Cellular adaption to repeated eccentric exercise-induced muscle damage

Eccentrically biased exercise results in skeletal muscle damage and stimulates adaptations in muscle, whereby indexes of damage are attenuated when the exercise is repeated. We hypothesized that changes in ultrastructural damage, inflammatory cell infiltration, and markers of proteolysis in skeletal muscle would come about as a result of repeated eccentric exercise and that gender may affect this adaptive response. Untrained male (n = 8) and female (n = 8) subjects performed two bouts (bout 1 and bout 2), separated by 5.5 wk, of 36 repetitions of unilateral, eccentric leg press and 100 repetitions of unilateral, eccentric knee extension exercises (at 120% of their concentric single repetition maximum), the subjects' contralateral nonexercised leg served as a control (rest). Biopsies were taken from the vastus lateralis from each leg 24 h postexercise. After bout 2, the postexercise force deficit and the rise in serum creatine kinase (CK) activity were attenuated. Women had lower serum CK activity compared with men at all times (P < 0.05), but there were no gender differences in the relative magnitude of the force deficit. Muscle Z-disk streaming, quantified by using light microscopy, was elevated vs. rest only after bout 1 (P < 0.05), with no gender difference. Muscle neutrophil counts were significantly greater in women 24 h after bout 2 vs. rest and bout 1 (P < 0.05) but were unchanged in men. Muscle macrophages were elevated in men and women after bout 1 andbout 2 (P < 0.05). Muscle protein content of the regulatory calpain subunit remained unchanged whereas ubiquitin-conjugated protein content was increased after both bouts (P < 0.05), with a greater increase after bout 2. We conclude that adaptations to eccentric exercise are associated with attenuated serum CK activity and, potentially, an increase in the activity of the ubiquitin proteosome proteolytic pathway.

Bone geometry in response to long-term tennis playing and its relationship with muscle volume : a quantitative magnetic resonance imaging study in tennis players

The benefit of impact-loading activity for bone strength depends on whether the additional bone mineral content (BMC) accrued at loaded sites is due to an increased bone size, volumetric bone mineral density (vBMD) or both. Using magnetic resonance imaging (MRI) and dual energy X-ray absorptiometry (DXA), the aim of this study was to characterize the geometric changes of the dominant radius in response to long-term tennis playing and to assess the influence of muscle forces on bone tissue by investigating the muscle–bone relationship. Twenty tennis players (10 men and 10 women, mean age: 23.1 ± 4.7 years, with 14.3 ± 3.4 years of playing) were recruited. The total bone volume, cortical volume, sub-cortical volume and muscle volume were measured at both distal radii by MRI. BMC was assessed by DXA and was divided by the total bone volume to derive vBMD. Grip strength was evaluated with a dynamometer. Significant side-to-side differences (P < 0.0001) were found in muscle volume (+9.7%), grip strength (+13.3%), BMC (+13.5%), total bone volume (+10.3%) and sub-cortical volume (+20.6%), but not in cortical volume (+2.6%, ns). The asymmetry in total bone volume explained 75% of the variance in BMC asymmetry (P < 0.0001). vBMD was slightly higher on the dominant side (+3.3%, P < 0.05). Grip strength and muscle volume correlated with all bone variables (except vBMD) on both sides (r = 0.48–0.86, P < 0.05–0.0001) but the asymmetries in muscle parameters did not correlate with those in bone parameters. After adjustment for muscle volume or grip strength, BMC was still greater on the dominant side. This study showed that the greater BMC induced by long-term tennis playing at the dominant radius was associated to a marked increase in bone size and a slight improvement in volumetric BMD, thereby improving bone strength. In addition to the muscle contractions, other mechanical stimuli seemed to exert a direct effect on bone tissue, contributing to the specific bone response to tennis playing.

