Recent-onset Diabetes Mellitus Research Articles

Effects of glimepiride on insulin secretion and sensitivity in patients with recently diagnosed type 2 diabetes mellitus

Background: The exact mechanism of the efficacy of glimepiride in the achievement of glycemic control has not yet been clearly defined. Objective: This study was conducted to examine the influence of glimepiride on insulin secretion and sensitivity in patients with type 2 diabetes mellitus (DM) of recent onset. Methods: This 24-week, open-label, controlled trial was conducted at the Division of Endocrinology and Metabolism, Veterans Affairs Medical Center (Phoenix, Arizona). Study participants were aged 32 to 75 years and had recent-onset (established by a short duration of symptoms 6 weeks to 6 months prior to the study) type 2 DM, or were age-matched healthy volunteers (control group). In the diabetic patients, glimepiride tablets were administered orally, initially at 2 mg once daily in the morning, with the dosage increased by 1 mg every 2 weeks until fasting plasma glucose (FPG) decreased to 6.7 mmol/L; the dosage was then maintained for the remainder of the 24-week study period. Oral glucose tolerance tests (OGTTs) were conducted in the control group and before treatment and at 24 weeks after the achievement and maintenance of glycemic control (glycosylated hemoglobin <7.0%) in the diabetic group. For OGTT, plasma insulin and glucose levels were determined after the subjects fasted overnight and then at every 15 minutes for 2 hours after glucose challenge. Results: Fourteen diabetic men (mean [SEM] age, 50 [6] years; range, 32–75 years) and 10 male healthy controls (mean [SD] age, 48 [5] years; range, 30–68 years) were enrolled. In the DM group, FPG decreased significantly after treatment ( P < 0.001); fasting plasma insulin was markedly elevated before treatment ( P < 0.001 vs controls) and decreased after treatment ( P < 0.01) but did not normalize; first-phase insulin secretion was markedly inhibited before treatment ( P < 0.001 vs controls) and normalized after treatment ( P < 0.001); total insulin secretion significantly improved after treatment ( P < 0.01) but did not normalize. Finally, the pretreatment insulin sensitivity index decreased significantly ( P < 0.01) after treatment and normalized in 6 of 14 patients (42.9%) with type 2 DM. Conclusions: In this study, glimepiride achieved desirable glycemic control in patients with recent-onset type 2 DM through improvement in insulin secretion and sensitivity.

Characterization of Monocyte-Derived Dendritic Cells in Recent-Onset Diabetes Mellitus Type 1

Dendritic cells (DC) may play an important role in the immunopathogenesis of type 1 diabetes mellitus (DM-1). In this study, we have analyzed phenotypical changes during cytokine-driven maturation from CD14 + monocytes to mature DC and DC-dependent T-cell stimulation in recent-onset pediatric DM-1 patients and healthy controls. DC maturation was monitored by flow cytometric analyses for the expression of surface markers (HLA-DR, CD1a, CD40, CD80, CD86, CD83, CD14, CD32, mannose-receptor, and CD11c). Flow cytometric analysis of isolated peripheral blood monocytes did not reveal apparent differences between patients and controls. During DC maturation no obvious differences in the expression patterns of surface markers over time or evidence for maturation impairments in DM-1 patients could be appreciated. Solely, a marginal, but significant, transient downregulation of CD1a on Day 3 (mean MDFI 3.82 vs 7.25; P = 0.021), which was accompanied by an increase of IL-6, could be observed. The comparison of mature DCs (Day 10) between patients and controls indicated no significant differences, except for CD83 (mean MDFI 1.7 vs 1.5; P = 0.042) and CD80 (mean MDFI 15.92 vs 12.73; P = 0.042). Moreover, no difference in T-cell stimulatory capacity was seen. In conclusion, our analysis of a cohort of recent-onset DM-1 patients and controls does not support a role for disease-related alterations in cytokine-driven maturation of monocyte-derived DC.

Frequency of islet cell autoantibodies (IA-2 and GAD) in young Brazilian type 1 diabetes patients.

