underwent modified Pereyra bladder neck suspension at our institution more than 5 years ago were reviewed. An attempt was made to contact all patients either by mail or telephone to complete a patient survey questionnaire. RESULTS: Of 177 patients 125 (71%) completed the patient survey with a mean followup of 9.8 years. At survey followup 20% of the patients reported no incontinence of any type and 51% reported stress urinary incontinence with or without urge incontinence. Of the patients 71% reported significant improvement in the incontinence and 73% were satisfied with the results of the procedure. No accurate preoperative predictors of long-term outcome were identified. The method and length of followup had significant impact on reported continence status after modified Pereyra bladder neck suspension. CONCLUSIONS: This long-term study using outcomes analysis reveals a high rate of recurrent stress incontinence after modified Pereyra bladder neck suspension. However, most patients maintain significant subjective improvement and remain satisfied with the results of the operation.

Female↗
PubMed1999

Metabolism of modified LDL and foam cell formation in murine macrophage-like RAW 264 cells.

The uptake of modified low density lipoprotein (LDL) by arterial macrophages is a key event in the atherogenesis. We studied 1) the uptake and degradation of modified LDL, 2) LDL recognition by specific receptors, and 3) the foam cell formation with murine macrophage-like RAW 264 cells in vitro. The cells took up and degraded effectively 125I-labeled acetylated LDL (Ac-LDL) and aggregated LDL (Aggr-LDL). Also oxidized LDL (Ox-LDL) was taken up but it was degraded poorly. The degradation of 125I-Ac-LDL was efficiently competed by both unlabeled Ac-LDL and Ox-LDL, whereas the degradation of 125I-Ox-LDL was partially competed by unlabeled Ox-LDL and Aggr-LDL but not at all by unlabeled Ac-LDL. The incubation with increasing concentrations of Ac-LDL, Aggr-LDL or Ox-LDL resulted in marked foam cell formation in the RAW 264 cells. Ox-LDL was cytotoxic at 500 to 1000 microg/ml concentrations. The results show that RAW 264 cells have at least two classes of receptors for modified lipoproteins: one that recognizes both Ox-LDL and Ac-LDL, and is similar to the scavenger receptors, and another that recognizes Ox-LDL but not Ac-LDL. RAW 264 cells are a convenient model cell line for examining the metabolism of modified lipoproteins, not only that of Ac-LDL but also that of Ox-LDL and Aggr-LDL, and cellular accumulation of lipids derived from modified LDL.

Animals↗
PubMedDate not supplied

Global assessment of functioning. A modified scale.

The modified Global Assessment of Functioning (GAF) scale has more detailed criteria and a more structured scoring system than the original GAF. The two scales were compared for reliability and validity. Raters who had different training levels assigned hospital admission and discharge GAF scores from patient charts. Intraclass correlation coefficients for admission GAF scores were higher for raters who used the modified GAF (0.81), compared with raters who used the original GAF (0.62). Validity studies showed a high correlation (0.80) between the two sets of scores. The modified GAF also correlated well with Zung Depression scores (-0.73). The modified GAF may be particularly useful when interrater reliability needs to be maximum and/or when persons with varying skills and employment backgrounds--and without much GAF training--must rate patients. Because of the increased structure, the modified GAF may also be more resistant to rater bias.

Activities of Daily Living↗
PubMed2002

Long-term complications related to the modified Indiana pouch.

