Analysis of Nkx3.1:Cre-driven Erk5 deletion reveals a profound spinal deformity which is linked to increased osteoclast activity

Extracellular signal-regulated protein kinase 5 (ERK5) has been implicated during development and carcinogenesis. Nkx3.1-mediated Cre expression is a useful strategy to genetically manipulate the mouse prostate. While grossly normal at birth, we observed an unexpected phenotype of spinal protrusion in Nkx3.1:Cre;Erk5fl/fl (Erk5fl/fl) mice by ~6–8 weeks of age. X-ray, histological and micro CT (µCT) analyses showed that 100% of male and female Erk5fl/fl mice had a severely deformed curved thoracic spine, with an associated loss of trabecular bone volume. Although sex-specific differences were observed, histomorphometry measurements revealed that both bone resorption and bone formation parameters were increased in male Erk5fl/fl mice compared to wild type (WT) littermates. Osteopenia occurs where the rate of bone resorption exceeds that of bone formation, so we investigated the role of the osteoclast compartment. We found that treatment of RANKL-stimulated primary bone marrow-derived macrophage (BMDM) cultures with small molecule ERK5 pathway inhibitors increased osteoclast numbers. Furthermore, osteoclast numbers and expression of osteoclast marker genes were increased in parallel with reduced Erk5 expression in cultures generated from Erk5fl/fl mice compared to WT mice. Collectively, these results reveal a novel role for Erk5 during bone maturation and homeostasis in vivo

OPA1 deficiency accelerates hippocampal synaptic remodelling and age-related deficits in learning and memory

A healthy mitochondrial network is essential for the maintenance of neuronal synaptic integrity. Mitochondrial and metabolic dysfunction contributes to the pathogenesis of many neurodegenerative diseases including dementia. OPA1 is the master regulator of mitochondrial fusion and fission and is likely to play an important role during neurodegenerative events. To explore this, we quantified hippocampal dendritic and synaptic integrity and the learning and memory performance of aged Opa1 haploinsufficient mice carrying the Opa1Q285X mutation (B6; C3-Opa1Q285STOP; Opa1+/−). We demonstrate that heterozygous loss of Opa1 results in premature age-related loss of spines in hippocampal pyramidal CA1 neurons and a reduction in synaptic density in the hippocampus. This loss is associated with subtle memory deficits in both spatial novelty and object recognition. We hypothesize that metabolic failure to maintain normal neuronal activity at the level of a single spine leads to premature age-related memory deficits. These results highlight the importance of mitochondrial homeostasis for maintenance of neuronal function during ageing

Examining the generalizability of research findings from archival data

This initiative examined systematically the extent to which a large set of archival research findings generalizes across contexts. We repeated the key analyses for 29 original strategic management effects in the same context (direct reproduction) as well as in 52 novel time periods and geographies; 45% of the reproductions returned results matching the original reports together with 55% of tests in different spans of years and 40% of tests in novel geographies. Some original findings were associated with multiple new tests. Reproducibility was the best predictor of generalizability—for the findings that proved directly reproducible, 84% emerged in other available time periods and 57% emerged in other geographies. Overall, only limited empirical evidence emerged for context sensitivity. In a forecasting survey, independent scientists were able to anticipate which effects would find support in tests in new samples

Increasing Community Resiliency through the Development of Microgrids in Cambridge, MA

In an era of increasing energy demand, climate change concerns, and a growing need for renewable energy sources, the use of local, self-contained energy systems, or microgrids, has emerged as a critical solution. The goal of this project was to identify the barriers keeping the communities from implementing microgrids. We accomplished this goal by working with our sponsor, Clean Energy Solutions Inc,. to investigate literature and conduct interviews with key stakeholders for a proposed microgrid site in Cambridge, MA. The main recommendations were to better inform the community on what microgrids are, overcome language barriers, and work as a team with the community, utilities, and policymakers to make the changes needed towards more renewable and resilient energy