Epidermal growth factor-induced ovarian carcinoma cell migration is associated with JAK2/STAT3 signals and changes in the abundance and localization of α6β1 integrin

Peritoneal dissemination of ovarian carcinoma is mediated by epithelial–mesenchymal interconversions leading to the disruption of cell–cell contact and modulation of cell–extracellular matrix (ECM) interactions. The present study was designed to evaluate the effects of epidermal growth factor (EGF) as a modulator of Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) signalling and changes in integrin expression during the process similar to EMT. A fibroblastic morphology with reduced intercellular cell contacts and increased cell motility was observed in ovarian cancer cell lines in response to EGF and was concomitant with the up regulation of EMT-associated N-cadherin and vimentin expression. These changes were accompanied by an increase in α2, α6 and β1 integrin subunits and activation of JAK2 and STAT3 signalling which was suppressed by a specific JAK2 inhibitor. Consistent with the suppression of STAT3 activity, N-cadherin and vimentin expression were abrogated and was coherent with the loss of cell motility and the expression of α6 and β1 integrin subunits. Neutralizing antibodies against α6 and β1 subunits inhibited cancer cell migration. A strong correlation between the expression of N-cadherin, vimentin and JAK2/STAT3 levels were detected in high-grade ovarian tumors and was consistent with the previously reported enhanced expression of α6 integrin subunit in advanced tumors [Ahmed N, Riley C, Oliva K, Rice G, Quinn M. Ascites induces modulation of α6β1 integrin and urokinase plasminogen activator receptor expression and associated functions in ovarian carcinoma. British Journal of Cancer 2005;92:1475–85]. Our data incorporating the clinical samples and the cancer cell lines is the first to demonstrate that JAK2/STAT3 pathway may be one of the downstream events in EMT-like process and α6β1 integrin-mediated signalling in ovarian carcinomas.

Exercise-induced histone modifications in human skeletal muscle

Skeletal muscle adaptations to exercise confer many of the health benefits of physical activity and occur partly through alterations in skeletal muscle gene expression. The exact mechanisms mediating altered skeletal muscle gene expression in response to exercise are unknown. However, in recent years, chromatin remodelling through epigenetic histone modifications has emerged as a key regulatory mechanism controlling gene expression in general. The purpose of this study was to examine the effect of exercise on global histone modifications that mediate chromatin remodelling and transcriptional activation in human skeletal muscle in response to exercise. In addition, we sought to examine the signalling mechanisms regulating these processes. Following 60 min of cycling, global histone 3 acetylation at lysine 9 and 14, a modification associated with transcriptional initiation, was unchanged from basal levels, but was increased at lysine 36, a site associated with transcriptional elongation. We examined the regulation of the class IIa histone deacetylases (HDACs), which are enzymes that suppress histone acetylation and have been implicated in the adaptations to exercise. While we found no evidence of proteasomal degradation of the class IIa HDACs, we found that HDAC4 and 5 were exported from the nucleus during exercise, thereby removing their transcriptional repressive function. We also observed activation of the AMP-activated protein kinase (AMPK) and the calcium–calmodulin-dependent protein kinase II (CaMKII) in response to exercise, which are two kinases that induce phosphorylation-dependent class IIa HDAC nuclear export. These data delineate a signalling pathway that might mediate skeletal muscle adaptations in response to exercise.

Unusual spectroscopic properties of PPE/TiO2 composite and its sensor response to TNT

A composite from a broad bandgap polymer, poly(phenylene ethylene) (PPE), and nano-sized TiO₂ particles was found to be able to sense 2,4,6-trinitrotoluene (TNT) for TNT sensor. Fluorescence quenching induced by charge transfer from PPE to nano-sized TiO₂ was observed in toluene solution. At high TiO₂ composition, a strong exciplex band occurred at 550 nm. Under prolonged light irradiation at 400 nm, unusual fluorescence gains took place at 460 nm, companied with a very small change in the UV–vis absorbance. After 30 min light irradiation, the fluorescence at 460 nm reached a maximum, but the peak at 550 nm disappeared. This composite showed amplified sensor response to TNT compared to the pristine PPE film, which can be potentially used as sensing material for detecting TNT based explosives.

Transient response analysis of steel in concrete

A new approach is presented for the analysis of galvanostatically induced transients allowing for the rapid evaluation of the corrosion activity of steel in concrete. This method of analysis is based on the iterative fitting of a non-exponential model based on a modified KWW (Kohlrausch–Williams–Watt) formalism. This analysis yields values for the parameters related to corrosion such as the concrete resistance, polarisation resistance, interfacial capacitance and β, the non-ideality exponent.

