A steep ramp test is valid for estimating maximal power and oxygen uptake during a standard ramp test in type 2 diabetes.

A short maximal steep ramp test (SRT, 25 W/10 s) has been proposed to guide exercise interventions in type 2 diabetes, but requires validation. This study aims to (a) determine the relationship between Wmax and V˙O2peak reached during SRT and the standard ramp test (RT); (b) obtain test-retest reliability; and (c) document electrocardiogram (ECG) abnormalities during SRT. Type 2 diabetes patients (35 men, 26 women) performed a cycle ergometer-based RT (women 1.2; men 1.8 W/6 s) and SRT on separate days. A random subgroup (n = 42) repeated the SRT. ECG, heart rate, and V˙O2 were monitored. Wmax during RT: 193 ± 63 (men) and 106 ± 33 W (women). Wmax during SRT: 193 ± 63 (men) and 188 ± 55 W (women). The relationship between RT and SRT was described by men RT V˙O2peak (mL/min) = 152 + 7.67 × Wmax SRT1 (r: 0.859); women RT V ˙ O 2 p e a k (mL/min) = 603 + 4.75 × Wmax SRT1 (r: 0.771); intraclass correlation coefficients between first (SRT1) and second SRT Wmax (SRT2) were men 0.951 [95% confidence interval (CI) 0.899-0.977] and women 0.908 (95% CI 0.727-0.971). No adverse events were noted during any of the exercise tests. This validation study indicates that the SRT is a low-risk, accurate, and reliable test to estimate maximal aerobic capacity during the RT to design exercise interventions in type 2 diabetes patients.

Consanguinity and founder effect for Gaucher disease mutation G377S in a population from Tabuleiro do Norte, Northeastern Brazil.

Gaucher's disease (GD) is caused by a ß-glucocerebrosidase deficiency, leading to the accumulation of glucocerebroside in the reticuloendothelial system. The prevalence of GD in Tabuleiro do Norte (TN) (1:4000) is the highest in Brazil. The purpose of this study was to present evidence of consanguinity and founder effect for the G377S mutation (c.1246G>A) among GD patients in TN based on enzyme, molecular and genealogical studies. Between March 2009 and December 2010, 131 subjects at risk for GD (GC in dried blood ≤2.19 nmol/h/ml) and 5 confirmed GD patients from the same community were submitted for molecular analysis to characterize the genetic profile of the population. Based on the enzymatic and molecular analysis, the subjects were classified into three categories: affected (n = 5), carrier (n = 20) and non-carrier (n = 111). All carriers were (G377S/wt). Affected subjects were homozygous (G377S/G377S). The identification of a single mutation in carriers and homozygotes from different generations, the history of the community and the genealogy study suggest that the high prevalence of GD in this population may be due to a combination of consanguinity and founder effect for the G377S mutation.

Do cervical spine X-rays for trauma have clinically significant incidental findings?

OBJECTIVES: About 800,000 cervical X-rays for trauma are taken every year in the USA. Those X-rays are reviewed by orthopedic specialists in the emergency room (ER) for traumatic findings. The quantity of incidental atraumatic findings in this very prevalent examination is unknown. We sought to determine the incidence of those findings. METHODS: We retrospectively reviewed 521 consecutive cervical X-rays of patients with a whiplash injury that visited our ER from February to July 2010. X-rays that were technically insufficient were excluded. This left 356 X-rays that met the inclusion criteria, which were analyzed for incidental findings. The examinations were reviewed by five staff radiologists for incidental findings. The findings were reviewed and classified. RESULTS: We identified incidental X-ray findings in 22 of the 356 patients (6.2 %) who underwent X-ray of the cervical spine during their visit to the ER. Stenosis with disk narrowing was the most common finding (2.8 %), followed by congenital anomaly of the cervical spine (2.2 %). Other findings were enlarged sella turcica (0.6 %), carotid atherosclerosis (0.3 %), and calcification of the stylomastoid ligament (0.3 %). Older age was found to be a risk factor for an incidental finding (p < 0.0001). CONCLUSION: Incidental findings in the cervical spine were associated with older age. Awareness of the prevalence of incidental findings is important in order to ensure that they are detected and managed appropriately.