OBJECTIVES: To describe a single-institution, single-surgeon experience with 125 modified Indiana pouches performed during a period of 14 years and their long-term complications. The modified Indiana pouch is a widely accepted and often used form of continent urinary diversion. Few studies have established the long-term complication rates associated with the procedure. METHODS: A retrospective chart review of 129 modified Indiana pouches constructed from March 1985 to August 1998 was performed, and the long-term complications and reoperation rates were tabulated. RESULTS: Complete information was obtained for 125 of the 129 charts, with a mean follow-up of 41.1 months (range 3 to 127). Complications occurred in 112 patients (89.6%; several patients had more than one complication), with a mean onset of 20.4 months (range 1 to 125) postoperatively. Seventy-three complications (58.4%) were due to the efferent limb, of which incontinence (defined as any leakage) was the most common (35 [28.0%]), followed by stomal stenosis in 19 (15.2%) and difficult catheterization in 12 (9.6%). Of the 26 pouch-related problems (21.8%) that occurred, the most common were stones in 13 (10.4%), perioperative leaks in 5 (4.0%), and perforations in 4 (3.2%). Ureteral anastomotic strictures were seen in 9 (7.2%). Other complications included gallstones in 32 (25.6%), kidney stones in 8 (6.4%), and small bowel obstruction in 6 (4.8%). Reoperation was performed in 65 patients (52.0%; several patients with more than one reoperation). Twenty-six (20.8%) of the patients required an open operation, and 39 (31.2%) received minimally invasive (percutaneous, endoscopic, extracorporeal shock wave lithotripsy) procedures. Sixty percent of the reoperations were minimally invasive. Reoperation was due to stomal stenosis in 18 (14.4%), pouch stones in 13 (10.4%), ureteral strictures in 9 (7.2%), and parastomal hernias in 6 (4.8%). Small bowel obstruction required reoperation in 5 patients (4.0%). CONCLUSIONS: In our experience, long-term complications of the modified Indiana pouch were mostly related to the efferent limb, and reoperations were usually due to stomal stenosis. Our data suggest that with longer follow-up, the complication and reoperation rates of the modified Indiana pouch appear to be higher than previously reported.

Constriction, Pathologic↗
PubMed2000

Gene transfer into hepatoma cells mediated by galactose-modified alpha-helical peptides.

To develop a receptor-mediated gene delivery system into hepatoma cells using the cationic alpha-helical peptide as the gene carrier molecule, we modified an alpha-helical peptide, which is known to have transfection abilities into cells, with a multi-antennary ligand containing several galactose residues that provide efficient binding to the asialoglycoprotein receptor. The galactose-modified peptides formed complexes with a plasmid DNA and showed gene transfer abilities into HuH-7 cells, a human hepatoma cell line. The transfection efficiency of the peptide was increased by increasing the number of modified galactose residues on the peptide. Furthermore, considerable inhibition of the transfection efficiency by the addition of asialofetuin, which is a ligand for the asialoglycoprotein receptor, was observed in all galactose-modified peptides. Based on this result, we could confirm that the internalization of the galactose-modified peptides occurred by the receptor-mediated endocytosis pathway. In addition, to understand the transport route of the peptide-DNA complex in the cell, the effects on the transfection efficiencies with several endocytosis inhibitors were examined. As a result, it was suggested that the translocation of the peptide-DNA complex from the endocytic compartments to the cytosol mainly occurred during an early endosome step.

Carcinoma, Hepatocellular↗
PubMed2002

Improved haemocompatibility of cysteine-modified polymers via endogenous nitric oxide.

A novel method for improving the haemocompatibility of biomedical materials through endogenous nitric oxide (NO) is presented. L-cysteine was covalently immobilized onto two biomedical polymers: polyurethane (PU) and polyethylene terephthalate (PET). The L-cysteine content on the polymers was approximately 5-8 nmol/cm2 as quantified via a chemiluminescence-based assay. The haemocompatibility of the modified polymers was evaluated in terms of the number of adhered platelets when exposed to a platelet suspension labeled with Cr51. Platelet adherence on the L-cysteine-modified polymers was reduced more than 50% as compared to the control (glycine-modified polymers) when the platelet suspension contained plasma constituents. No difference in platelet adhesion was observed in the absence of plasma constituents. Further experiments demonstrated that NO was easily transferred to the L-cysteine-modified polymers from S-nitroso-albumin in PBS buffer. The NO was then released from the polymer. NO transfer or release was not observed for the control. The results suggest that L-cysteine-modified polymers are effective in reducing platelet adhesion via the transfer of NO from endogenous S-nitrosoproteins in plasma to the polymer followed by the subsequent release of NO. Thus, exploiting endogenous NO is a viable option for improving the haemocompatibility of biomaterials.

Animals↗
PubMed2003

Immobilization of Chinese herbal medicine onto the surface-modified calcium hydrogenphosphate.

