Type and orientation of yielded trabeculae during overloading of trabecular bone along orthogonal directions

Trabecular architecture plays a major role in bone mechanics. Osteoporosis leads to a transition from a plate-like to a more rod-like trabecular morphology, which may contribute to fracture risk beyond that predicted by changes in density. In this study, microstructural finite element analysis results were analyzed using individual trabeculae segmentation (ITS) to identify the type and orientation of trabeculae where tissue yielded during compressive overloads in two orthogonal directions. For both apparent loading conditions, most of the yielded tissue was found in longitudinally oriented plates. However, the primary loading mode of yielded trabeculae was axial compression with superposed bending for on-axis loading in contrast to bending for transverse loading. For either loading direction, most plate-like trabeculae yielded in the same loading mode, regardless of their orientation. In contrast, rods oriented parallel to the loading axis yielded in compression, while rods oblique or perpendicular to the loading axis yielded in combined bending and tension. The predominance of tissue yielding in plates during both on-axis and transverse overloading explains why on-axis overloading is detrimental to the off-axis mechanical properties. At the same time, a large fraction of the tissue in rod-like trabeculae parallel to the loading direction yielded in both on-axis and transverse loading. Hence, rods may be more likely to be damaged and potentially resorbed by damage mediated remodeling.     

Proteomic analysis of early‐response to mechanical stress in neonatal rat mandibular condylar chondrocytes

The objectives of this study were to investigate the early response to mechanical stress in neonatal rat mandibular chondrocytes by proteomic analysis. To evaluate its molecular mechanism, chondrocytes were isolated and cultured in vitro, then loaded mechanical stress by four‐point bending system on different patterns. Morphological observation, flow cytometric analysis, and MTT assays indicated that 4,000 µstrain loading for 60 min was an appropriate mechanical stimulus for the following proteome analysis, which produced a transient but obvious inhibitory effect on the cell cycle. Therefore, we took a proteomic approach to identify significantly differential expression proteins in chondrocytes under this mechanical stress. Using 2‐DE and MALDI‐TOF, we identified seven differentially expressed proteins including the MAPK pathway inhibitor RKIP, cytoskeleton proteins, actin and vimentin, and other selected proteins. Some differentially expressed proteins were validated by both Western blot analysis and fluorescent staining of cytoskeleton at different loading times. The vimentin and RKIP responsive expression were also proven in vivo in oral orthopedic treatment rats, which was in line with the result in vitro. The histological changes in cartilage also showed the inhibition effect. Furthermore, the expressional level of phosphorylated ERK was increased, which demonstrates the changes in MAPK activity. Taken together, these data indicate that mechanical stress resulted in vimentin expression changes first and then led to the subsequent changes in actin expression, MAPK pathway regulated by RKIP and heat shock protein GRP75. All those changes contributed to the cytoskeleton remolding and cell cycle inhibition, finally led to condylar remodeling. J. Cell. Physiol. 223:610–622, 2010. © 2010 Wiley‐Liss, Inc.     

Ceramide plays a prominent role in MDA‐7/IL‐24‐induced cancer‐specific apoptosis

Melanoma differentiation associated gene‐7/interleukin‐24 (mda‐7/IL‐24) uniquely displays broad cancer‐specific apoptosis‐inducing activity through induction of endoplasmic reticulum (ER) stress. We hypothesize that ceramide, a promoter of apoptosis, might contribute to mda‐7/IL‐24 induction of apoptosis. Ad.mda‐7‐infected tumor cells, but not normal cells, showed increased ceramide accumulation. Infection with Ad.mda‐7 induced a marked increase in various ceramides (C16, C24, C24:1) selectively in prostate cancer cells. Inhibiting the enzyme serine palmitoyltransferase (SPT) using the potent SPT inhibitor myriocin (ISP1), impaired mda‐7/IL‐24‐induced apoptosis and ceramide production, suggesting that ceramide formation caused by Ad.mda‐7 occurs through de novo synthesis of ceramide and that ceramide is required for mda‐7/IL‐24‐induced cell death. Fumonisin B1 (FB1) elevated ceramide formation as well as apoptosis induced by Ad.mda‐7, suggesting that ceramide formation may also occur through the salvage pathway. Additionally, Ad.mda‐7 infection enhanced expression of acid sphingomyelinase (ASMase) with a concomitant increase in ASMase activity and decreased sphingomyelin in cancer cells. ASMase silencing by RNA interference inhibited the decreased cell viability and ceramide formation after Ad.mda‐7 infection. Ad.mda‐7 activated protein phosphatase 2A (PP2A) and promoted dephosphorylation of the anti‐apoptotic molecule BCL‐2, a downstream ceramide‐mediated pathway of mda‐7/IL‐24 action. Pretreatment of cells with FB1 or ISP‐1 abolished the induction of ER stress markers (BiP/GRP78, GADD153 and pospho‐eIF2α) triggered by Ad.mda‐7 infection indicating that ceramide mediates ER stress induction by Ad.mda‐7. Additionally, recombinant MDA‐7/IL‐24 protein induced cancer‐specific production of ceramide. These studies define ceramide as a key mediator of an ER stress pathway that may underlie mda‐7/IL‐24 induction of cancer‐specific killing. J. Cell. Physiol. 222: 546–555, 2010. © 2009 Wiley‐Liss, Inc.     

