Association of VO2 and VCO2 rate variability with serum glucose, insulin, and glucose intolerance.

Changes in the cellular metabolism assessed by the variability of oxygen consumption (VO(2) ) and carbon dioxide production (VCO(2) ) as well as the association of serum glucose and insulin to energy spectral density (ESD) of VO(2) and VCO(2) were evaluated. Ten nonglucose intolerant and 10 glucose intolerant subjects, aged 21-70 years, were included. Glucose and insulin concentrations and VO(2) and VCO(2) records were collected every 10 min during 3 h. ESD of VO(2) and VCO(2) was estimated and associated with glucose and insulin concentrations. Statistical significance in glucose levels, insulin, and ESD of VO(2) and VCO(2) among nonglucose intolerant subjects and glucose and insulin among glucose intolerance subjects at postload glucose (PLG) state compared with basal state was found. Moreover, glucose was significantly higher in glucose intolerance subjects than nonglucose intolerant subjects for basal and PLG states. These results show an increment in ESD of VO(2) and VCO(2) at PLG state among nonglucose intolerant subjects and suggest that their measurement may be a key indicator of the variability of cellular metabolic activity and contribute to confirm disturbances in glucose metabolism.

Resistant Starch Consumption Effects on Glycemic Control and Glycemic Variability in Patients with Type 2 Diabetes: A Randomized Crossover Study.

We previously observed beneficial effects of native banana starch (NBS) with a high resistant starch (RS) content on glycemic response in lean and obese participants. Here, we aimed to determine the effects of NBS and high-amylose maize starch (HMS) on glycemic control (GC) and glycemic variability (GV) in patients with type 2 diabetes (T2D) when treatments were matched for digestible starch content. In a randomized, crossover study, continuous glucose monitoring (CGM) was performed in 17 participants (aged 28-65 years, BMI ≥ 25 kg/m2, both genders) consuming HMS, NBS, or digestible maize starch (DMS) for 4 days. HMS and NBS induced an increase in 24 h mean blood glucose during days 2 to 4 (p < 0.05). CONGA, GRADE, and J-index values were higher in HMS compared with DMS only at day 4 (p < 0.05). Yet, NBS intake provoked a reduction in fasting glycemia changes from baseline compared with DMS (p = 0.0074). In conclusion, under the experimental conditions, RS from two sources did not improve GC or GV. Future longer studies are needed to determine whether these findings were affected by a different baseline microbiota or other environmental factors.

Molecular susceptibility to glycation and its implication in diabetes mellitus and related diseases.

The modification of free amino groups on proteins, lipids, and nucleic acids by non-enzymatic glycosylation produce a variety of complex structures named advanced glycation end products (AGEs). Glycation of these molecules participate in the development of diabetic complications and related diseases. Diabetes mellitus is characterized by short-term metabolic changes in lipid and protein metabolism, and long-term irreversible changes in vascular and connective tissue. AGEs are directly implicated in the development of chronic complications in diabetes such as nephropathy, rethinopathy, neuropathy, and other related diseases such as atherosclerosis, heart disease, stroke, and peripheral vascular disease. In this review, we aim to explain how glycation occurs in different molecules and what the pathological consequence of AGE formation in diabetes mellitus and other diseases are.

Trends in advanced glycation end products research in diabetes mellitus and its complications.

Advanced glycation end-products (AGEs) are heterogeneous groups of compounds that result from the non-enzymatic reaction of reducing sugars with free amino groups of biological molecules such as proteins, lipids, and nucleic acids. A large number of studies have been focused on AGEs metabolism, analysis, treatments, and their implications in the pathogenesis of diseases, especially in diabetes mellitus. Here, we review recent advances in the understanding of pathological complications caused by the production of AGEs. We provide an overview of the most important issues published within this area in last years; we also present the number of scientific papers related to AGEs available since 1950 until 2008 in the most important fields including metabolism, physiology, and pharmacology, thus as analytical methods for AGE detection and quantification and studies carried out in human body fluids. Data were collected from ovidSP.

Design, demonstration and performance of a versatile electrospray aerosol generator for nanomaterial research and applications.