To accelerate the healing of bone defects or for healing to take place, it is often necessary to fill them with suitable substance. Various artificial materials defects have been developed. Among these, calcium phosphates and bioactive glass have been proven to be biocompatibile and bioactive materials that can chemically bond with bone, and have been successfully used clinically for repair of bone defects and augmentation of osseous tissue. However, those bioceramics have only the property of osteoconduction without any osteoinduction. Many ligands have been physicochemically absorbed onto substrates to enhance cell-substrate interactions. Although widely developed, they are still limited to use in long-term implantation because of their half-life period. Thus, some interfacial modification will be required for enhancing the efficacy of the delivery system. These models involve the immobilization of biologically active ligands of natural and synthetic origin onto various substrates to produce an interface with stronger chemical bond between ligand and substrate. The advantage of covalently immobilizing a ligand is that a chemical bond is present to prevent ligand or medicine from desorption. In our study, a two-step chemical immobilization was performed to surface-modified calcium hydrogenphosphate powders. The first was to modify the surface of calcium hydrogen-phosphate (CHP) with a coupling agent of hexanmethylene diisocyanate (HMDI). CHP surface modified by HMDI is abbreviated as MCHP. The linkage between CHP and HMDI will be characterized by FTIR. The second step was to immobilize chemically Gusuibu onto MCHP. Moreover, the sorption and desorption of Gusuibu was evaluated and quantitatively analyzed by spectrophotometer and HPLC. Bioceramic CHP was surface-modified by a two-step chemical immobilization. First, the surface of calcium hydrogen-phosphate (CHP) was successfully modified with coupling agent of hexanmethylene diisocyanate (HMDI). The first step was also activated the surface of CHP to induce primary amine terminator. The reaction of this functional group with Gusuibu was the second step. We confirmed simultaneously that Gusuibu could be immobilized chemically onto the surface of MCHP. Although some immobilized Gusuibu was also released rapidly at the first 12h, the degree of the released Gusuibu was lower than both by Gusuibu-adsorbing MCHP and Gusuibu-adsorbing CHP.

Adsorption↗
PubMed2003

Quantification of ligand surface concentration of bulk-modified biomimetic hydrogels.

This study describes a method for the quantification of active ligand surface concentration for bulk-modified hydrogels. Two poly(propylene fumarate-co-ethylene glycol) (P(PF-co-EG)) block copolymers were synthesized with terminal poly(ethylene glycol) (PEG) chains of number average molecular weight 1960 and 5190 g/mol. Hydrogels were synthesized with bulk-modified biotin as a model ligand, making use of a PEG spacer arm with a molecular weight of 3400 g/mol. Bulk concentration of biotin was calculated from the initial concentration of biotin, sol fraction, equilibrium water content, and relative incorporation of the polymers to the hydrogel. Surface concentration of biotin bulk-modified hydrogels was quantified with an enzyme linked immunosorbent assay using mouse monoclonal anti-biotin antibody (IgG), horseradish peroxidase-conjugated anti-mouse IgG, and a chemiluminescent substrate. The larger size of the IgG relative to the mesh size of the hydrogels allowed for the quantification of the active biotin at the surface of the hydrogels. Luminescent imaging was used to qualitatively show the isolation of the horseradish peroxidase-conjugated antibodies to the surface of the bulk-modified hydrogel. The active biotin ligands at the surface of hydrogels synthesized with terminal PEG chains of 1960 g/mol were at the top 7.2 nm while for those synthesized with terminal PEG chains of 5190 g/mol were at the top 4.4 nm of the bulk-modified hydrogel. The relationship between bulk ligand concentration and the active ligand concentration at the surface was dependent on the hydrogel composition. The relative magnitude of the PEG spacer arm of the ligand compared to the PEG block length of the copolymer affected the surface availability of the ligand. The results suggest that steric hindrances caused by mobile PEG chains of the copolymer of molecular weight greater than that of the PEG spacer arm contributed to the decreased surface concentration of ligand. This work relates the bulk concentration of a ligand to its surface concentration, an important parameter for the adhesion, migration, and function of anchorage dependent cells.

Biomimetic Materials↗
PubMed2003

In vivo conjunctival reconstruction using modified PLGA grafts for decreased scar formation and contraction.

