Cathepsin D-deficient Drosophila recapitulate the key features of neuronal ceroid lipofuscinoses

Neuronal ceroid lipofuscinoses (NCLs) are a group of lysosomal storage disorders characterized pathologically by neuronal accumulation of autofluorescent storage material and neurodegeneration. An ovine NCL form is caused by a recessive point mutation in the cathepsin D gene, which encodes a lysosomal aspartyl protease. This mutation results in typical NCL pathology with neurodegeneration and characteristic neuronal storage material. We have generated a Drosophila NCL model by inactivating the conserved Drosophila cathepsin D homolog. We report here that cathepsin D mutant flies exhibit the key features of NCLs. They show progressive neuronal accumulation of autofluorescent storage inclusions, which are also positive for periodic acid Schiff and luxol fast blue stains. Ultrastructurally, the storage material is composed of membrane-bound granular electron-dense material, similar to the granular osmiophilic deposits found in the human infantile and ovine congenital NCL forms. In addition, cathepsin D mutant flies show modest age-dependent neurodegeneration. Our results suggest that the metabolic pathway leading to NCL pathology is highly conserved during evolution, and that cathepsin D mutant flies can be used to study the pathogenesis of NCLs.     

Alpha-synuclein pathology does not predict extrapyramidal symptoms or dementia

Intracytoplasmic aggregation of alpha-synuclein protein as Lewy bodies in the brainstem neurons is diagnostic for Parkinson's disease, whereas if this process also occurs in the cortical neurons, it is considered pathognomonic for dementia with Lewy bodies. However, the link between alpha-synuclein incorporation into inclusions, neuronal dysfunction, and clinical symptoms needs to be clarified. Another important issue of the pathogenetic puzzle is to understand where alpha-synuclein pathology begins and how it progresses in the brain. To study this, we collected all cases from autopsy material (N = 904) that had alpha-synuclein pathology in the dorsal motor nucleus of vagus, substantia nigra, and/or basal forebrain nuclei. In this way, our study has a unique design because the selection of material is entirely based on the presence of alpha-synuclein pathology regardless of clinical phenotype. Retrospective clinical assessment then showed that only 32 (30%) of 106 alpha-synuclein-positive cases were diagnosed with a neurodegenerative disorder. The distribution or load of alpha-synuclein pathology did not permit a dependable postmortem diagnosis of extrapyramidal symptoms or cognitive impairment. Some neurologically unimpaired cases had a reasonable burden of alpha-synuclein pathology in both brainstem and cortical areas, suggesting that alpha-synuclein-positive structures are not definite markers of neuronal dysfunction.     

Effect of fluoride varnishes on color stability of esthetic restorative materials

Fluoride varnish applications were applied to two hybrid resin composite materials, Z-100 (3M Dental Products, St Paul, MN, USA) and Esthet-X (Dentsply Caulk, Milford, DE, USA), shades A1 and A2 and a glass ionomer, GC Fuji IX GP Fast (GC Corporation, Tokyo, Japan), shade A2, to evaluate color stability. Specimens (12.6-mm dia x 2.3 mm) were prepared using a polyethylene frame, light-cured and polished through a 1-microm alumina finish. After the initial baseline color measurements, the discs were suspended in Fusayama artificial saliva (FAS) solution at 37 degrees C for 48 hours. Post immersion, the specimens were divided into five groups (n=15 each). The following fluoride varnishes were applied to four groups of test specimens: Duraphat (Colgate Oral Pharmaceutical, Inc, Canton MA, USA), Cavity Shield (OMNII Oral Pharmaceuticals, West Palm Beach, FL, USA), Duraflor (Pharmascience Inc, Montreal, Canada) and Fluor Protector (Vivadent, Ivoclar North America, Amherst, NY, USA). The varnish was allowed to dry for five minutes before immersion. The control group was not coated with varnish, although the specimens were immersed in FAS. All specimens were incubated in newly prepared FAS at 37 degrees C for 24 hours, cleaned with an electric toothbrush and the process repeated using newly prepared FAS. CIE L*a*b* color measurements were recorded five times: at baseline, after 48 hours FAS immersion, after cleaning the first and second fluoride varnish applications and after the final brushing using a commercial toothpaste (Crest). A Minolta CR-300 tristimulus colorimeter with an 8-mm aperture (Ramsey, NJ, USA) was used to record color measurements with the daylight (D65) setting. Calculations were performed for using CIE parameters deltaE*, deltaL*, delta a*, delta b*. Analysis of variance (ANOVA) and post-hoc test (Fisher's PLSD) were used for statistical analysis. After immersion in saliva, the tested glass ionomer (Fuji IX) produced the most significant color changes (deltaE*=1.19 and deltaL*=-1.03), indicating the effect of the color change was due to absorption. After fluoride varnish applications, Duraphat varnish produced significant changes in all tested materials and shades, resulting in color changes with deltaE greater than (>) 1 but less than (<) 3. These color changes are considered visually perceptible, yet have been reported in dental literature as clinically acceptable. Fluoride varnishes can be used without adversely affecting the color of restorative materials.     