Enzymatic assay of d-glucuronate using uronate dehydrogenase

d-Glucuronate is a key metabolite in the process of detoxification of xenobiotics and in a recently constructed synthetic pathway to produce d-glucaric acid, a “top value-added chemical” from biomass. A simple and specific assay of d-glucuronate would be useful for studying these processes, but existing assays are either time-consuming or nonspecific. Using uronate dehydrogenase cloned from Agrobacterium tumefaciens, we developed an assay for d-glucuronate with a detection limit of 5 μM. This method was shown to be more suitable for a system with many interfering compounds than previous methods and was also applied to assays for myo-inositol oxygenase activity.United States. Office of Naval Research. Young Investigator Program (N000140510656)National Science Foundation (U.S.) (EEC-0540879)Merck & Co., Inc. (Undergraduate Research Grant

Assessment of Bacterial <i>bph</i> Gene in Amazonian Dark Earth and Their Adjacent Soils

<div><p>Amazonian Anthrosols are known to harbour distinct and highly diverse microbial communities. As most of the current assessments of these communities are based on taxonomic profiles, the functional gene structure of these communities, such as those responsible for key steps in the carbon cycle, mostly remain elusive. To gain insights into the diversity of catabolic genes involved in the degradation of hydrocarbons in anthropogenic horizons, we analysed the bacterial <i>bph</i> gene community structure, composition and abundance using T-RFLP, 454-pyrosequencing and quantitative PCR essays, respectively. Soil samples were collected in two Brazilian Amazon Dark Earth (ADE) sites and at their corresponding non-anthropogenic adjacent soils (ADJ), under two different land use systems, secondary forest (SF) and manioc cultivation (M). Redundancy analysis of T-RFLP data revealed differences in <i>bph</i> gene structure according to both soil type and land use. Chemical properties of ADE soils, such as high organic carbon and organic matter, as well as effective cation exchange capacity and pH, were significantly correlated with the structure of <i>bph</i> communities. Also, the taxonomic affiliation of <i>bph</i> gene sequences revealed the segregation of community composition according to the soil type. Sequences at ADE sites were mostly affiliated to aromatic hydrocarbon degraders belonging to the genera <i>Streptomyces</i>, <i>Sphingomonas</i>, <i>Rhodococcus</i>, <i>Mycobacterium</i>, <i>Conexibacter</i> and <i>Burkholderia</i>. In both land use sites, shannon's diversity indices based on the <i>bph</i> gene data were higher in ADE than ADJ soils. Collectively, our findings provide evidence that specific properties in ADE soils shape the structure and composition of <i>bph</i> communities. These results provide a basis for further investigations focusing on the bio-exploration of novel enzymes with potential use in the biotechnology/biodegradation industry.</p></div

Bar charts representing the taxonomic affiliation of <i>bph</i> gene sequences.

<p>Sequences were affiliated using the TBlastX tool available in the GenBank database. ADE  =  Amazon Dark Earth; ADJ  =  Adjacent soils; SF  =  Secondary Forest; M =  Manioc cultivation.</p

Venn's diagram of <i>bph</i> data belonging to operational protein families (OPFs) for different soil types under different land uses.

<p>ADE  =  Amazon Dark Earth; ADJ  =  Adjacent soils; SF  =  Secondary Forest; M =  Manioc cultivation. Sequences were grouped into OPFs based on sequence identity of 94%.</p

Clustering analysis of T-RFLP data based on Bray-Curtis similarity for the <i>bph</i> gene.

<p>‘a’ and ‘b’ indicate the segregation patterns according to soil type. ADE  =  Amazon Dark Earth; ADJ  =  Adjacent soils; SF  =  Secondary Forest; M =  Manioc cultivation.</p

ANOSIM test for the bacterial 16S rRNA and <i>bph</i> gene based on T-RFLP data of the Amazonian Dark Earth (ADE) and Adjacent soil (ADJ) under secondary forest (SF) and under manioc cultivation (M).

a<p>Samples were compared using T-RF peak height as a measure of abundance.</p