Campath-1H does not alter bone marrow cell regulatory function

We have previously reported in laboratory volunteers (in vitro) and renal transplant recipients (ex vivo) that bone marrow cells (BMC) are potent downregulators of the immune response. Also, the use of alemtuzumab (Campath-1H, C1H) for immunodepletion is associated with the most potent lasting effects yet seen on T-cell immunity in renal transplantation. We questioned whether the administration of C1H to kidney allograft recipients of donor bone marrow cell (DBMC) infusions would lead to stronger or weaker immunoregulatory effects. Human BMC depleted of T cells (nT-BMC) were either untreated or treated with C1H and rabbit complement and compared for their ability to downregulate autologous or allogeneic T-cell responses and to generate T regulatory (T reg) cells. The proliferative responses to anti-CD3 monoclonal antibody of T cells derived from cocultures with C1H-treated or untreated autologous nT-BMC were equally suppressed, i.e., an equivalent alteration in CD3 complex signaling, not regained by the addition of interleukin 2. Adenosine triphosphate levels were also markedly reduced in T cells both from C1H-treated and untreated nT-BMC cocultures. The ability of C1H-treated or untreated nT-BMC to suppress autologous T-cell cytotoxic function was also equivalent, with a marked, but equivalent, capacity to induce CD4/CD25(high) T regs from CD3(+) cells, which effectively downregulated cytotoxic T cells. To mimic the clinical infusion of DBMC into (allogeneic) recipients, peripheral blood mononuclear cells were also cultured with allogeneic C1H-treated and untreated nT-BMC. T cells derived from these cultures secondarily stimulated with the same-donor mature antigen-presenting cells exhibited suppressed cytotoxicity by 85% and 54%, respectively. These in vitro studies suggest that C1H does not abrogate BMC immunoregulation and thus may allow its lympho-depleting effect to be synergistic.     

Acute hypoxia does not alter inter-alveolar perfusion distribution in unanesthetized rats

Effects of hypoxic vasoconstriction on inter-alveolar perfusion distribution (< or =1000 alveoli) have not been studied. To address this, we measured inter-alveolar perfusion distribution in the lungs of unanesthetized rats breathing 10% O(2). Perfusion distributions were measured by analyzing the trapping patterns of 4 microm diameter fluorescent latex particles infused into the pulmonary circulation. The trapping patterns were statistically quantified in confocal images of the dried lungs. Trapping patterns were measured in lung volumes that ranged between less than 1 and 1300 alveoli, and were expressed as the log of the dispersion index (logDI). A uniform (statistically random) perfusion distribution corresponds to a logDI value of zero. The more this value exceeds zero, the more the distribution is clustered (non-random). At the largest tissue volume (1300 alveoli) logDI reached a maximum value of 0.68+/-0.42 (mean+/-s.d.) in hypoxic rats (n = 6), 0.50+/-0.38 in hypercapnic rats (n.s.) and 0.48+/-0.25 in air-breathing controls (n.s.). Our results suggest that acute hypoxia did not cause significant changes in inter-alveolar perfusion distribution in unanesthetized, spontaneously breathing rats.     

Insulin therapy does not interfere with venous endothelial function evaluation in patients with type 2 diabetes mellitus

INTRODUCTION:

Endothelium-dependent dilation is improved in insulin-treated diabetic patients, but this effect is probably due to improved glycemic control. The objective of the present study was to compare endothelium-dependent dilation in patients with well-controlled type 2 diabetes who are or are not using insulin as part of their therapy.

METHODS:

We studied 27 patients with type 2 diabetes (11 women, 60.3 years ± 6 years, with HbA1c < 7% and no nephropathy), including 16 patients treated with anti-diabetic agents (No-Ins, 8 women) and 11 patients treated with insulin alone or in combination with anti-diabetic agents (Ins, 3 women). Endothelial function was evaluated by the dorsal hand vein technique, which measures changes in vein diameter in response to phenylephrine, acetylcholine (endothelium-dependent vasodilation) and sodium nitroprusside (endothelium-independent vasodilation).

