Evaluating suitability of saliva to measure steroid concentrations in grey seal pups.

Measurement of steroids in wild pinnipeds can facilitate assessment of breeding, nutritional and stress status, and is useful in understanding behavioral responses. Even in young animals, sex steroids may be important in behavioral interactions and immune modulation. Use of saliva can avoid the large fluctuations seen in some steroids in plasma, and can negate the need for venipuncture, making it a potentially useful matrix in the wildlife. However, its utility in estimating steroid levels in wild young pinnipeds has not been evaluated. Here, we investigated the suitability of saliva for steroid hormone analysis in wild grey seal pups during their suckling and post-weaning fast periods. We collected saliva (n = 38) and plasma (n = 71) samples during the breeding season on the Isle of May, Scotland, 2012. We investigated success of sample collection, ease of preparation, accuracy and precision of analysis, and, where possible, comparability of measurements (n = 27) from saliva and plasma. Plasma sampling was rapid, whereas sampling saliva took up to five times longer. Analytical performance criteria (parallelism, accuracy, and precision (intra and inter assay co-efficient of variation (% CV)) of commercial ELISA kits to measure estradiol, testosterone and cortisol in both matrices were assessed. Estradiol and cortisol assays performed well and can be used in plasma and saliva. However, we could not confidently validate testosterone for either matrix. Saliva estradiol correlated with levels in plasma. Saliva sample preparation was faster and simpler than plasma preparation because it did not require extraction. However, given the additional time taken to obtain saliva in the wild, the possibility of blood contamination from oral damage and the lower success rate in obtaining sufficient sample for analysis, we recommend that this matrix only be used as an alternative to plasma sampling measurement in pinnipeds when animals are anaesthetized, tolerate mouth swabbing, or have been trained to accept saliva sampling in captivity.

Sub-lytic C5b-9 induces functional changes in retinal pigment epithelial cells consistent with age-related macular degeneration.

PURPOSE: There is evidence for complement dysfunction in age-related macular degeneration (AMD). Complement activation leads to formation of the membrane attack complex (MAC), known to assemble on retinal pigment epithelial (RPE) cells. Therefore, the effect of sub-lytic MAC on RPE cells was examined with regard to pro-inflammatory or pro-angiogenic mediators relevant in AMD. METHODS: For sub-lytic MAC induction, RPE cells were incubated with an antiserum to complement regulatory protein CD59, followed by normal human serum (NHS) to induce 5% cell death, measured by a viability assay. MAC formation was evaluated by immunofluorescence and FACS analysis. Interleukin (IL)-6, -8, monocytic chemoattractant protein-1 (MCP-1), and vascular endothelial growth factor (VEGF) were quantified by enzyme-linked immunosorbent assay (ELISA). Intracellular MCP-1 was analysed by immunofluorescence, vitronectin by western blotting, and gelatinolytic matrix metalloproteinases (MMPs) by zymography. RESULTS: Incubation of RPE cells with the CD59 antiserum followed by 5% NHS induced sub-lytic amounts of MAC, verified by FACS and immunofluorescence. This treatment stimulated the cells to release IL-6, -8, MCP-1, and VEGF. MCP-1 staining, production of vitronectin, and gelatinolytic MMPs were also elevated in response to sub-lytic MAC. CONCLUSIONS: MAC assembly on RPE cells increases the IL-6, -8, and MCP-1 production. Therefore, sub-lytic MAC might have a significant role in generating a pro-inflammatory microenvironment, contributing to the development of AMD. Enhanced vitronectin might be a protective mechanism against MAC deposition. In addition, the increased expression of gelatinolytic MMPs and pro-angiogenic VEGF may be associated with neovascular processes and late AMD.

Effect of induction dose, lactation stage and body condition on tiletamine-zolazepam anaesthesia in adult female grey seals (Halichoerus grypus) under field conditions.

