Maghemite nanoparticles coated with human serum albumin: combining targeting by the iron-acquisition pathway and potential in photothermal therapies.

Human serum albumin (HSA), the most abundant plasma protein in human blood, is a natural transport vehicle with multiple ligand binding sites. It, therefore, constitutes an attractive candidate for drug delivery. Targeting may occur via the most known interaction of the protein with the neonatal Fc receptor (FcRn). Here, we investigate another HSA delivery path, involving the transferrin receptor, and we elaborate a maghemite-HSA nanohybrid, opening up new opportunities for medical applications. Fluorescence spectrophotometric titration and size-exclusion chromatography were used to substantiate, in cell-free assays, an interaction between HSA and the transferrin receptor R1. This occurs with a dissociation constant, KD of 6.7 nM. This interaction was confirmed in HeLa cell culture where, by confocal microscopy, rhodamine-labeled HSA is shown to be internalized. HSA was then covalently conjugated onto maghemite nanoparticles (NPs) to give a NP-HSA nanohybrid. The therapeutic potential of this hybrid was demonstrated through its heating capacity in magnetic hyperthermia (MH) and near-infrared (NIR) photothermia (PT). In particular, the Specific Absorption Rate (SAR) in the PT Therapy (PTT) mode, using a 808 nm NIR-LASER (1 W cm-2) and at iron concentration as low as 2.5 mM, was found to be very high, equal to 1870 W g-1 with a temperature increment of 9.2 °C. The nanohybrids incubated with HeLa cells were mainly localized at the cell surface. When the PTT mode was applied under the same conditions as in vitro, mortality was higher in HeLa cells than in fibroblasts (non-malignant cells). Cytotoxicity was checked in both cell lines without the PTT mode; the nanohybrids do not seem to affect cell viability. These results make the nanohybrids very promising agents for NIR-PT and for targeting in cancer therapy, since non-malignant cells were not damaged.

Disfunción tiroidea y corazón / Thyroid dysfunction and heart

Los trastornos de la función tiroidea afectan profundamente al sistema cardiovascular. En esta revisión se presentan algunos aspectos fisiológicos de la interrelación entre tiroides y corazón, como también las consecuencias de la tirotoxicosis e hipotiroidismo sobre el aparato cardiovascular. Se analiza la influencia del hipertiroidismo en la gèc)nesis de la fibrilación auricular y del hipotiroidismo en el metabolismo de las lipoproteínas. Adicionalmente, el artículo se referirá a los potenciales efectos adversos del antiarrítmico amiodarona sobre la función tiroidea y cómo se investigan y tratan. Finalmente, se expone un caso clínico real para ilustrar con mayor claridad la enorme importancia que pueden alcanzar las relaciones fisiopatológicas entre el corazón y las afecciones de esta glándula endocrina.

Disorders of thyroid function profoundly affect the cardiovascular system. Inthisreviewsomephysiologicalaspectsoftherelationship between thyroid and the heart as well as the consequences of thyrotoxicosis and hypothyroidism on the cardiovascular system are presented. The influence of hyperthyroidism is analyzed in the genesis of atrial fibrillation and of hypothyroidism on lipoprotein metabolism. Furthermore, we refer to the potential adverse effects of the antiarrhythmic amiodarone on thyroid function and how they are investigated and treated. Finally, a real clinical case is exposed to more clearly illustrate the enormous importance that can reach the pathophysiological relationships between the heart and the diseases of the thyroid gland.

[Irreversible monoamine oxidase inhibitors (IMAOI) to treat depressive disorders - limited use at present in Flanders]. / Irreversibele monoamineoxidaseremmers bij depressieve stoornissen: snel afgenomen gebruik in Vlaanderen.

