and untreated. Depending on the type of diabetes i.e 1 or 2, treatment consists of insulin injections [1] or anti diabetic drugs that lower blood glucose and hopefully hemoglobin A1C as well more vigorously as well and more frequently. Previously, using a diabetic-streptozotocin (STZ) rat model, we demonstrated that diabetes and cataracts could be entirely prevented using equimolar amounts of STZ and a nitric oxide oxidizing compound called sodium carboxy-PTIO. STZ from increasing blood glucose and all the signs of diabetesincluding cataracts were prevented. Later work indicated how the STZ-diabetes pathway was likely created. Now, we demonstrate that STZ actually generates the oxidizing and nitrating peroxide known as peroxynitrite (OONO-). In the cases of diabetes 1,2 most or all of many anti-diabetic drugs only control the level of blood in diabetic children over a three year period that even with tight diabetic damage stems from peroxynitrite. Using STZ to generate peroxynitrite in animals, we demonstrate that a variety of nitration targets can diminish peroxynitite which is the most important step in controlling both diabetes and possibly Alzheimer’s disease since recent data links diabetes and Alzheimer’s disease. 2 diabetes is the utilization of injected streptozotocin (STZ) which was originally developed as an antibiotic for gram negative bacteria by the Upjohn company of Kalamazoo, Michigan and later found to cause diabetes [1]. Depending on the dose of STZ injectedeither type 1 (high dose) or type 2 (low dose) causes these diseases. It is well understood that STZ causes the death of the beta cells of the cells and lower doses create the killing of some beta cells which produce insulin, but spares some of the beta cells. STZ injected rats increases blood glucose from normal levels (100 milligrams/ dl) to about 500 mg/dl displaying type 1 diabetes in mice or rats with blood glucose 300 mg/dl in the type 2 diabetic animals [2,3]. In a previous publication in Luminescence [4], we demonstrated that diabetes caused by streptozotocin could be completely blocked or abrogated using an equimolar concentration of Carboxy-PTIO (C-PTIO) and STZ. Since it is known that C-PTIO oxidizes nitric oxide (.NO) to NO2, it prevents the nitric oxide from being utilized to form the nitric oxide-based toxin (OONO-) in vivo. Naturally, if the toxin cannot form, diabetes does not develop. One of the reviewers of this original paper commented that this was the beginning of the end for diabetes and naturally we believe this comment to be quite correct. Figure 1. methylnitrosourea. In later work, using targets of nitration, we demonstrated that these could also prevent streptozotocin-based diabetes (3.5). Knox Van Dyke (2017) Luminescence Reveals the Possible Cause of Diabetes 1,2,3 (Alzheimer’s Disease). BAOJ Diabet 3: 025. Page 2 of 9 BAOJ Diabet, an open access journal Volume 3; Issue 2; 025 nitrosourea. (Figure1) It is known that tissues that have substantial amounts of the sugar transport mechanism-(glut-2) tend to concentrate glucose type molecules like 2 deoxyglucose and be more damaged by STZ eg. beta cells of the pancreas, liver and kidney cells. Once the STZ with 2-deoxyglucose bonded to the methylated nitrosourea gets into the beta cell, the STZ is split into 2 deoxy the nitric oxide free radical which reacts with superoxide (.O2-) producing (OONOor later its carbonate derivative) and MNU can methylate DNA bases, break both single stranded or double stranded DNA or even RNA, activate poly-ADP-ribose polymerase (PARP) depleting NADH and ATP killing mitochondria and cells dependent on it and causing nitration, nitrosylation and nitrosation of key proteins and damaging their important enzymatic activity [6]. In a recent JBC paper [7], it was suggested that STZ could evidence that free radicals are produced-which we already knew, since STZ can clearly generate the nitric oxide radical as was both indicated from our previous studies [3,5] and studies by others Tsuji and Skurai [8] Furthermore, a similar set of diabetic conditions of impaired glucose utilization [9] and insulin resistance occurs in (FDG), a sugar which is transported into brain like glucose but this radioactive remnant remains long enough to produce superb early-stage dementia or pre-Alzheimer’s disease (AD) compared to When streptozotocin is intra-articulary injected into mouse brain, glucose utilization in the mouse brain is depressed and insulin resistance occurs similar to when STZ is injected into rodents peripherally damaging Beta cells causing diabetes [11,12]. Further, best animal model of Alzheimer’s disease ever developed. Many investigators have clearly demonstrated that STZ injection into the brain is a superior method compared to the gene defect models in mice which do not demonstrate many of the complete characteristics of human Alzheimer’s disease. It should not be too surprising that some investigators have termed Alzheimer’s disease as diabetes 3 because of its similarity to diabetes [9]. Many investigators have switched to this STZ-ALZ based superior standard mouse model to study possible inhibitors of the disease Kimura and Lecanu and Papadopoulous [13,14]. In this manuscript, we will develop what we believe is clear evidence that STZ does generate a peroxide namely peroxynitrite (PN) (OON=O-) and show that the reaction of STZ producing PN generates luminescence via its reaction with L-012. Furthermore, STZ based luminescence is abrogated by micromolar doses of acetaminophen or tyrosine hydrochloride or L-cysteine hydrochloride, etc. acting as targets of oxidation or nitration is very similar to the peroxynitrite generation reaction of SIN-1 (sydnoimine) with L-012, whose luminescent reaction generates similar blue luminescence reaction kinetics as we have previously published [15]. A compound must be very strong oxidizer to react with L-012 to produce blue light production eg because hydrogen peroxide, hypochlorite or superoxide do not react with L-012. L-012 luminescence from peroxynitrite or streptozotocin both produce blue light which is extinguished by tyrosine. HCl, acetaminophen or L-cysteine-HCl/n-acetylcysteine(NAC) or mono or polyphenols depending on the polyphenolic structure published in [16].