Type 1 diabetes, as an autoimmune disease, presents several islet cell-specific autoantibodies such as islet cell antibody (ICA), anti-insulin, anti-glutamic acid decarboxylase (GAD) and the antibody (Ab) against tyrosine phosphatase (PTP)-like protein known as ICA-512 (IA-2). In order to determine the frequency of the anti-GAD and anti-IA-2 autoantibodies in Brazilian type 1 diabetes patients we studied 35 diabetes mellitus (DM) type 1 patients with recent-onset disease (</=12 months) and 37 type 1 diabetes patients with long-duration diabetes (>12 months) who were compared to 12 children with normal fasting glucose. Anti-GAD65 and anti-IA-2 autoantibodies were detected with commercial immunoprecipitation assays. The frequency of positive results in recent-onset DM type 1 patients was 80.0% for GADAb, 62.9% for IA-2Ab and 82.9% for GADAb and/or IA-2Ab. The long-duration type 1 diabetes subjects presented frequencies of 54.1% for GADAb and IA-2Ab, and 67.5% for GAD and/or IA-2 antibodies. The control group showed no positive cases. Anti-GAD and IA-2 assays showed a high frequency of positivity in these Brazilian type 1 diabetes patients, who presented the same prevalence as a Caucasian population.

Large incidence variation of Type I diabetes in central-southern Italy 1990-1995: lower risk in rural areas.

To evaluate the relation between the incidence of childhood Type I (insulin-dependent) diabetes mellitus and the degree of urbanization in the central-southern part of Italy. The incidence was determined in two areas: area A encompasses 3 regions of central-eastern Italy (Marche, Abruzzo, Umbria), whereas area B encompasses one southern region (Campania). During 1990-1995, 706 children aged 14 or under with insulin-dependent diabetes mellitus of recent onset were registered. The completeness of the case ascertainment in the registries analysed separately for each region was high, ranging from 96.3 % to 99%. The age-standardized incidence was higher in area A (9.6 per 100000 person per year; 95 % confidence interval: 8.5-10.8) than in area B (5.4 per 100000 person per year; 95% confidence interval: 4.9-6.0). In both areas the standardized incidence ratios increased with the degree of urbanization (chi-squared for trend: area A= 140, p < 0.0001; area B = 79, p < 0.0001). The highest standardized incidence ratios were in the most urban communities. This study showed a statistically significant difference in incidence of childhood insulin-dependent diabetes mellitus among different areas of the continental peninsula of Italy. People living in the rural communities appear to have a lower risk.

Increased CD69 and human leukocyte antigen-DR expression on T lymphocytes in insulin-dependent diabetes mellitus of long standing.

To better define prevailing activation of circulating T cell subsets in insulin-dependent diabetes mellitus (IDDM) of recent onset (DM; n = 31; median age +/- SD, 28 +/- 6.9 yr) and of long standing (DML; n = 27; age, 33 +/- 10.4 yr; median duration of disease, 105 months), CD4+ and CD8+ T cells were analyzed to determine their naive and memory subsets as well as their expression of human leukocyte antigen (HLA)-DR, interleukin-2 receptor alpha-chain (CD25), and CD69 by three-color flow cytometry. Twenty-six healthy subjects (HS; age, 32.0 +/- 8.2 yr) served as controls. No deviation was seen in either IDDM group compared to HS in CD25 expression on CD4+ or CD8+ cells or in their CD45RA+ or CD45RA- subsets. HLA-DR expression, however, was increased (P < 0.05) in total CD8+ cells and CD45RA+ cells, with CD45RA- CD8+ cells joining the prevailing pattern only in DML. Among CD4+ cells, increased expression of HLA-DR molecules was restricted to total and CD45RA- cells in DML. CD69 expression did not differ between IDDM and HS, but differed between DML (CD4+, CD8+, and CD45RA- CD4+) and DM only. In conclusion, our data demonstrate that HLA-DR expression in IDDM is restricted to memory cells (CD45RA-) among CD4+ cells in DML and is more markedly confined to naive (CD45RA+) than to memory CD8+ cells, whereas the early activation antigen CD69 is more readily expressed in DML than in DM. The observed activation of circulating T cells suggests an ongoing immune process in IDDM both at clinical manifestation and after long duration.

Heterogeneity of islet pathology in two infants with recent onset diabetes mellitus

The mechanisms by which the beta cells of pancreatic islets are destroyed in insulin-dependent diabetes mellitus (IDDM) are poorly understood. In this report the pancreatic histo- and immunopathology of two children, both HLA-DR 3/4, DQ 2/8 positive and who both died from cerebral oedema within a day of clinical diagnosis of IDDM, were investigated. Patient 1, a 14-month-old girl, had a 4-week history of polydipsia and polyuria. Patient 2, a 3-year-old boy, had 2 days of illness. Both patients had a similarly severe loss of insulin cells but differed markedly as to the extent of lymphocytic islet infiltration (insulitis). Apart from insulitis, marked islet macrophage infiltration was demonstrated in both patients with the HAM-56 monoclonal antibody. Neither patient showed aberrant expression of HLA class II antigens on insulin-immunoreactive cells, but allele-specific HLA-DQ8 expression was evident on endothelial cells. Glutamic acid decarboxylase immunoreactivity was detected in both insulin- and glucagon-immunoreactive cells. It is concluded that the heterogeneity of islet pathology, especially insulitis, may reflect different dynamics and extent rather than different pathomechanisms of immune destruction of islets in IDDM.

