Effects of working posture and roof slope on activation of lower limb muscles during shingle installation.

Awkward and extreme kneeling during roofing generates high muscular tension which can lead to knee musculoskeletal disorders (MSDs) among roofers. However, the combined impact of roof slope and kneeling posture on the activation of the knee postural muscles and their association to potential knee MSD risks among roofers have not been studied. The current study evaluated the effects of kneeling posture and roof slope on the activation of major knee postural muscles during shingle installation via a laboratory assessment. Maximum normalized electromyography (EMG) data were collected from knee flexor and extensor muscles of seven subjects, who mimicked the shingle installation process on a slope-configurable wooden platform. The results revealed a significant increase in knee muscle activation during simulated shingle installation on sloped rooftops. Given the fact that increased muscle activation of knee postural muscles has been associated with knee MSDs, roof slope and awkward kneeling posture can be considered as potential knee MSD risk factors. Practitioner Summary: This study demonstrated significant effects of roof slope and kneeling posture on the peak activation of knee postural muscles. The findings of this study suggested that residential roofers could be exposed to a greater risk of developing knee MSDs with the increase of roof slope during shingle installation due to increased muscle loading. Abbreviations: MSDs: musculoskeletal disorders; EMG: electromyography; ANOVA: analysis of variance; MNMA: maximum normalized muscle activation; RF: rectus femoris; VL: vastus lateralis; VM: vastus medialis; BF: biceps femoris; S: semitendinosus.

Kneeling trunk kinematics during simulated sloped roof shingle installation.

Trunk musculoskeletal disorders are common among residential roofers. Addressing this problem requires a better understanding of the movements required to complete working tasks, such as affixing shingles on a sloped residential roof. We analyzed the extent to which the trunk kinematics during a shingling process are altered due to different angles of roof slope. Eight male subjects completed a kneeling shingle installation process on three differently sloped roof surfaces. The magnitude of the trunk kinematics was significantly influenced by both slope and task phase of the shingling process, depending on the metric. The results unequivocally point to roof slope and task phase as significant factors altering trunk kinematics. However, extension of the results to roofing workers should be done carefully, depending on the degree to which the study protocol represents the natural setting. Future studies on shingle installation in residential roofing should absolutely consider capturing a wider array of shingling procedures in order to encapsulate all the possible methods that are used due to the lack of a standardized procedure.

Are knee savers and knee pads a viable intervention to reduce lower extremity musculoskeletal disorder risk in residential roofers?

One factor commonly associated with musculoskeletal disorder risk is extreme postures. To lessen this risk, individuals must be in an as neutral posture as possible while working. We analyzed how the inclusion of different combinations of two interventions-knee pads and knee savers-can alter lower extremity kinematics during deep or near full flexion kneeling occurs while on different sloped surfaces. Nine male subjects were requested to keep a typical resting posture while kneeling on sloped roofing simulator. We observed that the introduction of a wearable third party device considerably altered lower extremity full flexion kneeling kinematics compared to level deep kneeling. This study provided a sound base for the use of third party devices to reduce musculoskeletal disorder risk on a sloped surface, however further testing with other musculoskeletal disorder risk factors is needed prior to conclusive recommendation.

Intermediaries of branched chain amino acid metabolism induce fetal hemoglobin, and repress SOX6 and BCL11A, in definitive erythroid cells.

High levels of fetal hemoglobin (HbF) can ameliorate human ß-globin gene disorders. The short chain fatty acid butyrate is the paradigmatic metabolic intermediary that induces HbF. Inherited disorders of branched-chain amino acid (BCAA) metabolism have been associated with supranormal HbF levels beyond infancy, e.g., propionic acidemia (PA) and methylmalonic acidemia (MMA). We tested intermediaries of BCAA metabolism for their effects on definitive erythropoiesis. Like butyrate, the elevated BCAA intermediaries isovalerate, isobutyrate, and propionate, induce fetal globin gene expression in murine EryD in vitro, are associated with bulk histone H3 hyperacylation, and repress the transcription of key gamma globin regulatory factors, notably BCL11A and SOX6. Metabolic intermediaries that are elevated in Maple Syrup Urine Disease (MSUD) affect none of these processes. Percent HbF and gamma (γ) chain isoforms were also measured in non-anemic, therapeutically optimized subjects with MSUD (Group I, n=6) or with Isovaleric Acidemia (IVA), MMA, or PA (Group II, n=5). Mean HbF was 0.24 ± 0.15% in Group I and 0.87 ± 0.13% in Group II (p=.01); only the Gγ isoform was detected. We conclude that a family of biochemically related intermediaries of branched chain amino acid metabolism induces fetal hemoglobin during definitive erythropoiesis, with mechanisms that mirror those so far identified for butyrate.