Multicenter analysis of glucocerebrosidase mutations in Parkinson's disease.

BACKGROUND: Recent studies indicate an increased frequency of mutations in the gene encoding glucocerebrosidase (GBA), a deficiency of which causes Gaucher's disease, among patients with Parkinson's disease. We aimed to ascertain the frequency of GBA mutations in an ethnically diverse group of patients with Parkinson's disease. METHODS: Sixteen centers participated in our international, collaborative study: five from the Americas, six from Europe, two from Israel, and three from Asia. Each center genotyped a standard DNA panel to permit comparison of the genotyping results across centers. Genotypes and phenotypic data from a total of 5691 patients with Parkinson's disease (780 Ashkenazi Jews) and 4898 controls (387 Ashkenazi Jews) were analyzed, with multivariate logistic-regression models and the Mantel-Haenszel procedure used to estimate odds ratios across centers. RESULTS: All 16 centers could detect two GBA mutations, L444P and N370S. Among Ashkenazi Jewish subjects, either mutation was found in 15% of patients and 3% of controls, and among non-Ashkenazi Jewish subjects, either mutation was found in 3% of patients and less than 1% of controls. GBA was fully sequenced for 1883 non-Ashkenazi Jewish patients, and mutations were identified in 7%, showing that limited mutation screening can miss half the mutant alleles. The odds ratio for any GBA mutation in patients versus controls was 5.43 across centers. As compared with patients who did not carry a GBA mutation, those with a GBA mutation presented earlier with the disease, were more likely to have affected relatives, and were more likely to have atypical clinical manifestations. CONCLUSIONS: Data collected from 16 centers demonstrate that there is a strong association between GBA mutations and Parkinson's disease.

Glomerular filtration rate estimation and mortality in an elderly population.

BACKGROUND: Few studies have addressed the link between minor renal dysfunction and mortality in the elderly. AIM: To compare three equations for estimated GFR (eGFR) in assessing renal dysfunction and predicting mortality in an elderly population. DESIGN: Longitudinal observational study. METHODS: We studied 441 people from the Jerusalem Seventy Year Olds Longitudinal Study who had measurements of serum creatinine, all of whom were aged 70 years at study initiation and were living in the community. GFR was estimated based on serum creatinine and using the Cockcroft-Gault (CG), the abbreviated Modification of Diet in Renal Disease (MDRD) and the Mayo Clinic equations. Twelve-year mortality was the main outcome measure. RESULTS: The prevalence of reduced eGFR was 51% using the CG, 34% using MDRD and 16% using the Mayo Clinic equation. eGFR dichotomized by the definition of CKD significantly predicted mortality only with the Mayo Clinic equation (hazard ratio 1.56, 95%CI 1.01-2.39). When eGFR was divided into quartiles and the lowest compared to the highest, all equations predicted mortality. Hazard ratios (95%CI) were 5.48 (1.27-23.65), 7.47 (2.74-20.3), and 7.375 (3.13-17.36), for CG, MDRD, and Mayo Clinic, respectively. DISCUSSION: Reduced eGFR was prevalent in this study group, and associated with mortality. This association was strongest using the Mayo Clinic equation.

Detection of 12 new mutations in Gaucher disease Brazilian patients.

Gaucher disease is the most frequent lysosome storage disease and presents an autosomal recessive mode of inheritance. It is caused by mutations at the GBA gene leading to deficient activity of the glucocerebrosidase enzyme. This report describes 12 new mutations [c.38A>G (K-27R), c.220G>A (G35S), c.448G>A (E111K), IVS4+1G>A, c.746C>T (A210V), c.776A>G (Y220C), c.793delC (Q226_fs4X), c.1102C>T (R329C), c.1300C>T (R395C), c.1309G>A (V398I), c.1324-1326delATT (delI403) and c.1583T>C (I489T)] and 4 novel silent alterations [c.342C>T (F75), c.528C>T (D137), c.1011C>T (D298) and c.1092G>A (G325)] detected among 40 unrelated Brazilian type 1 Gaucher disease patients by a combination of RFLP, dHPLC and DNA sequencing procedures. The R329C mutation, previously described in a Parkinson's disease patient (A. Lwin, E. Orvisky, O. Goker-Alpan, M.E. LaMarca, E. Sidransky. Glucocerebrosidase mutations in subjects with Parkinsonism. Mol. Genet. Metab. 81 (2004) 70-73), is described here for the first time in a Gaucher disease patient. Several genotype-phenotype correlations could be established, contributing significantly to the panel of reported mutations and conferring predictive value to their detection.