Does Endothelin Play a Role in the Pathogenesis of Early Diabetic Nephropathy?

Endothelin-1 (ET-1) causes dramatic vasoconstriction and reduction of renal blood flow, with a decreased glomerular filtration rate (GFR). Early diabetic nephropathy is characterized by elevation of GFR and the formation of intrarenal microaneurysms. However, the mechanisms are unclear. To elucidate the pathophysiologic significance of urinary ET-1 in early diabetic nephropathy, the 24-h urinary excretion of ET-1 in 12 normal subjects and 20 patients with newly diagnosed type 2 diabetes mellitus were determined by a highly sensitive radioimmunoassay. The 24-h urinary ET-1 excretion in patients with diabetes mellitus (14.2 +/- 3.1 pmol, mean +/- SEM) was significantly lower (p < 0.05) than that of normal subjects (25.0 +/- 3.7 pmol). This decrease in urinary excretion of ET-1 in patients with recent-onset diabetes mellitus suggests a possible role of ET-1 in the pathogenesis of early diabetic nephropathy.

Clinical Presentation and Ultrasonography in the Diagnosis of Pancreatic Cancer

One thousand twenty patients consecutively admitted because of a clinical suspicion of pancreatic cancer were investigated to evaluate the accuracy of simple clinical, laboratory, and ultrasonographic data in the diagnosis of pancreatic cancer. Age, weight loss, recent-onset diabetes mellitus, palpable abdominal mass or gallbladder, elevated serum bilirubin or alkaline phosphatase levels, and ultrasonography were significant criteria in discriminating 80 pancreatic cancers from 940 controls. The most sensitive criteria were ultrasonography (83%), weight loss (66%), and bilirubin level of > 3 mg/dl (61%); the most specific were ultrasonography (99%), recent-onset diabetes (97%), and a distended palpable gallbladder (94%). Only ultrasonography demonstrated an elevated positive predictive value (86%), while weight loss, elevated bilirubin and alkaline phosphatase, besides ultrasonography had an elevated negative predictive value (95%). These results show that advanced pancreatic cancer may be excluded with simple clinical and laboratory data; ultrasonography can confirm the diagnosis with a high degree of accuracy. We suggest that the results of any new diagnostic tests for pancreatic cancer be compared with these clinical findings.

Autoantibodies to the GLUT-2 glucose transporter of beta cells in insulin-dependent diabetes mellitus of recent onset.

Purified immunoglobulin G (IgG) from the serum of patients with insulin-dependent diabetes mellitus (IDDM) of recent onset inhibits high-Km uptake of 3-O-methyl-beta-D-glucose by rat pancreatic islets. To determine if the inhibition is the result of antibodies against GLUT-2, the high-Km glucose transporter of beta cells, we incubated IDDM sera with rat islet cells and with AtT-20ins cells transfected to express GLUT-2. IDDM sera inhibited glucose uptake in islet cells and in GLUT-2-expressing AtT-20ins cells but not in AtT-20ins cells transfected to express the low-Km isoform, GLUT-1. In 24 of 30 (77%) patients with newly diagnosed IDDM, IgG binding as measured by immunofluorescence and flow cytometry of the cells transfected to express GLUT-2 was > 2 standard deviations from the mean of the nondiabetic population; 29 of 31 (96%) of nondiabetic children were negative (P < 0.0001). Increased IgG binding could be removed by absorption with GLUT-2-expressing cells but not with GLUT-1-expressing cells. We conclude that most patients with IDDM of recent onset have autoantibodies to GLUT-2.

Risk factors for cyclosporine-induced nephropathy in patients with autoimmune diseases. International Kidney Biopsy Registry of Cyclosporine in Autoimmune Diseases.