Targeting RET to induce medullary thyroid cancer cell apoptosis: an antagonistic interplay between PI3K/Akt and p38MAPK/caspase-8 pathways.

Mutations in REarranged during Transfection (RET) receptor tyrosine, followed by the oncogenic activation of RET kinase is responsible for the development of medullary thyroid carcinoma (MTC) that responds poorly to conventional chemotherapy. Targeting RET, therefore, might be useful in tailoring surveillance of MTC patients. Here we showed that theaflavins, the bioactive components of black tea, successfully induced apoptosis in human MTC cell line, TT, by inversely modulating two molecular pathways: (i) stalling PI3K/Akt/Bad pathway that resulted in mitochondrial transmembrane potential (MTP) loss, cytochrome-c release and activation of the executioner caspases-9 and -3, and (ii) upholding p38MAPK/caspase-8/caspase-3 pathway via inhibition of Ras/Raf/ERK. Over-expression of either constitutively active myristoylated-Akt-cDNA (Myr-Akt-cDNA) or dominant-negative-caspase-8-cDNA (Dn-caspase-8-cDNA) partially blocked theaflavin-induced apoptosis, while co-transfection of Myr-Akt-cDNA and Dn-caspase-8-cDNA completely eradicated the effect of theaflavins thereby negating the possibility of existence of other pathways. A search for the upstream signaling revealed that theaflavin-induced disruption of lipid raft caused interference in anchorage of RET in lipid raft that in turn stalled phosphorylation of Ras and PI3Kinase. In such anti-survival cellular micro-environment, pro-apoptotic signals were triggered to culminate into programmed death of MTC cell. These findings not only unveil a hitherto unexplained mechanism underlying theaflavin-induced MTC death, but also validate RET as a promising and potential target for MTC therapy.

Induction of fetal/embryonic globin gene expression depends on intact cell signaling in definitive erythroid cells.

OBJECTIVE: The induction of fetal hemoglobin during definitive erythropoiesis is a major therapeutic goal in ß-globin gene disorders. Butyrate induces fetal hemoglobin, and p38 phosphorylation has been implicated in this process. We studied p38 and the effect of its inhibitors in a physiologic primary cell model of fetal/embryonic globin gene induction during definitive erythropoiesis. METHODS: p38 phosphorylation was evaluated in a short-term culture of definitive erythroid precursor (EryD) cells following butyrate induction, absent prolonged exposure to cytokines. The impact of p38 inhibitors on embryonic/fetal globin gene induction by butyrate and on normal erythroid processes, including proliferation, differentiation, cell cycle occupancy, and RNA transcription, was also examined. RESULTS: p38 phosphorylation, maximal at harvest of murine fetal liver-derived EryD (FL EryD), when minimal embryonic/fetal globin gene expression is seen, is suppressed by EPO (as reported by others). Butyrate initially delays EPO-mediated suppression of p38 phosphorylation, but p38 phosphorylation thereafter, at 30 minutes to 48 hours, is equivalent and at low levels in EPO-treated FL EryD, with or without butyrate. Inhibitors of p38, at 10-50 µM, prevent butyrate-mediated induction of embryonic/fetal globin gene expression. We found that p38 inhibitors, which also disrupt non-p38 signaling pathways, perturb cell division, erythroid differentiation, transit through the cell cycle, and RNA transcription in primary EryD. CONCLUSION: p38 inhibitors interrupt normal erythropoiesis and the capacity for embryonic/fetal globin gene induction. However, p38 signaling is maximal in primary EryD at harvest, when embryonic globin genes are minimally expressed, and diminishes thereafter. We conclude that p38 inhibitors disrupt cellular pathways that are essential to butyrate-induced embryonic/fetal globin gene expression. However, levels of p38 phosphorylation are not coordinate with embryonic/fetal globin gene expression in EryD, and increased signaling through p38 may not be the sine qua non for embryonic/fetal globin gene induction.

Short-chain fatty acid-mediated effects on erythropoiesis in primary definitive erythroid cells.