Carbon nanotubes are difficult to aerosolize in a controlled manner. We present a method for generating aerosols not only of carbon nanotubes, but also of many reference and proprietary materials including quantum dots, diesel particulate matter, urban dust, and their mixtures, using electrospraying. This method can be used as a teaching tool, or as the starting point for advanced research, or to deliver nanomaterials in animal exposure studies. This electrospray system generates 180 microg of nanotubes per m(3) of carrier gas, and thus aerosolizes an occupationally relevant mass concentration of nanotubes. The efficiency achievable for single-walled carbon nanotubes is 9.4%. This system is simple and quick to construct using ordinary lab techniques and affordable materials. Since it is easy to replace soiled parts with clean ones, experiments on different types of nanomaterial can be performed back to back without contamination from previous experiments. In this paper, the design, fabrication, operation and characterization of our versatile electrospray method are presented. Also, the morphological changes that carbon nanotubes undergo as they make the transition from dry powders to aerosol particles are presented.

Effect of Preoperative Administration of Oral Arginine and Glutamine in Patients with Enterocutaneous Fistula Submitted to Definitive Surgery: a Prospective Randomized Trial.

BACKGROUND: The use of glutamine and arginine has shown several advantages in postoperative outcomes in patients after gastrointestinal surgery. We determined the effects of its use in patients with enterocutaneous fistula after operative treatment. PATIENTS AND METHODS: Forty patients with enterocutaneous fistula were randomly assigned to one of two groups. The control group received the standard medical care while the patients of the experimental group were supplemented with enteral administration of 4.5 g of arginine and 10 g of glutamine per day for 7 days prior to the surgery. The primary outcome variable was the recurrence of the fistula and the secondary outcomes were preoperative and postoperative serum concentrations of interleukin 6 and C-reactive protein and postoperative infectious complications. RESULTS: Twenty patients were assigned to each group. The fistula recurred in two patients (10%) of the experimental group and in nine patients (45%) of the control group (P < 0.001). We found a total of 13 infectious complications in six patients of the control group (all with fistula recurrence) and none in the experimental group. Mean preoperative serum concentrations of interleukin 6 and C-reactive protein were lower in patients from the experimental group. In addition, these levels were lower in patients who had recurrence if compared to patients that did not recur. CONCLUSION: Preoperative administration of oral arginine and glutamine could be valuable in the postoperative recovery of patients with enterocutaneous fistulas submitted to definitive surgery.

[Is vitamin-E supplementation beneficial or harmful?]. / Suplementación con vitamina E, benéfica o dañina?

Even though the beneficial effects of vitamin E have been experimentally observed, some clinical trials failed to verify a consistent benefit. One reason for this situation has been the difficulty to make comparisons among different studies. There are differences due to subjects, chemical forms of vitamin E, stages of the disease and others. The intake of high doses of vitamin E as a supplement has increased in many countries. Novel studies, have informed that vitamin E not only has antioxidant properties but can acts through precise molecular actions interacting with proteins and enzymes and modulating cellular signaling, transcriptional regulation and apoptosis induction. However, some recent clinical and meta analysis studies have found that daily supplementation with vitamin E 400 IU or higher is associated to increased mortality. In conclusion, a complete understanding of vitamin E actions at the cell does not exist yet and the controversy about its clinical effects is still present. This paper offers current knowledge on the characteristics, metabolism, properties, beneficial effect as well as the potential toxicity of vitamin E.

In vitro glycation of brain aminophospholipids by acetoacetate and its inhibition by urea.