Since oxidative/nitrosative stress cause diabetes, can we prevent this chemistry generating the disease? Streptozotocin causes diabetes by entering the pancreatic beta cell generating excessive nitric oxide which reacts with oxygen creating a toxin possibly peroxynitrite, dinitrogen trioxide, dinitrogen tetraoxide and so forth. The toxic compounds damage the DNA causing beta cell death. This prevents insulin synthesis, storage and release. By using antioxidant substances that destroy the nitric-oxide-based toxins (e.g., carboxy-PTIO (oxidizes nitric oxide), polyphenolic-quercetin and monophenolic acetaminophen (Tylenol)) which are oxidation and nitration targets can the diabetes I causing toxins in animals be destroyed? Will this tri-drug combination completely prevent the deleterious effects of diabetes namely poor blood glucose control and blindness from cataracts for the entire length of the experiment (one year). These disease reversal experiments were accomplished in rats where the streptozotocin-diabetic effects were completely thwarted. In vitro experiments were accomplished to provide the scientific basis for the experimental results in animals.

Recently we demonstrated that streptozotocin (STZ) diabetes (type I) in rats is preventable using a simultaneous equimolar injection of carboxy-PTIO (c-PTIO). Both changes in blood sugar and cataracts are prevented. This apparently occurs because the nitric oxide (NO) (from STZ) generated in the beta cells is oxidized to nitrite by c-PTIO preventing diabetes. STZ generates NO producing a NO-based toxin. The toxin damages DNA by nicking and activates poly-ADP-ribose causing necrosis and triggering inflammation. Is there evidence that O/N stress occurs in early human type I diabetes? We studied 40 children with or without early type I diabetes and observed that urate is decreased 25% in all these diabetic children each over the age of 3 years. Urate is a major portion of blood-antioxidant load. Surely this decrease in urate indicates ongoing O/N stress. Does O/N stress initiate disease? STZ studies in rats indicates that this is correct.

