Mutation analysis of NR0B2 among 1545 Danish men identifies a novel c.278G>A (p.G93D) variant with reduced functional activity

Variations of the small heterodimer partner (SHP, NR0B2) gene, an atypical nuclear receptor that inhibits transactivation by hepatocyte nuclear factor (HNF)-4alpha, are associated with obesity among Japanese. The purpose of the study was to evaluate the prevalence of SHP variants among obese Danish men. Using combined SSCP and heteroduplex analysis, we analyzed the entire coding region of SHP for variants in a cohort of 750 Danish men with early-onset obesity and genotyped a cohort of 795 nonobese control subjects using PCR-RFLP. Functional analyses of the identified coding region variants were performed in both MIN6-m9 and HepG2 cell lines. A total of five novel variants, including three missense variants (c.100C>G [p.R34G], c.278G>A [p.G93D], and c.415C>A [p.P139H]) and two silent variants (c.65C>T [p.Y22Y] and c.339G>A [p.P113P]) were identified. Moreover, the previously reported c.512G>C [p.G171A] polymorphism was identified. The 171A allele was not associated with obesity (p = 0.07). The 34G, 93D, and 139H-alleles were rare variants, which were found only among obese subjects. Among the four coding region variants, the 93D-allele showed a reduced in vitro inhibition of the HNF-4alpha transactivation of the HNF-1alpha promoter expression when expressed in MIN6-m9 and HepG2 cell lines (p<0.01). In contrast to reported findings among obese Japanese, functional variants are rare among Danish men. A functional 93D variant of SHP was identified in 1 out of 750 obese and in none of 795 nonobese control subjects. Further large-scale population studies are necessary to assess the clinical impact of this rare variant on obesity risk among European subjects.     

Histological evaluation of healing after transalveolar maxillary sinus augmentation with bioglass and autogenous bone

Objectives: The aim was to evaluate histologically the outcome of a bioglass and autogenous bone (at 1 : 1 ratio) composite implantation for transalveolar sinus augmentation. Methods: In 31 patients, during implant installation ca. 4 months after sinus augmentation, biopsies were harvested through the transalveolar osteotomy by means of a trephine bur and non‐decalcified sections through the long axis of the cylinder were produced. After a strict selection process, taking into account the presurgical residual bone height and biopsy length, 8 and 15 biopsies representing the new tissues formed inside the sinus and the transalveolar osteotomy, respectively, qualified for analysis. The tissue fractions occupied by newly formed bone (mineralized tissue+bone marrow), soft connective tissue, residual biomaterial+empty spaces, and debris inside the sinus cavity or the transalveolar osteotomy were estimated. Results: Bone and connective tissue fraction in the newly formed tissues inside the sinus cavity averaged 23.4 ± 13.2% and 54.1 ± 23.5%, respectively. Residual biomaterial, empty spaces, and debris averaged 1.9 ± 3.5%, 10.5 ± 6.3%, and 8.4 ± 14.5%, respectively. In the transalveolar osteotomy, bone and connective tissue fraction averaged 41.6 ± 14.3% and 46.1 ± 13%, respectively, while the amount of residual biomaterial, empty spaces, and debris was 2.8 ± 5%, 4.7 ± 1.9%, and 3.2 ± 2.6%, respectively. Statistically significant differences between the sinus cavity and the transalveolar osteotomy were found only for bone and empty spaces' values (P=0.02 and 0.04, respectively). Conclusion: Sinus augmentation with a bioglass and autogenous bone composite is compatible with bone formation that, in a short distance from the floor of the sinus, shows similar density as that reported previously for other commonly used bone substitutes. New bone fraction inside the transalveolar osteotomy was almost twice as much as in the sinus cavity, while the amount of residual biomaterial was much less than that inside the sinus. To cite this article :
Stavropoulos A, Sima C, Sima A, Nyengaard J, Karring T, Sculean A. Histological evaluation of healing after transalveolar maxillary sinus augmentation with bioglass and autogenous bone.
Clin. Oral Impl. Res. 23 , 2012; 125–131.
doi: 10.1111/j.1600‐0501.2011.02161.x     

Change in Proportional Protein Intake in a 10-Week Energy-Restricted Low- or High-Fat Diet,in Relation to Changes in Body Size and Metabolic Factors

ObjectiveTo investigate in a secondary analysis of a randomised trial the effects of a low-/high-fat diet and reported change from baseline in energy% from protein (prot%), in relation to changes in body size and metabolic factors.MethodsObese adults (n = 771) were randomised to a 600 kcal energy-deficient low-fat (20-25 fat%) or high-fat (40-45 fat%) diet over 10 weeks. Dietary intake data at baseline and during the intervention were available in 585 completers. We used linear regression to calculate the combined effects of randomised group and groups of prot% change (<−2 /−2 to 2/>2) on outcomes.ResultsThe low-fat group with >2 prot% increase lost 1.1 kg more weight (p = 0.03) and reduced cholesterol by 0.25 mmol/l more (p = 0.003) than the high-fat group with >2 prot% decrease. These differences were 2.5-fold and 1.8-fold greater than the differences between the low-fat and high-fat groups while not considering prot% change. The high-fat group reduced plasma triglycerides more than the low-fat group, but not compared to those in the low-fat group with >2 units prot% increase (p fat-protein interaction = 0.01).ConclusionsUnder energy restriction, participants on a low-fat diet who had increased the percentage energy intake from protein showed the greatest reduction in weight and cholesterol, and a triglyceride reduction equally large to that of participants on a high-fat diet.Key Words: Obesity, Dietary fats, Dietary proteins, Lipids, Blood glucose     