Tiletamine-zolazepam (TZ) was used at a mean (sd) dose of 1.18 (0.15) mg/kg administered intramuscularly to anaesthetise adult female grey seals (Halichoerus grypus) under field conditions at three different stages during their lactation period. A significant correlation was observed between the induction dose and time to induction (r=-0.582, P=0.011). Stage of lactation had a significant effect on condition index (CI), calculated as axial girth divided by length (P<0.001), and time to induction (P=0.009). No effect of CI on induction or recovery time was demonstrated. Respiratory rate decreased during induction and increased significantly (P<0.001) during surgical biopsy of blubber. Recovery occurred after 32.5 (11.9) minutes. Minor complications (tremor, vocalisation and mild dyspnoea) were observed in a small number of cases, none of which required treatment.

Annexinopathies.

Annexins comprise a conserved family of proteins characterised by their ability to bind and order charged phospholipids in membranes, often in response to elevated intracellular calcium. The family members (there are at least 12 in humans) have become specialised over evolutionary time and are involved in a diverse range of cellular functions both inside the cell and extracellularly Although a mutation in an annexin has never been categorically proven to be the cause of a disease state, they have been implicated in pathologies as diverse as autoimmunity, infection, heart disease, diabetes and cancer. 'Annexinopathies' were first described by Jacob H. Rand to describe the pathological sequelae in two disease states, the overexpression of annexin 2 in a patients with a haemorrhagic form of acute promyelocytic leukaemia, and the under-expression of annexin 5 on placental trophoblasts in the antiphospholipid syndrome. In this chapter we will outline some of the more recent observations in regard to these conditions, and describe the involvement of annexins in some other major causes of human morbidity.

Peak expiratory flow rate: relationship to risk variables and mortality: the Wisconsin Epidemiologic Study of diabetic retinopathy.

OBJECTIVE: To examine correlates of peak expiratory flow rate in people with type 1 diabetes and to evaluate the relationship of peak expiratory flow rate to mortality. RESEARCH DESIGN AND METHODS: A cohort study that was originally designed to determine the prevalence, incidence, and severity of diabetic retinopathy also provided the opportunity to measure peak expiratory flow rate. This was first measured at a 10-year follow-up and was evaluated in regard to risk factors for microvascular complications of diabetes. Mortality during 6 years of follow-up after the measurement was also ascertained. RESULTS: In multivariable analysis, peak expiratory flow rate was associated with sex, age, height, BMI, history of cardiovascular disease, pulse rate, duration of diabetes, glycosylated hemoglobin, and end-stage renal disease. Peak expiratory flow rate was significantly associated with survival in categorical analyses. Even after considering age, sex, renal disease, history of cardiovascular disease, respiratory symptoms, duration of diabetes, cigarette smoking, and hypertension, peak expiratory flow rate was still significantly related to survival (hazard ratio 0.61 [95% CI 0.46-0.82]). CONCLUSIONS: These data indicate that peak expiratory flow rate is associated with risk factors for other complications of diabetes. In addition, peak expiratory flow rate is a significant predictor of survival over even a relatively short period of time (6 years) in patients with younger-onset diabetes.

Annexin 2 has an essential role in actin-based macropinocytic rocketing.

Annexin 2 is a Ca(2+) binding protein that binds to and aggregates secretory vesicles at physiological Ca(2+) levels [1] and that also associates Ca(2+) independently with early endosomes [2, 3]. These properties suggest roles in both exocytosis and endocytosis, but little is known of the dynamics of Annexin 2 distribution in live cells during these processes. We have used evanescent field microscopy to image Annexin 2-GFP in live, secreting rat basophilic leukemia cells and in cells performing pinocytosis. Although we found no evidence of Annexin 2 involvement in exocytosis, we observed an enrichment of Annexin 2-GFP in actin tails propeling macropinosomes. The association of Annexin 2-GFP with rocketing macropinosomes was specific because Annexin 2-GFP was absent from the actin tails of rocketing Listeria. This finding suggests that the association of Annexin 2 with macropinocytic rockets requires native pinosomal membrane. Annexin 2 is necessary for the formation of macropinocytic rockets since overexpression of a dominant-negative Annexin 2 construct abolished the formation of these structures. The same construct did not prevent the movement of Listeria in infected cells. These results show that recruitment of Annexin 2 to nascent macropinosome membranes 16656is an essential prerequisite for actin polymerization-dependent vesicle locomotion.