RESULTS:

Age, systolic blood pressure (No-Ins: 129.4 mmHg ± 11.8 mmHg, Ins: 134.8 mmHg ± 12.0 mmHg; P =  0.257), HbA_1c, lipids and urinary albumin excretion rate [No-Ins: 9 mg/24 h (0-14.1 mg/24 h) vs. Ins: 10.6 mg/24 h (7.5-14.4 mg/24 h), P = 0.398] were similar between groups. There was no difference between endothelium-dependent vasodilation of the No-Ins group (59.3% ± 26.5%) vs. the Ins group (54.0% ± 16.3%; P = 0.526). Endothelium-independent vasodilation was also similar between the No-Ins (113.7% ± 35.3%) and Ins groups (111.9% ± 28.5%; P = 0.888).

CONCLUSIONS:

Subcutaneous insulin therapy does not interfere with venous endothelial function in type 2 diabetes when glycemic and blood pressure control are stable.     

NFkappaB1 gene does not affect type 1 diabetes predisposition in a Spanish population

The chromosomal location of the NFkappaB1 gene on 4q, a region linked to type 1 diabetes (T1D), together with the observed resistance to T1D of NFkappaB1-deficient mice, suggests its potential role as candidate gene increasing diabetes predisposition. Previous association studies in diverse populations yielded inconclusive results. Two polymorphisms in the promoter region of the NFkappaB1 gene have been studied: a functional -94ins/delATTG regulating the gene expression and a very informative CA-repeat microsatellite. A strong association with the latter was reported in British population but could not be replicated in Danish families. No evidence of association was detected for those genetic markers in 270 Spanish T1D patients and 484 healthy ethnically matched controls. Therefore, it seems that this gene plays no major role in T1D predisposition.     

Quality of life of adolescents with type 1 diabetes

INTRODUCTION:

Diabetes mellitus is a highly prevalent chronic disease. Type 1 diabetes mellitus usually develops during infancy and adolescence and may affect the quality of life of adolescents.

OBJECTIVE:

To evaluate the quality of life of adolescents with type 1 diabetes mellitus in a metropolitan region of western central Brazil.

METHODS:

Adolescents aged 10–19 years who had been diagnosed with type 1 diabetes mellitus at least 1 year previously were included. Patients with verbal communication difficulties, severe disease, and symptomatic hypo- or hyperglycemic crisis as well as those without an adult companion and who were <18 years of age were excluded. The self-administered Diabetes Quality of Life for Youths instrument was applied.

RESULTS:

Among 96 adolescents (57% females; 47% white, and 53% nonwhite), 81% had an HbA1c level of >7%. In general, the adolescents consistently reported having a good quality of life. The median scores for the domains of the instrument were as follows: “satisfaction”: 35; “impact”: 51; and “worries“: 26. The total score for all domains was 112. Bivariate analysis showed significant associations among a lower family income, public health assistance, and insulin type in the “satisfaction” domain; and a lower family income, public health assistance, public school attendance, and a low parental education level in the “worries“ domain and for the total score. A longer time since diagnosis was associated with a lower total score. Multivariable analysis confirmed the association of a worse quality of life with public health assistance, time since diagnosis, and sedentary lifestyle in the “satisfaction” domain; female gender in the “worries” domain; and public health assistance for the total score.

CONCLUSIONS:

Overall, the adolescents evaluated in this study viewed their quality of life as good. Specific factors that led to the deterioration of quality of life, including public assistance, time since diagnosis, sedentary lifestyle, and female gender, were identified.     

Maternal trait anxiety and diabetes control in adolescents with type 1 diabetes

OBJECTIVE: To examine the relationship of maternal trait anxiety with diabetes regulation among adolescents with type 1 diabetes. METHODS: Adolescents and their mothers completed surveys assessing trait anxiety, maternal involvement in diabetes care, adolescent management skills, autonomous motivations, mood state, and absenteeism due to diabetes. HbA1c readings, used to assess metabolic control, were obtained from medical records. RESULTS: Trait-anxious mothers reported taking more responsibility for diabetes management tasks and perceived their adolescents as having poorer management skills. Adolescents with high-anxious mothers reported stronger beliefs that their mothers had high control over their diabetes and their parents were over-protective. For younger adolescents, maternal trait anxiety was associated with higher HbA1c levels and greater absenteeism. For older adolescents, maternal trait anxiety was associated with lower autonomous motivations for diabetes care and lower positive affect. CONCLUSIONS: Interventions for adolescents with diabetes may benefit from addressing these maternal anxiety dynamics in ways that improve diabetes control.     