Genetic and environmental influences on plasma homocysteine: results from a Danish twin study

BACKGROUND: Increased plasma homocysteine has been linked to many clinical conditions including atherosclerosis and ischemic stroke. We assessed the genetic and environmental influences on homocysteine in adult twins and tested the influence of 3 candidate polymorphisms. METHODS: Homocysteine was analyzed in 1206 healthy twins, who were genotyped for 3 polymorphisms: MTHFR 677C>T, MTR 2756A>G, and NNMT (dbSNP: rs694539). To perform quantitative trait linkage analysis of the MTHFR locus, the genotyping was supplemented with 2 genetic markers localized on each site of the MTHFR locus. The twin data were analyzed using biometric structural equation models as well as a combined association and linkage analysis in 2 age cohorts. RESULTS: Age, sex, and MTHFR genotype have a significant impact on homocysteine concentrations, whereas the other genotypes were not associated with homocysteine concentrations. The variance in homocysteine could be solely ascribed to additive genetic and nonshared environmental factors, with an estimated additive genetic proportion of total variation at age 18-39 years of 0.63 (95% CI, 0.53-0.71) and at age 40-65 years of 0.27 (95% CI, 0.10-0.41). The impact of the MTHFR locus is estimated to explain 53% (95% CI, 0.07-0.67) of the total phenotypic variation in persons 18-39 years old and 24% (95% CI, 0.00-0.39) in persons 40-65 years old, i.e., almost all additive genetic variance. CONCLUSIONS: Homocysteine concentrations have a high heritability that decreases with age. The MTHFR gene locus is responsible for almost all the variation attributable to genetic factors, leaving very little influence of other genetic variations.     

An educational intervention to update health workers about HIV and infant feeding

Clinical guidelines are used to translate research findings into evidence‐based clinical practice but are frequently not comprehensively adopted by health workers (HWs). HIV and infant feeding guidelines were revised by the World Health Organization to align feeding advice for HIV‐exposed and unexposed infants, and these were adopted in South Africa in 2017. We describe an innovative, team‐based, mentoring programme developed to update HWs on these guidelines. The intervention was underpinned by strong theoretical frameworks and aimed to improve HWs' attitudes, knowledge, confidence, and skills about breastfeeding in the context of HIV. On‐site workshops and clinical mentoring used interactive participatory methods and a simple low‐tech approach, guided by participants' self‐reported knowledge gaps. Workshops were conducted at 24 participating clinics over three sessions, each lasting 1–2 hr. Evaluation data were collected using a self‐administered questionnaire. Of 303 participating HWs, 249/303 (82.2%) attended all workshops. Achieving high workshop attendance was challenging and “catch‐up” sessions were required to achieve good coverage. Common knowledge gaps identified included antiretroviral therapy adherence monitoring during breastfeeding and management of viral load results (173 participants), management of breast conditions (79), and advice about expressing and storing breastmilk (64). Most participants reported all their knowledge gaps were addressed and anticipated that their practice would change. We describe a feasible, sustainable approach to updating HWs on HIV and infant feeding guidelines and improving skills in breastfeeding counselling in resource‐constrained settings. This approach could be adapted to other topics and, with further evaluation, implemented at scale using existing resources.     

Regeneration of surgically removed buccal alveolar bone in dogs

The aim of the present study was to examine if the establishment of a new fibrous attachment to the root surfaces is accompanied by regeneration of surgically romoved buccal alveolar bone.
The experiments were carried out in two dogs with healthy periodontal conditions. The marginal 5–7 mm of the buccal alveolar bone between the mesial and distal line angles of each root of one maxillary second and one mandibular third premolar in both dogs was removed after elevation of a muco-peristeal flap. During this procedure attempts were made to minimize the mechanica injury to the root cementum and the supraalveolar connective tissue attachment. A notch was prepared in the root surfaces at the level of teh surgically esablished bone crest. This notch served as a landmark for measurements to be made in histological sections. The flaps were repositioned and sutured with interrupted mattress sutures. During the entire experimental period, meticulous plaque control was maintained. In each dog. the contralateral teeth served as non-treated controls. The animals were sacrificed 8 months after sugery and sections of all test-and control areas prepared for light microscopic examination.
The results of the study demonstrated that in all experimental areas, new cementum formation and fibrous reattachment occurred in the supraalveolar area between the marginal bone crest and teh junctional epithlium. In only two out of eight test areas did the alveolar bone regenerate to or close ot the normal distance from the apical portion of the junctional epithelium whereas in the remaining six areas, a long supracrestal connective tissue attachment was present.     

The Validation and Assessment of Machine Learning: A Game of Prediction from High-Dimensional Data

In applied statistics, tools from machine learning are popular for analyzing complex and high-dimensional data. However, few theoretical results are available that could guide to the appropriate machine learning tool in a new application. Initial development of an overall strategy thus often implies that multiple methods are tested and compared on the same set of data. This is particularly difficult in situations that are prone to over-fitting where the number of subjects is low compared to the number of potential predictors. The article presents a game which provides some grounds for conducting a fair model comparison. Each player selects a modeling strategy for predicting individual response from potential predictors. A strictly proper scoring rule, bootstrap cross-validation, and a set of rules are used to make the results obtained with different strategies comparable. To illustrate the ideas, the game is applied to data from the Nugenob Study where the aim is to predict the fat oxidation capacity based on conventional factors and high-dimensional metabolomics data. Three players have chosen to use support vector machines, LASSO, and random forests, respectively.