Diagnosis and molecular characterization of non-classic forms of Tay-Sachs disease in Brazil.

Molecular analysis of five Brazilian families, including eight patients presenting with nonclassic Tay-Sachs disease, was performed to identify frequent causative mutations and their correlation with clinical course. Three patients were affected by the B1 subacute variant and were shown to carry the R178H mutation (the DN allele), which is also common among Portuguese patients. Two of them were compound heterozygotes, whereas the third presented with the mutation in both alleles. Since Brazil was a Portuguese colony for over two centuries, common ancestry might be the probable explanation. The fourth patient presented with a juvenile phenotype and carries the R499H mutation, which has been reported only once worldwide and is associated with residual enzyme activity, responsible for a slower clinical course. The fifth family, of an Ashkenazi Jewish background, showed an extensive intrafamilial clinical variability among three affected sibs presenting with muscle atrophy, ataxia, and psychiatric symptoms. They were first diagnosed as having atypical spinal muscular atrophy and, subsequently, spinocerebellar ataxia, but, recently, the diagnosis of late-onset Tay-Sachs disease was confirmed based on reduced plasma hexosaminidase A activity and the G269S/InsTATC1278 genotype. It is therefore highly recommended to test patients with a similar clinical history for Tay-Sachs disease. In the same family, one first cousin committed suicide at the age of 24 years, presenting with a clinical phenotype that suggested an undiagnosed case and highlighting the effect of the intrafamilial clinical variability in delaying a prompt diagnosis. It is now recognized that his parents are, in fact, a carrier couple. Additionally, another relative had been previously identified as a heterozygote in a Tay-Sachs disease screening program, but the information was not shared among the family. Since this information might anticipate diagnosis and genetic counseling, it is advisable that heterozygote screening programs encourage families to share genetic information.

High frequency of mutation G377S in Brazilian type 3 Gaucher disease patients.

Gaucher disease (GD), the most prevalent lysosome storage disorder, presents an autosomal recessive mode of inheritance. It is a paradigm for therapeutic intervention in medical genetics due to the existence of effective enzyme replacement therapy. We report here the analysis of GD in 262 unrelated Brazilian patients, carried out in order to establish the frequency of the most common mutations and to provide prognostic information based on genotype-phenotype correlations. Among 247 type 1 GD patients, mutation N370S was detected in 47% of all the alleles, but N370S/N370S homozygosity was found in only 10% of the patients, a much lower frequency than expected, suggesting that most individuals presenting this genotype may not receive medical attention. Recombinant alleles were detected at a high frequency: 44% of the chromosomes bearing mutation L444P had other mutations derived from the pseudogene sequence, present in 25% of patients. Three neuronopathic type 2 patients were homozygous for L444P, all presenting additional mutations (E326K or recombinant alleles) that probably lead to the more severe phenotypes. Six children, classified as type 1 GD patients, had a L444P/L444P genotype, showing that neuronopathic symptoms may only manifest later in life. This would indicate the need for a higher treatment dose during enzyme replacement therapy. Finally, mutation G377S was present in 4 homozygous type 1 patients and also in compound heterozygosity in 5 (42%) type 3 patients. These findings indicate that G377S cannot be unambiguously classified as mild and suggest an allele-dose effect for this mutation.