Amino groups of amino acids, nucleic acids and lipids can react non-enzymatically with reducing sugars to form unstable Schiff bases that can then undergo the Amadori rearrangement to form irreversible advanced glycation end products (AGEs). Ketoacidosis is a life-threatening complication in patients with untreated diabetes mellitus and it is characterized by increased circulating ketone body concentrations. Recently, the in vitro glycation of hemoglobin by beta-hydroxybutyrate and acetone was described by our laboratory. This study was designed to evaluate the in vitro effect of acetoacetate on brain aminophospholipids at similar concentrations to that observed in ketoacidosis (16.13 mM total ketone bodies). The effect of acetoacetate was compared to that of glucose and the other ketone bodies; beta-hydroxybutyrate and acetone. The antiglycating activity of urea and glycylglycine was also investigated. The incubation of aminophospholipids with acetoacetate results in the formation of a new compound with an absorption peak at 280 nm. When this reaction product was analyzed by thin layer chromatography using an elusion system of methanol:chloroform:acetic acid:water (8:1:1:0.4), the R(f) value obtained (0.24-0.26) was similar to that of the compound formed by aminophospholipids with glucose. In contrast, this reaction product was not detected in those samples containing beta-hydroxybutyrate and acetone. The formation of this new compound was inhibited by urea more effectively than glycylglycine. In conclusion, this study provides the evidence that brain aminophospholipids react with acetoacetate forming AGEs and that this glycating effect of acetoacetate was remarkably decreased by urea, suggesting a protective physiological role for urea in the body as it was previously stated. Finally, this information adds knowledge about the contribution of ketoacidosis in the pathophysiology of diabetic complications, especially in type 1 diabetic patients.

New insights in endothelial and smooth muscle cell communication.

Based on immunohistochemical techniques against connexins and the intercellular flux of staining molecules, it has previously been shown that electrotonic communication occurs among endothelial and vascular smooth muscle cells, this due to the presence of myoendothelial gap junctions. The aim of this study was to evaluate the density of myoendothelial contacts in the left coronary and internal mammary arteries as well as in the left saphenous vein by means of electron microscopy, the distance between both cells participating in an myoendothelial contact with a semi-automatic image analysis system and the presence of homocellular and heterocellular gap junctions between endothelial and smooth muscle cells by using the immunohistochemical technique and confocal microscopy in thoracic aorta were also analyzed. The results are that all blood vessels studied present myoendothelial contacts, while density studies show that they are more abundant in the saphenous vein. The myoendothelial contact distance is constant and in no case the cytoplasmic processes reach the plasma membrane of the partner cell toward which they are advanced. Homocellular gap junctions were found between smooth muscle cells and between endothelial cells. Heterocellular gap junctions were absent, evidencing the possibility that signaling molecules between endothelial and smooth muscle cells may be transferred through plasma membranes as was once thought and not necessarily by electrotonic communication.

Inhibition by L-arginine and spermidine of hemoglobin glycation and lipid peroxidation in rats with induced diabetes.

The effect of L-arginine and spermidine on hemoglobin glycation and lipid peroxidation in serum of normal and diabetic rats was studied. Five groups of 40 rats were studied during 20 days and compared with a control group (Group I) that consisted of normal rats (N = 6) not treated with L-arginine or spermidine. Group II, diabetic rats (alloxan 120 mg/kg, i.p. at the day 0 and alloxan 60 mg/kg, i.p. at the day 10) were considered as diabetic control. Group III, diabetic rats treated with 10 mM L-arginine (i.p.). Group IV, diabetic rats treated with 10 microM spermidine (i.p.). Group V, normal rats treated with 10 mM L-arginine (i.p.). Group VI, normal rats treated with 10 microM spermidine (i.p.). The rats of each group were divided in subgroups of four each. Rats were anesthetized and blood was taken from aorta to determine glucose, triglycerides (TGs), total cholesterol (TC), low- and high-density lipoproteins (LDL and HDL), glycated hemoglobin (HbA(1C)), and thiobarbituric acid-reactive substances (TBARS). We observed that the alloxan concentrations used in this study reproduced the clinical manifestations of disease including hyperglycemia (from 116 +/- 7 mg/dl to 435 +/- 80 mg/dl) in 96 hours. As a consequence the levels of TGs, TC, LDL, TBARS, and HbA(1C) were increased, whereas HDL diminished. HbA(1C) concentration was significantly correlated with the concentration of TBARS. The L-arginine and spermidine injection tended to normalize the glycemia from 24 hours, similarly, hyperlipidemia, TBARS, and HbA(1C). From these results, we conclude that l-arginine and spermidine exerted an inhibitory effect of hemoglobin glycation and lipid peroxidation in vivo, which may be relevant in preventing diabetic complications.