Recessive tolerance to preproinsulin 2 reduces but does not abolish type 1 diabetes

Although autoimmune diseases can be initiated by immunization with a single antigen, it is not clear whether a single self antigen is essential for the initiation and, perhaps, the perpetuation of spontaneous autoimmunity. Some studies have suggested that insulin may represent an essential autoantigen in type 1 diabetes. Here we show that unlike tolerance to glutamic acid decarboxylase, tolerance to transgenically overexpressed preproinsulin 2 substantially reduced the onset and severity of type 1 diabetes in nonobese diabetic mice. However, some mice still developed type 1 diabetes, suggesting that insulin is a key, but not absolutely essential, autoantigen. The results are consistent with the idea that the human IDDM2 locus controls susceptibility to type 1 diabetes by regulating intrathymic preproinsulin expression.     

Acute hypoxic exercise does not alter post-exercise iron metabolism in moderately trained endurance athletes

Purpose

This study measured the influence of acute hypoxic exercise on Interleukin-6 (IL-6), hepcidin, and iron biomarkers in athletes.

Methods

In a repeated measures design, 13 moderately trained endurance athletes performed 5 × 4 min intervals at 90 % of their peak oxygen consumption velocity (vVO_2peak) in both normoxic [NORM, fraction of inspired oxygen (F _IO₂) = 0.2093, 15.3 ± 1.7 km h^?1] and simulated hypoxic (HYP, F _IO₂ = 0.1450, 13.2 ± 1.5 km h^?1) conditions. Venous blood samples were obtained pre-, post-, and 3 h post-exercise, and analysed for serum hepcidin, IL-6, ferritin, iron, soluble transferrin receptor (sTfR), and transferrin saturation.

Results

Peak heart rate was significantly lower in HYP compared with NORM (p = 0.01); however, the rating of perceived exertion was similar between trials (p = 0.24). Ferritin (p = 0.02), transferrin (p = 0.03), and IL-6 (p = 0.01) significantly increased immediately post-exercise in both conditions, but returned to baseline 3 h later. Hepcidin levels significantly increased in both conditions 3 h post-exercise (p = 0.05), with no significant differences between trials. A significant treatment effect was observed between trials for sTfR (p = 0.01), but not iron and transferrin saturation.

Conclusion

Acute exercise in hypoxia did not influence post-exercise IL-6 production, hepcidin activity or iron metabolism compared with exercise at the same relative intensity in normoxia. Hence, acute exercise performed at the same relative intensity in hypoxia poses no further risk to an athlete’s iron status, as compared with exercise in normoxia.     

Intradermal skin test with diabetes specific antigens in patients with type 1 diabetes

Cell mediated immune response in vitro to a number of antigens has been reported in patients with Type 1 diabetes. The aim of the present study was to develop an in vivo intradermal (delayed type hypersensitivity) skin test using antigens known to be recognized by lymphocytes of patients with Type 1 diabetes and to compare, where possible, the in vivo response to the in vitro T cell proliferation to the same antigens. The skin test was performed in the following group of patients: 55 with recent onset Type 1 diabetes; 16 patients with Type 1 diabetes of longer duration; 10 patients with autoimmune thyroid disease and 20 patients with Latent Autoimmune Diabetes in Adults (LADA). Type 1 diabetes specific antigens for the skin test included glutamic acid decarboxilase (GAD65), insulin and beta casein, whereas diabetes non specific antigens included tetanus toxoid, diphteria, proteus, tubercolin, streptococcus, and glycerol as control. A multitest device consisting of heads delivering intradermally 10 microl of solution containing the antigens was applied to the forearms; the specific antigens were injected in one forearm whereas the non specific antigens were injected in the other forearm. Reading of the reaction, which was considered positive in the presence of a nodule of 2 mm diameter was performed 48 h after the multitest application. The in vitro T cell response to diabetes specific antigens used in the multitest was studied using conventional proliferation assays in patients with recent onset Type 1 diabetes and in age matched normal subjects. Only recent onset Type 1 diabetes patients showed an in vivo positive response to GAD65, such response being detectable in 10 patients (18%). Two patients reacted also to beta casein and insulin, all other patient groups resulted negative but 2 patients with longer duration of Type 1 diabetes. There was no apparent link between the in vivo skin test and in vitro T cell proliferation to GAD65. We conclude that in vivo cell mediated immune reaction to GAD65, insulin and beta casein can be visualized in a minority of patients with recent onset Type 1 diabetes. Further studies are required to determine specificity and whether altering the dose can improve the sensitivity of the test.     