Cyclosporine is an immunosuppressive drug that is used to treat patients with autoimmune disease as well as patients who have received allografts. The drug can cause renal damage, but the incidence of and risk factors for nephropathy in patients treated with cyclosporine for autoimmune or inflammatory diseases are not known. We analyzed data from renal biopsies performed in 192 patients (129 adults and 63 children) who had been treated with cyclosporine for insulin-dependent diabetes mellitus of recent onset, uveitis, psoriasis, Sjögren's syndrome, or polychondritis. The mean (+/- SD) initial dose of cyclosporine was 8.2 +/- 2.8 mg per kilogram of body weight per day, and the duration of treatment was 4 to 39 months (median, 13). Forty-one patients (37 adults and 4 children) had cyclosporine-induced nephropathy, defined as at least moderate focal interstitial fibrosis with tubular atrophy, arteriolar alterations, or both. As compared with patients in whom nephropathy did not develop, these patients received a larger initial dose of cyclosporine (9.3 +/- 2.8 vs. 8.0 +/- 2.8 mg per kilogram per day), had a larger maximal increase in the serum creatinine concentration above base-line values (101 +/- 77 percent vs. 50 +/- 33 percent), and were older (31 +/- 13 vs. 23 +/- 12 years). These three variables were shown by multivariate logistic-regression analysis to be significant risk factors. The duration of the elevation in the serum creatinine concentration and the occurrence of elevated blood pressure were not additional risk factors. Nephropathy is an important potential effect of cyclosporine therapy. The risk of its development in patients with autoimmune diseases who are treated with cyclosporine can be minimized by allowing a dose no higher than 5 mg per kilogram per day and avoiding increases in serum creatinine of more than 30 percent above the patient's base-line value.

Autoantibodies to the insulin receptor are infrequent findings in Type 1 (insulin-dependent) diabetes mellitus of recent onset

To determine whether autoantibodies to the insulin receptor may represent markers of Type 1 (insulin-dependent) diabetes, the prevalence of such antibodies was investigated in sera of 60 newly diagnosed untreated Type 1 diabetic patients. A sensitive assay, based on enzyme linked immunosorbent assay has been set up which detects antibodies to the insulin receptor irrespective of their potentially inhibiting effect on insulin binding. Moreover, this method allows easy determination of the immunoglobulin class involved in the anti-receptor activity. Among the 60 sera examined, only one was found to contain anti-insulin receptor autoantibodies (IgG class). In view of our data, we conclude that autoantibodies to the insulin receptor are infrequent findings in Type 1 diabetes of recent onset.

Phagocyte oxidative metabolism in cyclosporine-or placebo-treated patients with insulin-dependent (Type I) diabetes mellitus of recent onset

Several lines of evidence suggest that phagocyte-mediated oxidative processes are involved in damage to pancreatic islet cells of Type I insulin-dependent diabetes mellitus (IDDM). This hypothesis, however, has not yet been explored at the clinical onset of IDDM. Similarly, the possibility that cyclosporine A (Cy-A) might exert a selective influence on these phagocyte-mediated oxidative reactions has also not yet been investigated as compared to a placebo. The present study tested both hypotheses in 32 patients with recently diagnosed IDDM who were part of the recent French multicenter randomized therapeutic trial of Cy-A. The production of reactive oxygen intermediates (ROI) by circulating polymorphonuclear (PMN) and mononuclear (MN) phagocytes was determined by luminol-dependent chemiluminescence (CL), both directly within microamounts of whole blood and in purified PMN or MN phagocyte suspensions. Lastly, CL production was measured in the absence (resting CL) and the presence of a panel of particular and soluble phagocyte membrane-stimulating agents. We found that on entry into the trial, i.e. within less than 2 months of the clinical onset of IDDM, patients had normal whole blood CL production in the absence of a stimulating agent and upon phagocytic challenge with latex or opsonised zymosan particles. By contrast, whole blood CL responses to soluble stimuli such as phorbol myristate acetate (PMA), concanavalin A (Con-A) and F Met-Leu-Phe (FMLP) were significantly higher than in the control group of 52 normal subjects ( P<0.01). In purified PMN and MN phagocyte suspensions, both resting and stimulated CL productions were normal, regardless of the type of stimulating agent. After 3 months of treatment, whole blood CL responses to Con-A and FMLP returned to almost normal levels in patients treated with Cy-A (15 cases) but not in those receiving the placebo (17 cases); PMA-induced CL responses were also decreased, but this was found in both groups of patients. In purified phagocyte suspensions we detected no effect of Cy-A on PMN, whereas MN phagocytes from Cy-A-treated patients showed reduced CL responses to FMLP but not to other stimuli. Altogether, these results demonstrate for the first time that the capacity of circulating PMN and MN phagocytes to generate ROI is normal at the clinical onset of IDDM and suggest that circulating substances increase oxidative responses to soluble, but not particulate, stimuli. With regard to the in vivo effect of Cy-A, it appears that Cy-A does not influence PMN oxidative response-potential, whereas it selectively alters MN oxidative responses to chemotactic agents such as FMLP. This latter finding suggests that, as already described for T lymphocytes, Cy-A may interfere with calcium-dependent activation pathways of monocytes.