Testosterone manipulation post-castration does not alter cloacal gland growth differences in male quail selected for divergent plasma corticosterone stress response

Japanese quail selected for reduced (low-stress, LS) rather than exaggerated (high-stress, HS) plasma corticosterone response to brief restraint have consistently shown greater cloacal gland (CG) development, an androgen-dependent trait. In this study, the effects of testosterone implants on levels of plasma testosterone and CG development in castrated LS and HS quail were determined. Stress-line males were castrated and randomly allocated to 1 of 3 testosterone treatments: the empty testosterone (ET), low testosterone (LT), or high testosterone (HT) implant group. Cloacal gland volume was determined at 4 weekly intervals that represented ranges of 1 to 9 d, 8 to 17 d, 15 to 24 d, and 22 to 31 d after castration and testosterone implantation. Levels of plasma testosterone were also assessed at the end of the study. Development of the CG was affected by quail line (LS > HS), testosterone treatment (HT > LT > ET), and time of measurement (1 to 9 d < 8 to 17 d < 15 to 24 d = 22 to 31 d after castration and testosterone implantation). A significant interaction between testosterone treatment and time of measurement on CG volume was also detected (with CG volume generally increasing with time in LT- and HT-treated quail, but not in ET-treated quail). However, even though HT implant treatments induced higher CG development than did LT treatments beyond the first interval of CG volume measurement, and despite the finding of greater CG volumes in LS than HS quail during the last 2 measurement intervals within each of the LT and HT groups, no interaction was observed between testosterone implant dosages and quail stress line on CG volume. Thus, by the end of the study, regardless of testosterone dose, CG volume was consistently greater in LS quail than in their HS counterparts. In addition, although, as expected, the testosterone implant treatment significantly altered levels of plasma testosterone (HT > LT > ET), neither quail line nor its interaction with testosterone treatment affected plasma testosterone. The present findings suggest that the often-observed depressed CG development in the HS line may be independent of testosterone effects

Dorsal and ventral medullary catecholamine cell groups contribute differentially to systemic interleukin-1beta-induced hypothalamic pituitary adrenal axis responses

Medial parvocellular paraventricular corticotropin-releasing hormone (mPVN CRH) cells are critical in generating hypothalamic-pituitary-adrenal (HPA) axis responses to systemic interleukin-1beta (IL-1beta). However, although it is understood that catecholamine inputs are important in initiating mPVN CRH cell responses to IL-1beta, the contributions of distinct brainstem catecholamine cell groups are not known. We examined the role of nucleus tractus solitarius (NTS) and ventrolateral medulla (VLM) catecholamine cells in the activation of mPVN CRH, hypothalamic oxytocin (OT) and central amygdala cells in response to IL-1beta (1 microg/kg, i.a.). Immunolabelling for the expression of c-fos was used as a marker of neuronal activation in combination with appropriate cytoplasmic phenotypic markers. First we confirmed that PVN 6-hydroxydopamine lesions, which selectively depleted catecholaminergic terminals, significantly reduced IL-1beta-induced mPVN CRH cell activation. The contribution of VLM (A1/C1 cells) versus NTS (A2 cells) catecholamine cells to mPVN CRH cell responses was then examined by placing ibotenic acid lesions in either the VLM or NTS. The precise positioning of these lesions was guided by prior retrograde tracing studies in which we mapped the location of IL-1beta-activated VLM and NTS cells that project to the mPVN. Both VLM and NTS lesions reduced the mPVN CRH and OT cell responses to IL-1beta. Unlike VLM lesions, NTS lesions also suppressed the recruitment of central amygdala neurons. These studies provide novel evidence that both the NTS and VLM catecholamine cells have important, but differential, contributions to the generation of IL-1beta-induced HPA axis responses.

Medial prefrontal cortex suppression of the hypothalamic–pituitary–adrenal axis response to a physical stressor, systemic delivery of interleukin-1β