High frequency of mutation G377S in Brazilian type 3 Gaucher disease patients

Gaucher disease (GD), the most prevalent lysosome storage disorder, presents an autosomal recessive mode of inheritance. It is a paradigm for therapeutic intervention in medical genetics due to the existence of effective enzyme replacement therapy. We report here the analysis of GD in 262 unrelated Brazilian patients, carried out in order to establish the frequency of the most common mutations and to provide prognostic information based on genotype-phenotype correlations. Among 247 type 1 GD patients, mutation N370S was detected in 47 percent of all the alleles, but N370S/N370S homozygosity was found in only 10 percent of the patients, a much lower frequency than expected, suggesting that most individuals presenting this genotype may not receive medical attention. Recombinant alleles were detected at a high frequency: 44 percent of the chromosomes bearing mutation L444P had other mutations derived from the pseudogene sequence, present in 25 percent of patients. Three neuronopathic type 2 patients were homozygous for L444P, all presenting additional mutations (E326K or recombinant alleles) that probably lead to the more severe phenotypes. Six children, classified as type 1 GD patients, had a L444P/L444P genotype, showing that neuronopathic symptoms may only manifest later in life. This would indicate the need for a higher treatment dose during enzyme replacement therapy. Finally, mutation G377S was present in 4 homozygous type 1 patients and also in compound heterozygosity in 5 (42 percent) type 3 patients. These findings indicate that G377S cannot be unambiguously classified as mild and suggest an allele-dose effect for this mutation.

Moderate renal insufficiency at 70 years predicts mortality.

BACKGROUND: Glomerular filtration rate (GFR) diminishes with age. Kidney function in the elderly is often assessed by serum creatinine alone, although it is insensitive in this age group. Formulae for predicting GFR are not widely used. AIM: To study the effect of low predicted GFR on mortality. DESIGN: Longitudinal cohort study. SETTING: The community-based Jerusalem Seventy Year Olds Longitudinal Study. METHODS: We studied 445 subjects, all aged 70 years, using questionnaires, a medical examination with history-taking, and standard laboratory tests. Moderate renal insufficiency was defined as a predicted GFR of <60 ml/min, based on the Cockcroft-Gault (CG) and the Modification of Diet in Renal Disease (MDRD) equations. RESULTS: Predicted GFR was normally distributed, with a mean +/- SD of 62.4 +/- 15.27 ml/min. Predicted GFR was <60 ml/min in 221 (46%), most of whom had normal serum creatinine. Twelve-year mortality was 38.7% in these 221 vs. 27% in the other 204. The survival advantage was already evident after 3 years. Under Cox proportional hazard analysis using numerous common risk factors as independent variables, lower predicted GFR had a significant mortality risk (hazard ratio 2.108, 95%CI 1.43-3.12, p = 0.0002). DISCUSSION: In community-dwelling elderly people, moderate renal insufficiency as assessed using the CG equation is a strong and independent predictor of mortality. Most of these at-risk patients have 'normal' serum creatinine.

Tay-Sachs disease in Brazilian patients: prevalence of the IVS7+1g>c mutation.

Seven Brazilian Tay-Sachs disease cases were screened for the most frequent causative mutations. They all presented at least one copy of the IVS7+1g>c mutation. Three patients were homozygotes, three were compound heterozygotes, and in one case only the mother was tested and shown to carry the IVS7+1g>c mutation. In the second allele the compound heterozygotes presented: R178H (the DN allele), InsTATC1278 and an unidentified mutation. The IVS7+1g>c mutation has already been described in three Portuguese patients. In this study, all families were unaware of any Portuguese ancestry. Since Brazil was a Portuguese colony, the mutation most probably came from ancient common ancestry. The initial molecular analysis of Tay-Sachs disease patients in Brazil indicated a prevalence of the IVS7+1g>c mutation, possibly as a result of genetic drift.

Ochratoxin A in domestic and imported beers in Belgium: occurrence and exposure assessment.