Effects of cyclosporine in recent-onset juvenile type 1 diabetes: Impact of age and duration of disease

Administration of cyclosporine resulted in reduced insulin requirements and improved glycemic control in patients with insulin-dependent diabetes mellitus of recent onset, but the drug was less effective in young children. Renal toxic effects and other problems related to therapy resolved after discontinuation of the drug. Sustained remission seemed dependent on continued administration of cyclosporine. Although short-term control of diabetes may be achieved in some patients, more studies are needed to determine whether cyclosporine can be given safely as maintenance therapy to maintain glycemic control and prevent the long-term consequences of the disease.

The immune response against therapeutic monoclonal antibodies

Autoimmune diseases have been increasing in incidence in recent decades, especially in industrialized countries. As of today these represent a major public health problem that has called for complex therapeutic approaches. These include the development of biotechnology-derived products such as humanized monoclonal antibodies or fusion proteins that target membrane receptors of immune cells or cytokines involved in the pathogenesis of autoimmune diseases. Major disease-modifying drugs have emerged that have revolutionized the management of rheumatoid arthritis, multiple sclerosis and others. Presented here are the premises that led to the development of these agents, and the conditions of their current use in autoimmune disease. Despite the major advances, most of these therapies clearly do not provide a real cure of autoimmune diseases as their effect is not specific for the autoantigen-induced responses and, importantly, their time span of activity is limited, and self-tolerance is not restored. In other words, our newly-enriched drug armamentarium of immunomodulatory and anti-inflammatory molecules still does not provide for a radical and durable solution to autoimmunity. However gratifying exceptions to this do exist, in particular monoclonal antibodies specific for the CD3 complex, the transducing element of the T cell antigen receptor, with experimental data having been successfully translated to the clinic for therapy of recent-onset autoimmune diabetes mellitus, with an ensuing and sustained remission (up to 4 years) after just a single short course of treatment. Other such successes can be anticipated.

Clinical utility of oral hypoglycemic agents in the management of patients with noninsulin-dependent diabetes mellitus

Clinical usage of the sulfonylurea drugs is based on experience in well-controlled long-term studies in large numbers of patients. The results of such studies indicate that approximately 60 to 70 percent of the patients with noninsulin-dependent diabetes mellitus of recent onset will respond initially to sulfonylurea therapy with satisfactory control of glycemia, about 15 to 20 percent will not respond initially (primary failures), and another 15 to 20 percent who respond initially will lose their responsiveness during the first few years of treatment (secondary failures). In extensive studies in small numbers of patients, we found that successful management of noninsulin-dependent diabetes mellitus with glipizide was correlated with increased insulin responsiveness and that patients who did not have an increase in their insulin-mediated glucose disposal during glipizide therapy were primary failures. Successful treatment of noninsulin-dependent diabetes mellitus with sulfonylureas is associated with onset of of the disease at 40 years of age or later, normal or increased body weight, duration of disease less than five years, and a history of either no previous insulin therapy or therapy with less than 20 units of insulin per day. The use of specific sulfonylurea drugs is predicated on differences in their metabolism, side effects, and perhaps potency.

Hypoglycemic therapy and hemostasis in maturity-onset diabetes mellitus: Existence of an early prethrombotic state

We studied 115 patients with uncomplicated maturity-onset diabetes mellitus, 55 males and 60 females; patients were divided into 7 groups according to the hypoglycemic therapy (diet alone, tolbutamide, glibenclamide, phenformin, combination of the last two drugs, insulin, combination of insulin and phenformin). All patients were tested for a prethrombotic state by the simultaneous determination of the following parameters: ADP-induced platelet aggregation, Factor VIII as antigen and procoagulant activity and the plasma levels of fibrinogen and antithrobin III. Our results show that these parameters were markedly altered in diabetic patients when compared to apparently normal subjects of the same age; on the contrary, differences between groups of diabetic patients were very slight and hardly ever statistically significant. These results suggest that the prethrombotic alterations of hemostatic system are very similar in all groups of diabetic patients; this is true also for patients submitted to diet alone, i.e. with slight and usually recent-onset diabetes mellitus. In conclusion, our study suggests the opportunity, at least from the theoretical point of view, of systematic antiaggregant and/or anticoagulant treatment(s) in maturity-onset diabetes mellitus, whose severe and precocious prethrombotic alterations seem to be independent of the hypoglycemic treatment applied.

