Cytogenetic analysis reveals clonal proliferation of smooth muscle cells in atherosclerotic plaques

Summary Cytogenetic analysis of primary cell cultures from human atherosclerotic fibrous plaques revealed clonal chromosome abnormalities in 13 of the 18 cases studied. Loss of the Y chromosome and del(13)(q14) were present as single clonal abnormalities in eight cases; in five cases separate clones were found involving loss of the Y and a XXY karyotype, trisomy 10 and 18, loss of the Y and trisomy 7. A variety of single numerical and structural abnormalities were present in all but two of the 18 cases. Immunocytochemical studies were performed on cells from the same cultures used for cytogenetic analysis using monoclonal antibodies to human leucocyte common antigen, to human vimentin and to muscle actin. The immunoreactivity was positive for actin in 70–80% of the cells; 100% of the cells were positive for vimentin and all cells were ALC negative. These results indicated that the chromosomal abnormalities are present in the smooth muscle cells of the plaque. The hypothesis is proposed that the proliferation leading to the atherosclerotic lesion may primarily represent a hyperplastic response to mechanical and biological injuries and that this reactive proliferation is, in turn, associated with a tendency to chromosome instability.     

Distribution and abundance of fish larvae in the northern Ionian Sea (Eastern Mediterranean)

The purpose of this paper was to study the spatial distribution, abundance and composition of fish larvae in the northern Ionian Sea. Samples were collected to the 600 m depth with an electronic multinet BIONESS during the “INTERREG Italia-Grecia” oceanographic cruise carried out in March 2000 off the Apulian Italian coast. A total of 46 species of teleost early stages were collected, belonging to 38 genera and 22 families. Over 52% of the larvae identified were mesopelagic species, almost 27% were demersal and about 21% pelagic. A total of 307 myctophids, 69 clupeids and 61 gadid post-larvae dominated the community. Benthosema glaciale (mean 6.1 mm SL) was the most abundant species (21.6%), the most frequent in the samples (28.8%), and dominant in the whole study area (mean 1.4 ind/100 m³). Particular attention was given to the horizontal and vertical distribution and abundance of the three dominant post-larval species: Benthosema glaciale, Sprattus sprattus sprattus and Notoscopelus elongatus. The Pearson coefficient (R = 0.734) showed a high correlation between total zooplankton and fish larval assemblages in terms of spatial distribution abundance values. Regarding the vertical distribution of fish larvae, Sorensen’s index (S = 0.69) showed that fish larvae and total zooplankton abundance peaks co-occurred along the water column.     

Dystrophin is required for the normal function of the cardio-protective K(ATP) channel in cardiomyocytes

Duchenne and Becker muscular dystrophy patients often develop a cardiomyopathy for which the pathogenesis is still unknown. We have employed the murine animal model of Duchenne muscular dystrophy (mdx), which develops a cardiomyopathy that includes some characteristics of the human disease, to study the molecular basis of this pathology. Here we show that the mdx mouse heart has defects consistent with alteration in compounds that regulate energy homeostasis including a marked decrease in creatine-phosphate (PC). In addition, the mdx heart is more susceptible to anoxia than controls. Since the cardio-protective ATP sensitive potassium channel (K(ATP)) complex and PC have been shown to interact we investigated whether deficits in PC levels correlate with other molecular events including K(ATP) ion channel complex presence, its functionality and interaction with dystrophin. We found that this channel complex is present in the dystrophic cardiac cell membrane but its ability to sense a drop in the intracellular ATP concentration and consequently open is compromised by the absence of dystrophin. We further demonstrate that the creatine kinase muscle isoform (CKm) is displaced from the plasma membrane of the mdx cardiac cells. Considering that CKm is a determinant of K(ATP) channel complex function we hypothesize that dystrophin acts as a scaffolding protein organizing the K(ATP) channel complex and the enzymes necessary for its correct functioning. Therefore, the lack of proper functioning of the cardio-protective K(ATP) system in the mdx cardiomyocytes may be part of the mechanism contributing to development of cardiac disease in dystrophic patients.     

