Pietro Compagnucci

The importance of determining irreversibly glycosylated hemoglobin in diabetics.

The concentration of glycohemoglobins (HbA1(a+b+c), HbA1,) was measured before and after incubation of normal and diabetic erythrocytes for 6 h at 37°C in saline. This procedure removes as much as 80-90% of the labile glucose-HbA0 adduct (labile HbA1,), thus allowing accurate estimation of irreversibly glycosylated hemoglobin (stable HbA1,). The concentration of HbA1 measured before such an incubation is total HbA1, (stable + labile). We determined the concentration of total, stable, and labile HbA1, in the same blood samples used to measure fasting plasma glucose (FPG) every day, for 4 consecutive days, in two groups of hospitalized insulin-treated diabetics. Group A subjects (N = 7) were type I, C-peptide negative, unstable diabetics, while group B subjects (N = 15) were type II, C-peptide positive, stable diabetics. Individual day-to-day variations of total HbA1, were wide in group A (Δ = 1.58 ± 0.14%), and slight in group B (Δ = 0.12 ± 0.01%; P < 0.001), paralleling similar plasma glucose fluctuations. Day-to-day variations of stable HbA1, were virtually absent not only in group B subjects with stable glycemic values (Δ = 0.08 ± 0.01%), but also in those of group A with marked glycemic instability (Δ = 0.07 ± 0.01%; P = NS). Day-to-day variations of labile HbA1, were marked in group A (Δ = 1.31 ± 0.14%), but negligible in group B (Δ = 0.15 ± 0.03%; P < 0.001). On admission, FPG correlated with labile HbA1, in group A (r = 0.89) and B (r = 0.71). FPG correlated with stable HbA1, in group B (r = 0.73) but not in group A subjects and with total HbA1, more closely in group B (r = 0.73) than in A (r = 0.61). A very close correlation was found between the concentration of total and labile HbA1 in subjects of group B (r = 0.82). In group B, fasting, post-breakfast, and mean daily plasma glucose values, determined every 3-6 days during the 2 mo before admission, significantly correlated both with total and stable HbA1 determined on admission, while in group A they did not. In group A, the correlation was significant when stable instead of total HbA1 was considered. We conclude that the significant fluctuations of total HbA1 reduce its value as an index of long-term control in unstable diabetics. On the other hand, a single determination of stable HbA1, totally independent of simultaneous blood glucose values, closely reflects blood glucose control over the previous 2 mo. We propose routine estimation of stable HbA1, which is simple and straightforward, to carry out follow-up studies of unstable diabetics.

Mechanisms of glucagon secretion during insulin-induced hypoglycemia in man. Role of the beta cell and arterial hyperinsulinemia.

To elucidate the mechanisms controlling the response of glucagon to hypoglycemia, a vital component of the counterregulatory hormonal response, the role of intraislet insulin was studied in seven normal subjects and five subjects with insulin-dependent diabetes mellitus (IDDM) (of less than 15-mo duration). In the normal subjects, hypoglycemia (arterial plasma glucose [PG] 53 +/- 3 mg/dl) induced by an intravenous insulin infusion (30 mU/m2 X min for 1 h, free immunoreactive insulin [FIRI] 58 +/- 2 microU/ml) elicited a 100% fall in insulin secretion and an integrated rise in glucagon of 7.5 ng/ml per 120 min. When endogenous insulin secretion was suppressed by congruent to 50 or congruent to 85% by a hyperinsulinemic-euglycemic clamp (FIRI 63 +/- 1.5 or 147 +/- 0.3 microU/ml, respectively) before hypoglycemia, the alpha cell responses to hypoglycemia were identical to those of the control study. When the endogenous insulin secretion was stimulated by congruent to 100% (hyperinsulinemic-hyperglycemic clamp, FIRI 145 +/- 1.5 microU/ml, PG 132 +/- 2 mg/dl) before hypoglycemia, the alpha cell responses to the hypoglycemia were also superimposable on those of the control study. Finally, in C-peptide negative diabetic subjects made euglycemic by a continuous overnight intravenous insulin infusion, the alpha cell responses to hypoglycemia were comparable to those of normal subjects despite absent beta cell secretion, and were not affected by antecedent hyperinsulinemia (hyperinsulinemic-euglycemic clamp for 2 h, FIRI 61 +/- 2 microU/ml). These results indicate that the glucagon response to insulin-induced hypoglycemia is independent of the level of both endogenous intraislet and exogenous arterial insulin concentration in normal man, and that this response may be normal in the absence of endogenous insulin secretion, in contrast to earlier reports. Thus, loss of beta cell function is not responsible for alpha cell failure during insulin-induced hypoglycemia in IDDM.