Short-chain fatty acids (SCFAs; butyrate and propionate) up-regulate embryonic/fetal globin gene expression through unclear mechanisms. In a murine model of definitive erythropoiesis, SCFAs increased embryonic beta-type globin gene expression in primary erythroid fetal liver cells (eFLCs) after 72 hours in culture, from 1.7% (+/- 1.2%) of total beta-globin gene expression at day 0 to 4.9% (+/- 2.2%) in propionate and 5.4% (+/- 3.4%) in butyrate; this effect was greater in butyrate plus insulin/erythropoietin (BIE), at 19.5% (+/- 8.3%) compared with 0.1% (+/- 0.1%) in ins/EPO alone (P < .05). Fetal gamma-globin gene expression was increased in human transgene-containing eFLCs, to 35.9% (+/- 7.0%) in BIE compared with 4.4% (+/- 4.2%) in ins/EPO only (P < .05). Embryonic globin gene expression was detectable in 11 of 15 single eFLCs treated with BIE, but in0 of 15 ins/EPO-only treated cells. Butyrate-treated [65.5% (+/- 9.9%)] and 77.5% (+/- 4.0%) propionate-treated eFLCs were highly differentiated in culture, compared with 21.5% (+/- 3.5%) in ins/EPO (P < .005). Importantly, signaling intermediaries, previously implicated in induced embryonic/fetal globin gene expression (STAT5, p42/44, and p38), were not differentially activated by SCFAs in eFLCs; but increased bulk histone (H3) acetylation was seen in SCFA-treated eFLCs. SCFAs induce embryonic globin gene expression in eFLCS, which are a useful short-term and physiologic primary cell model of embryonic/fetal globin gene induction during definitive erythropoiesis.

Fusing imperfect experimental data for risk assessment of musculoskeletal disorders in construction using canonical polyadic decomposition.

Field or laboratory data collected for work-related musculoskeletal disorder (WMSD) risk assessment in construction often becomes unreliable as a large amount of data go missing due to technology-induced errors, instrument failures or sometimes at random. Missing data can adversely affect the assessment conclusions. This study proposes a method that applies Canonical Polyadic Decomposition (CPD) tensor decomposition to fuse multiple sparse risk-related datasets and fill in missing data by leveraging the correlation among multiple risk indicators within those datasets. Two knee WMSD risk-related datasets-3D knee rotation (kinematics) and electromyography (EMG) of five knee postural muscles-collected from previous studies were used for the validation and demonstration of the proposed method. The analysis results revealed that for a large portion of missing values (40%), the proposed method can generate a fused dataset that provides reliable risk assessment results highly consistent (70%-87%) with those obtained from the original experimental datasets. This signified the usefulness of the proposed method for use in WMSD risk assessment studies when data collection is affected by a significant amount of missing data, which will facilitate reliable assessment of WMSD risks among construction workers. In the future, findings of this study will be implemented to explore whether, and to what extent, the fused dataset outperforms the datasets with missing values by comparing consistencies of the risk assessment results obtained from these datasets for further investigation of the fusion performance.

Assessing work-related risk factors for musculoskeletal knee disorders in construction roofing tasks.

Roofers often suffer from musculoskeletal disorders (MSDs) to their knees due to spending a large amount of time kneeling while performing work-related roofing activities on sloped rooftops. Several ergonomic studies have identified kneeling as a potential risk factor for knee injuries and disorders. Existing biomechanical models and sensor technologies used to assess work-related risk factors for different construction trades are not applicable in roof work settings especially on slanted rooftop surfaces. This work assesses the impacts of work-related factors, namely working posture and roof slope, on the potential risk of developing knee MSDs due to residential roofing tasks in a laboratory setting. Nine human subjects participated in the experiment and mimicked shingle installation on a slope-configurable wooden platform. Maximum angles of right and left knee flexion, abduction, adduction, and axial rotation (internal and external) were measured as risk indicators using a motion capture system under different roof slope settings. The results demonstrated that roof slope, working posture and their interaction may have significant impacts on developing knee MSDs during roofing activities. Knees are likely to be exposed to increased risk of MSDs due to working in a dynamic kneeling posture during shingle installation. In our study, flexion in both knees and adduction in the right knee were found lower in high-pitched rooftops; however, abduction in the left knee and internal rotation in the right knee were found higher during shingle installation. Hence proper attention is needed for these situations. This study provides useful information about the impact of roof work settings on knee MSDs development, which may facilitate effective interventions such as education, training, and tools to prevent knee injuries in construction roofing tasks.

Validation of using gene expression in mononuclear cells as a marker for hepatic cholesterol metabolism.