Spermine increases arginase activity in the liver after carbon tetrachloride-induced hepatic injury in Long-Evans rats.

Arginase is the enzyme which synthesizes urea and ornithine, a precursor from which putrescine, spermidine and spermine are formed. These natural polyamines have been implicated in cell growth, replication and wound healing. The present study evaluated the possibility that spermine increases arginase activity and reduces liver damage caused by carbon tetrachloride. Intraperitoneally injected spermine at a dose of 1 mg/kg after a single intragastric administration of carbon tetrachloride (1.6 ml/kg) increased arginase activity (6.30-7.79 microg urea/mg protein per min) (P<0.05) as well as total protein content (0.29-0.37 mg/mg dry weight) in hepatic tissue, compared to the group which only received carbon tetrachloride. When liver cell damage was biochemically assessed, the carbon tetrachloride-treated group showed a 20-fold increase in serum glutamic oxaloacetate transaminase, compared to the control group (P<0.05), and this was significantly diminished by the administration of spermine (P<0.05). Serum triglycerides increased four times compared to the control group as a result of the carbon tetrachloride treatment and were diminished by spermine as well. These results indicate that spermine may play a role in the recovery of liver tissue after carbon tetrachloride-induced liver injury, maybe by increasing the synthesis of putrescine, a polyamine which has been found out to participate in the recovery of the hepatic tissue after an insult with carbon tetrachloride.

Increased uptake of oxidized LDL by macrophages from type 2 diabetics is inhibited by polyamines.

The aim of this study was to evaluate the effect of polyamines putrescine, spermidine and spermine on human LDL oxidation and to assess the ability of macrophages derived from type 2 diabetic patients to uptake oxLDL. Polyamine effect was compared with α-tocopherol. Four healthy subjects and eight type 2 diabetic patients were included in this study. To characterize type 2 diabetic patients and non-diabetic subjects, laboratory test were carried out. Glucose, glycated haemoglobin (HbA1C), triglycerides, low (LDL) and high density lipoproteins (HDL) and serum lipid peroxidation were measured in blood. The study was performed in three stages. For each stage, ten experimental conditions comparing the effect of polyamines with α-tocopherol (10µM solutions) on LDL oxidation and the uptake of oxLDL by macrophages were analyzed. MDA concentration was found to be significantly higher in type 2 diabetic patients compared to healthy subjects (5.6±0.58 vs. 2.66±0.31µM MDA, respectively, (P<0.05)). Percent of macrophages containing oxLDL was determined by means of red oil staining. The uptake of oxLDL by macrophages derived from diabetic patients was clear. The uptake of oxLDL was inhibited when the oxidation was prevented by polyamines or α-tocopherol. Spermine showed high antioxidant capacity (96.67±1.53% vs. 25.67±2.30%) compared to α-tocopherol (96.67±1.53% vs. 47.00±7.20%) at the concentration tested. In conclusion, polyamines especially spermine, has a potent antioxidant effect compared to α-tocopherol on human LDL oxidation, followed by spermidine and putrescine. The results have clinical relevance in the diabetic complications and add knowledge on the role of polyamines as natural antioxidants. This research is not a clinical evaluation rather a functional analysis utilizing clinical samples.

Oral infections and glycemic control in pregnant type 2 diabetics.