Overexpression of FOXO1 in skeletal muscle does not alter longevity in mice

Caloric restriction (CR) is the most robust and reproducible intervention that can extend lifespan in rodents. Studies in invertebrates have led to the identification of genes that regulate lifespan, some of which encode components of the insulin or insulin-like signaling pathway, including DAF-16 (C. elegans) and dFOXO (Drosophila). Mice subjected to CR for 8 weeks showed an increase in FOXO1 mRNA and other longevity-related genes: Gadd 45α, glutamine synthase, and catalase in skeletal muscle. To investigate whether FOXO1 expression affects longevity in mammals, transgenic mice were studied that over-express FOXO1 in their skeletal muscle (FOXO1 mice), and in which muscle atrophy occurs. FOXO1 mice showed increases in Gadd 45α, and glutamine synthase proteins in skeletal muscle. In FOXO1 mice, the phosphorylation/dephosphorylation state of the p70 S6K and 4E-BP1 proteins were not altered, suggesting that translation initiation of protein synthesis might not be suppressed. The lifespan of FOXO1 mice was similar to their wild-type littermates. FOXO1 overexpression could not prevent aging-induced reduction in catalase, CuZu-SOD, and Mn-SOD mRNA in skeletal muscle. These data suggest that an increase in FOXO1 protein and its activation in skeletal muscle does not extend lifespan in mice.     

Endothelial dysfunction and microvascular complications in type 1 diabetes mellitus

We examined whether alterations in vascular endothelial function and early structural changes in atherosclerosis are associated with microvascular complications in patients with type 1 diabetes mellitus (DM). Flow-mediated dilation (FMD) of the brachial artery and carotid intima-media thickness (IMT) measurement were performed in 70 young adults (aged 19 to 35 yr), 48 with type 1 DM, and 22 normal controls. Patients with diabetes had a lower peak FMD response (7.8+/-3.9 vs. 11.1+/-1.9%, p<0.001) and increased IMT (0.51+/-0.10 vs. 0.42+/-0.07 mm, p<0.001) compared with controls. Twenty (41.7%) of the patients had microvascular complications including neuropathy, nephropathy, or retinopathy. In these complicated diabetic patients, we found a lower FMD response (6.1+/-2.5 vs. 9.9+/-3.5%, p=0.001) compared with diabetics without microvascular complications. The presence of microvascular complications was also associated with older age and longer duration of the disease. However, no differences were observed in IMT, body size, blood pressure, HbA1c, C-reactive protein, low-density lipoprotein or high-density lipoprotein cholesterol levels between complicated and non-complicated patients. Endothelial dysfunction and early structural atherosclerotic changes are common manifestations in type 1 DM, and endothelial dysfunction is thought to be an early event in the atherosclerotic process and important in the pathogenesis of microvascular complications.     

Hepatic uptake of small-latticed immune complexes does not alter mononuclear phagocyte system function.