Important Role of Adrenergic Mechanisms in Acute Glucose Counterregulation Following Insulin-induced Hypoglycemia in Type I Diabetes: Evidence for an Effect Mediated by Beta-Adrenoreceptors

During hypoglycemia induced by an i.v. insulin infusion for 60 min, rates of plasma glucose (PG) decrease and recovery, PG nadir, and plasma counter-regulatory hormone and free fatty acid responses were studied in eight type I uncomplicated diabetic subjects and eight nondiabetic subjects. Each subject was tested three times at two different rates of insulin infusion (25 and 32 mU/m2/min): (1) during infusion of saline, (2) during infusion of phentolamine + propranolol (combined alpha, beta-blockade), and (3) during infusion of propranolol alone (isolated beta-blockade) for 150 min. At the time of the studies, the diabetic subjects had been made euglycemic by an overnight i.v. insulin infusion. During infusion of insulin (25 mU/m2/min) and saline, the rates of PG decrease and recovery were slower (P < 0.01) and PG nadir was delayed in the diabetic subjects. Moreover, their plasma glucagon response was blunted while plasma epinephrine, norepinephrine, growth hormone, and cortisol responses were similar in both groups. Infusion of insulin at 32 mU/m2/min caused larger decreases in PG than had been observed when insulin was infused at 25 mU/m2/min. Plasma glucagon responses increased in the nondiabetic subjects (P < 0.05) but not in the diabetic subjects. However, in the diabetic subjects, plasma epinephrine increased more than in the nondiabetic subjects (P < 0.05). There was an inverse correlation between the individual plasma epinephrine responses and the plasma glucagon responses in the diabetic subjects (r = −0.72) but not in the nondiabetic subjects. Alpha, beta-adrenergic blockade decreased the plasma glucose nadir and impaired the rate at which normoglycemia was restored in the diabetic subjects (P < 0.005 vs. saline) but not in the nondiabetic subjects. Plasma catecholamine and growth hormone responses were increased and plasma FFA recovery was suppressed in both groups (P < 0.05 vs. saline), while the cortisol responses were unaltered. During isolated beta-adrenergic blockade, changes in plasma glucose, counterregulatory hormones and FFA were essentially identical to those observed during combined alpha, beta-adrenergic blockade in both groups except that the augmented plasma norepinephrine responses were no longer apparent. Conclusions although epinephrine is not essential for prompt restoration of normoglycemia in normal man following insulin-induced hypoglycemia, it plays a major role in glucose counterregulation in diabetics who have an impaired glucagon secretion in response to hypoglycemia. These counterregulatory effects of epinephrine are mediated by beta-adrenoreceptors.

Lack of glucagon response in glucose counter-regulation in Type 1 (insulin-dependent) diabetics: Absence of recovery after prolonged optimal insulin therapy

SummaryMild hypoglycaemia was induced using an artificial pancreas in five normal subjects (from 5.00 ±0.15 to 2.83±0.15 mmol/l) by infusing 28 mU/m2 per min soluble insulin for 60 min. Six Type 1 (insulin-dependent) diabetic patients were stabilized for 14h using an artificial pancreas. They were then rendered hypoglycaemic (from 4.94±0.09 to 2.89±0.11 mmol/l) by infusing 28mU/m2 per min plus 16 ±3.8mU/min insulin for 60 min. Before the study, the diabetic patients were in optimal blood glucose control (mean blood glucose 6.72±0.11 mmol/l over the previous 14–20 days; HbA1 8.3±0.1%). During the insulin infusion test, blood glucose decrement was slower in the diabetic patients than in the control subjects. The blood glucose nadir was delayed in the diabetics until 75 min compared with 55 min in the control subjects. Blood glucose recovery rate in the diabetic subjects was severely impaired. In Type 1 diabetes, the counter-regulatory hormonal response to insulin induced hypoglycaemia is similar to that of non-diabetics, except for that of glucagon, the blunted response of which is not reversed by prolonged optimisation of blood glucose control. This impaired response of the A cell does not seem to be a consequence of insulin deficiency.