BACKGROUND: Irreversible monoamine oxidase inhibitors (imaoi) are rarely used in Flanders. Such an anti-imaoi policy is not in keeping with the role that imaoi now play in the general guidelines for the treatment of depressive disorders. AIM: To provide an overview of the history and the current use of imaoi in Flanders. METHOD: We searched the literature and the literature used in the psychiatric courses taught at Ghent University and the Catholic University of Leuven and we consulted the Acta (Neurologica et) Psychiatrica Belgica. The information we collected was supplemented by personal communications from experts and by data about the period of commercialisation, the pharmaceutical companies producing imaoi and the use of imaoi. RESULTS: imaoi were introduced rapidly onto the Flemish market but their popularity was short-lived. University courses did not give much attention to imaoi and the attitude to these inhibitors was negative. At the moment, phenelzine is the only imaoi available on the Flemish market and is only rarely prescribed. CONCLUSION: Following the international trend, imaoi in Flanders initially enjoyed a short period of popularity. However, the limited use of phenelzine at present is not in line with the current guidelines for the treatment of depressive disorders. Practitioners and health professionals need to be better informed. Better education and wider use of imaoi in Flanders are recommended.

Design and synthesis of 3-isoxazolidone derivatives as new Chlamydia trachomatis inhibitors.

Chlamydia trachomatis (Ct) is a bacterial human pathogen responsible for the development of trachoma, the worldwide infection leading to blindness, and is also a major cause of sexually transmitted diseases. As iron is an essential metabolite for this bacterium, iron depletion presents a promising strategy to limit Ct proliferation. The aim of this study is to synthesize 3-isoxazolidone derivatives bearing known chelating moieties in an attempt to develop new bactericidal anti-Chlamydiaceae molecules. We have investigated the paths by which these new compounds affect Ct serovar L2 development in HeLa cells, in the presence or absence of exogenously added iron. The iron-chelating properties of these molecules were also determined. Our data reveal important bactericidal effects which are distinguishable from those due to iron chelation.

Porphyromonas gingivalis infection sequesters pro-apoptotic Bad through Akt in primary gingival epithelial cells.

Porphyromonas gingivalis, a self-limiting oral pathogen, can colonize and replicate in gingival epithelial cells (GECs). P. gingivalis-infected GECs are protected from mitochondrion-dependent apoptosis, partially through activation of phosphatidyl inositol 3-kinase/Akt signaling. Biochemical events associated with P. gingivalis-induced inhibition of apoptosis include the blocking of mitochondrial membrane permeability and cytochrome-c release. We studied functional importance of Akt and the status of associated key mitochondrial molecules, pro-apoptotic Bad and caspase-9, during infection of GECs. We found that P. gingivalis infection caused significant phosphorylation of Bad progressively, while messenger RNA levels for Bad slowly decreased. Fluorescence microscopy showed translocation of the mitochondrial Bad to the cytosol post-infection. Conversely, P. gingivalis lost the ability to promote phosphorylation and translocation of Bad in Akt-deficient GECs. Caspase-9 activation induced by a chemical inducer of apoptosis was significantly inhibited by infection over time. However, Akt depletion by small interfering RNA did not reverse inhibition of caspase-9 activation by infection. Hence, P. gingivalis inactivates pro-apoptotic Bad through Akt. The inhibition of caspase-9 activation appears to be independent of Akt. Overall, our findings suggest that Akt is a key component of anti-apoptotic pathways stimulated by P. gingivalis. The P. gingivalis uses other mitochondrial pathways to protect host cells from cell-death and to ensure its survival in gingival epithelium.

Tiempo de exposición al gluten y marcadores de riesgo de diabetes mellitus insulino dependiente en pacientes celíacos / Risk markers for insulin-dependent diabetes mellitus and duration of exposure to gluten in celiac patients