Asteroid hyalosis in a population: the Beaver Dam eye study.

PURPOSE: To estimate the prevalence of asteroid hyalosis and to examine correlates of asteroid hyalosis in a population-based cohort. METHODS: The population of Beaver Dam, Wisconsin, that was 43 to 86 years of age was examined from 1988 to 1990 (n = 4926). The population is predominantly white (99.4%) northern European. Asteroid hyalosis was determined from stereoscopic fundus photographs of three standard fields. RESULTS: Fundus photographs were gradable in 4747 subjects. Asteroid hyalosis was present in 1.2% (95% confidence interval, 0.9,1.5%). In subjects in which it was present, asteroid hyalosis was bilateral in 9%. Prevalence increased significantly (P <.001) with age from 0.2% in subjects 43 to 54 years to 2.9% in subjects 75 to 86 years. After adjusting for age, men were more likely (1.8%) to have asteroid hyalosis than women (0.8%). After adjusting for age and sex, asteroid hyalosis was significantly more likely to be found in subjects with greater body mass (P =.02) and higher alcohol consumption (P =.03). There were nonsignificant trends with systolic blood pressure (P =.07), serum cholesterol (P =.09), and serum albumin (P =.09). It was not significantly associated (P >.05) with diastolic blood pressure, hypertension, diabetes, cardiovascular disease history, high-density-lipoprotein cholesterol, serum calcium, cigarette smoking, physical activity, intraocular pressure, or refractive error. These relationships were confirmed in a multivariable logistic model. CONCLUSIONS: The current study documents the infrequency of asteroid hyalosis in the population as graded from three photographic fields of the fundus. It does not support previous observations of an association of asteroid hyalosis with diabetes or refractive error. The relevance of the new associations reported (body mass, alcohol) remains to be determined.

The relation of socioeconomic factors to the incidence of early age-related maculopathy: the Beaver Dam eye study.

PURPOSE: To investigate the relationship between socioeconomic factors and the 5-year incidence of age-related maculopathy. METHODS: The Beaver Dam Eye Study, a population-based cohort study, examined 3681 adults (range, 43-86 years of age at baseline) living in Beaver Dam, Wisconsin, at baseline and 5 years later. Standardized protocols for physical examination, including administration of a questionnaire that included questions regarding income, education level, and status and type of employment, and fundus photography to determine age-related maculopathy, were performed. RESULTS: While controlling for age and sex, less education, and being in a service-related occupation compared with a white collar professional occupation, was associated (P <.05) with the incidence of early age-related maculopathy. CONCLUSION: These data show an association of education and occupation but not income with the incidence of early age-related maculopathy that appears independent of smoking or vitamin supplement use status.

Is the pulse rate useful in assessing risk of diabetic retinopathy and macular oedema? The Wisconsin Epidemiological Study of Diabetic Retinopathy.

AIM: To describe the relation between pulse rate and incident diabetic retinopathy. METHODS: Population based cohort study of people with diabetes. Resting pulse rate was measured in 30 second intervals. Diabetic retinopathy was evaluated from masked gradings of fundus photographs. RESULTS: People with higher pulse rates were more likely to have 4 year progression of retinopathy, progression to proliferative retinopathy, and incident macular oedema than those with lower pulse rates. However, these associations were attenuated after controlling for blood pressure, glycosylated haemoglobin, and other risk factors. CONCLUSION: Pulse rate may be a clinical indicator of overall risk of diabetic retinopathy, but is not independently associated with the condition.