Determination of ochratoxin A (OTA) concentration was performed in commercial beer in Belgium using immunoaffinity column (OchraTest) clean-up and liquid chromatography. The procedure was validated and fulfilled the European Committee for Standardization's criteria. It offered a detection limit of 3 ng l(-1) and a quantification limit of 10 ng l(-1). Recovery experiments carried out with the spiked samples in the range 50-200 ng OTA l(-1) showed an overall average recovery rate of 97% (RSD = 2.8%). The validated method was applied to the analysis of 62 Belgian beers and 20 commercial beers imported from Denmark, France, Germany, Ireland, Mexico, The Netherlands and Scotland. None of these beers exceeded the previously suggested EU limit of 200 ng l(-1). However, OTA was detected in 60 Belgian beers and in all imported beers. The average levels of contamination were 33 ng l(-1) (RSD = 112%) and 32 ng l(-1) (RSD = 81%), respectively. The highest level found was 185 ng l(-1). On the basis of the established tolerable daily intake (TDI) of 5 ng kg(-1) body weight, accepted by the scientific committee on food of the EU, this study indicates that beer consumption in Belgium is not likely to contribute to more than a few per cent of the TDI based on the average consumption. This study also shows variability of the OTA contamination in beer with time. Thus, there is a potential risk of having highly contaminated batches from time to time. We therefore recommend to control further the OTA contamination in brewery products and to take precautionary measures during harvest, transport and storage of the raw materials to maintain the OTA intake at the lowest achievable level.

Structure characterization of flavonoid O-diglycosides by positive and negative nano-electrospray ionization ion trap mass spectrometry.

Isomeric flavonoid O-diglycosides were analyzed by positive and negative nano-electrospray ionization (ESI) ion trap mass spectrometry (ITMS) in order to evaluate whether the two most common interglycosidic linkage types, i.e. 1 --> 2 and 1 --> 6, found for glycosides containing a rhamnosylglucose glycan part can be differentiated. In the positive ion mode the degree of internal glucose residue loss was found to be strongly dependent on the aglycone type and was very pronounced for aglycones of the flavanone type. The relative abundance of the Y-type ions formed by fragmentation at glycosidic bonds only allows one to infer the interglycosidic linkage types in the case of flavone O-diglycosides. In contrast, the negative ion mode makes a clear differentiation between a rutinoside (1 --> 6) and a neohesperidoside (1 --> 2) glycan residue possible for all aglycone types. The neohesperidose-containing compounds could be characterized by additional product ions. When the compounds were dissolved in pure methanol a molecular radical ion was found to be the base peak in nano-ESI.

The frequency of Tay-Sachs disease causing mutations in the Brazilian Jewish population justifies a carrier screening program.

CONTEXT: Tay-Sachs disease is an autosomal recessive disease characterized by progressive neurologic degeneration, fatal in early childhood. In the Ashkenazi Jewish population the disease incidence is about 1 in every 3,500 newborns and the carrier frequency is 1 in every 29 individuals. Carrier screening programs for Tay-Sachs disease have reduced disease incidence by 90% in high-risk populations in several countries. The Brazilian Jewish population is estimated at 90,000 individuals. Currently, there is no screening program for Tay-Sachs disease in this population. OBJECTIVE: To evaluate the importance of a Tay-Sachs disease carrier screening program in the Brazilian Jewish population by determining the frequency of heterozygotes and the acceptance of the program by the community. SETTING: Laboratory of Molecular Genetics--Institute of Biosciences--Universidade de São Paulo. PARTICIPANTS: 581 senior students from selected Jewish high schools. PROCEDURE: Molecular analysis of Tay-Sachs disease causing mutations by PCR amplification of genomic DNA, followed by restriction enzyme digestion. RESULTS: Among 581 students that attended educational classes, 404 (70%) elected to be tested for Tay-Sachs disease mutations. Of these, approximately 65% were of Ashkenazi Jewish origin. Eight carriers were detected corresponding to a carrier frequency of 1 in every 33 individuals in the Ashkenazi Jewish fraction of the sample. CONCLUSION: The frequency of Tay-Sachs disease carriers among the Ashkenazi Jewish population of Brazil is similar to that of other countries where carrier screening programs have led to a significant decrease in disease incidence. Therefore, it is justifiable to implement a Tay-Sachs disease carrier screening program for the Brazilian Jewish population.

The effects of physical activity on mortality in the Jerusalem 70-Year-Olds Longitudinal Study.