The in vivo reconstruction of conjunctiva was investigated by using modified poly(lactide-co-glycolide) (PLGA) 50/50 scaffolds. The porous PLGA matrices were prepared by a solvent-casting particulate-leaching method with NaCl, then modified with collagen, hyaluronic acid (HA) or/and human amniotic membrane (AM) component. The growth of corneal epithelial cells and human stromal fibroblasts on the scaffolds was investigated in vitro. All the modified PLGA scaffolds demonstrated enhanced cell adhesion and proliferation as compared to PLGA untreated, and the number of cells proliferated after 1 week was increased in the order of PLGA<PLGA/collagen=PLGA/collagen/AM<PLGA/collagen/HA=PLGA/collagen/AM/HA, which was the same order as the adhesion of the cells onto the surfaces. Conjunctival wounds were created on the eyes of white rabbits bilaterally and PLGA/collagen/HA scaffolds were grafted for 4 weeks to evaluate the regeneration of ocular surface tissue at the wounds. At postoperative 4 weeks, all the wounds were completely covered with epithelial membranes. However, the contraction was minimal (6%) in the wound grafted with the modified PLGA, while that of the ungrafted wound was substantially large (25%). In addition, the collagen fibers regenerated within the scaffolds were characterized by a random array of a loose network that resembled natural conjunctiva. On the contrary, an aligned array of dense collagen fibers formed in the ungrafted wounds. Our results indicated that the modified PLGA graft as an acellular bed might allow the reconstruction of the damaged conjunctival tissue with less scar formation and contraction.

Animals↗
PubMed1999

Osteoblast population migration characteristics on substrates modified with immobilized adhesive peptides.

The process of cell migration is inextricably linked with the process of cell adhesion and, therefore, with cell/substrate adhesiveness. The present study adapted an under-agarose cell migration assay to quantitatively examine population migration characteristics of osteoblasts, on substrates modified with adhesive peptides, in the absence and presence of growth factors. Short-term, that is, 48 h osteoblast migration distances on substrates modified with adhesive Arg-Gly-Asp-Ser peptides were significantly (P < 0.05) less than migration distances on substrates modified with non-adhesive Arg-Asp-Gly-Ser peptides, demonstrating that osteoblast population haptokinesis was significantly decreased on substrates modified with adhesive peptides. Random motility coefficients calculated in the present study for osteoblast populations were an order of magnitude lower than a published random motility coefficient for leukocytes, proving quantitatively that, compared to leukocytes, osteoblasts migrate via haptokinesis more slowly. The 48 and 72 h osteoblast population migration differentials in the presence of an initial mass of 60 ng of basic Fibroblast Growth Factor, on substrates modified with Arg-Gly-Asp-Ser or with Arg-Asp-Gly-Ser, were larger than all other chemotactic differentials on these substrates. Quantitative investigations (such as the present study) of cell population migration characteristics on model biomaterial surfaces will become increasingly necessary as the discipline of cell/tissue engineering matures.

Adhesiveness↗
PubMed1999

An assessment of the strength of NG108-15 cell adhesion to chemically modified surfaces.

The strength of adhesion of NG108-15 cells to glass substrates modified with adsorbed proteins (laminin and poly-ornithine) or modified with covalently bound peptides (tri-ornithine and Tyr-Ile-Gly-Ser-Arg) was quantitatively assessed, by determining the shear stresses necessary to denude the cells from substrates using a spinning disk device. The shear stresses required to detach NG108-15 cells from glass modified with either adsorbed poly-ornithine or with both poly-ornithine and laminin were significantly (P < 0.05) higher than the shear stresses required to detach the cells from plain glass substrates. Covalent surface modifications resulted in higher strengths of NG108-15 adhesion than were exhibited on surfaces modified with adsorbed proteins. NG108-15 cell adhesion strength was maximal on surfaces covalently modified with only amine groups (without any peptides or proteins). These results indicate that general (i.e., not necessarily receptor-specific) surface modification strategies, which increase the net surface charge of a substrate, will elicit strong adhesion of NG108-15 cells.

Cell Adhesion↗
PubMedDate not supplied

Effects of modified positioning and mobilization of back pain and delayed bleeding in patients who had received heparin and undergone angiography: a pilot study.