The NEI-VFQ-25 in people with long-term type 1 diabetes mellitus: the Wisconsin Epidemiologic Study of Diabetic Retinopathy.

OBJECTIVES: To examine the association of the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) overall and specific scale scores with visual acuity, diabetic retinopathy, and other characteristics, in a cohort of persons with type 1 diabetes. DESIGN: Population-based cohort study. SETTING: An 11-county area in southern Wisconsin. PARTICIPANTS: Six hundred two persons with diabetes whose conditions were diagnosed when they were younger than 30 years and who were currently taking insulin participated in baseline, 4-year, 10-year, and 14-year follow-up examinations. MAIN OUTCOME MEASURES: An interview that consisted of the 25-item NEI-VFQ was completed. Visual acuity was measured by the Early Treatment of Diabetic Retinopathy Study (ETDRS) protocol and the presence and severity of retinopathy and macular edema were detected by masked grading of stereoscopic color fundus photographs using the modified Airlie House classification and the ETDRS retinopathy severity scheme. RESULTS: Univariate analyses revealed that the total NEI-VFQ-25 score was lower in persons who were older, had a longer duration of diabetes, higher glycosylated hemoglobin, were in renal failure, had a history of cardiovascular disease, hypertension, or amputation of a lower limb, had poorer visual acuity, more severe diabetic retinopathy, macular edema, glaucoma, cataract, abnormalities in tactile sensation or temperature sensitivity, smoked more total pack-years, led a more sedentary lifestyle, and had poor peak expiratory flow. In multivariate analyses, while controlling for the physical and mental component scores from the Medical Outcomes Survey 36-Item Short-Form Health Survey as measures of comorbidity, lower total NEI-VFQ-25 scores were independently associated with poorer visual acuity, more severe retinopathy, older age, history of loss of tactile sensation, and more total pack-years of cigarettes smoked. CONCLUSIONS: In this cross-sectional study, the 25-item NEI-VFQ seems to be strongly associated with vision, independent of severity of retinopathy and other complications associated with type 1 diabetes. It may be a useful measure of health-related quality of life as it relates to vision in epidemiological studies and clinical trials in persons with diabetes.

How many steps of progression of diabetic retinopathy are meaningful? The Wisconsin epidemiologic study of diabetic retinopathy.

OBJECTIVE: To determine whether a 1-step or more or 2-step or more progression on the Early Treatment Diabetic Retinopathy Study retinopathy severity scale over a 4-year period is meaningful in predicting the subsequent incidence of proliferative diabetic retinopathy (PDR) and clinically significant macular edema (CSME) over the following 6 years. DESIGN: Population-based study of diabetic persons with 10 years of follow-up. SETTING AND PATIENTS: Eleven-county area in southern Wisconsin. There were 1025 persons with diabetes who had fundus photographs at baseline and at 4- and 10-year follow-up examinations. MAIN OUTCOME MEASURES: Incidence of PDR or CSME between the 4- and 10-year follow-up examinations as determined by masked grading of color stereoscopic fundus photographs of 7 standard fields. RESULTS: In a univariate analysis, those with 1 or more steps of progression (n = 551) over the first 4 years of the study were significantly (P<.0001) more likely to develop PDR over the next 6 years than those with no progression (n = 474) (26% vs 4%) (relative risk, 5.85; 95% confidence interval, 4.05-8.47). Similarly, those with 2 or more (n = 364) (33%) or 3 or more (n = 231) (41%) steps of progression over the first 4 years of the study were significantly (P<.0001) more likely to develop PDR over the next 6 years than those with lesser progression (n = 661 [7%] and n = 794 [9%], respectively) (relative risk, 5.10; 95% confidence interval, 3.83-6.80; and relative risk, 4.61; 95% confidence interval, 3.57-5.99, respectively). Similar associations were apparent at every level of retinopathy, duration of diabetes, and glycosylated hemoglobin, and by type of diabetes at baseline. There were also associations between retinopathy progression and incidence of CSME. CONCLUSIONS: It seems that 1 or more or 2 or more steps of progression of retinopathy over a 4-year period strongly predict the development of PDR over the next 6 years. Therefore, using these end points of progression would result in the need for fewer subjects or shorter follow-up in some clinical trials.