Microsatellite marker association at chromosome region 2p13 in Finnish patients with preeclampsia and obstetric cholestasis suggests a common risk locus

The pathophysiology of preeclampsia is incompletely understood, but the familial nature of the disease has long been recognized. Recent genome-scan studies have indicated linkage at the p23 region of chromosome 2. We have previously reported microsatellite marker association at chromosome region 2p13 in patients with obstetric cholestasis. We conducted population-based association screening with microsatellite markers to find potential preeclampsia-associated loci on chromosome region 2p13-p12 and to test whether preeclampsia and obstetric cholestasis share a single risk locus. The study was carried out among 115 unrelated control women, 133 preeclamptic women and 57 cholestatic women. Screening with microsatellite markers at the 2p13-p12 region revealed that the marker D2S286 was significantly associated with obstetric cholestasis in the overall association analysis (P=0.03), while it revealed only borderline association with preeclampsia (P=0.08). However, single allele association analysis indicated that both preeclampsia and obstetric cholestasis showed a statistically significant association with a common allele (P < 0.05), which was overrepresented in both the obstetric cholestasis (0.42) and preeclamptic (0.37) groups when compared with the control group (0.28). In conclusion, These findings suggest a possible genetic link between chromosome region 2p13-p12, preeclampsia and obstetric cholestasis. More specifically, these data suggest that there may be a common risk locus associated with both obstetric complications located in the vicinity of the 2p13-p12 association region.     

Mucosal pathology of the foregut associated with food allergy and recurrent abdominal pains in children

To determine whether children with recurrent abdominal pain (RAP) include an excess of children with food allergy (FA), this study examined a consecutive series of 84 children (43M, 41F, mean age 7.9 y, range 1.6-15 y) referred during 1 y to 2 university hospitals. In addition to a clinical examination, the patients underwent gastroduodenoscopy with three biopsy specimens, skin-prick and patch tests, and comprehensive laboratory tests for atopic allergy. Based on an open elimination-challenge test, a total of 28 (33%) subjects were diagnosed for FA. In the whole material, specific endoscopic abnormalities were found in 38 (45%) subjects: oesophagitis in 17, gastric erosions in 8, lymphonodular duodenitis in 12 and erosive duodenitis in 5. FA showed a close relationship with duodenal lesions, but no significant association with oesophagitis and gastritis. The histological findings were mild, although some alterations could be observed in up to 66 (79%) subjects, equally often in patients with and without FA. None showed villous atrophy or severe infiltration of eosinophilic or mononuclear cells. Slightly increased densities of eosinophilic cells were significantly associated with endoscopic findings, especially oesophagitis. At least one positive skin-prick test with food allergens was found in 11 subjects and a positive patch test in 21 subjects, but neither showed an association with the endoscopic or histological findings, or even with clinical FA. Conclusion: Since the children with FA showed significantly more often concomitant mucosal pathology of the foregut than those without FA, FA may be considered one of the major factors underlying RAP. The report suggests the trial of an elimination diet in cases with RAP if lymphonodular hyperplasia or duodenitis is seen on gastroduodenoscopy.     