OBJECTIVE: To examine the effects that a modified positioning and mobilization routine had no back pain and delayed bleeding in patients who had received heparin and undergone cardiac angiography. DESIGN: An experimental research design was used. Each patient was assigned randomly to either the control group, which required 6 hours of bed rest after cardiac angiography, or the experimental group. The experimental group had modified positioning, in which the head of the bed was elevated to a maximum of 45 degrees, and modified mobilization, in which they were ambulated briefly at the bedside 4 hours after angiography. SETTING: Two cardiology units of a 700-bed urban teaching hospital in western Canada. SAMPLE: All patients admitted for nonemergent cardiac angiography were approached for consent, to attain a sample of 29 patients, and were randomly assigned to the experimental or the control group. METHOD: Each patient was randomly assigned before cardiac angiography. The assignment was confidential until the patient was admitted to the cardiac unit after angiography. A demographic tool and the McGill Present Pain Intensity Scale were used to collect data. Perception of pain was evaluated over four observation periods. A research assistant monitored sanguineous drainage on the dressing and hematoma to evaluate the presence of delayed bleeding. DATA ANALYSIS: Demographic information was analyzed primarily through descriptive statistics. Results were analyzed to compare back pain and delayed bleeding between the two groups. Wilcoxon scores and t tests both were used for analysis and correlated well with each other. RESULTS: The group with the modified positioning and mobilization routine experienced significantly less pain overall (p = 0.02), less pain at each interval, and significantly less pain intensity (p < 0.05). There was no difference in bleeding. One person in each group had an estimated blood loss of more than 100 ml through the pressure dressing. CONCLUSION: This pilot study supports our hypothesis that modifying the immobilization of patients after cardiac angiography is associated with a reduction in back pain and with no increase of delayed bleeding at the femoral access site. The results support the need for further investigation of ambulation interventions after cardiac angiography.

Adult↗
PubMed1990

Modified Allen pictures to assess amblyopia in young children.

To produce a test of visual acuity for young children that is more sensitive to amblyopia than current preschool vision tests, the authors surrounded four Allen pictures with "crowding" bars. This modified Allen test was evaluated by measuring acuity of amblyopic children (n = 28) and children (n = 10) and adults (n = 5) with normal eyes. Mean acuities of amblyopic eyes for the modified pictures was 0.8 octaves or nearly three logMar Snellen lines poorer than for the isolated pictures. For nonamblyopic and normal eyes modified picture acuity averaged 0.1 to 0.4 octaves poorer than isolated picture acuity. Average acuities of amblyopic eyes (n = 22) for the modified pictures agreed with line letter acuities. These results suggest that the modified picture test induces contour interaction similar to that of line letter tests, and thus, is a more sensitive test of amblyopia in the preschool child than isolated symbols.

Adolescent↗
PubMed1997

Reactive oxygen species modified thymine and poly(dT) present unique epitope for human anti-DNA autoantibodies.

Hydroxyl radical, one of the most potent of all reactive oxygen species has been implicated in many human degenerative diseases and is known to modify adenine and thymine in cellular DNA. In the present studies, adenine, thymine and their synthetic homopolymers poly(dA), poly(dT) were ROS-modified and subsequently used as inhibitors of native DNA binding to human anti-DNA autoantibodies. Besides nDNA, modified thymine and poly(dT) were effective inhibitors of DNA-anti-DNA antibody interaction. The relative affinity of ROS-modified poly(dT) was better than that of native DNA. Visual detection of modified thymine and poly(dT) binding to affinity purified anti-DNA IgG by an indirect band shift assay support competition inhibition data. The enhanced recognition of ROS-DNA by anti-DNA autoantibodies, as reported earlier, could be due to the ROS-induced modification of thymine.

Adenine↗
PubMed2000

A modified mercury saturation assay for measuring metallothionein.

A thorough re-evaluation of assay procedures for a mercury displacement analytical method for metallothionein (MT) described in 1993, was conducted using an analytical standard of rabbit liver MT-II and MT determinations, in liver, kidney, gill and intestine from lake trout (Salvelinus namaycush). These studies indicated that: (1) loss of MT by adherence to polypropylene material occurs with the dilution series; (2) use of a calibration curve using a rabbit liver MT-II analytical standard, is superior to the dilution series for determining unknown MT concentrations; (3) Hg incubation time requirements are less in 20% TCA than in 10% TCA; and (4) employing both 20% TCA and heat at 95 degrees C for 5 min to denature non MT proteins results in the most accurate MT spike recoveries, or recoveries that did not differ significantly from those closest to 100%, in liver, kidney, gill and intestine. Calibration curves with an average r(2) of 0.9998 for 69 trials over different days, while passing through the origin, provided evidence of negligible non-specific binding of mercury. Use of water or saline for preparing tissue homogenates, produced slight, but significant, differences in MT estimates in the intestine and kidney. Comparisons between the original and the modified methods were made on liver, kidney and gill from control, sham-injected and Cd-treated lake trout. Differences in MT estimates obtained between the two methods depended upon the tissue. MT estimates in liver by the modified method were 9, 13, and 10% higher in control, sham-injected and Cd-treated groups, respectively, than those resulting from the original assay. MT estimates in gill by the modified method, were 3.2-fold, 4.0-fold and 69% higher in control, sham-injected, and Cd-treated groups, respectively. Corresponding values for kidney were 37, 75, and 57% higher. Because percent recoveries of MT-II spikes were virtually 100%, in all four tissues, using the modified method, MT estimates obtained using these modifications were more accurate than those obtained using the procedures of the original assay. Application of the modified assay to fish tissues in a Canadian biomonitoring program for metal mining, demonstrated that in 1 week an experienced technologist can complete 90 MT analyses of liver and kidney and 75 analyses of gill and intestine.