Prevalence of and risk factors for dry eye syndrome.

OBJECTIVE: To examine risk factors for the prevalence of dry eye syndrome in a population-based cohort. METHODS: The prevalence of dry eye was determined by history at the second examination (1993-1995) of the Beaver Dam Eye Study cohort (N = 3722). RESULTS: The cohort was aged 48 to 91 years (mean +/- SD, 65 +/- 10 years) and 43% male. The overall prevalence of dry eye was 14.4%. Prevalence varied from 8.4% in subjects younger than 60 years to 19. 0% in those older than 80 years (P<.001 for test of trend). Age-adjusted prevalence in men was 11.4% compared with 16.7% in women (P<.001). After controlling for age and sex, the following factors were independently and significantly associated with dry eye in a logistic model: history of arthritis (odds ratio [OR], 1.91; 95% confidence interval [CI], 1.56-2.33), smoking status (past, OR, 1.22; 95% CI, 0.97-1.52; current, OR, 1.82; 95% CI, 1.36-2.46), caffeine use (OR, 0.75; 95% CI, 0.61-0.91), history of thyroid disease (OR, 1.41; 95% CI, 1.09-1.84), history of gout (OR, 1.42; 95% CI, 1.02-1.96), total to high-density lipoprotein cholesterol ratio (OR, for 1 unit, 0.93; 95% CI, 0.88-0.99), diabetes (OR, 1.38; 95% CI, 1.03-1.86), and multivitamin use (past, OR, 1.35; 95% CI, 1. 01-1.81; current, OR, 1.41; 95% CI, 1.09-1.82). Nonsignificant variables included body mass; blood pressure; white blood cell count; hematocrit; history of osteoporosis, stroke, or cardiovascular disease; history of allergies; use of antihistamines, parasympathetics, antidepressants, diuretics, antiemetics, or other drying drugs; alcohol consumption; time spent outdoors; maculopathy; central cataract; and lens surgery. CONCLUSION: The results suggest several factors, such as smoking, caffeine use, and multivitamin use, could be studied for preventive or therapeutic efficacy. Arch Ophthalmol. 2000;118:1264-1268

The epidemiology of retinal vein occlusion: the Beaver Dam Eye Study.

PURPOSE: To describe the prevalence and the 5-year incidence of retinal central and branch vein occlusion and associated risk factors. METHODS: The Beaver Dam Eye Study (n = 4,926) is a population-based study in which retinal vein occlusions were detected at baseline (1988-1990) and at a 5-year follow-up examination (1993-1995) by grading of 30 degrees color fundus photographs. RESULTS: The prevalence and 5-year incidence of retinal branch vein occlusion were each 0.6%. The prevalence of retinal central vein occlusion was 0.1%, and the 5-year incidence was 0.2%. While adjusting for age, the prevalence of branch vein occlusion was associated with hypertension (odds ratio [OR] 5.42, 95% confidence interval [CI] 2.18, 13.47), diabetes mellitus (OR 2.43, 95% CI 1.04, 5.70), pulse pressure (OR 1.24 for 10 mm Hg, 95% CI 1.03, 1.48), ocular perfusion pressure (OR 2.09 for 10 mm Hg, 95% CI 1.45, 3.01), arteriovenous nicking (OR 16.75, 95% CI 7.33, 38.24), and focal arteriolar narrowing (OR 22.86, 95% CI 8.43, 62.03). The age-adjusted incidence of retinal branch vein occlusion was associated with current smoking (OR 4.43 95%, CI 1.53, 12.84) compared with nonsmokers and to focal arteriolar narrowing (OR 5.24, 95% CI 1.97, 13.94) at baseline. While controlling for age, the incidence of branch vein occlusion was not associated with serum lipid levels, body mass index, white blood cell count, alcohol consumption, aspirin use, glaucoma, intraocular pressure, or ocular hypertension. CONCLUSIONS: Retinal vein occlusion is infrequent in the population. These data suggest a strong association between retinal branch vein occlusion and retinal arteriolar changes. Data from larger populations are needed to further assess associations between risk factors and the incidence of retinal vein occlusions.