The biochemical mechanisms by which hyperglycemia causes microvascular disease and neuropathy are poorly understood. Experimental studies have established that oxidative stress is present in diabetic rodents with neuropathy, and that antioxidant therapy is protective. Oxidative stress is also present in human diabetes, but its clinical importance is uncertain.

We have previously documented that octreotide therapy suppresses sweating and palpitations in patients with the postural tachycardia syndrome. We now report that octreotide also suppresses these and related symptoms in patients with postmenopausal hot flushes.

Oxidative stress damages DNA in experimental diabetes, and in vitro studies have suggested that it is linked to lipid peroxidation. The objective of the study was to determine whether lipid peroxidation, as assessed with malondialdehyde excretion in recent-onset type 1 diabetes mellitus, is associated with oxidative damage to DNA, as assessed from 8-hydroxydeoxyguanosine excretion. A 3-year longitudinal study of recent-onset type 1 diabetes mellitus was performed. Age- and sex-matched control subjects were studied once. Patients were studied as inpatients at West Virginia University Hospitals. Thirty-seven patients with recent-onset (2-22 months) type 1 diabetes mellitus (male, 10; female, 27) were enrolled in a longitudinal study of oxidative stress. The mean age of the patients was 20 years. None of the patients had hyperlipidemia or were treated with lipid-lowering drugs. Only 1 patient had hypertension and was being treated with beta-adrenergic blocking therapy. Thirty-six patients completed the study; one withdrew after the second evaluation. Lipid peroxidation was assessed by measuring malondialdehyde excretion. Oxidative damage to DNA was assessed from 8-hydroxydeoxyguanosine excretion. Malondialdehyde excretion was increased in the diabetic patients at the first evaluation (2.43 +/- 0.31 micromol/g creatinine), second evaluation (2.34 +/- 0.24), and third evaluation (1.93 +/- 0.15) compared with control subjects (1.51 +/- 0.11) (P < .005). 8-Hydroxydeoxyguanosine excretion, however, was not increased in the diabetic patients. There was no correlation between malondialdehyde and 8-hydroxydeoxyguanosine excretion. We confirmed the presence of oxidative stress in early diabetes as assessed from malondialdehyde excretion. We were unable, however, to confirm oxidative damage to DNA in this cohort of patients; and there was no evidence of a correlation between lipid peroxidation and DNA damage.

Streptozotocin (STZ)-induced diabetes is linked to excessive nitric oxide (NO), and possibly peroxynitrite (OONO(-)) and/or other nitrogen oxides, e.g. nitrogen trioxide (N(2)O(3)), which damages DNA of pancreatic beta cells, causing death and loss of insulin. Simultaneous injection of carboxy-PTIO (CPTIO) and STZ prevents diabetes and cataract formation in rats, whereas 4-hydroxy-Tempo (4HT) does not. CPTIO oxidizes nitric oxide to nitrite, which prevents production of the diabetogenic toxin. Peroxynitrite may not be involved, since 4HT (converts O(2)(-) to H(2)O(2)) injected with STZ produces diabetes. All six of the control rats injected with STZ became diabetic and developed cataracts after 3 months. Eight rats injected with STZ and CPTIO were non-diabetic with no cataracts up to a year. This work establishes the idea that excessive nitric oxide is a primary initiator in STZ diabetes. Luminescence experiments using OONO(-) generation from SIN-1 with L-012 indicates that 4HT is an effective inhibitor, while CPTIO is ineffective. Experiments with dilute solutions of nitrogen trioxide added to ladder or plasmid DNA reveal extensive nicking of DNA, thereby raising the possibility that other oxides of nitrogen could be involved with the damage to DNA. It can be concluded that diabetes can be prevented by oxidizing excessive NO from STZ.

