The Family’s Voice: Caregiving for an Older Adult

The purpose of this research was to explore the family as a unit, in particular the effects on the multigenerational family when at least one person is giving care to an adult over the age of 65. While, most prior research focused on the caregiver, this study looked at family as a whole single unit. The respondents were asked to describe how the caregiving affected them personally as well as how it affected the family. The sample for this study included 16 adult family members of caregivers for a person over age 65. Data was collected utilizing an online survey. Respondents were recruited with the assistance of local senior assisted living and apartment communities and by some postings in local social work groups. Respondents voiced that life was now stressful and frustrating. Although some revealed there were benefits, many stated the caregiving affected the family in an adverse way. This study confirmed the idea that having a family member caring for an older adult member does affect the dynamics of the whole family

The Family’s Voice: Caregiving for an Older Adult

The purpose of this research was to explore the family as a unit, in particular the effects on the multigenerational family when at least one person is giving care to an adult over the age of 65. While, most prior research focused on the caregiver, this study looked at family as a whole single unit. The respondents were asked to describe how the caregiving affected them personally as well as how it affected the family. The sample for this study included 16 adult family members of caregivers for a person over age 65. Data was collected utilizing an online survey. Respondents were recruited with the assistance of local senior assisted living and apartment communities and by some postings in local social work groups. Respondents voiced that life was now stressful and frustrating. Although some revealed there were benefits, many stated the caregiving affected the family in an adverse way. This study confirmed the idea that having a family member caring for an older adult member does affect the dynamics of the whole family

Does protein kinase R mediate TNF-α- and ceramide-induced increases in expression and activation of matrix metalloproteinases in articular cartilage by a novel mechanism?

We investigated the role of the proinflammatory cytokine TNF-α, the second messenger C(2)-ceramide, and protein kinase R (PKR) in bovine articular cartilage degradation. Bovine articular cartilage explants were stimulated with C(2)-ceramide or TNF-α for 24 hours. To inhibit the activation of PKR, 2-aminopurine was added to duplicate cultures. Matrix metalloproteinase (MMP) expression and activation in the medium were analysed by gelatin zymography, proteoglycan release by the dimethylmethylene blue assay, and cell viability by the Cytotox 96(® )assay. C(2)-ceramide treatment of cartilage explants resulted in a significant release of both pro- and active MMP-2 into the medium. Small increases were also seen with TNF-α treatment. Incubation of explants with 2-aminopurine before TNF-α or C(2)-ceramide treatment resulted in a marked reduction in expression and activation of both MMP-2 and MMP-9. TNF-α and C(2)-ceramide significantly increased proteoglycan release into the medium, which was also inhibited by cotreatment with 2-aminopurine. A loss of cell viability was observed when explants were treated with TNF-α and C(2)-ceramide, which was found to be regulated by PKR. We have shown that C(2)-ceramide and TNF-α treatment of articular cartilage result in the increased synthesis and activation of MMPs, increased release of proteoglycan, and increased cell death. These effects are abrogated by treatment with the PKR inhibitor 2-aminopurine. Collectively, these results suggest a novel role for PKR in the synthesis and activation of MMPs and support our hypothesis that PKR and its activator, PACT, are implicated in the cartilage degradation that occurs in arthritic disease

Considerations for accurate gene expression measurement by reverse transcription quantitative PCR when analysing clinical samples

Reverse transcription quantitative PCR is an established, simple and effective method for RNA measurement. However, technical standardisation challenges combined with frequent insufficient experimental detail render replication of many published findings challenging. Consequently, without adequate consideration of experimental standardisation, such findings may be sufficient for a given publication but cannot be translated to wider clinical application. This article builds on earlier standardisation work and the MIQE guidelines, discussing processes that need consideration for accurate, reproducible analysis when dealing with patient samples. By applying considerations common to the science of measurement (metrology), one can maximise the impact of gene expression studies, increasing the likelihood of their translation to clinical tools

Exogenous sphingomyelinase increases collagen and sulphated glycosaminoglycan production by primary articular chondrocytes: an in vitro study

We previously established a role for the second messenger ceramide in protein kinase R (PKR)-mediated articular cartilage degradation. Ceramide is known to play a dual role in collagen gene regulation, with the effect of ceramide on collagen promoter activity being dependent on its concentration. Treatment of cells with low doses of sphingomyelinase produces small increases in endogenous ceramide. We investigated whether ceramide influences articular chondrocyte matrix homeostasis and, if so, the role of PKR in this process. Bovine articular chondrocytes were stimulated for 7 days with sphingomyelinase to increase endogenous levels of ceramide. To inhibit PKR, 2-aminopurine was added to duplicate cultures. De novo sulphated glycosaminoglycan and collagen synthesis were measured by adding [(35)S]-sulphate and [(3)H]-proline to the media, respectively. Chondrocyte phenotype was investigated using RT-PCR and Western blot analysis. Over 7 days, sphingomyelinase increased the release of newly synthesized sulphated glycosaminoglycan and collagen into the media, whereas inhibition of PKR in sphingomyelinase-treated cells reduced the level of newly synthesized sulphated glycosaminoglycan and collagen. Sphingomyelinase treated chondrocytes expressed col2a1 mRNA, which is indicative of a normal chondrocyte phenotype; however, a significant reduction in type II collagen protein was detected. Therefore, small increments in endogenous ceramide in chondrocytes appear to push the homeostatic balance toward extracellular matrix synthesis but at the expense of the chondrocytic phenotype, which was, in part, mediated by PKR