Background: Celiac patients are at high risk of developing insulin-dependent diabetes mellitus, a condition that has a long pre-diabetic period. During this lapse, anti-islet cell antibodies serve as markers for future disease. This may be related with the duration of the exposure to gluten. Aim: To test the hypothesis that long term adherence to a gluten free diet decreases the frequency of risk markers for insulin dependent diabetes mellitus during adolescence and early adulthood. Patients and methods: 158 celiac patients were classified as: G1, (n=30 patients) studied at the time of diagnosis; G2 (n=97 patients) exposed to gluten as a result of non compliance with the gluten free diet and, G3 (n=31 patients) who had maintained a long term, strict gluten free diet. Isotype IgG anti-islet cell antibodies were detected by indirect immunofluorescence using monkey pancreas; results were reported in Juvenile Diabetes Foundation (JDF) units. Results: Celiac patients exposed to a gluten containing diet had a significantly higher prevalence of anti-islet cell antibodies than those who had been exposed only briefly (p <0.017). In addition, a significantly higher prevalence of anti-islet cell antibodies was observed in those patients whose exposure to gluten was longer than 5 years than in those whose exposure was shorter (p <0.02). Conclusions: Celiac patients long exposed to gluten have a significantly higher prevalence of anti-islet cell antibodies than those exposed for a short period. This fact supports the hypothesis that the development of these antibodies is associated with the length of the exposure to gluten (Rev Méd Chile 2004; 132: 979-84).

Dieta libre de gluten y disminución del riesgo de asociación de enfermedades autoinmunes en el paciente celiaco / Gluten free diet and decrease in the risk of association of autoinmune disease in celiac patient

Estudios publicados concluyen que el desarrollo de enfermedades autoinmunes en pacientes celiacos se incrementa al aumentar el tiempo de exposición al gluten y cuanto mayor sea la edad al diagnóstico. La adolescencia constituye un periodo de la vida en el cual una dieta libre de gluten frecuentemente se abandona. Por esto es de suma importancia investigar marcadores inmunológicos endocrinos en pacientes celiacos que cumplen o no la dieta libre de gluten.

Oligomeric and polymeric aggregates formed by proteins containing expanded polyglutamine.

Neurological diseases resulting from proteins containing expanded polyglutamine (polyQ) are characteristically associated with insoluble neuronal inclusions, usually intranuclear, and neuronal death. We describe here oligomeric and polymeric aggregates formed in cells by expanded polyQ. These aggregates are not dissociated by concentrated formic acid, an extremely effective solvent for otherwise insoluble proteins. Perinuclear inclusions formed in cultured cells by expanded polyQ can be completely dissolved in concentrated formic acid, but a soluble protein oligomer containing the expanded polyQ and released by the formic acid is not dissociated to monomer. In Huntington's disease, a formic acid-resistant oligomer is present in cerebral cortex, but not in cerebellum. Cortical nuclei contain a polymeric aggregate of expanded polyQ that is insoluble in formic acid, does not enter polyacrylamide gels, but is retained on filters. This finding shows that the process of polymerization is more advanced in the cerebral cortex than in cultured cells. The resistance of oligomer and polymer to formic acid suggests the participation of covalent bonds in their stabilization.

Anticuerpos anti-transglutaminasa tisular: ¿una mejor prueba para detectar transgresiones menores de la dieta en pacientes celíacos? / Tissue transglutaminase antibodies: a better test to detec minor dietary trasgressions in celiac patients?

Background: Endomysium antibodies (EMA) do not detect minor dietary transgressions in patients with celiac disease. Aim : To compare the sensitivity and specificity of tissue transglutaminase antibodies (tTGA) and endomysium antibodies (EMA) in biopsy proven celiac patients at the time of diagnosis and during gluten free diet (GFD). Patients and methods : One hundred fifty three subjects were studied: a) 30 healthy controls; b) 9 cases with cow's milk allergy; c) 24 celiac patients at time of diagnosis; d) 25 celiac patients adhering to the GFD; e) 65 celiac patients with poor/no adhesion to GFD. EMA and tTGA IgA were measured by immunofluorescence and ELISA, respectively. Results: Sensitivity and specificity were 100 percent and 97.4 percent for tTGA, respectively. All patients with cow's milk allergy were EMA (-) and 8 of 9 (88.9 percent) were tTGA (-). In celiac patients not adhering to the GFD, EMA and tTGA positivity were similar (80 percent and 81,5 percent, respectively); 95,4 percent of the subjects tested positive for at least one of them. All patients adhering to GFD were EMA (-) but tTGA were (+) in 28 percent of them. Conclusions : EMA and tTGA have similar sensitivity and specificity at the time of diagnosis of celiac disease. Positive tTGA in 28 percent of patients that adhered strictly to the GFD and whose EMA were negative suggest that tTGA may be helpful in detecting minor dietary transgressions and should be further evaluated