Previous studies have shown that the medial prefrontal cortex can suppress the hypothalamic–pituitary–adrenal axis response to stress. However, this effect appears to vary with the type of stressor. Furthermore, the absence of direct projections between the medial prefrontal cortex and corticotropin-releasing factor cells at the apex of the hypothalamic–pituitary–adrenal axis suggest that other brain regions must act as a relay when this inhibitory mechanism is activated. In the present study, we first established that electrolytic lesions involving the prelimbic and infralimbic medial prefrontal cortex increased plasma adrenocorticotropic hormone levels seen in response to a physical stressor, the systemic delivery of interleukin-1β. However, medial prefrontal cortex lesions did not alter plasma adrenocorticotropic hormone levels seen in response to a psychological stressor, noise. To identify brain regions that might mediate the effect of medial prefrontal cortex lesions on hypothalamic–pituitary–adrenal axis responses to systemic interleukin-1β, we next mapped the effects of similar lesions on interleukin-1β-induced Fos expression in regions previously shown to regulate the hypothalamic–pituitary–adrenal axis response to this stressor. It was found that medial prefrontal cortex lesions reduced the number of Fos-positive cells in the ventral aspect of the bed nucleus of the stria terminalis. However, the final experiment, which involved combining retrograde tracing with Fos immunolabelling, revealed that bed nucleus of the stria terminalis-projecting medial prefrontal cortex neurons were largely separate from medial prefrontal cortex neurons recruited by systemic interleukin-1β, an outcome that is difficult to reconcile with a simple medial prefrontal cortex–bed nucleus of the stria terminalis–corticotropin-releasing factor cell control circuit.

Fructose-1,6-Bisphosphatase overexpression in pancreatic [beta]-cells results in reduced insulin secretion : a new mechanism for fat-induced impairment of [beta]-cell function

Fructose-1,6-bisphosphatase (FBPase) is a gluconeogenic enzyme that is upregulated in islets or pancreatic beta-cell lines exposed to high fat. However, whether specific beta-cell upregulation of FBPase can impair insulin secretory function is not known. The objective of this study therefore is to determine whether a specific increase in islet beta-cell FBPase can result in reduced glucose-mediated insulin secretion.

To test this hypothesis, we have generated three transgenic mouse lines overexpressing the human FBPase (huFBPase) gene specifically in pancreatic islet beta-cells. In addition, to investigate the biochemical mechanism by which elevated FBPase affects insulin secretion, we made two pancreatic beta-cell lines (MIN6) stably overexpressing huFBPase.

FBPase transgenic mice showed reduced insulin secretion in response to an intravenous glucose bolus. Compared with the untransfected parental MIN6, FBPase-overexpressing cells showed a decreased cell proliferation rate and significantly depressed glucose-induced insulin secretion. These defects were associated with a decrease in the rate of glucose utilization, resulting in reduced cellular ATP levels.

Taken together, these results suggest that upregulation of FBPase in pancreatic islet beta-cells, as occurs in states of lipid oversupply and type 2 diabetes, contributes to insulin secretory dysfunction.

Corneal staining as a response to contact lens wear

Objective: To review the effects of contact lenses on the corneal surface.

Methods: A review of the literature and in-house research of corneal staining and its various forms of presentation.

Results: Corneal staining manifests in many different forms. The severity of staining or insult of the cornea is usually determined by the extent (area of coverage), density, and depth. The cause of staining is multifactorial, and its location is often linked to the type of lens that is being worn, the solution used to clean/disinfect the lens, the state of hydration of the soft lens, and the state of the cornea that has been affected by the lens.

Conclusions: Sodium fluorescein dye effectively highlights corneal integrity changes referred to as corneal staining. This review describes the manifestations, the cause, the mechanisms, and the methods of remediation of corneal staining.

1H-NMR and mass spectrometric characterization of the metabolic response of juvenile Atlantic salmon (Salmo salar) to long-term handling stress

Stressors of various kinds constantly affect fish both in the wild and in culture, examples being acute water temperature and quality changes, predation, handling, and confinement. Known physiological responses of fish to stress such as increases in plasma cortisol and glucose levels, are considered to be adaptive, allowing the animal to cope in the short term. Prolonged exposure to stressors however, has the potential to affect growth, immune function, and survival. Nonetheless, little is known about the mechanisms underlying the long-term stress response. We have investigated the metabolic response of juvenile Atlantic salmon (Salmo salar) to long-term handling stress by analyzing fish plasma via ¹H nuclear magnetic resonance spectroscopy and ultra high performance liquid chromatography–mass spectrometry (UPLC–MS), and comparing results with controls. Analysis of NMR data indicated a difference in the metabolic profiles of control and stressed fish after 1 week of stress with a maximum difference observed after 2 weeks. These differences were associated with stress-induced increases in phosphatidyl choline, lactate, carbohydrates, alanine, valine and trimethylamine-N-oxide, and decreases in low density lipoprotein, very low density lipoprotein, and lipid. UPLC-MS data showed differences at week 2, associated with another set of compounds, tentatively identified on the basis of their mass/charge. Overall the results provided a multi-faceted view of the response of fish to long-term handling stress, indicating that the metabolic disparity between the control and stress groups increased to week 2, but declined by weeks 3 and 4, and revealed several new molecular indicators of long-term stress.

