Feeding Blueberry Diets in Early Life Prevent Senescence of Osteoblasts and Bone Loss in Ovariectomized Adult Female Rats

Appropriate nutrition during early development is essential for maximal bone mass accretion; however, linkage between early nutrition, childhood bone mass, peak bone mass in adulthood, and prevention of bone loss later in life has not been studied.In this report, we show that feeding a high quality diet supplemented with blueberries (BB) to pre-pubertal rats throughout development or only between postnatal day 20 (PND20) and PND34 prevented ovariectomy (OVX)-induced bone loss in adult life. This protective effect of BB is due to suppression of osteoblastic cell senescence associated with acute loss of myosin expression after OVX. Early exposure of pre-osteoblasts to serum from BB-fed rats was found to consistently increase myosin expression. This led to maintenance osteoblastic cell development and differentiation and delay of cellular entrance into senescence through regulation of the Runx2 gene. High bone turnover after OVX results in insufficient collagenous matrix support for new osteoblasts and their precursors to express myosin and other cytoskeletal elements required for osteoblast activity and differentiation.These results indicate: 1) a significant prevention of OVX-induced bone loss from adult rats can occur with only 14 days consumption of a BB-containing diet immediately prior to puberty; and 2) the molecular mechanisms underlying these effects involves increased myosin production which stimulates osteoblast differentiation and reduces mesenchymal stromal cell senescence

Analysis of RNA splicing defects in PITX2 mutants supports a gene dosage model of Axenfeld-Rieger syndrome

BACKGROUND: Axenfeld-Rieger syndrome (ARS) is associated with mutations in the PITX2 gene that encodes a homeobox transcription factor. Several intronic PITX2 mutations have been reported in Axenfeld-Rieger patients but their effects on gene expression have not been tested. METHODS: We present two new families with recurrent PITX2 intronic mutations and use PITX2c minigenes and transfected cells to address the hypothesis that intronic mutations effect RNA splicing. Three PITX2 mutations have been analyzed: a G>T mutation within the AG 3' splice site (ss) junction associated with exon 4 (IVS4-1G>T), a G>C mutation at position +5 of the 5' (ss) of exon 4 (IVS4+5G>C), and a previously reported A>G substitution at position -11 of 3'ss of exon 5 (IVS5-11A>G). RESULTS: Mutation IVS4+5G>C showed 71% retention of the intron between exons 4 and 5, and poorly expressed protein. Wild-type protein levels were proportionally expressed from correctly spliced mRNA. The G>T mutation within the exon 4 AG 3'ss junction shifted splicing exclusively to a new AG and resulted in a severely truncated, poorly expressed protein. Finally, the A>G substitution at position -11 of the 3'ss of exon 5 shifted splicing exclusively to a newly created upstream AG and resulted in generation of a protein with a truncated homeodomain. CONCLUSION: This is the first direct evidence to support aberrant RNA splicing as the mechanism underlying the disorder in some patients and suggests that the magnitude of the splicing defect may contribute to the variability of ARS phenotypes, in support of a gene dosage model of Axenfeld-Rieger syndrome

Foxf2: A Novel Locus for Anterior Segment Dysgenesis Adjacent to the Foxc1 Gene

Anterior segment dysgenesis (ASD) is characterised by an abnormal migration of neural crest cells or an aberrant differentiation of the mesenchymal cells during the formation of the eye's anterior segment. These abnormalities result in multiple tissue defects affecting the iris, cornea and drainage structures of the iridocorneal angle including the ciliary body, trabecular meshwork and Schlemm's canal. In some cases, abnormal ASD development leads to glaucoma, which is usually associated with increased intraocular pressure. Haploinsufficiency through mutation or chromosomal deletion of the human FOXC1 transcription factor gene or duplications of the 6p25 region is associated with a spectrum of ocular abnormalities including ASD. However, mapping data and phenotype analysis of human deletions suggests that an additional locus for this condition may be present in the same chromosomal region as FOXC1. DHPLC screening of ENU mutagenised mouse archival tissue revealed five novel mouse Foxf2 mutations. Re-derivation of one of these (the Foxf2W174R mouse lineage) resulted in heterozygote mice that exhibited thinning of the iris stroma, hyperplasia of the trabecular meshwork, small or absent Schlemm's canal and a reduction in the iridocorneal angle. Homozygous E18.5 mice showed absence of ciliary body projections, demonstrating a critical role for Foxf2 in the developing eye. These data provide evidence that the Foxf2 gene, separated from Foxc1 by less than 70 kb of genomic sequence (250 kb in human DNA), may explain human abnormalities in some cases of ASD where FOXC1 has been excluded genetically

