Stabilization of the Particle-Hole Pfaffian Order by Landau-Level Mixing and Impurities That Break Particle-Hole Symmetry.

Numerical results suggest that the quantum Hall effect at ν=5/2 is described by the Pfaffian or anti-Pfaffian state in the absence of disorder and Landau-level mixing. Those states are incompatible with the observed transport properties of GaAs heterostructures, where disorder and Landau-level mixing are strong. We show that the recent proposal of a particle-hole (PH)-Pfaffian topological order by Son is consistent with all experiments. The absence of particle-hole symmetry at ν=5/2 is not an obstacle to the existence of the PH-Pfaffian order since the order is robust to symmetry breaking.

Activation of natural killer T cells by alpha-galactosylceramide treatment prevents the onset and recurrence of autoimmune Type 1 diabetes.

Type 1 diabetes (T1D) in non-obese diabetic (NOD) mice may be favored by immune dysregulation leading to the hyporesponsiveness of regulatory T cells and activation of effector T-helper type 1 (Th1) cells. The immunoregulatory activity of natural killer T (NKT) cells is well documented, and both interleukin (IL)-4 and IL-10 secreted by NKT cells have important roles in mediating this activity. NKT cells are less frequent and display deficient IL-4 responses in both NOD mice and individuals at risk for T1D (ref. 8), and this deficiency may lead to T1D (refs. 1,6-9). Thus, given that NKT cells respond to the alpha-galactosylceramide (alpha-GalCer) glycolipid in a CD1d-restricted manner by secretion of Th2 cytokines, we reasoned that activation of NKT cells by alpha-GalCer might prevent the onset and/or recurrence of T1D. Here we show that alpha-GalCer treatment, even when initiated after the onset of insulitis, protects female NOD mice from T1D and prolongs the survival of pancreatic islets transplanted into newly diabetic NOD mice. In addition, when administered after the onset of insulitis, alpha-GalCer and IL-7 displayed synergistic effects, possibly via the ability of IL-7 to render NKT cells fully responsive to alpha-GalCer. Protection from T1D by alpha-GalCer was associated with the suppression of both T- and B-cell autoimmunity to islet beta cells and with a polarized Th2-like response in spleen and pancreas of these mice. These findings raise the possibility that alpha-GalCer treatment might be used therapeutically to prevent the onset and recurrence of human T1D.

Neonatal activation of CD28 signaling overcomes T cell anergy and prevents autoimmune diabetes by an IL-4-dependent mechanism.

Optimal T cell responsiveness requires signaling through the T cell receptor (TCR) and CD28 costimulatory receptors. Previously, we showed that T cells from autoimmune nonobese diabetic (NOD) mice display proliferative hyporesponsiveness to TCR stimulation, which may be causal to the development of insulin-dependent diabetes mellitus (IDDM). Here, we demonstrate that anti-CD28 mAb stimulation restores complete NOD T cell proliferative responsiveness by augmentation of IL-4 production. Whereas neonatal treatment of NOD mice with anti-CD28 beginning at 2 wk of age inhibits destructive insulitis and protects against IDDM by enhancement of IL-4 production by islet-infiltrating T cells, administration of anti-CD28 beginning at 5-6 wk of age does not prevent IDDM. Simultaneous anti-IL-4 treatment abrogates the preventative effect of anti-CD28 treatment. Thus, neonatal CD28 costimulation during 2-4 wk of age is required to prevent IDDM, and is mediated by the generation of a Th2 cell-enriched nondestructive environment in the pancreatic islets of treated NOD mice. Our data support the hypothesis that a CD28 signal is requisite for activation of IL-4-producing cells and protection from IDDM.

Cellular toxicity of pancreatic carcinogens.

Azaserine (CAS: 115-02-6), streptozocin (CAS: 18883-66-4; streptozotocin), and N-nitrosobis(2-oxopropyl)amine [(BOP) CAS: 60599-38-4] produce different types of pancreatic tumors in rodents. We have investigated the toxic effects of these compounds on pancreatic tissues from Wistar rats and Syrian hamsters. Inhibition of protein synthesis was used as a measure of toxicity. Pancreatic islets and acinar cells from rat and hamster were labeled with [3H]leucine for 60 minutes in vitro in the presence of the various carcinogens. Azaserine, which produces acinar cell tumors in the rat, inhibited synthesis by all tissues; rat acinar cells, however, were most sensitive. Glutamine, but not serine, provided some protection against azaserine toxicity. Streptozocin inhibited synthesis by islets of both species and acinar cells from hamster; islets were the most sensitive. BOP and N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine, which induce ductal tumors in the hamster, had no effect on any of the tissues examined. These results indicate that the specificities of the cellular toxicities of the pancreatic carcinogens parallel, to some degree, their tumorigenic effects.