BACKGROUND: The relationship between diabetes mellitus and infection is bidirectional. Diabetes favors infection, while infections make controlling diabetes much more difficult. The most frequent infections encountered in pregnant diabetics are those of the urinary tract (UTI) and cervicovaginal (CVI) area. Periodontal diseases (PD) and active caries (AC) are infectious diseases frequently found in these cases often remaining as hidden entities of low intensity. The aim of this study was to assess whether there is an association between PD and caries with lack of glycemic control often encountered in pregnant type 2 diabetic women. METHODS: A single skilled researcher performed the periodontal evaluation of PD and AC cases. Glycated alpha hemoglobin (HbA(1c)) over 6.5% was used to diagnose metabolic glucose disturbances. The controlled variables were UTI, CVI and adherence to treatment. The statistical tests used were chi-square adjusted for continuity, analysis of variance, odds ratio with 95% confidence intervals and logistic regression with a significance level of p < 0.05. RESULTS: One hundred twenty-seven type 2 diabetic women were seen during the second trimester of their pregnancies. Mean age was 31.3 years; 55% had lack of glycemic control associated with CVI (chi2 21.07, p < 0.000), PD (chi2 5.72, p < 0.005) and UTI (chi2 13.77, p < 0.000) with therapeutic adherence (TA) (chi2 14.80, p < 0.00). No association was found with AC. The logistic regression results showed that UTI, CIV, PD and TA are associated with lack of glycemic control. CONCLUSIONS: Periodontal diseases may act as "hidden" infections in pregnant diabetics and be associated with lack of glycemic control. This situation should be taken into consideration by healthcare teams in charge of prenatal care.

L-arginine and polyamine administration protect beta-cells against alloxan diabetogenic effect in Sprague-Dawley rats.

In the searching for new substances with the capacity to protect beta-cells from the toxic effects of alloxan, we evaluated the effect of L-arginine and the polyamines putrescine, spermidine and spermine in a murine experimental model of diabetes. Diabetes was induced by the i.p. injection of either 200 mg/kg (24-h experiments) or 120 mg/kg (12 days experiments) body weight. L-Arginine and polyamines were administered 10 min before or 10 min after alloxan administration, once its half-life had elapsed, respectively. In the 24-h study, serum glucose (199.8+/-27.6 mg/dl) and triglyceride (54.6+/-4.9 mg/dl) concentrations showed a protective effect of spermine, as these parameters were not too high (P < or = 0.05), compared to the alloxan-treated group (415.4+/-47.8 and 90.2+/-11.6 mg/dl, respectively), and were closer to glucose (132.3+/-6.0 mg/dl) and similar to triglycerides (63.8+/-7.1 mg/dl) of the control group. A similar pattern was observed on the parameters measured when L-arginine and polyamines were administered daily for 12 days, starting 10 min after a single alloxan administration, which provides evidence that L-arginine and polyamines are effective in impeding the increase in serum glucose, triglyceride and cholesterol concentration showed on day 3 by the alloxan-treated group, as well as a higher acinar cell regenerative capacity as determined by immunohistochemical techniques. Spermine turning out to be more effective than L-arginine, putrescine or spermidine in counteracting the marked hyperglycemia and triglyceridemia showed by the alloxan-treated group and similar in effect when evaluating cholesterolemia. These results show a clear protective role of L-arginine and polyamines over the pancreatic beta-cell, in addition to the induction of neogenesis from both ductal and acinar cells that leads to the recovery of endocrine pancreatic function in rats with experimental diabetes.

Effect of alpha-tocopherol on the metabolic control and oxidative stress in female type 2 diabetics.

In this study we evaluate the effects of alpha-tocopherol on the metabolic control and oxidative stress in female patients with type 2 diabetes mellitus. Thirty-four female type 2 diabetics 40-70 years old up to 14 years with diabetes, under medical treatment, were randomly divided in two groups. One group received placebo (Control group, n = 21) and the other received alpha-tocopherol (800 IU/day, n = 13) during 6 weeks. Blood samples were collected at the beginning and at the end of the study to measure malondialdehyde production, glycated hemoglobin, selenium dependent-glutathione peroxidase, Cu,Zn-superoxide dismutase in erythrocytes and total antioxidant status, glucose, lipid and lipoproteins in serum. Erythrocyte malondialdehyde decreased and serum-total antioxidant status increased after alpha-tocopherol treatment (P < 0.0001). However, an unexpected increase on cholesterol levels and a reduced erythrocyte-Cu,Zn-superoxide dismutase activity was observed after alpha-tocopherol treatment. alpha-Tocopherol administration did not affect glucose, glycated hemoglobin, triacylglycerides, lipoprotein levels and serum malondialdehyde. A minor oxidative stress was observed in female type 2 diabetic patients after alpha-tocopherol treatment inferred from the reduced levels of erythrocyte malondialdehyde and the increased values of total antioxidant status. On the other hand, no beneficial changes were observed on glycemic control or lipid metabolism.