Reduced levels of oxidized and glycoxidized proteins in human fibroblasts exposed to repeated mild heat shock during serial passaging in vitro.

Repeated mild heat shock (RMHS) has beneficial hormesis-like effects on various characteristics of human skin fibroblasts undergoing replicative senescence in vitro. We have tested whether RMHS could reduce the accumulation of oxidized and glycoxidized proteins, which is a major age-related change. Levels of carbonylated proteins, furosine, N(epsilon)-carboxymethyl-lysine-rich proteins and advanced glycation end products increased during serial passaging of fibroblasts in culture. However, the extent of accumulation of oxidized and glycoxidized proteins was significantly reduced in RMHS cells. The basal concentration of reduced glutathione was higher and that of oxidized glutathione was lower in RMHS cells. Whereas the basal level of heat shock protein HSP27 decreased in both RMHS and control cells during serial passaging, the increase of the basal level of HSP70 with increasing passage level was significantly higher in RMHS cells. These results show that the slower accumulation of damaged proteins in fibroblasts exposed to RMHS results partly from the increased ability of these cells to cope with oxidative stress, and to synthesize HSP responsible for protein capping and refolding.

Papel del tejido conectivo en la morfología y función de la mucosa intestinal: su importancia en la patogenia de la enfermedad celíaca / Role of the connective tissue in the morphology and function of intestinal mucosa: its participation in the pathogenesis of celiac disease

The differentiation, architecture and function of the mucosa of the digestive tract is the result of close interactions between the epithelial cells and their underlying connective tissue. This interaction not only plays a role in the normal morphological organization of the intestinal mucosa but also in the organization and maturation of the enterocytes, the development of some of their enzymatic activities, the transport of nutriens and the restitution of the epithelium following denudation of the surface of villi due to loss of enterocytes. It is thought that disturbances of the cells of the connective tissue, especially of the myofibroblasts as well as of some of the above mentioned mechanisms, participate in the pathogenesis of some gastrointestinal diseases, celiac disease among them. This review summarizes some of the knowledge in this area

Proteasome inhibition in glyoxal-treated fibroblasts and resistance of glycated glucose-6-phosphate dehydrogenase to 20 S proteasome degradation in vitro.

Glycation and glycoxidation protein products are formed upon binding of sugars to NH(2) groups of lysine and arginine residues and have been shown to accumulate during aging and in pathologies such as Alzheimer's disease and diabetes. Because the proteasome is the major intracellular proteolytic system involved in the removal of altered proteins, the effect of intracellular glycation on proteasome function has been analyzed in human dermal fibroblasts subjected to treatment with glyoxal that promotes the formation of N epsilon-carboxymethyl-lysine adducts on proteins. The three proteasome peptidase activities were decreased in glyoxal-treated cells as compared with control cells, and glyoxal was also found to inhibit these peptidase activities in vitro. In addition, the activity of glucose-6-phosphate dehydrogenase, a crucial enzyme for the regulation of the intracellular redox status, was dramatically reduced in glyoxal-treated cells. Further analysis was performed to determine whether glycated proteins are substrates for proteasome degradation. In contrast to the oxidized glucose-6-phosphate dehydrogenase, both N epsilon-carboxymethyl-lysine- and fluorescent-glycated enzymes were resistant to degradation by the 20 S proteasome in vitro, and this resistance was correlated with an increased conformational stability of the glycated proteins. These results provide one explanation for why glycated proteins build up both as a function of disease and aging. Finally, N epsilon-carboxymethyl-lysine-modified proteins were found to be ubiquitinated in glyoxal-treated cells suggesting a potential mechanism by which these modified proteins may be marked for degradation.