HMG-CoA reductase and the LDL receptor are ubiquitously expressed in major tissues. Since the liver plays a major role in regulating circulating LDL, it is usually of interest to measure the effects of drug or dietary interventions on these proteins in liver. In humans, peripheral blood mononuclear cells have been used as a surrogate for liver to assess regulation of these genes, although there is concern regarding the validity of this approach. The purpose of this study was to evaluate the relationship between liver and mononuclear cell expression of HMG-CoA reductase and the LDL receptor in guinea pigs, a well established model for human cholesterol and lipoprotein metabolism. We extracted RNA from liver and mononuclear cells of guinea pigs from a previous study where the effects of rapamycin, an immunosuppresant drug used for transplant patients, on lipid metabolism were evaluated. Guinea pigs were assigned to three different diets containing the same amount of fat (15 g/100 g) and cholesterol (0.08 g/100 g) for a period of 3 weeks. The only difference among diets was the concentration of rapamycin: 0, 0.0028 or 0.028 g/100 g. There were no differences in plasma LDL cholesterol (LDL-C) among groups. Values were 78.4 +/- 14.3, 65.8 +/- 17.2 and 68.4 +/- 45.4 mg/dL (P > 0.05) for guinea pigs treated with 0, low or high doses of rapamycin, respectively. The mRNA abundance for the LDL receptor and HMG-CoA reductase was measured both in liver (n = 30) and mononuclear cells (n = 22) using reverse transcriptase PCR. In agreement with the finding of no changes in plasma LDL-C, there were also no differences for the expression of HMG-CoA reductase or the LDL receptor among groups. However, a positive correlation was found between liver and mononuclear cells for both HMG-CoA reductase (r = 0.613, P < 0.01) and the LDL receptor (r = 0.622, P < 0.01). These correlations suggest that monocytes can be used in humans as an index for liver to assess diet and drug effects on the expression of HMG-CoA reductase and the LDL receptor.

Rapid homeostatic response of H4IIE cells to diethylenetriaminepentaacetic acid is not due to changes in the amount or localization of ZnT-1 protein.

We have demonstrated that reducing zinc availability with the extracellular chelator diethylenetriaminepentaacetic acid (DTPA) causes rapid inhibition of cellular zinc efflux in H4IIE hepatoma cells but increases zinc efflux in primary hepatocytes. Similar differences were also observed between the rat anterior pituitary cell line GH3 and primary anterior pituitary cells. We hypothesized that the difference between the transformed and primary cells is due to differential regulation of ZnT-1 or SLC-30A-1 because this is the only zinc efflux transporter localized to the plasma membrane. The effects of DTPA (50 µM) and zinc (100 µM) treatment on messenger RNA (mRNA) and protein expression and protein localization of ZnT-1 were studied in H4IIE cells and primary hepatocytes. Although zinc tended to increase ZnT-1 mRNA in H4IIE cells, DTPA had no effect on ZnT-1 mRNA and protein expression in either hepatoma cells or hepatocytes. Although ZnT-1 is thought to be localized on the plasma membrane, this localization was not seen in these liver cells where ZnT-1 was distributed throughout the cytoplasm. Vesicular localization of ZnT-1 appeared to increase with zinc treatment. Total zinc content was reduced by DTPA in H4IIE cells. Diethylenetriaminepentaacetic acid also reduced metallothionein 1 mRNA, reflecting this reduction in cellular zinc. We conclude that the rapid homeostatic response of cells to altered zinc availability must be attributed to a transporter other than ZnT-1 or to changes in the activity of ZnT-1 by a novel mechanism.

Zinc retention differs between primary and transformed cells in response to zinc deprivation.

Previous studies in our laboratory have demonstrated that reducing the availability of zinc with the extracellular metal chelator DTPA (diethylenetriaminepentaacetate) enhances, rather than inhibits, the thyroid hormone induction of growth hormone mRNA in GH3 rat anterior pituitary tumor cells. To understand the actions of the chelator on cellular zinc status, we observed the effects of DTPA on (65)Zn uptake and retention. DTPA reduced the uptake of (65)Zn by GH3 cells from the medium, but when GH3 cells were prelabeled with (65)Zn, it resulted in greater retention of the isotope. In primary hepatocytes, DTPA both reduced the uptake of (65)Zn from the medium and increased efflux from prelabeled cells. To investigate this difference, we studied the effects of DTPA on radioactive zinc flux in the H4IIE (rat hepatoma), MCF-7 (human breast cancer) and Hs578Bst (nontransformed human mammary) cell lines and in rat primary anterior pituitary cells. DTPA reduced the uptake of (65)Zn in all cell lines examined. DTPA increased the retention of (65)Zn in prelabeled H4IIE, MCF-7 and Hs578Bst cells but reduced it in primary pituitary cells. Time course experiments showed that (65)Zn efflux is shut down rapidly by DTPA in transformed cells, whereas the chelator causes greater efflux from primary hepatocytes over the first 6 h. Experiments with (14)C-labeled DTPA confirmed that this chelator does not cross cell membranes, showing that it operates entirely within the medium. Expression of ZnT-1, the efflux transporter, was not affected by DTPA in H4IIE cells. Thus, zinc deprivation enhanced zinc retention in established cell lines but increased efflux from primary cells, perhaps reflecting differing requirements for this mineral.