OBJECTIVE: To study the association between physical activity and mortality in older men and women. DESIGN: A community-based cohort study: the Jerusalem 70-Year-Olds Longitudinal Study. PARTICIPANTS: A systematically selected and representative sample of all residents of the western part of Jerusalem born in 1920-1921: 456 subjects, 25% of the total population. MEASUREMENTS: An extensive social and medical profile was developed by extensive interview and physical and ancillary examination. Medical diagnoses were established and subjects reported their level of regular physical activity. RESULTS: Unadjusted mortality at 6-year follow-up was significantly greater for subjects reporting no regular exercise than for those walking as little as 4 hours weekly (23.4% vs 9.9%, odds ratio (OR) = 2.77; 95% confidence interval (CI), 1.64-4.69). The significance of these benefits was demonstrated for males as well as for females (30.28% vs 12.14%, P < .001, 16.19% vs 6.86%, P = .036, respectively). Logistic regression analysis demonstrated the survival advantage to be independent of gender, smoking, subjective economic hardship, or preexisting medical conditions (hypertension; diabetes; coronary artery, cerebrovascular, renal, and respiratory diseases; anemia; and malignancy). Increased regularity of activity correlated with declining mortality. The odds ratios for mortality compared to the sedentary group were 0.73 (CI, 0.33-1.62) for those doing sports activity at least twice weekly, 0.41 (CI, 0.19-0.91) for those walking at least 4 hours weekly, 0.14 (CI, 0.04-0.50) for those exercising daily, and 0.40 (CI, 0.22-0.72) for all levels of physical activity combined. CONCLUSIONS: These results suggest that regular physical activity confers increased survival in the aged. It is proposed that older people be encouraged to engage in regular, moderate physical activity.

Independence in instrumental activities of daily living and its effect on mortality.

Factors relating to six-year mortality in a representative sample of seventy-year-old Jerusalem residents (N = 605) were investigated using logistic regression techniques. Around 16.3% of the study population died during the six-year post-interview period. Bivariate analysis found elevated mortality related to being male, having more than one IADL dysfunction, more than two ADL dysfunctions, financial problems, no social support in times of emergencies, bad self-rated health status, cognitive impairment, confinement to bed during the fortnight prior to interview, and lack of regular exercise. Logistic regression controlling for gender, various clinical diagnoses, financial state, social support and smoking status showed IADL (ROR = 4.57, 95% CI 1.51, 13.90), cognitive impairment (ROR = 3.99, 95% CI 1.85, 8.59) and having been bed-sick a week or more during the preceding fortnight (ROR = 6.60, 95% CI 1.00, 43.86) to be independent predictors of mortality. All persons who had a cognitive problem and were dysfunctional in more than two IADL categories, and 93.8% of persons who had been bed-sick and had more than one IADL dysfunction died during the study period. Combined measures of these three easily obtainable variables could prove a cheap and efficient method of identifying at-risk elderly persons in order to provide them with specific programs aimed at decreasing functional decline, and hence mortality.

The siesta in the elderly: risk factor for mortality?

BACKGROUND: During the siesta, blood pressure declines like it does during night sleep. Because cardiovascular and cerebrovascular events cluster during the morning hours, when hemodynamic changes from nocturnal baseline are maximal, we hypothesized that an additional sleep period during the day (the siesta) may increase cardiovascular and cerebrovascular events, and thus mortality. METHODS: A prospective population-based cohort study of 455 70-year-old residents of Jerusalem, Israel, using self-reported siesta at baseline and 6 1/2 years of total mortality data. RESULTS: The prevalence of the practice of the siesta was 60.7%. It was more prevalent among men than women (68% vs 51%, P<.001) and in survivors of previous myocardial infarction than in those without previous myocardial infarction (78% vs 58%, P = .009). After 6 1/2 years of follow-up (1990-1996), 75 subjects died. For those who practiced the siesta, total mortality was 20% vs 11% for those who did not (P = .01; risk odds ratio, 2.0; 95% confidence interval, 1.1-3.4). In a multiple logistic regression model that included several lifestyle descriptors, risk factors, and diseases, the siesta remained predictive of mortality (P = .03; risk odds ratio, 2.1; 95% confidence interval, 1.1-3.9).