Heat shock response and ageing: mechanisms and applications.

Ageing is associated with a decrease in the ability of cells to cope with environmental challenges. This is due partly to the attenuation of a primordial stress response, the so-called heat shock (HS) response, which induces the expression of heat shock proteins (HSPs), composed of chaperones and proteases. The attenuation of the HS response during ageing may be responsible for the accumulation of damaged proteins as well as abnormal regulation of cell death. Maintenance of the HS response by repeated mild heat stress causes anti-ageing hormetic effects on cells and organisms. Here, we describe the molecular mechanism and the state of the HS response as well as the role of specific HSPs during ageing, and discuss the possibility of hormetic modulation of ageing and longevity by repeated mild stress.

N-phenacylthiazolium bromide decreases renal and increases urinary advanced glycation end products excretion without ameliorating diabetic nephropathy in C57BL/6 mice.

AIMS: Advanced glycation end products (AGE), which form from the non-enzymatic reaction of proteins and sugars, have been implicated in the pathogenesis of diabetic nephropathy. Recently, a compound [N-phenacylthiazolium bromide (PTB)] has been described which cleaves alpha,beta-dicarbonyl compounds. In the present study we used diabetic C57BL/6 mice to determine if PTB altered renal AGE levels and reduced diabetic glomerulosclerosis. METHODS: Mice with stable hyperglycaemia induced by streptozotocin were given daily subcutaneous injections of either PTB (10 microg/g) or saline for 12 weeks. Renal-collagen bound AGE and urinary AGE-peptides were measured by ELISA using an anti-AGE-RNase antibody. Renal collagen-released Nepsilon(carboxymethyl)lysine (CML) and pentosidine were determined by high pressure liquid chromatography (HPLC). Glomerular lesions (volume and mesangial/total surface area) were evaluated by computer-assisted image analysis. We determined urinary protein/creatinine ratio as a functional parameter. AGE localization was examined by immunohistochemistry using the anti-AGE-RNase antibody. RESULTS: Renal collagen-bound AGE were decreased and urinary AGE excretion was increased in PTB-treated diabetic mice. However, collagen-released CML and pentosidine were similar in both groups. Glomerular histology and morphometric analysis revealed also no differences between PTB-and saline-treated diabetic mice. The urinary protein/creatinine ratio was unaffected by PTB-treatment. AGE staining by anti-AGE-RNase antibody was present in Bowman's capsules, glomerular basement membranes and cortical tubules. It was decreased in all structures in PTB-treated diabetic mice. CONCLUSION: In summary, PTB decreased renal AGE accumulation but did not ameliorate glomerular lesions or proteinuria. Thus, cleavage of AGE by PTB is not sufficient to prevent development of diabetic nephropathy in C57BL/6 mice.

Inhibition of nitric oxide synthase activity by early and advanced glycation end products in cultured rabbit proximal tubular epithelial cells.

Nitric oxide (NO) is important in the regulation of renal tubular function. We have investigated whether glycated proteins could impair the NO production by examining the effects of Amadori products (AP-BSA) and advanced glycation end products (AGE-BSA) on primary cultures of rabbit proximal tubular epithelial (PTE) cells. Nitric oxide synthase activity was assessed by measurement of the conversion of L-arginine to L-citrulline and by production of NO, after short-term (30 min) or long-term (1 or 3 days) incubation. Short incubations of PTE cells with either 200 microg/ml AP-BSA or 40 microg/ml AGE-BSA significantly decreased NO production. AP-BSA (3000 microg/ml) inhibited the Ca(2+)-dependent NOS activity even though above 50 microg/ml it increased Ca(2+)-independent NOS activity. In contrast, 40 microg/ml AGE-BSA inhibited both isoforms of NOS. Longer incubations with 200 microg/ml AP-BSA or 250 microg/ml AGE-BSA decreased NO release and inhibited Ca(2+)-dependent and -independent NOS activities. APs did not affect NO release by S-nitroso-N-acetyl-penicillamine (SNAP), while 250 microg/ml AGEs decreased it. After 3 days incubation, glycation products had no effect on the NOS cell content. Cell viability and proliferation were not modified under these experimental conditions, suggesting that the fall in NO production was not due to there being fewer cells. These data indicate that APs and AGEs directly inhibit NOS activity, and additionally that AGEs quench released NO. Thus, both types of glycated proteins alter the production of NO by PTE cells and could participate in the renal tubule dysfunction associated with aging and diabetes.