AMPA/kainate glutamate receptor antagonists prevent posttraumatic osteoarthritis

Musculoskeletal disorders represent the 3rd greatest burden on health in the developed world. Osteoarthritis is the single greatest cause of chronic pain, has no cure, and affects 8.5 and 27 million in the UK and US respectively. Osteoarthritis commonly occurs after joint injury, particularly affecting younger patients. Painful joints are often treated with injections of steroid or hyaluronic acid (HA), but treatments to prevent subsequent joint degeneration remain elusive. In animals, joint injury increases glutamate release into the joint, acting on nerves to cause pain, and joint tissues to cause inflammation and degeneration. This study investigated synovial fluid glutamate concentrations and glutamate receptor (GluR) expression in injured human joints and compared efficacy of GluR antagonists with current treatments in a mouse model of injury-induced osteoarthritis (ACL rupture). GluRs were expressed in ligament and meniscus after knee injury and synovial fluid glutamate concentrations ranged from 19–129 µM. Intra-articular injection of NBQX (GluR antagonist), administered at the time of injury, substantially reduced swelling and degeneration in the mouse ACL rupture model. HA had no effect and depo-medrone reduced swelling for 1 day, but increased degeneration by 50%. Intra-articular administration of NBQX was both symptom and disease modifying to a greater extent than current treatments. There is an opportunity for repurposing related drugs, developed for CNS disorders, with proven safety in man, to prevent injury-induced osteoarthritis. This could quickly reduce the substantial burden associated with osteoarthritis

CTL epitope distribution patterns in the Gag and Nef proteins of HIV-1 from subtype A infected subjects in Kenya: Use of multiple peptide sets increases the detectable breadth of the CTL response

BACKGROUND: Subtype A is a major strain in the HIV-1 pandemic in eastern Europe, central Asia and in certain regions of east Africa, notably in rural Kenya. While considerable effort has been focused upon mapping and defining immunodominant CTL epitopes in HIV-1 subtype B and subtype C infections, few epitope mapping studies have focused upon subtype A. RESULTS: We have used the IFN-γ ELIspot assay and overlapping peptide pools to show that the pattern of CTL recognition of the Gag and Nef proteins in subtype A infection is similar to that seen in subtypes B and C. The p17 and p24 proteins of Gag and the central conserved region of Nef were targeted by CTL from HIV-1-infected Kenyans. Several epitope/HLA associations commonly seen in subtype B and C infection were also observed in subtype A infections. Notably, an immunodominant HLA-C restricted epitope (Gag 296–304; YL9) was observed, with 8/9 HLA-C(W)0304 subjects responding to this epitope. Screening the cohort with peptide sets representing subtypes A, C and D (the three most prevalent HIV-1 subtypes in east Africa), revealed that peptide sets based upon an homologous subtype (either isolate or consensus) only marginally improved the capacity to detect CTL responses. While the different peptide sets detected a similar number of responses (particularly in the Gag protein), each set was capable of detecting unique responses not identified with the other peptide sets. CONCLUSION: Hence, screening with multiple peptide sets representing different sequences, and by extension different epitope variants, can increase the detectable breadth of the HIV-1-specific CTL response. Interpreting the true extent of cross-reactivity may be hampered by the use of 15-mer peptides at a single concentration and a lack of knowledge of the sequence that primed any given CTL response. Therefore, reagent choice and knowledge of the exact sequences that prime CTL responses will be important factors in experimentally defining cross-reactive CTL responses and their role in HIV-1 disease pathogenesis and validating vaccines aimed at generating broadly cross-reactive CTL responses

Caught in a Trap? Proteomic Analysis of Neutrophil Extracellular Traps in Rheumatoid Arthritis and Systemic Lupus Erythematosus