The effects of D- and L-glyceraldehyde on glucose oxidation, insulin secretion and insulin biosynthesis by pancreatic islets of the rat.

D-glyceraldehyde stimulated insulin secretion from isolated rat pancreatic islets in static incubation and perifusion systems. At low concentrations (2-4 mM) D-glyceraldehyde was a more potent secretagogue than glucose. The insulinotropic action of 15 mM D-glyceraldehyde was not affected by D-mannoheptulose, was potentiated by cytochalasin B (5 mug/ml) and theophylline (4 mM), and was inhibited by both adrenalin (2 muM) and somatostatin (10 mug/ml). D-glyceraldehyde at a concentration of 1.5 mM produced a 10-fold increase of L-[4,5-3H]leucine incorporation into proinsulin and insulin without a significant increase into other islet proteins. Glucose at 1.5 mM did not stimulate proinsulin biosynthesis. D-Glyceraldehyde at concentrations higher than 1.5 mM, in marked contrast to glucose, progressively inhibited incorporation of labelled leucine into proinsulin + insulin and other islet proteins. D-Glyceraldehyde also inhibited the oxidation of glucose. L-Glyceraldehyde did not stimulate proinsulin biosynthesis and had less effect than the D-isomer on insulin release and glucose oxidation. The results strongly suggest that metabolites below D-glyceraldehyde-3-P are signals for insulin biosynthesis and release. Interaction of D-glyceraldehyde with a "membrane receptor" cannot, however, be excluded with certainty.

Persisting enhanced proinsulin-insulin and protein biosynthesis (3H-leucine incorporation) by pancreatic islets of the rat after glucose exposure.

Pancreatic islets isolated from rats infused with glucose for twenty-four hours incorporated 3H-leucine into protein at higher rates than islets isolated from normoglycemic rats. Incorporation into proinsulin-insulin showed a thirteenfold increase. The effect on other islet proteins was fourfold. Exposure of islets from normoglycemic rats to high glucose in vitro for twenty and ninety minutes and subsequent incubation with 3H-leucine at low glucose showed a twofold and fivefold increase in proinsulin biosynthesis. In the in vitro system pre-exposure of the islets to mannose and dibutyryl-cyclic AMP induced a much smaller increase in proinsulin biosyntheses.

Insulin resistance prevented by portal delivery of insulin in rats with renal subcapsular islet grafts.

We determined the metabolic effects of insulin derived from renal subcapsular islet grafts, either with systemic delivery of insulin through renal venous drainage (REN) or with portal delivery of insulin after renal vein-to-superior mesenteric vein anastomosis (RMA), in streptozotocin-induced diabetic Lewis rats, in comparison with normal rats. After gavage glucose, the plasma glucose responses were similar to normal in REN and RMA rats; however, hyperinsulinemia occurred in REN rats (area under the concentration curves [AUCs] of insulin, 27 +/- 3 nmol x 1(-l) min) in comparison with RMA (14 +/- 2) and normal rats (19 +/- 2), P < 0.003, with no difference in C-peptide responses. The ratio of AUC C-peptide to AUC insulin was lower in REN (2.0 +/- 0.2) than in RMA (3.4 +/- 0.3) and normal animals (3.2 +/- 0.3), P < 0.0005. In euglycemic-hyperinsulinemic clamp studies using the same insulin infusion rate (10 pmol x kg(-1) x min(-1), insulin resistance was found in REN animals (mean glucose infusion rate [GIR], REN: 7.5 +/- 1.2; RMA: 12.0 +/- 1.2; normal: 12.7 +/- 1.0 mg x kg(-1) x min(-1); P < 0.008), with higher steady-state insulin levels in REN (554 +/- 63 pmol/l) than in RMA (291 +/- 26) and normal rats (269 +/- 60), P < 0.0001. With matching steady-state insulin levels in RMA and REN rats during infusion of insulin at 20 pmol x kg(-1) x min(-1) in RMA rats (steady-state insulin 623 +/- 64 pmol/l), GIR was 15.7 +/- 0.7 mg x kg(-1) x min(-1). Thus, systemic delivery of insulin from islet grafts is associated with hyperinsulinemia, insulin resistance, and decreased metabolic clearance of insulin. These abnormalities are prevented by portal delivery of insulin from islet grafts in the same site. The findings are consistent with the hypothesis that portal delivery of insulin is important in maintenance of normal whole-body insulin sensitivity.