Kinetin inhibits protein oxidation and glycoxidation in vitro.

We tested the ability of N(6)-furfuryladenine (kinetin) to protect against oxidative and glycoxidative protein damage generated in vitro by sugars and by an iron/ascorbate system. At 50 microM, kinetin was more efficient (82% inhibition) than adenine (49% inhibition) to inhibit the bovine serum albumin (BSA)-pentosidine formation in slow and fast glycation/glycoxidation models. Kinetin also inhibited the formation of BSA-carbonyls after oxidation significantly more than adenine did. However both compounds inhibited the advanced glycation end product (AGE) formation to the same extent (59-68% inhibition). At 200 microM, kinetin but not adenine, limited the aggregation of BSA during glycation. These data suggest that kinetin is a strong inhibitor of oxidative and glycoxidative protein-damage generated in vitro.

Modulating cellular aging in vitro: hormetic effects of repeated mild heat stress on protein oxidation and glycation.

Intracellular and extracellular proteins are subject to a variety of spontaneous non-enzymatic modifications which affect their structure, function and stability. Protein oxidation and glycation are tightly linked and are implicated in the development of many pathological consequences of aging. Although multiple endogenous pathways in the cell can prevent the formation of oxidized and glycated proteins, and repair and degrade abnormal proteins, such abnormal proteins do accumulate during aging. The heat shock response involving the family of stress-proteins or the so-called heat shock proteins (HSP), represents the quickest and highly conserved response to proteotoxic insults. Since repeated mild heat stress is able to prevent the onset of various age-related changes during cellular aging in vitro, we suggest that treatments which increase HSP expression should reduce the extent of accumulation of abnormal proteins during aging. Such modulation of aging is an example of hormesis, which is characterized by the beneficial effects resulting from the cellular responses to mild repeated stress.

Accumulation of advanced glycation endproducts in the rat nephron: link with circulating AGEs during aging.

The accumulation of advanced glycosylation end products (AGEs) is believed to be a factor in the development of aging nephropathy. We have attempted to establish a link between the formation of AGEs and the onset of renal impairment with aging, indicated by albuminuria, using a fluorescence assay and immunohistochemical detection of AGEs in the renal extracellular matrix in rats. The fluorescence of collagenase-digested Type IV collagen from GBM increased with age, from 1.65 +/- 0.05 AU/mM OHPro (3 months) and 1.58 +/- 0.04 (10 months) to 2.16 +/- 0.06 (26 months) (p < 0.001) and 2.53 +/- 0.18 (30 months) (p < 0.001). In contrast, the extent of early glycation products significantly decreased from 5.35 +/- 0.25 nmol HCHO/nmol OHPro at 3 months to 3.14 +/- 0.19 at 10 months (p < 0.001), 3.42 +/- 0.38 at 26 months, and 0.74 +/- 0.08 at 30 months (p < 0.001). The urinary fluorescence of circulating AGE rose from 2.42 +/- 0.15 AU/mg protein (3 months), 1.69 +/- 0.07 (10 months), to 4.63 +/- 0.35 (26 months) (p < 0.01) and 4.73 +/- 0.72 (30 months), while the serum fluorescence increased from 0.39 +/- 0.02 AU/mg protein at 3 months and 0.43 +/- 0.02 at 10 months to 0.59 +/- 0.04 at 26 months (p < 0.001) and 0.54 +/- 0.03 at 30 months (p < 0.04). Polyclonal antibodies raised against AGE RNase showed faint areas of AGE immunoreactivity in mesangial areas in the nephrons of young rats. The immunolabeling of Bowman's capsule, the mesangial matrices, and the peripheral loops of glomerular and tubule basement membranes increased with rat age. The increase in circulating AGE peptides parallels the accumulation of AGEs in the nephron, and this parallels the pattern of extracellular matrix deposition, suggesting a close link between AGE accumulation and renal impairment in aging rats.