Neutrophil Extracellular Traps (NETs) are implicated in the development of auto-immunity in diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) through the externalization of intracellular neoepitopes e.g., dsDNA and nuclear proteins in SLE and citrullinated peptides in RA. The aim of this work was to use quantitative proteomics to identify and measure NET proteins produced by neutrophils from healthy controls, and from patients with RA and SLE to determine if NETs can be differentially-generated to expose different sets of neoepitopes. Ultra-pure neutrophils (>99%) from healthy individuals (n = 3) and patients with RA or SLE (n = 6 each) were incubated ± PMA (50 nM, PKC super-activator) or A23187 (3.8 μM, calcium ionophore) for 4 h. NETs were liberated by nuclease digestion and concentrated onto Strataclean beads prior to on-bead digestion with trypsin. Data-dependent LC-MS/MS analyses were conducted on a QExactive HF quadrupole-Orbitrap mass spectrometer, and label-free protein quantification was carried out using Progenesis QI. PMA-induced NETs were decorated with annexins, azurocidin and histone H3, whereas A23187-induced NETs were decorated with granule proteins including CAMP/LL37, CRISP3, lipocalin and MMP8, histones H1.0, H1.4, and H1.5, interleukin-8, protein-arginine deiminase-4 (PADI4), and α-enolase. Four proteins were significantly different between PMA-NETs from RA and SLE neutrophils (p < 0.05): RNASE2 was higher in RA, whereas MPO, leukocyte elastase inhibitor and thymidine phosphorylase were higher in SLE. For A23187-NETs, six NET proteins were higher in RA (p < 0.05), including CAMP/LL37, CRISP3, interleukin-8, MMP8; Thirteen proteins were higher in SLE, including histones H1.0, H2B, and H4. This work provides the first, direct comparison of NOX2-dependent (PMA) and NOX2-independent (A23187) NETs using quantitative proteomics, and the first direct comparison of RA and SLE NETs using quantitative proteomics. We show that it is the nature of the stimulant rather than neutrophil physiology that determines NET protein profiles in disease, since stimulation of NETosis in either a NOX2-dependent or a NOX2-independent manner generates broadly similar NET proteins irrespective of the disease background. We also use our proteomics pipeline to identify an extensive range of post-translationally modified proteins in RA and SLE, including histones and granule proteins, many of which are known targets of auto-antibodies in each disease

Strategies for Engaging Engineering Faculty in Continuing Education

This presentation was part of the session : Practical Strategies for Engaging Engineering Faculty in Continuing Education ITerry J. Reed: Director, Engineering Continuing & Distance Education, Penn State University, University, University Park, PA, USA. Mr. Reed received B.S. in Electrical Engineering from Penn State, an M.S. in Electrical Engineering and an MBA from the University of Pittsburgh. Thomas M. Iwinski: eLearning Specialist, Engineering Continuing & Distance Education, Penn State University, University, University Park, PA, USA. Mr. Iwinski received a B.S. in Communications Media from Indiana University of Pennsylvania and an M.S. in Instructional Technologies from Bloomsburg University of Pennsylvania. Deborah L. Zimmerman: Program Manager, Engineering Continuing & Distance Education, Penn State University, University, University Park, PA, USA. Ms. Zimmerman has over 30 years of experience working with faculty on continuing and distance programs. John M. Mason: Associate Dean for Research, Graduate Studies, and Outreach; Professor of Civil Engineering; and Director of the Pennsylvania Transportation Institute in the College of Engineering, Penn State University, University, University Park, PA, USA. Dr. Mason received a B.S. in Transportation from Penn State, an M.S. in Transportation Engineering from Villanova University, and a Ph.D. in Civil Engineering from Texas A&M University.IACEE 11th World Conference on Continuing Engineering EducationEngaging engineering faculty at a large research university in continuing education is a challenge because faculty choose to invest their time and talents in other activities which are perceived to have higher benefits. This paper describes strategies that can be employed to increase the benefits of continuing education activities to make them more attractive. These strategies include reducing the faculty time requirements and increasing the rewards. The paper illustrates how these strategies have been applied to the activities necessary for the design, production and delivery a graduate level credit course taught to distant students.Distance Learning and Professional Education ; International Association for Continuing Engineering Educatio

Vegetation and floristics of a lowland tropical rainforest in northeast Australia

Background: Full floristic data, tree demography, and biomass estimates incorporating non-tree life forms are seldom collected and reported for forest plots in the tropics. Established research stations serve as important repositories of such biodiversity and ecological data. With a canopy crane setup within a tropical lowland rainforest estate, the 42-ha Daintree Rainforest Observatory (DRO) in Cape Tribulation, northern Australia is a research facility of international significance. We obtained an estimate of the vascular plant species richness for the site, by surveying all vascular plant species from various mature-phase, remnant and open vegetation patches within the site. We also integrate and report the demography and basal areas of trees ≥ 10 cm diameter at breast height (dbh) in a new 1- ha core plot, an extension to the pre-existing forest 1-ha plot under the canopy crane. In addition, we report for the canopy crane plot new demography and basal areas for smaller size shrubs and treelets subsampled from nine 20 m quadrats, and liana basal area and abundance from the whole plot. The DRO site has an estimated total vascular plant species richness of 441 species, of which 172 species (39%) are endemic to Australia, and 4 species are endemics to the Daintree region. The 2 x 1-ha plots contains a total of 262 vascular plant species of which 116 (1531 individuals) are tree species ≥ 10 cm dbh. We estimate a stem basal area of 34.9 m ha, of which small stems (tree saplings and shrubs <10cm dbh) and lianas collectively contribute c.4.2%. Comparing the stem density-diversity patterns of the DRO forest with other tropical rainforests globally, our meta-analysis shows that DRO forests has a comparatively high stem density and moderate species diversity, due to the influence of cyclones. These data will provide an important foundation for ecological and conservation studies in lowland tropical forest. New information: We present a floristic checklist, a life form breakdown, and demography data from two 1-ha rainforest plots from a lowland tropical rainforest study site. We also present a meta-analysis of stem densities and species diversity from comparable-sized plots across the tropics