Role of voltage‐gated potassium channels in the fate determination of embryonic stem cells

Embryonic stem cells (ESCs) possess two unique characteristics: self‐renewal and pluripotency. In this study, roles of voltage‐gated potassium channels (K_v) in maintaining mouse (m) ESC characteristics were investigated. Tetraethylammonium (TEA⁺), a K_v blocker, attenuated cell proliferation in a concentration‐dependent manner. Possible reasons for this attenuation, including cytotoxicity, cell cycle arrest and differentiation, were examined. Blocking K_v did not change the viability of mESCs. Interestingly, K_v inhibition increased the proportion of cells in G₀/G₁ phase and decreased that in S phase. This change in cell cycle distribution can be attributed to cell cycle arrest or differentiation. Loss of pluripotency as determined at both molecular and functional levels was detected in mESCs with K_v blockade, indicating that K_v inhibition in undifferentiated mESCs directs cells to differentiate instead of to self‐renew and progress through the cell cycle. Membrane potential measurement revealed that K_v blockade led to depolarization, consistent with the role of K_v as the key determinant of membrane potential. The present results suggest that membrane potential changes may act as a “switch” for ESCs to decide whether to proliferate or to differentiate: hyperpolarization at G₁ phase would favor ESCs to enter S phase while depolarization would favor ESCs to differentiate. Consistent with this notion, S‐phase‐synchronized mESCs were found to be more hyperpolarized than G₀/G₁‐phase‐synchronized mESCs. Moreover, when mESCs differentiated, the differentiation derivatives depolarized at the initial stage of differentiation. This investigation is the first study to provide evidence that K_v and membrane potential affect the fate determination of ESCs. J. Cell. Physiol. 224:165–177, 2010 © 2010 Wiley‐Liss, Inc.     

Identification and characterization of the pathogenic effect of a Vibrio parahaemolyticus-related bacterium isolated from clam Meretrix meretrix with mass mortality

A mass mortality of clam, Meretrix meretrix, occurred in Jiangsu Province of China in the late September of 2007. Of the isolates obtained from the diseased clams, MM21 had the strongest virulence to the clam in the virulence test, with a LD50 value of ∼6 × 10⁶ CFU ml⁻¹. MM21 was identified as Vibrio parahaemolyticus by the VITEK 2 Compact system and 16S rDNA sequencing. Detection of virulence-associated genes by PCR indicated that MM21 was positive for toxR and tlh, and negative for tdh. Compared with control group, histiocytes from MM21-infected clams displayed a variety of cytopathological changes by transmission electron microscopy examination, which included increased lipid droplets in hepatocytes, deposition of granules in the mantle, excessive secretion in the gill. The results of our study suggested that MM21 may have been an etiological element in the mass mortalities of hard clam (M. meretrix) in Jiangsu Province of China in 2007.     