The hepatic and splenic uptake of circulating, small-latticed immune complexes and the effect of these complexes on the hepatic mononuclear phagocyte system (MPS) were examined in mice. The small-latticed immune complexes were prepared at fifty-fold antigen excess. The clearance from circulation and uptake by the liver and spleen of two probes of MPS function, aggregated human IgG and aggregated mouse albumin, were quantified. The hepatic uptake of a dose of small-latticed complexes, containing 5 mg of antibodies, at 1 hr was comparable with the uptake of a similar dose of complexes that contained large-latticed complexes. At later time points, the hepatic uptake of the small complexes was significantly less than that of the larger complexes. The splenic uptake of the small-latticed complexes was less at all time points. Doses of the small-latticed complexes, ranging from 1 to 5 mg antibody in the complexes, produced no significant inhibition of the clearance or organ uptake of the MPS probes when administered 1 hr after the preload injections. In contrast, large-latticed complexes produced a dose-dependent delay in clearance due to a decreased hepatic uptake of the probes. These observations showed that small-latticed immune complexes were ineffectively removed by the hepatic MPS and that the presence of large quantities of small-latticed complexes in circulation did not alter MPS function.     

Prophylactic fenbendazole therapy does not affect the incidence and onset of type 1 diabetes in non-obese diabetic mice

Fenbendazole (FBZ) is a common, highly efficacious broad-spectrum anthelmintic drug used to treat and limit rodent pinworm infections. However, the effect of its prophylactic use on the immune response of rodents is largely undefined. The non-obese diabetic (NOD) mouse is a model commonly used to study type 1 diabetes (T1D). Parasitic infections will inhibit diabetes development in NOD mice; thus, in the presence of contamination, prophylactic treatment with anthelmintics must be considered to maintain experimental research. Herein, we investigated the prophylactic use of FBZ in NOD mice to determine its effect on the incidence and onset of diabetes, lymphocyte sub-populations and T cell proliferative responses. NOD mice were separated into control and treatment groups. The treatment group received a diet containing FBZ. Animals were monitored for the incidence and onset of T1D. At matched time points, diabetic and non-diabetic mice were killed and splenic lymphocytes analyzed for various cell sub-populations and mitogen-induced proliferative responses using flow cytometry. Treated and control mice were monitored >23 weeks with no detectable effects on the incidence or onset of diabetes. Moreover, no significant differences were detected in lymphocyte sub-populations and mitogen-induced CD4(+) and CD8(+) proliferative responses between control and treatment groups. These results suggest that prophylactic FBZ treatment does not significantly alter the incidence or onset of diabetes in NOD mice. The prophylactic use of FBZ, therefore, presents a viable approach for the prevention of pinworm infection in precious experimental animals with substantial scientific and economic benefits.     

Peer and family support in children and adolescents with type 1 diabetes

OBJECTIVE: To examine social support and peer and family involvement in relation to diabetes management within a developmental context. METHODS: Sixty-eight youths ages 8 to 17 diagnosed with type 1 diabetes participated. This study represents the phase 1 data from a multisystemic, home-based intervention. Data included parent and youth report of disease management and conflict, youth-reported perceptions of support, peer participation in the intervention, and HbA1c. RESULTS: Adolescents perceived greater diabetes-related peer support than did school-age children. Perceived peer and family support were not correlated with metabolic control. Peer participation in the intervention was correlated with metabolic control. CONCLUSIONS: There is a developmental shift in perceptions of peer support. Increased perceptions of peer and family support overall may not result in improved metabolic control. Social support interventions should focus on the types of support that are most highly associated with positive health outcomes.     

Health beliefs and regimen adherence in minority adolescents with type 1 diabetes

OBJECTIVE: To examine the appraisal of short- and long-term diabetes health risk and adherence, determine whether health risk predicts adherence and glycemic control in an ethnic minority sample, and determine whether perceptions of personal risk differ from risk to others. METHODS: Seventy-four youths with type 1 diabetes (ages 11-16) completed measures of risk perception and regimen adherence during their clinic visit; parents completed a measure of their children's adherence. Glycosylated hemoglobin A1c level was measured as part of the clinic visit. RESULTS: Regression analyses predicting parental report and self-reported adherence from appraisal of risk yielded nonsignificant results; perceived short-term complications to self predicted glycemic control. Appraisal of risk was higher for short- and long-term complications occurring to someone else with diabetes than to self. Greater risk for short-term complications than for long-term complications to self and other was found. CONCLUSIONS: The distinction between long-term and short-term complications and complications occurring to ones' self or someone else with diabetes was supported. Assessment of perceived risks for short-term complications is important for this age group and should be addressed in interventions to improve adherence.