Modification of glycosylated haemoglobin concentration during artificial endocrine pancreas treatment of diabetics

SummaryIn order to verify whether or not insulininduced blood glucose control can acutely lower glycosylated haemoglobin levels, HbAI (a+b+c) (HbAI) was measured in 11 diabetics before, during and after 3 days of treatment with an “artificial endocrine pancreas” (Biostator). Initially 5 patients were in fair glycaemic control (group A), while the other 6 showed poor control (group B). HbAI levels decreased significantly after 3 days in both groups A (from 9.6 ± 0.2% to 8.5 ± 0.3%, mean ± SEM, p < 0.05) and B (from 13.7 ± 0.2% to 12.6 ± 0.3%, p < 0.05). A further HbAI decrease was observed until day 60 following Biostator treatment, during which period glycaemic control inproved, as assessed by fasting and post-lunch plasma glucose values and daily glycosuria determined every 10 days.These results suggest that increased HbAI levels may be reversed early by strict blood glucose control during a 3 day period. It is concluded that HbAI levels not only reflect long-term glycaemic control, but also recent acute variations in mean blood glucose values.

HbA1 in Subjects with Abnormal Glucose Tolerance but Normal Fasting Plasma Glucose

HbA1(a+b+c)(HbA1) was determined chromatographically in 107 subjects with normal fasting plasma glucose (FPG) and 112 patients with overt diabetes. Subjects with normal FPG were divided into two groups based on their response to two oral glucose tolerance tests (OGTTs), at an interval of 2 mo. In 40 subjects with normal OGTT (group I), HbA1 ranged from 5.2% to 7.2%, while in 67 subjects with abnormal OGTT (group II), it ranged from 6.3% to 9.6%. HbA1 levels were significantly higher in group II than in group I (7.7 ± 0.09% versus 6.4 ± 0.08%, mean ± SEM, P < 0.0005), but 14 subjects of group II had HbA1 levels less than 7.2%. No correlation was found between HbA1 and FPG, OGTT peak, and curve area in either group. However, the correlation became significant in all 107 subjects with normal FPG (groups I + II). In patients with overt diabetes, HbA1 ranged from 6.3% to 18% (11.9 ± 0.22%) and correlated with FPG (r = 0.78, P < 0.0005). The traditional OGTT seems more sensitive than the HbA1 measurement in detecting subjects with reduced carbohydrate tolerance. HbA1 level, on the other hand, is known to be a more specific indicator of structural abnormalities following long-term hyperglycemia. Thus HbA1 determination might be a helpful test along with OGTT to improve both selection and follow-up of subjects with true borderline diabetes.

A Risk-Benefit Appraisal of Acarbose in the Management of Non-Insulin-Dependent Diabetes Mellitus

SummaryAcarbose is an α-glucosidase inhibitor proposed for the treatment of diabetic patients. It acts by competitively inhibiting the α-glucosidases in the intestinal brush border. The principal action of these enzymes is to convert nonabsorbable dietary starch and sucrose into absorbable monosaccharides (e.g. glucose). Enzyme inhibitors delay this conversion, slowing the formation and consequently the absorption of monosaccharides, and thus reducing the concentration of postprandial blood glucose. Both starch and sucrose are influenced, whereas lactose and glucose are not.Many studies in experimental animals, healthy volunteers and patients with non-insulin-dependent diabetes mellitus (NIDDM) have shown that acarbose decreases postprandial blood glucose, with a lesser reduction of fasting blood glucose, plasma triglycerides and postprandial insulin levels. In long term studies in NIDDM patients, acarbose significantly reduced glycosylated haemoglobin levels.Acarbose is only minimally absorbed from the gut and no systemic adverse effects have been demonstrated after long term administration. The drug allows undigested carbohydrates to pass into the large bowel where they are fermented causing flatulence, bloating and diarrhoea. These symptoms, which occur in approximately 30 to 60% of patients, tend to decrease with time and seem to be dose-dependent. They are minimised by starting therapy with low doses (such as 50mg 3 times daily) which may be effective in many patients. An increase in serum hepatic transaminases observed in earlier studies in the US, where doses of acarbose up to 900mg daily were used, has been not reported with the lower doses of the drug actually recommended [150 to 300mg (up to 600mg) daily].In conclusion, acarbose may be useful in patients with NIDDM when diet alone is no longer able to maintain satisfactory blood glucose control. Furthermore, it may be a valid alternative to sulphonylurea or biguanide therapy when these drugs are contraindicated and insulin administration may be delayed. Acarbose seems also to be a useful adjunct to hypoglycaemic oral agents but its precise role in this field has not been fully clarified.