Antibodies to the smaller isoform of glutamic acid decarboxylase (GAD65Ab) have been linked to the presence of neuropathy in Type 1 diabetes in several small studies. We attempted to confirm this association by measuring GAD65Ab, GAD65Ab epitopes and IA-2Ab in 511 patients who participated in the Diabetes Control and Complications Trial (DCCT). We also tested for correlations between these autoantibodies and C-peptide and glycemic control. We only included patients for whom serum was available from the first 4 years of their illness. The presence or absence of neuropathy was determined by electrophysiological studies, autonomic testing and clinical evaluation at baseline and 5 years into the trial or at close out. Samples from controls (patients without neuropathy at 5 years) were selected for patients who had similar C-peptide responses to a standardized meal at baseline. The GAD65Ab index correlated with HgbA(1c) only in the adult participants and only at baseline. The adults initially in poor control (upper tertile for glycemia) had higher GAD65Ab and lower C-peptides. The GAD65Ab index was not significantly different in patients with confirmed clinical neuropathy at 5 years versus controls matched for C-peptide (.248 +/- 03 versus .278 +/- 03). Epitope analysis, based on the blocking of conformational epitopes by recombinant Fab, revealed that the binding to multiple epitopes was decreased in the patients with neuropathy. (c) 2007 Elsevier B.V. All rights reserved.

The purpose of this study was to determine whether autonomic neuropathy and the postural tachycardia syndrome can be treated with octreotide LAR (Long Acting Release). This was an open-label pilot project. Protocol 1 Patients with autonomic neuropathy (n = 4) were given increasing doses of octreotide LAR once a month for three months. Blood pressure was measured in the sitting posture every two weeks. Pretreatment mean blood pressure averaged 83.8 ± 7.1 mm Hg. After four, six and eight weeks of therapy the blood pressures averaged 96.3 ± 6.4, 98.2 ± 6.1 (p < .025), and 104.1 ± 3.1 (p < .025) respectively. Therapy led to a dramatic improvement in symptoms in one patient but another had an unacceptable elevation in supine blood pressure. Protocol 2 Patients with POTS or orthostatic intolerance were given 10, 20, or 30 mg of octreotide LAR over three months. Seven patients entered and five completed the study. After two months treatment, standing time increased from 36.0 ± 9.2 to 59.2 ± .8 minutes (p < .01). Heart rate in the standing position was suppressed from 106 ± .83 to 93.2 ± .8 beats per minute (p < .05). Orthostatic dizziness and chronic fatigue improved. We conclude that octreotide LAR can be used to treat autonomic neuropathy but there is a risk of an excessive pressor response. Octreotide LAR improved standing time and suppressed tachycardia in patients with orthostatic intolerance.

We assessed the potency of octreotide and midodrine, and their combination, in the treatment of the postural tachycardia syndrome (POTS) and orthostatic intolerance (OI). Nine patients with POTS and six patients with OI stood for up to 1 hour while their HR and BP were monitored. Patients received on separate days, midodrine 10 mg 1 hour before testing, octreotide 0.9 µg/kg 8 minutes before testing or combination therapy. Standing time in the patients with POTS was 41.2 ± 8.4 minutes and not improved by midodrine or octreotide, but increased to 56.3 ± 2.7 (P < 0.01) minutes following combination therapy. The standing heart rate in POTS, 114 ± 0.7 bpm, was suppressed by midodrine 92.8 ± 0.7 (P < 0.001), octreotide 90.6 ± 0.78 (P < 0.001), and combination therapy 84.7 ± 0.7 (P < 0.001). Combination therapy was better than monotherapy (P < 0.001) but only for the first 10 minutes of standing.