Glucose turnover and insulin sensitivity in rats with pancreatic islet transplants.

To study the metabolic effects of insulin derived from islet grafts, oral glucose tolerance (OGT) and glucose turnover were examined in streptozotocin-induced diabetic Lewis rats rendered normoglycemic by syngeneic islet grafts in the renal subcapsular space (REN), in REN with renal vein-to-mesenteric vein anastomosis (REN-RMA), in the liver (intrahepatic [IH]), or in a parahepatic omental pouch (POP) and compared with normal rats. Normal OGT was found at 1 month posttransplant in all animals receiving approximately 3,000 islets, with hyperinsulinemic responses in the REN group compared with the other groups, and with higher C-peptide responses in the IH group than in the other groups (P < 0.05 by one-way analysis of variance). Glucose turnover studies in the insulin-stimulated steady state (INS-SS; infusion of insulin at 10 pmol x kg(-1) x min(-1)) at 2 months posttransplant showed that whole body glucose disappearance rates (Rd) were similar in all groups, but the REN group had higher steady-state insulin levels than the other groups. Glucose infusion rates (GIRs) were lower in the REN and IH groups than in the other groups. Apparent endogenous glucose production (EGP) was not completely inhibited in the REN and IH groups, while complete inhibition was observed in the other groups. When INS-SS insulin levels were matched to the level in REN rats by increasing the insulin infusion rate to 20 pmol x kg(-1) x min(-1) in REN-RMA, IH, and normal rats, GIR and Rd were elevated, exceeding those values in REN rats, but GIR in IH rats was still lower than in REN-RMA and normal rats. Thus, 1) in the REN group, impairment of inhibition of EGP and of stimulation of Rd by exogenous insulin contribute to insulin resistance; 2) in the IH group, incomplete inhibition of EGP is the major determinant of insulin resistance; and 3) with portal delivery of insulin in the REN-RMA and POP groups, normal insulin sensitivity is preserved. The present study confirms that hepatic portal delivery of islet secretions is necessary for physiological regulation of glucose metabolism. The study also suggests the IH grafts do not provide physiological regulation of glucose metabolism, raising the question of whether the liver is an appropriate site for insulin-secreting tissue replacement therapy in diabetes.

Continued insulin dependence despite normal range insulin sensitivity and insulin connecting peptide levels in a kidney/islet transplant patient.

OBJECTIVE: The majority of islet transplant recipients remain insulin-requiring, although many have near-normal connecting peptide (CP) levels. Insulin resistance may be one possible cause of the continuing need for exogenous insulin in islet transplant recipients. To assess this, we have studied the insulin sensitivity index (S1) in one patient with near-normal CP levels after islet transplant who remained insulin-requiring. RESEARCH DESIGN AND METHODS: The islet transplant recipient is a 36-year-old woman with no residual CP who received a kidney transplant, followed 7 days later by an islet transplant. The islets were infused into the liver via the umbilical vein. Induction immunosuppression consisted of OKT3, prednisone, cyclosporin A, and azathioprine, with maintenance on the latter three. RESULTS: Maximum CP levels after a standardized Sustacal meal were 2.09, 1.18, 0.85, and 0.81 nmol/l at 1,6,18, and 24 months posttransplant, respectively. Insulin requirements at the same times were 0.27, 0.45, 0.49, and 0.62 U.kg(-1).d(-1), while S1 was 36.3, 53.3, and 13.2 min (-1).nmol(-1).ml at 6,18, and 24 months, respectively. This compares with S1 values of 43.3+/- 10.0 min (-1).nmol(-1).ml for normal subjects. CONCLUSIONS: This patient had near-normal S1 and CP levels, but she was unable to discontinue insulin therapy, suggesting that other factors are critical. Despite this, she maintained normal or near-normal glycated hemoglobins, indicating metabolic benefit from the islet transplant.

Ipecac-induced myopathy simulating dermatomyositis.

We studied a young woman with an eating disorder. To induce vomiting, she took syrup of ipecac daily for 2 years, and then developed insidious, progressive muscle weakness. Skin findings were similar to those of dermatomyositis. Muscle biopsy, however, was similar to experimental emetine myopathy and lacked inflammatory features. Upon cessation of ipecac abuse, strength returned. We believe that this patient had ipecac-induced muscle weakness.