Journal Article↗
PubMed1997

Methylglyoxal-modified arginine residues--a signal for receptor-mediated endocytosis and degradation of proteins by monocytic THP-1 cells.

Non-enzymatic glycosylation or glycation of proteins to form advanced glycation endproducts (AGE) has been proposed as a process which provides a signal for the degradation of proteins. Despite this, the AGE which act a recognition factor for receptor-mediated endocytosis and degradation of glycated proteins by monocytes and macrophages has not been identified. Methylglyoxal, a reactive alpha-oxoaldehyde and physiological metabolite, reacted irreversibly with arginine residues in proteins to form Ndelta-(5-hydro-5-methyl-4-imidazolon-2-yl)ornithine and Ndelta-(5-methyl-4-imidazolon-2-yl)ornithine residues. Human serum albumin minimally-modified with methylglyoxal (MG(min)-HSA) was bound by cell surface receptors of human monocytic THP-1 cells in vitro at 4 degrees C: the binding constant K(d) value was 377 +/- 35 nM and the number of receptors per cell was 5.9 +/- 0.2 X 10(5) (n = 12). N alpha-Acetyl-Ndelta-(5-hydro-5-methyl-4-imidazolon-2-yl)orni thine displaced MG(min)-HSA from THP-1 cells, suggesting that the Ndelta-(5-hydro-5-methyl-4-imidazolon-2-yl)ornithine residue was the receptor recognition factor. At 37 degrees C, MG(min)-HSA was internalised by THP-1 cells and degraded. Similar binding and degradation of human serum albumin modified by glucose-derived AGE was found but only when highly modified. MG(min)-HSA, therefore, is the first example of a protein minimally-modified by AGE-like compounds that binds specifically to monocyte receptors. The irreversible modification of proteins by methylglyoxal is a potent signal for the degradation of proteins by monocytic cells in which the arginine derivative, Ndelta-(5-hydro-5-methyl-4-imidazolon-2-yl)ornithine, is the receptor recognition factor. This factor is not present in glucose-modified proteins.

Arginine↗
PubMed2002

Effect of poly(ethylene glycol) grafts on temperature-sensitivity of thermosensitive polymer-modified liposomes.

A copolymer of N-isopropylacrylamide and N-acryloylpyrrolidine, which exhibits a lower critical solution temperature (LCST) at 38 degrees C, having a didodecyl group at the chain terminal was synthesized and calcein-loaded liposomes were prepared from a mixture of dioleoylphosphatidylethanolamine and the copolymer. While the contents release from the copolymer-modified liposomes was strongly accelerated around the LCST of the copolymer, some extent of the release was also observed below the LCST, probably because the partly-dehydrated copolymer chains interacted with the liposome membrane. When poly(ethylene glycol) with the number average molecular weight of 550 having a didodecyl group at the chain end was introduced into the copolymer-modified liposomes, the contents release from the copolymer-modified liposomes was effectively suppressed below the LCST but hardly affected above the LCST. As a result, the liposomes exhibited much sharper enhancement of the contents release in a narrow temperature region. The temperature-sensitive contents release property of the copolymer-modified liposomes became less significant in the presence of serum. However, inclusion of the anchor-bearing poly(ethylene glycol) derivative improved the contents release behavior of the copolymer-modified liposomes in the presence of serum.

Chemistry, Pharmaceutical↗