The present study was performed to determine whether increased lipid peroxidation, as assessed from malondialdehyde (MDA) excretion, is associated with deterioration in peripheral nerve function in early type 1 diabetes mellitus. These parameters were measured annually for 3 years in 36 patients who entered the study less than 2 years after the diagnosis of diabetes. Malondialdehyde excretion was 1.51 ± 0.20 μmol/g creatinine in the controls, and 2.43 ± 0.21, 2.39 ± 0.22, and 1.93 ± 0.21 μmol/g creatinine at the first, second, and third evaluations, respectively (P < .005). The increased MDA was seen only in the female participants. Malondialdehyde excretion was increased in those with high vs low hemoglobin Alc across all years (P < .05). Malondialdehyde excretion correlated negatively with sudomotor function below the waist. The mean sweat production from the 3 evaluations correlated with mean MDA excretion across all years in the proximal leg (r = −0.42, P < .005) and distal leg (r = −0.40, P < .01). Below the waist, sweating correlated with MDA (r = −0.40, P < .01) as did total sweat (r = −0.38, P < .01). The response amplitudes of the peroneal nerves correlated negatively with MDA excretion (for the mean values at the second 2 evaluations, P < .005, r = −0.45). Tests of sensory function correlated inconsistently with MDA excretion. In summary, lipid peroxidation, as assessed from malondialdehyde excretion, is associated with sudomotor dysfunction in early diabetes.

The results of ablation of the parasympathetic nervous system in fat deposits in mice suggest that activity of this system increases the rate of synthesis of fat and may therefore lead to different rates of fat accumulation in various parts of the body.

Although hyperglycemia has been shown to cause peripheral nerve dysfunction in patients with diabetes, the biochemical mechanisms for this effect are poorly understood. The excessive production of reactive oxygen species and reactive nitrogen species has been proven to be detrimental in experimental diabetes, but there is little evidence that these metabolic events take place clinically and are physiologically important in man. To assess this we measured nitrite and nitrate (indices of nitric oxide production), nitrotyrosine (an index of peroxynitrite), 8-isoprostaglandin F-2 alpha, an isoprostane reflective of oxidative stress and lipid peroxidation, and uric acid, an index of antioxidant defense in patients with recently diagnosed Type 1 diabetes and aged-matched controls. The diabetic patients were followed for three years. We documented the overproduction of nitric oxide and increased lipid peroxidation in early diabetes and showed these changes had detectable adverse effects on peripheral nerve function especially sympathetic sudomotor nerves. We documented the suppression of uric acid and showed this was associated with multiple abnormalities in autonomic function. In addition, we present indirect evidence that overproduction of reactive oxygen species and reactive nitrogen species have adverse effects on beta cell function and blood pressure.

Background: Peroxynitrite is a toxic compound formed during the inactivation of nitric oxide (NO) by the superoxide anion. The physiologic significance of this pathway of NO metabolism has never been documented in vivo. Because peroxynitrite provides a pathway for the inactivation of NO we postulated that peroxyrtitrite's correlation with physiologic parameters would be the opposite of those associated with NO, which is a vasodilator and suppresses sudomotor function. We assessed the significance of peroxynitrite by comparing its associations with blood pressure (BP) and sudomotor responses with those of NO.Methods: Thirty-seven patients with type I diabetes enrolled in a longitudinal study of oxidative stress. Nitric C oxide was assessed from nitrite and nitrate (collectively NOx) and peroxynitrite was assessed from the nitrotyrosine (nTy) content of protein.Results: nTy was 13.3 +/- 2.0 mumol/L in the control subjects and 26.8 +/- 4.4 mumol/L, 26.1+/- 4.3 mumol/L, and 32.7 +/- 4.3 mumol/L in the diabetic patients (P <.01) at the time of the first, second, and third evaluations, respectively. Patients with increased nitrotyrosine/tyrosine (nTy/Ty) had higher mean BP than those with low nTy/Ty (81.1 +/- 1.9 mm Hg v 75.5 +/- 1.7 at the third evaluation, P <.025). The ratio of nTy/NOx correlated with BP at the first (P <.05), second (P <.05), and third (P <.01) evaluations. Patients with high nTy/Ty had increased sudomotor responses (5.85 +/- 0.75 &mu;L of total sweat) at the third evaluation compared to those with low nTy/Ty (3.32 +/- 0.43 &mu;L, P <.005) and normal controls (3.90 +/- 0.41 muL, P <.05). The associations of nTy with BP and sudomotor responses were the opposite of those with NOx.Conclusions: The conversion of NO oxide to peroxynitrite is physiologically significant in humans.