Effect of glycation of albumin on its binding to renal brush-border membrane vesicles: influence of aging in rats.

Aging is associated with the loss of preferential urinary excretion of Amadori-product glycated albumin. We have measured the binding of 125I-labeled glycated albumin to the renal brush-border membrane vesicles from young and old rats to determine whether a specific receptor-mediated endocytosis system may be involved. 125I-Glycated albumin was specifically bound by renal brush-border membrane vesicles in a time- and temperature-dependent manner; the binding was concentration-dependent, saturable and reversible. Scatchard plots gave an apparent dissociation constant Km of 488 +/- 17 nM, and a number of binding sites N of 33.5 +/- 3.4 pmol/mg protein/min in membrane vesicles from young (3 months old) rats; the binding of native [125I]albumin, gave a Km of 1194 +/- 200 nM (P < 2%) and N of 82.4 +/- 16.3 pmol/mg protein/min (P < 3%). Vesicles from 10-month-old rats had a similar Km (619.6 +/- 135.3 nM) and N (21.91 +/- 2.98 pmol/mg protein/min), while those from older (30 months old) rats had significantly increased Km (1344 +/- 237 nM, P < 3%) and N (81.3 +/- 10.9 pmol/mg protein/min, P < 1%) for 125I-glycated albumin binding. 125I-Glycated HSA was not displaced by unlabeled native HSA in less than 100-fold excess and native [125I]HSA was only displaced by a 10-fold excess of unlabeled glycated HSA. The binding of native [125I]HSA was partly inhibited (85%) by unlabeled glycated HSA. Thus, there appear to be two different binding sites, one for glycated and the other for native albumin, lying close together; and the glycation site on albumin is the discriminatory recognition factor.

Glycation of albumin with aging and diabetes in rats: changes in its renal handling.

Albumin glycation was investigated in old rats to elucidate the link between the preferential excretion of glycated albumin and age-related microalbuminuria. Postprandial blood glucose and the glycated albumin in the serum and urine of 3-, 10- and 30-month-old Wistar rats and in streptozotocin diabetic rats were determined. Blood glucose increased from 1.46 +/- 0.046 g l-1 in 3-month-old rats to 2.08 +/- 0.06 (10 months) and 1.75 +/- 0.23 (30 months) (P < 0.05). Albumin glycation level in the serum increased from 0.79 +/- 0.07 nmol HCHO/nmol albumin (3 months) to 1.41 +/- 0.14 (10 months) and 1.73 +/- 0.21 (30 months) (P < 0.05); urinary level increased from 1.63 +/- 0.39 nmol HCHO/nmol albumin (3 months) to 2.92 +/- 0.57 (10 months) and 2.39 +/- 0.36 (30 months) (P < 0.01). The percent glycated albumin in serum rose from 3.33 +/- 0.64 to 6.81 +/- 0.63 and 6.99 +/- 1.79% of total albumin (P < 0.05), whereas the urine percentage decreased from 12.81 +/- 3.97 to 12.64 +/- 2.87 and 2.63 +/- 0.97% (P < 0.05) in 3-, 10- and 30-month-old rats, respectively. Editing decreased with aging from 4.28 +/- 0.83 (3 months) to 1.84 +/- 0.32 (10 months) and 0.52 +/- 0.14 (30 months) (P < 0.01). Editing in microproteinuric diabetic rats was lower (0.95 +/- 0.08) than in 3-month-old control rats (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)