Idiopathic membranous glomerulonephritis in diabetic patients: report of three cases and review of the literature.

Three adults with diabetes mellitus and nephrotic syndrome of recent onset underwent renal biopsies because of certain unusual clinical features. Despite heavy proteinuria, one patient had no evidence of diabetic retinopathy, one had preservation of normal renal function, while the third had sudden onset of renal failure. Microscopic examination of renal biopsy material disclosed pure membranous nephropathy in one, membranous nephropathy and nodular glomerulosclerosis in two. Because of significant differences in the natural history of these two glomerulopathies and the possible beneficial effects of steroid therapy in membranous nephropathy, we suggest that renal biopsies be performed in diabetic patients having persistent hematuria, sudden onset of renal failure, massive proteinuria without azotemia, retinopathy, or other evidence of microvascular disease, to uncover superimposed and treatable disorders that may influence the course of renal disease.

Idiopathic Membranous Glomerulonephritis in Diabetic Patients

Three adults with diabetes mellitus and nephrotic syndrome of recent onset underwent renal biopsies because of certain unusual clinical features. Despite heavy proteinuria, one patient had no evidence of diabetic retinopathy, one had preservation of normal renal function, while the third had sudden onset of renal failure. Microscopic examination of renal biopsy material disclosed pure membranous nephropathy in one, membranous nephropathy and nodular glomerulosclerosis in two. Because of significant differences in the natural history of these two glomerulopathies and the possible beneficial effects of steroid therapy in membranous nephropathy, we suggest that renal biopsies be performed in diabetic patients having persistent hematuria, sudden onset of renal failure, massive proteinuria without azotemia, retinopathy, or other evidence of microvascular disease, to uncover superimposed and treatable disorders that may influence the course of renal disease.

Islet-cell antibodies and HLA type in Japanese insulin-dependent diabetics.

Pancreatic islet-cell antibodies (I.C.Ab.) were detected in the sera of 6 of 123 patients with insulin-dependent diabetes mellitus of recent onset, compared with only 2 positive results among the 434 control sera. The prevalence of humoral I.C.Ab. was strongly dependent on the duration of the diabetes, being 4 of 6 I.C.Ab. positive patients during the first year from diagnosis and falling to 2 of 6 I.C.Ab. positive patients at one to three years. Over four years from the time of diagnosis, there was no I.C.Ab. in the sera of the diabetics. Six I.C.Ab. positive in our group of 123 diabetic patients showed no association with any particular B locus antigen or DYT.

878 FAILURE TO ISOLATE A TRANSMISSABLE AGENT FROM THE PANCREAS OF A CHILD WITH NEWLY DIAGNOSED DIABETES MELLITUS

A 9 year old girl died in ketoacidotic coma that developed less than two weeks after her first symptoms of diabetes mellitus. Electron microscopic examination of pancreatic tissue showed acute beta cell necrosis associated with myxoparamyxovirus-like particles in islet, acinar and ductal cells. We attempted to isolate a virus from the pancreas of this patient by several techniques. Primary inoculation and passage of pancreas onto Hep 2, cerco (CMK) and rhesus monkey kidney, and human embryonic lung (HEL) tissue culture were negative for cytopathic effect (CPE), hemadsorption with guinea pig red blood cells (HA), challenge with echovirus 11 and electron microscopy (EM). Whole islet and pancreatic suspension cultures were adapted to monolayer cultures and maintained for 20 passages. These materials were also co-cultivated with CMK Hep 2 and HEL for 6 passages and with Hela and Vero cells for 20 passages. All passages (cell contents and supernatant) of pancreas and co-cultivations remained negative for CPE, HA, hemagglutination, echovirus 11 challenge and EM after ultracentrifugation. Late passages treated with 5-iodo-2-deoxyuridine for 24 and 48 hours also remained negative. Conventional and “rescue” techniques failed to detect complete or latent viruses in the pancreas of a child with recent onset diabetes mellitus.