Population consequences of a predator-induced habitat shift by trout in whole-lake experiments

In a replicated whole-lake experiment, we (a) tested for the existence of a flexible habitat shift in response to predator presence in age-0 rainbow trout (Oncorhynchus mykiss) at risk of cannibalism and (b) evaluated the population-level consequences of habitat shifts in terms of growth and survival over their first growing season. Daphnid food and adult trout predators were substantially more abundant in pelagic than in littoral habitats. Age-0 trout used all habitats in populations without adult trout predators, whereas age-0 trout were observed only in the less profitable littoral habitat in populations with adult trout. Consequently, mean fall mass of age-0 trout in the presence of predators was almost half that observed in populations without adult trout. Despite the shift in habitat use, age-0 trout experienced 90% mortality when adult trout predators were present, in comparison to only 36% mortality when absent. We conclude that the commonly observed habitat shifts by fish at risk of predation, observed at smaller scales, do in fact occur at the whole-system scale over long time intervals. These results suggest that fish are able to perceive risk at large spatial scales and thus take advantage of profitable (but normally risky) habitats when predators are absent, or move to less profitable refuge habitats when predators are present.

cJUN N-terminal kinase (JNK) activation mediates islet amyloid-induced beta cell apoptosis in cultured human islet amyloid polypeptide transgenic mouse islets

Aims/hypothesis

Aggregation of human islet amyloid polypeptide (hIAPP) as islet amyloid is associated with increased beta cell apoptosis and reduced beta cell mass in type 2 diabetes. Islet amyloid formation induces oxidative stress, which contributes to beta cell apoptosis. The cJUN N-terminal kinase (JNK) pathway is a critical mediator of beta cell apoptosis in response to stress stimuli including oxidative stress and exogenous application of hIAPP. We determined whether amyloid formation by endogenous hIAPP mediates beta cell apoptosis through JNK activation and downstream signalling pathways.
Methods

hIAPP transgenic and non-transgenic mouse islets were cultured for up to 144 h in 16.7 mmol/l glucose to induce islet amyloid in the presence or absence of the amyloid inhibitor Congo Red or a cell-permeable JNK inhibitor. Amyloid, beta cell apoptosis, JNK signalling and activation of downstream targets in the intrinsic and extrinsic apoptotic pathways were measured.
Results

JNK activation occurred with islet amyloid formation in hIAPP transgenic islets after 48 and 144 h in culture. Neither high glucose nor the hIAPP transgene alone was sufficient to activate JNK independent of islet amyloid. Inhibition of islet amyloid formation with Congo Red reduced beta cell apoptosis and partially decreased JNK activation. JNK inhibitor treatment reduced beta cell apoptosis without affecting islet amyloid. Islet amyloid increased mRNA levels of markers of the extrinsic (Fas, Fadd) and intrinsic (Bim [also known as Bcl2l11]) apoptotic pathways, caspase 3 and the anti-apoptotic molecule Bclxl (also known as Bcl2l1) in a JNK-dependent manner.
Conclusions/interpretation

Islet amyloid formation induces JNK activation, which upregulates predominantly pro-apoptotic signals in both extrinsic and intrinsic pathways, resulting in beta cell apoptosis.

Acclimation to captivity and GnRHa-induced spawning of meagre (Argyrosomus regius)

Twelve meagre (Argyrosomus regius) of mean weight 20 ± 7 kg were captured and acclimatized to captivity using environmental control and de-parasitizing protocols. An active feeding response and the occurrence of sexual maturation to advanced stages indicated that the fish adapted well to captivity. Spawning was induced with hormonal treatments, two GnRHa-injected females spawned 7,221,000 floating eggs (in 3 spawns) and two GnRHa-implanted females spawned 9,515,000 eggs (in 9 spawns), with a mean ± SD hatching percentage of 45 ± 29 and 64 ± 28, respectively.