Clinical evolution of diabetic rats after transplant of electrofused pancreatic islet cells and dermic cells.

This work was designed to study an alternative treatment of diabetes mellitus by using a transplant of hybrid cells obtained by the electrofusion of pancreatic islet cells from a healthy donor with dermic cells obtained from a recipient. The hybrid cells kept the capacity of insulin production, its regulation, and the natural control of glycemia, as well as the factors of histocompatibility to avoid the rejection. Four groups of four rats each were established: Group 1. Healthy animals (healthy control), Group 2. Diabetized non-treated animals (diabetic control), Group 3. Transplant recipient rats with extraction of dermic cells which were mixed with pancreatic insular cells from a healthy donor (transplant without fusion), and Group 4. Transplant recipient rats, with extraction of dermic cells which were electrofused with pancreatic insular cells from a healthy donor (transplant with fusion). For the Group 4, the cells were combined and they were submitted to dielectrophoresis conditions with an alternating current pulse of 15 s of 10 V RMS of 0.5 MHz. The fusion was made with a direct current pulse of 1 ms of 300 V. Clinical signs were registered (weight, diuresis, food and water intake), and several biochemical parameters in blood which included basal glycemia, uric acid, cholesterol, triglycerides, glutamate oxalacetate transaminase, glutamate pyruvate transaminase, urea, creatinine, insulin, glycated hemoglobin were registered. Additionally, ketone bodies and glucose were also measured in urine. All determinations were made at 30, 60, and 90 days. Animals of Group 1 maintained its parameters within the normal ranges. Rats of Group 2 presented alterations corresponding to a diabetic state in almost all the parameters measured, none of the animals showed a tendency to improve spontaneously, two of the rats died at 66 and 72 days. The Group 3 showed a clinical profile similar to the diabetic control group without improvement, only one rat died at day 33, while in the rats transplanted with fusion (Group 4) an improvement was observed on some parameters including body weight, water intake and glycemia. Although insulin concentration was under the normal range, it was higher than in the Group 3. None rat died. These results indicate that it is possible to improve the diabetic profile by the transplant of dermic cells from a diabetic animal fused with insular cells from a healthy donor in the recipient animal.

Effects of Acute Ingestion of Native Banana Starch on Glycemic Response Evaluated by Continuous Glucose Monitoring in Obese and Lean Subjects.

An abnormal glycemic profile, including postprandial glycemia and acute glucose spikes, precedes the onset of overt diabetes in obese subjects. Previous studies have shown the beneficial effects of chronic native banana starch (NBS) supplementation. In this study, we examined the effects of acute ingestion of NBS on glycemic profiles by means of continuous glucose monitoring in obese and lean subjects. In a crossover study, obese and lean subjects consumed beverages containing either 38.3 g of NBS or 38.3 g of digestible corn starch (DCS) twice daily during 4 days. On day 5, a 3-h meal tolerance test (MTT) was performed to evaluate glucose and insulin responses. After 1 week of washout period, treatments were inverted. NBS supplementation reduced the 48-h glycemia AUC in lean, obese, and in the combined group of lean and obese subjects in comparison with DCS. Postprandial glucose and insulin responses at MTT were reduced after NBS in comparison with DCS in all groups. However, no changes were observed in glycemic variability (GV) indexes between groups. In conclusion, acute NBS supplementation improved postprandial glucose and insulin responses in obese and lean subjects during 48 h of everyday life and at MTT. Further research to elucidate the mechanism behind these changes is required.

Regeneration of beta-cells and neogenesis from small ducts or acinar cells promote recovery of endocrine pancreatic function in alloxan-treated rats.