Lens thickness and five-year cumulative incidence of cataracts: The Beaver Dam Eye Study.

PURPOSE: To evaluate whether lens thickness is related to incidence of cataracts. METHODS: Lens thickness was measured from slit-lamp photographs of the lens at the time of the prevalence evaluation in the Beaver Dam Eye Study. Incident cataract was determined by grading standard slit-lamp and retroillumination photographs of the lens at the baseline and five-year follow-up examinations. Medical history was obtained and blood pressures, height and weight were measured according to protocol. RESULTS: Lens thickness was positively associated with incident nuclear cataract and inversely associated with incident cortical cataract after accounting for age, sex, diabetes status, hypertension, heavy drinking and cigarette smoking. CONCLUSIONS: Lens thickness is related to incidence of cataracts. Mechanisms to explain these relationships require further laboratory and epidemiologic investigation.

Association of ocular disease and mortality in a diabetic population.

OBJECTIVE: To investigate the association of ocular disease with all-cause and cause-specific mortality in a diabetic population. DESIGN: Geographically defined population-based cohort study. SETTING: An 11-county area in Wisconsin. STUDY POPULATION: Participants were all younger-onset diabetic persons (diagnosed as having diabetes at <30 years of age and taking insulin) and a random sample of older-onset diabetic persons (diagnosed as having diabetes at > or =30 years of age). Diabetic retinopathy, macular edema, visual acuity, and cataract were measured using standardized protocols at baseline examinations from 1980 to 1982, in which 996 younger-onset and 1370 older-onset persons participated. Participants were followed up for 16 years. MAIN OUTCOME MEASURE: All-cause and cause-specific mortality as determined from death certificates. RESULTS: In the younger-onset group, after controlling for age and sex, retinopathy severity, macular edema, cataract, history of cataract surgery, and history of glaucoma at baseline were associated with all-cause and ischemic heart disease mortality. In the older-onset group, after controlling for age and sex, retinopathy and visual impairment were related to all-cause, ischemic heart disease, and stroke mortality. No ocular variable under study was related to cancer mortality in the older-onset group. After controlling for systemic risk factors, visual impairment was associated with all-cause and ischemic heart disease mortality in the younger-onset group. In the older-onset group, retinopathy severity was related to all-cause and stroke mortality, and visual impairment was related to all-cause, ischemic heart disease, and stroke mortality. CONCLUSIONS: Presence of more severe retinopathy or visual impairment in diabetic patients is a risk indicator for increased risk of ischemic heart disease death. Presence of these ocular conditions may identify individuals who should be under care for cardiovascular disease.

Risk factors for hospitalization in people with diabetes.

OBJECTIVE: To determine factors predicting hospitalization in people with diabetes. METHODS: Two population-based groups with diabetes were examined at baseline (1980-1982), 4 years (1984-1986), and 10 years (1990-1992). The younger-onset group (n = 777) consisted of all persons diagnosed as having diabetes before age 30 years who were taking insulin, and the older-onset group (n = 542) consisted of a sample of persons diagnosed after age 30 years. At the 10-year examination, participants were asked if they had been hospitalized in the previous year. Factors from the 4-year examination were examined for their ability to predict hospitalization at the 10-year examination. RESULTS: In the younger-onset group, 25.5% reported being hospitalized. In logistic models, glycosylated hemoglobin level (odds ratio [OR], 1.27; 95% confidence interval [CI], 1.16-1.39 for a 1% increment) and hypertension (OR, 1.60; 95% CI, 1.08-2.38) predicted hospitalization. Factors that were not significant included age, sex, systolic and diastolic blood pressures, body mass, smoking status, and alcohol consumption. In the older-onset group, 30.8% reported being hospitalized. In logistic models, only glycosylated hemoglobin level (OR, 1.16; 95% CI, 1.06-1.29 for a 1% increment) predicted hospitalization. CONCLUSIONS: Glycemic control is subject to intervention. Better control may decrease hospitalization among people with diabetes. Thus, there is considerable potential for reducing health care costs.