Model-Guided Mutagenesis Drives Functional Studies of Human NHA2, Implicated in Hypertension

Human NHA2 is a poorly characterized Na⁺/H⁺ antiporter recently implicated in essential hypertension. We used a range of computational tools and evolutionary conservation analysis to build and validate a three-dimensional model of NHA2 based on the crystal structure of a distantly related bacterial transporter, NhaA. The model guided mutagenic evaluation of transport function, ion selectivity, and pH dependence of NHA2 by phenotype screening in yeast. We describe a cluster of essential, highly conserved titratable residues located in an assembly region made of two discontinuous helices of inverted topology, each interrupted by an extended chain. Whereas in NhaA, oppositely charged residues compensate for partial dipoles generated within this assembly, in NHA2, polar but uncharged residues suffice. Our findings led to a model for transport mechanism that was compared to the well-known electroneutral NHE1 and electrogenic NhaA subtypes. This study establishes NHA2 as a prototype for the poorly understood, yet ubiquitous, CPA2 antiporter family recently recognized in plants and metazoans and illustrates a structure-driven approach to derive functional information on a newly discovered transporter.     

The urban heat island and its impact on heat waves and human health in Shanghai

With global warming forecast to continue into the foreseeable future, heat waves are very likely to increase in both frequency and intensity. In urban regions, these future heat waves will be exacerbated by the urban heat island effect, and will have the potential to negatively influence the health and welfare of urban residents. In order to investigate the health effects of the urban heat island (UHI) in Shanghai, China, 30 years of meteorological records (1975–2004) were examined for 11 first- and second-order weather stations in and around Shanghai. Additionally, automatic weather observation data recorded in recent years as well as daily all-cause summer mortality counts in 11 urban, suburban, and exurban regions (1998–2004) in Shanghai have been used. The results show that different sites (city center or surroundings) have experienced different degrees of warming as a result of increasing urbanization. In turn, this has resulted in a more extensive urban heat island effect, causing additional hot days and heat waves in urban regions compared to rural locales. An examination of summer mortality rates in and around Shanghai yields heightened heat-related mortality in urban regions, and we conclude that the UHI is directly responsible, acting to worsen the adverse health effects from exposure to extreme thermal conditions.     

The expression and function of β-1,4-galactosyltransferase-I in dendritic cells

β-1,4-Galactosyltransferase-I (GalTI) is unusual among the galactosyltransferase family, which has two isoforms that differ only in the length of their cytoplasmic domains [1]. In this study, we found that both the long and short isoforms of GalTI were expressed in human monocyte-derived dendritic cells (MoDCs), and localized in the cytoplasm near nucleus and cytomembrane. The expression level of GalTI and cellular adhesion ability was increased when DCs continued to mature. We also demonstrated that the cellular adhesion ability of DCs was inhibited by α-lactalbumin (α-LA) via interference with cell surface GalTI function, suggesting that the adhesion ability was positively correlated with the expression of cell surface long GalTI. α-LA also could inhibit DC-T clustering and CD4⁺ T cell proliferation. Collectively, the data suggests that GalTI might act as a key adhesion molecular participating in T cells-DCs contacts.     

An energy budget approach for evaluating the biocontrol potential of cotton aphid (Aphis gossypii) by the ladybeetle Propylaea japonica

Biological control of economically important crop pests is an important component of integrated pest management (IPM) strategies. Predator–prey energy relationships are critical to the success of biocontrol strategies; however, these relationships are often ignored in many IPM programs. In this study, the biocontrol potential of cotton aphid, Aphis gossypii (Glover) (Hemiptera: Aphididae), by the ladybeetle Propylaea japonica (Thunberg) (Coleoptera: Coccinellidae) was estimated in terms of energy budgets calculated at 27 ± 1 °C. The energy equivalent of prey subjects (aphids) consumed was estimated from bomb calorimetry and partitioned into the energy associated with ingestion, assimilation, respiration, reproduction, and waste for each developmental stage of the lady beetle. The average assimilation efficiencies for larval and adult ladybeetles were 88.2 and 91.1%, respectively, whereas net ecological efficiencies were 17.6% for larvae and 2.6% for adults. Similarly, assimilation efficiencies of cotton aphids were 71.5 and 74.4% for nymphs and adults, respectively. Based on energy budget calculations, approximately 520, 3‐day‐old aphids and 5 356, 3‐day‐old aphids were estimated to be consumed by the ladybeetle larval stage and the female adult stage, respectively. These estimates were similar to the actual number of aphids consumed by the ladybeetles, based on actual counts. The current data demonstrate that P. japonica is an important natural enemy of the cotton aphid, and that predator–prey energy relationships can play a critical role in biocontrol strategies and IPM programs.