Effects of Recent, Short-Term Hyperglycemia on Responses to Hypoglycemia in Humans: Relevance to the Pathogenesis of Hypoglycemia Unawareness and Hyperglycemia-Induced Insulin Resistance

A single episode of recent hypoglycemia increases, whereas long-term hyperglycemia decreases, the glycemic thresholds of responses of counterregulatory hormone and symptoms to subsequent hypoglycemia in humans. To assess whether short-term, antecedent hyperglycemia exerts effects opposite to those observed after acute hypoglycemia, seven normal, nondiabetic subjects and eight insulin-dependent diabetes mellitus (IDDM) patients were studied during hyperinsulinemic-hypoglycemic clamp (sequential, 90-min plateaus of plasma glucose [PG] of 4.3, 3.7, 3.0, and 2.4 mmol/l). Nondiabetic subjects were studied the morning after either 6-h clamped hyperglycemia (PG ∼ 13.5 mmol/l) or euglycemia (PG ∼5 mmol/l) between 1600 and 2200 the previous day (glucose and insulin infused on both occasions), as well as after nocturnal hyperglycemia (PG ∼ 13.5 mmol/l) or euglycemia between 2300 and 0500. The IDDM patients were studied after 15 h of euglycemia or hyperglycemia (∼ 17 mmol/l) but identical hyperinsulinemia (∼ 225 pmol/l) between 1600 and 0700. Neither PG thresholds of counterregulatory hormone, symptoms, onset of cognitive dysfunction to hypoglycemia, nor maximal responses were affected by antecedent, short-term hyperglycemia in normal nondiabetic subjects and IDDM patients (NS). However, the rate of glucose infusion required to maintain hypoglycemic plateaus during hypoglycemia was lower after hyperglycemia (nondiabetic subjects 31.2 ± 3.4 vs. 36.7 ± 4 ³mol · kg−1·min−1, IDDM patients 33 ± 3.1 vs. 42.5 ± 3.9 ³mol · kg−1 · min−1; P < 0.05) indicating greater insulin resistance induced by antecedent hyperglycemia. In conclusion, in contrast to acute hypoglycemia and long-term hyperglycemia, recent, short-term hyperglycemia does not affect physiological responses to hypoglycemia. However, recent, short-term hyperglycemia induces insulin resistance that contributes to glucose counterregulation. This is relevant to IDDM patients who have deficient glucagon and adrenaline responses to hypoglycemia.

Analysis of short-term changes in reversibly and irreversibly glycosylated haemoglobin AI: Relevance to diabetes mellitus

SummaryWe have determined the stable (irreversibly glycosylated) fraction of haemoglobin AI (HbAI) on Bio-Rex 70 after incubation of red blood cells in 0.9% saline solution for 6 h at 37 °C. The total (reversibly + irreversibly glycosylated fractions) HbAI was determined before each incubation. Labile (reversibly glycosylated) tHbAI represented the difference between total and stable HbAI fractions. Total and stable HbAI fractions were determined during insulin- or meal-induced blood glucose fluctuations in 24 insulin-dependent diabetics and in seven subjects with impaired glucose tolerance. In the diabetics, the maxmial fluctuation of total HbAI was 1.47% over 2–12 h, while the simultaneous plasma glucose variation was 21.5 mmol/1. The stable HbAI fraction did not change significantly. In diabetics the differences between the maximal and minimal values of plasma glucose and total HbAI were significantly correlated. Plasma glucose correlated with simultaneously determined total and labile HbAI fractions, but not with stable HbAI In subjects with impaired glucose tolerance, similar changes in total but not in stable HbAI were observed during an oral glucose tolerance test. We conclude that, although rapid changes in chromatographically determined HbAI are relatively small, the determination of stable HbAI should be performed to circumvent this problem and to ensure a more accurate index of blood glucose control.

Urinary excretion and plasma levels of norepinephrine and epinephrine during diabetic ketoacidosis.

Sympathetic activity was determined in 13 ketoacidotic diabetics by evaluation of plasma and urinary catecholamines, before and in the course of medical management. Patients were divided into two groups. Group A (severe ketoacidosis, n = 5) and Group B (moderate ketoacidosis, = 8), depending on plasma glucose, pH and plasma bicarbonate levels. The results showed an enhanced sympathetic activity in all patients before treatment, with significant decrease during therapy. In Group A plasma catecholamines were higher than in Group B, both before and in the course of therapy. A significant correlation was found between basal plasma catecholamines and initial plasma glucose, plasma bicarbonate, hours of therapy and insulin dosage required to obtain plasma glucose levels below 150 mg/100 ml .These results, suggesting a close correlation between glycometabolic control and adrenergic activity, emphasize the role of the sympathetic nervous system as a powerful contrainsular factor in the pathogenesis and metabolic derangement of diabetic ketoacidosis.