Transplanted MHC class I-deficient nonobese diabetic mouse islets are protected from autoimmune injury in diabetic nonobese recipients.

The injury of transplanted islets may occur by both autoimmune and alloimmune processes directed against MHC targets. To examine the role of MHC class I in islet graft injury, we transplanted syngeneic and allogeneic beta2-microglobulin-deficient islets into diabetic nonobese diabetic (NOD) mice. Loss of graft function was observed within 14 days using allogeneic C57BL/6 and BALB/c MHC class I deficient as well as wild-type MHC class I-bearing NOD donor islets. However, islets isolated from MHC class I-deficient NOD mice (NOD-B2 m-/-) survived indefinitely when transplanted under the kidney capsule of diabetic NOD recipients. Transplanted NOD-B2 m-/- islets were surrounded by a nondestructive periinsular infiltrate that expressed interleukin-4 in addition to interferon-gamma. These studies demonstrate the primary role of MHC class I molecules in causing autoimmune destruction or recurrent diabetes in transplanted islets.

Isolation of pancreatic islets from dogs. Semiautomated purification on albumin gradients.

We describe a procedure for the isolation of large numbers of canine pancreatic islets with a high level of purity. The method utilizes enzymatic and mechanical disruption of the pancreas, followed by islet purification on discontinuous density gradients of bovine serum albumin. Purification has been semiautomated through use of the Cobe 2991 cell processor. Islet preparations have a purity of 86.5% and contain 36.8 microliters of endocrine tissue. Islets isolated by this procedure are functionally intact, and restore fasting euglycemia following autotransplantation.

Micronucleus formation induced in rat liver and esophagus by nitrosamines.

The rat hepatocarcinogen nitrosodimethylamine and the esophageal carcinogens nitrosomethylbenzylamine and nitrosomethylamylamine were shown to produce chromosomal damage, as manifested by micronucleus formation, in their target tissues. There was cross-reactivity in the two tissues, however, at high dose levels. Nitrosodiethylamine, which produces tumors in both the liver and esophagus in the rat, also produced micronuclei in both tissues.

Oral contraceptives and neoplasia: 1987 update.

PIP: This article reviews the research evidence on the relationship between oral contraceptives (OCs) and neoplasia. More recent epidemiologic studies in this area are considered to have greater validity than earlier studies, largely because of improved assessment of confounding factors. Encouraging has been the finding of a protective effect of OCs on endometrial and ovarian neoplasia: about 2000 cases of endometrial cancer and 1700 cases of ovarian cancer are averted in the US each year as a result of OC use. No consistent association, either adverse or beneficial, has emerged between OCs and breast cancer; however, high-dose combined OCs exert a protective effect on the development of benign breast disease after 2 years of use. Longterm combined OC use appears to be related to the development of benign liver lesions, but the research evidence on the association between OCs and hepatocellular carcinoma remains inconclusive. Of concern is the finding that longterm OC use is associated with an increased risk of cervical neoplasia. The mechanisms by which OCs might exert adverse effects on cervical epithelium are unclear.^ieng

The issue of microinvasive squamous cell carcinoma of the vulva: an evaluation of the criteria of diagnosis and methods of therapy.

Current literature on definition, metastatic potential, and treatment of microinvasive squamous cell carcinoma of the vulva is reviewed. There is marked disparity among the various reports about the features of this lesion that are most crucial in predicting the propensity to recur or metastasize. Nonuniformity of the techniques for measuring stromal invasion is noted among many of them as well. The issue of conservative versus more aggressive surgical treatment of microinvasive squamous cell carcinoma of the vulva is discussed within the context of the reviewed data. One should exercise prudence in determining which affected patients are appropriate candidates for conservative surgical therapy.

Using client satisfaction surveys to evaluate and improve services in locked and unlocked adult inpatient facilities.

This article describes the implementation, in five inpatient subacute treatment facilities, of a satisfaction survey designed especially for adults with serious and persistent mental illnesses. The survey measures not only global satisfaction but also client perceptions about different treatment modalities and services, important treatment goals, and the philosophy of treatment. Data are presented from 770 completed surveys, illustrating patterns of satisfaction across facilities and services and patterns over time of stability and change in satisfaction. Data are also presented showing how the surveys were used to facilitate and measure improvements in clinical services. Finally, the implications for mental health services delivery are summarized.