The diabetes risk score: a practical tool to predict type 2 diabetes risk

OBJECTIVE: Interventions to prevent type 2 diabetes should be directed toward individuals at increased risk for the disease. To identify such individuals without laboratory tests, we developed the Diabetes Risk Score. RESEARCH DESIGN AND METHODS: A random population sample of 35- to 64-year-old men and women with no antidiabetic drug treatment at baseline were followed for 10 years. New cases of drug-treated type 2 diabetes were ascertained from the National Drug Registry. Multivariate logistic regression model coefficients were used to assign each variable category a score. The Diabetes Risk Score was composed as the sum of these individual scores. The validity of the score was tested in an independent population survey performed in 1992 with prospective follow-up for 5 years. RESULTS: Age, BMI, waist circumference, history of antihypertensive drug treatment and high blood glucose, physical activity, and daily consumption of fruits, berries, or vegetables were selected as categorical variables. Complete baseline risk data were found in 4435 subjects with 182 incident cases of diabetes. The Diabetes Risk Score value varied from 0 to 20. To predict drug-treated diabetes, the score value >or=9 had sensitivity of 0.78 and 0.81, specificity of 0.77 and 0.76, and positive predictive value of 0.13 and 0.05 in the 1987 and 1992 cohorts, respectively. CONCLUSIONS: The Diabetes Risk Score is a simple, fast, inexpensive, noninvasive, and reliable tool to identify individuals at high risk for type 2 diabetes.     

Lung volumes in infants who had mild to moderate bronchopulmonary dysplasia

New bronchopulmonary dysplasia (BPD) has been suggested to be a maldevelopment sequence with reduced alveolarisation of the lungs; affected infants then would be predicted to have low lung volumes. The aim of this study was to test that hypothesis by comparing the lung volumes of infants who had had mild-moderate BPD with those without BPD of similar postmenstrual age. Lung volumes of 17 infants who had mild-moderate BPD (oxygen dependent beyond 28 days, but not past term) (BPD infants) were compared to those of 17 infants without BPD (non-BPD infants). All were born at less than 33 weeks of gestation and studied at postmenstrual ages of 33 to 39 weeks. Lung volume was assessed by measurement of functional residual capacity (FRC). The BPD infants had lower lung volumes (median 19.1 ml/kg) than the non-BPD infants (median 26.5 ml/kg) (p=0.0001). The BPD compared to the non-BPD infants were of greater postnatal age (p=0.0003), born at a lower gestational age (p=0.0001) and of lighter birthweight (p=0.0001). Regression analysis, however, demonstrated that lung volume was significantly related to BPD status (p=0.005), independently of postnatal age, birthweight and gestational age. It is concluded that the lower lung volumes of the infants who had had mild-moderate BPD support the hypothesis that new BPD is associated with poor alveolarisation.     

Enhanced film-forming properties for ethyl cellulose and starch acetate using n-alkenyl succinic anhydrides as novel plasticizers

Purpose: The aim of this study was to investigate the ability of n-alkenyl succinic anhydrides (n-ASAs) to improve the film-forming characteristics of a novel coating polymer, potato starch acetate degree of substitution 2.8 (SA). n-ASAs were also applied to improve the otherwise brittle properties of ethyl cellulose (EC) aqueous dispersion (Aquacoat^®) and EC solvent-based films. Methods: The effectiveness of two n-ASAs, 2-octenyl succinic anhydride (OSA) and 2-dodecen-1-ylsuccinic anhydride were evaluated as plasticizers. Mechanical properties, both water vapor and drug permeabilities, and glass transition temperatures of the cast free films were measured. Triethyl citrate and dibutyl sebacate were used as reference plasticizers. Results: The long hydrocarbon chain of n-ASA, with its accessible carbonyl groups, enabled a strong plasticization effect on the tested polymers. Due to the excellent mechanical properties (i.e., a tough film structure with considerable flexibility) and low permeability of the plasticized films, n-ASAs, and especially OSA proved to be an ideal plasticizer particularly for EC based coatings. Also, the EC aqueous dispersion plasticized with n-ASAs resulted in a markedly enhanced coalescence of the colloidal polymer particles, even at low drying temperatures. Conclusions: In applications where a coating with high flexibility is required, n-ASAs can be used as plasticizers at moderately high concentrations (up to 60–70%, w/w) without losing the high tensile strength, excellent toughness and low permeability of EC and SA films.