Retinal emboli and stroke: the Beaver Dam Eye Study.

OBJECTIVE: To describe the prevalence at baseline and the 5-year incidence of retinal emboli, associated risk factors, and the relationship of retinal emboli at baseline to stroke and ischemic heart disease mortality. METHODS: The Beaver Dam Eye Study is a large (N = 4926) population-based study of persons aged 43 to 86 years at the baseline examination. Retinal emboli were detected at baseline (1988-1990) and at a 5-year follow-up (1993-1995) by grading of stereoscopic 30 degrees color fundus photographs using standardized protocols. Cause-specific mortality was determined from death certificates. RESULTS: The prevalence of retinal arteriolar emboli was 1.3%, and the 5-year incidence was 0.9%. After adjustments were made for age and sex, the prevalence of retinal emboli was associated with higher pulse pressure, hypertension, diabetes mellitus, past and current smoking, cardiovascular disease, and the presence of retinopathy. After adjustments were made for age and sex, the incidence of retinal emboli was associated with past and current smoking and a history of coronary artery bypass surgery. After age, sex, and systemic factors were controlled for, people with retinal emboli had a significantly higher hazard of dying with a mention of stroke on the death certificate (hazard ratio = 2.61, 95% confidence interval = 1.12-6.08) than those without retinal emboli. CONCLUSIONS: Persons with retinal emboli are at an increased risk of stroke-related death. Data also show an association of smoking, hypertension, and cardiovascular disease with the prevalence of retinal emboli. CLINICAL RELEVANCE: Data from this population-based study suggest that after discovery of retinal emboli in the asymptomatic patient, referral for possible medical intervention to control hypertension, if present, may be beneficial.

Inhibition of EGF-dependent calcium influx by annexin VI is splice form-specific.

Annexin VI is a widely expressed calcium- and phospholipid-binding protein that lacks a clear physiological role. We now report that A431 cells expressing annexin VI are defective in their ability to sustain elevated levels of cytosolic Ca(2+) following stimulation with EGF. Other aspects of EGF receptor signaling, such as protein tyrosine phosphorylation and induction of c-fos are normal in these cells. However, EGF-mediated membrane hyperpolarization is attenuated and Ca(2+) entry abolished in cells expressing annexin VI. This effect of annexin VI was only observed for the larger of the two annexin VI splice forms, the smaller splice variant had no discernable effect on either cellular phenotype or growth rate. Inhibition of Ca(2+) influx was specific for the EGF-induced pathway; capacitative Ca(2+) influx initiated by emptying of intracellular stores was unaffected. These results provide the first evidence that the two splice forms of annexin VI have different functions.

Alcohol intake and the risk of coronary heart disease mortality in persons with older-onset diabetes mellitus.