BACKGROUND: We previously showed by using biochemical parameters that male Sprague-Dawley rats receiving a single intraperitoneal (i.p.) administration of alloxan (120 mg/kg body weight) with no further treatment recovered endocrine pancreatic function after 12 days. METHODS: Male Sprague-Dawley rats received an i.p. injection of alloxan (120 mg/kg body wt), were killed at 3, 6, 9, or 12 days (n=7), and their capacity to recover endocrine function was evaluated by means of a) biochemical parameters, which included glucose, triglyceride, and total cholesterol measurements and b) nuclear incorporation of 5'-bromodeoxyuridine (BrdU) by beta and acinar cells as well as presence of neogenesis from either ductal or acinar cells using double-staining BrdU-insulin immunohistochemical technique. RESULTS: Three days after receiving a single i.p. administration of alloxan, rats showed increase in serum glucose, triglyceride, and total cholesterol concentrations, reaching levels of 542.4+/-63.1, 907.6+/-154.9, and 106.0+/-2.7 mg/dL (mean+/-standard deviation [SD]), respectively. At this time, increase in beta-cell replication was also observed, although this reached maximum by day 6 (p <0.001). Replication was also present in acinar cells, but these cells showed their maximum at day 3 (p <0.001) and subsequently decreased, as did beta-cells, almost steadily to normal values by day 12. Neogenesis of beta-cells was observed mainly as transdifferentiation from acinar cells at day 3 and from ductal cells at day 6, after which it tended to be normal. CONCLUSIONS: Male Sprague-Dawley rats receiving a single i.p. alloxan dose tended to normalize their endocrine function by day 12 after alloxan administration. This process included both regeneration and neogenesis of pancreatic beta-cells from either ductal or acinar cells.

Inhibition of in vitro pyrraline formation by L-arginine and polyamines.

Glycation of biomolecules such as proteins, nucleic acids and lipids leading to the formation of advanced glycation end products (AGEs) may be a major contributor to the pathological manifestations of diabetes mellitus. Several studies have shown that the chemical inhibition of AGEs formation results in attenuation of diabetic complications. We tested the in vitro inhibition of pyrraline formation on bovine serum albumin and L-lysine by L-arginine and the polyamines putrescine, cadaverine, spermidine and spermine. Among the inhibitors, L-arginine and spermine potently inhibited pyrraline formation. This effect could be related to the presence of the guanidino group in L-arginine and four amino groups in spermine, but this inhibitory effect was also shown by putrescine, cadaverine and spermidine, suggesting that these natural compounds may have a novel therapeutic potential in preventing diabetic complications. A significant unexpected observation emerged when experiments were carried out with aminoguanidine. It showed increased absorbance produced by a non-identified compound whose peak appears at 285 nm, but this aspect remains to be investigated.

Healing of diabetic foot ulcers in L-arginine-treated patients.

Experimentally, we demonstrated the beneficial effects of L-arginine on regulation of hyperglycemia and dyslipidemia in experimental diabetes, in addition to a positive anti-aggregating effect in platelets in animals and humans. Here, the effect of L-arginine on foot ulcers from diabetic patients was studied. Three groups of diabetic patients were included: 11 patients without ulcer received neither treatment and served as controls. Eleven patients with diabetic ulcer received the standard treatment, this group served as diabetic control with diabetic ulcer. Eleven remain patients with diabetic ulcer received 10 mM L-arginine subcutaneously on the site of the wound. Biopsy with punch number 5 on wound site comprising both ulcerative and contiguous undamaged skin were performed in all patients with ulcerative lesions before any treatment. Patients with intact skin had biopsy performed with punch number 5 on external malleolar region of right lower limb. Biopsies were examined by light and confocal microscopy utilizing histochemical and immunohistochemical methods. Initial and final blood samples were collected to determine glucose, triglycerides, total cholesterol, glycated hemoglobin (HbA(1c)), low (LDL), and high density lipoproteins (HDL). Significant differences (P < 0.05) were observed between initial and final serum glucose levels for treated patients, and initial serum glucose levels between treated and control patients without diabetic ulcer. Glycated hemoglobin, triglycerides, cholesterol, and lipoprotein levels showed no significant changes. Eight patients treated with L-arginine reached total wound healing and the remaining three who abandoned the study because of change of residence showed relevant improvement. Histochemistry and immunohistochemistry methods have shown vascular impairment in both patients with diabetic ulcer (prior to treatment) and control patients without diabetic ulcer. Our observations strongly support efficacy of L-arginine for successful wound healing of diabetic ulcers.