Avaliação da densidade mineral óssea em pacientes com doença inflamatória intestinal Bone mineral density evaluation in inflammatory bowel disease patients

RACIONAL: Pacientes com doença inflamatória intestinal têm maior prevalência de redução da densidade mineral óssea em comparação às pessoas saudáveis. OBJETIVO: Avaliar a densidade mineral óssea em uma população de pacientes com doença inflamatória intestinal. MÉTODOS: Noventa pacientes de 20 a 50 anos de idade, do ambulatório de doença inflamatória intestinal do Serviço de Gastroenterologia do Hospital de Clínicas da Universidade Federal do Paraná, Curitiba, PR, foram selecionados para avaliação. Desses, 76 completaram todas as etapas de avaliação. A densitometria foi realizada da coluna lombar e fêmur direito em aparelho Hologyc QDR 1000/W. RESULTADOS: Os pacientes com doença inflamatória intestinal tiveram significativa redução da densidade mineral óssea, avaliada por massa óssea em números absolutos (g/cm²), em todas as regiões avaliadas, colo do fêmur, fêmur total e coluna lombar. As variáveis analisadas como, índice de atividade de doença, uso de corticóide, cirurgias prévias, índice de massa corpórea e falta de atividade física, não demonstraram correlação com a massa óssea, ou seja, não influenciaram os resultados da densidade mineral óssea no grupo estudado de doentes com doença inflamatória intestinal. CONCLUSÃO: Densidade mineral óssea reduzida foi encontrada nos pacientes com doença inflamatória intestinal do Ambulatório de Doença Inflamatória Intestinal do Serviço de Gastroenterologia do Hospital de Clínicas da Universidade Federal do Paraná, mais pronunciadamente nos pacientes com doença de Crohn, semelhante ao descrito na literatura. Nenhuma das variáveis analisadas mostrou correlação com a densidade mineral óssea.<br>BACKGROUND: Inflammatory bowel disease patients have shown greater reduction of the bone mineral density compared to healthy people. AIM: To evaluate the bone mineral density in a population of patients with inflammatory bowel disease. METHODS: Ninety patients from 20 to 50 years old, of the Inflammatory Bowel Disease Ambulatory of the Gastroenterology Service of the Clinics Hospital, Curitiba, PR, Brazil, were selected for the evaluation. From those, 76 completed all the stages of the evaluation. The densitometry was made from lumbar column and right femur with a dual-energy x-ray absortiometry (Hologyc QDR 1000/W) device. RESULTS: The inflammatory bowel disease patients had a significant reduction of the bone mineral density in all the evaluated parts, femur neck, total femur and lumbar column. The analysed variables, disease activity index, usage of corticoids, the lack of physical activities, the index body mass and previous surgeries did not have influence in the results. CONCLUSION: Reduced bone mineral density was founded in inflammatory bowel disease patients of the Clinics Hospital, mainly in the Crohn's disease patients, as described in literature. None analyzed variables had significant correlation to the bone mineral density

Climate change mitigation potential in sanitation via off-site composting of human waste

Approximately 4.5 billion people lack access to safely managed sanitation globally, and 1 billion live in slums, often relying on anaerobic waste containment in pit latrines. Providing access to safely managed sanitation may lead to reduced GHG emissions and thus simultaneously address both Sustainable Development Goals. Here we measure cumulative GHG emissions of carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) during the off-site composting of human waste to estimate scalable emission factors. We find that CH4 emission factors are one to two orders of magnitude smaller than IPCC values for other excreta collection, treatment and disposal processes. After accounting for GHG emissions throughout the sanitation cycle, including transport, urine and compost end-use, the climate change mitigation potential is 126 kg of CO2-equivalent per capita per year for slum inhabitants. If scaled to global slum populations, composting could mitigate 3.97 Tg CH4 yr−1, representing 13-44% of sanitation sector CH4 emissions