CONTEXT: Despite nutrition information and guidelines that advise against depriving diabetic patients of the potential benefit of moderate alcohol intake against cardiovascular events, the association between alcohol consumption and risk of cardiovascular outcomes in diabetic individuals has not been determined. OBJECTIVE: To examine the relationship between alcohol intake and coronary heart disease (CHD) mortality in persons with older-onset diabetes. DESIGN: Population-based, prospective cohort study conducted from 1984 through 1996, with a follow-up of up to 12.3 years. SETTING AND PARTICIPANTS: A total of 983 older-onset diabetic individuals (mean [SD] age, 68.6 [11.0] years; 45.2% male; 98.5% white) were interviewed about their past-year intake of alcoholic beverages during the 1984-1986 follow-up examination of a population-based study of diabetic persons in southern Wisconsin. MAIN OUTCOME MEASURE: Time to mortality from CHD by category alcohol intake. RESULTS: Alcohol use was inversely associated with risk of CHD mortality in older-onset diabetic subjects. The CHD mortality rates for never and former drinkers were 43.9 and 38.5 per 1000 person-years, respectively, while the rates for those with alcohol intakes of less than 2, 2 to 13, and 14 or more g/d were 25.3, 20.8, and 10.0 per 1000 person-years, respectively. Compared with never drinkers and controlling for age, sex, cigarette smoking, glycosylated hemoglobin level, insulin use, plasma C-peptide level, history of angina or myocardial infarction, digoxin use, and the presence and severity of diabetic retinopathy, former drinkers had a relative risk (RR) of 0.69 (95% confidence interval [CI], 0.43-1.12); for those who drank less than 2 g/d (less frequent than 1 drink a week), the RR was 0.54 (95% CI, 0.33-0.90); for 2 to 13 g/d, it was 0.44 (95% CI, 0.23-0.84); and for 14 or more g/d (about 1 drink or more a day), it was 0.21 (95% CI, 0.09-0.48). Further adjustments for blood pressure, body mass index, education, physical activity, diabetes duration, hypertension history, overt nephropathy, peripheral neuropathy, lipid measures, or intake of medications such as aspirin and antihypertensive agents did not change the associations observed. CONCLUSION: Our results suggest an overall beneficial effect of alcohol consumption in decreasing the risk of death due to CHD in people with older-onset diabetes.

The 10-year incidence of renal insufficiency in people with type 1 diabetes.

OBJECTIVE: To describe the 10-year decrease in estimated creatinine clearance and the incidence of renal insufficiency and end-stage renal disease in a cohort of people with type 1 diabetes. RESEARCH DESIGN AND METHODS: A population-based cohort of individuals with younger-onset diabetes (diagnosed at < 30 years old and taking insulin) participated in an examination during 1984-1986 (n = 891), a 6-year follow-up examination during 1990-1992 (n = 765), and a 10-year follow-up examination during 1995-1996 (n = 634). Serum creatinine and risk factors were measured during standardized protocols at each examination. Estimated adjusted creatinine clearance was computed by a modification of the Cockroft-Gault formula. A clinically meaningful change was defined as a decrease in the estimated annual creatinine clearance of > or = 3 ml.min-1.1.73 m-2.year-1. Renal insufficiency was defined by the development of a serum creatinine of 2.0 mg/dl or greater after the 1984-1986 examination. RESULTS: The 10-year estimated incidence of an annual decrease in the creatinine clearance of > or = 3 ml.min-1.1.73 m-2 for the cohort was 52.5%, and the cumulative 10-year incidence of renal insufficiency and end-stage renal failure was 14.4%. In univariate analyses, incidence of a decrease in the estimated creatinine clearance of > or = 3 ml.min-1.1.73 m-2.year-1 and the incidence of renal insufficiency were both related to higher glycosylated hemoglobin; higher diastolic blood pressure; the presence of microalbuminuria and gross proteinuria; more severe retinopathy; and a history of loss of tactile sensation or temperature sensitivity at baseline. In logistic regression analysis, after adjusting for the presence of microalbuminuria and gross proteinuria at baseline, higher glycosylated hemoglobin and higher diastolic blood pressure were associated with decreasing estimated creatinine clearance. In logistic regression analyses, after adjusting for the presence of microalbuminuria and gross proteinuria at baseline, the incidence of renal insufficiency was independently associated with age, glycosylated hemoglobin, hypertension, and serum HDL cholesterol. CONCLUSIONS: These data suggest that a public health approach aimed at controlling glycemia, blood pressure, and serum lipids might result in reducing the rate of decline in renal function and development of renal insufficiency in people with type 1 diabetes.