Short-term changes in zooplankton community in Paso Ancho basin (Strait of Magellan): functional trophic structure and diel vertical migration

Knowledge on community structure oriented to describe energy flow during late summer season in Paso Ancho basin (Strait of Magellan) is scarce and particularly affected by vertical diel migration (VDM). The main aim of this work is to identify the VDM patterns of selected species and functional feeding groups of mesozooplankton, collected by the electronic multinet BIONESS in 1995. Detailed studies were carried out on keystone components of the community: 7 species of copepods (Ctenocalanus citer, Drepanopus forcipatus, Metridia lucens, Clausocalanus brevipes, Scolecithricella minor, Paraeuchaeta antarctica, Calanus simillimus), one ostracod (Discoconchoecia elegans), one chaetognath (Sagitta tasmanica), one euphausiid (Euphausia vallentini), and two polychaetes (Pelagobia longicirrata, Tomopteris planktonis). Unexpected deviations from the classic pattern reported in literature were uncovered. The shallow layer mesozooplankton at night, although dominated by exclusively or preferentially herbivores, particularly by E. vallentini, was not represented by several species known as herbivores that remained in the deep layer throughout the day cycle. The deep-layer zooplankton throughout the day cycle was well represented by carnivores, detritivores, and omnivores. It is suggested that during low-chlorophyll summer conditions, the composition of functional groups and diet, and VDM patterns changed to take advantage of sinking phytoplankton and picoplankton in deep layers. Pelagic-benthic coupling would be strengthened due to animals that suppressed their vertical daily rise to the shallow layer at nights but remained in the deep layers to feed on a rain of particulate organic matter and other non-migrant zooplankton.     

Validity and reliability of a new in vivo ankle stiffness measurement device

This investigation was designed to test the validity and reliability of a new measure of inversion/eversion ankle stiffness on a unique medial/lateral swaying cradle device utilizing a test/retest with comparison to a known standard. Ankle stiffness is essential to maintaining joint stability. Most ankle injuries occur via an inversion mechanism. To date, very little information is available regarding stiffness of the evertor muscles in the prevention of excessive inversion joint rotation. Transient oscillation data representing inversion/eversion stiffness was obtained in a bipedal weight-bearing stance with an upright posture. Using commercially available springs with stiffness of 4.80N/cm the measured value recorded by the cradle was 4.87N/cm. Mean active stiffness values of the ankle were 35.70Nm/cm (SD 9.45). The trial-to-trial reliability ICC (2,1) coefficient was 0.96 with an SEM of 2.05Nm/rad, and the day-to-day reliability ICC (2,k) coefficient was 0.93 and an SEM of 3.00Nm/rad. The results demonstrate that inversion/eversion ankle stiffness measures on this device are a valid, repeatable and consistent measure. This is relevant because the ability to accurately quantify inversion/eversion ankle stiffness will improve our understanding of biomechanical stability and factors that influence it. It will also enable identification of ankle injury risk factors that will lead to more efficient rehabilitation programs and injury prevention strategies.     

The dystonia-associated protein torsinA modulates synaptic vesicle recycling

The loss of a glutamic acid residue in the AAA-ATPase (ATPases associated with diverse cellular activities) torsinA is responsible for most cases of early onset autosomal dominant primary dystonia. In this study, we found that snapin, which binds SNAP-25 (synaptosome-associated protein of 25,000 Da) and enhances the association of the SNARE complex with synaptotagmin, is an interacting partner for both wild type and mutant torsinA. Snapin co-localized with endogenous torsinA on dense core granules in PC12 cells and was recruited to perinuclear inclusions containing mutant DeltaE-torsinA in neuroblastoma SH-SY5Y cells. In view of these observations, synaptic vesicle recycling was analyzed using the lipophilic dye FM1-43 and an antibody directed against an intravesicular epitope of synaptotagmin I. We found that overexpression of wild type torsinA negatively affects synaptic vesicle endocytosis. Conversely, overexpression of DeltaE-torsinA in neuroblastoma cells increases FM1-43 uptake. Knockdown of snapin and/or torsinA using small interfering RNAs had a similar inhibitory effect on the exo-endocytic process. In addition, down-regulation of torsinA causes the persistence of synaptotagmin I on the plasma membrane, which closely resembles the effect observed by the overexpression of the DeltaE-torsinA mutant. Altogether, these findings suggest that torsinA plays a role together with snapin in regulated exocytosis and that DeltaE-torsinA exerts its pathological effects through a loss of function mechanism. This may affect neuronal uptake of neurotransmitters, such as dopamine, playing a role in the development of dystonic movements.     

Comparative,double-blind,controlled study of intra-articular hyaluronic acid (Hyalubrix?) injections versus local anesthetic in osteoarthritis of the hip

Introduction

Comparison of intra-articular bacterial-derived hyaluronic acid (Hyalubrix^®) (HA) with local analgesia (mepivacaine) for osteoarthritis (OA) of the hip.

Methods

A pilot prospective, double-blind, 6-month randomized trial of 42 patients with hip OA. HA or mepivacaine was administered twice (once a month) under ultrasound guidance. Efficacy measurements included the Lequesne''s algofunctional index, a visual analog scale for pain, concomitant use of analgesia, patient and physician global measurement, and safety.

Results

Patients in the HA group exhibited a significantly reduced Lequesne''s algofunctional index 3 and 6 months after treatment (P < 0.001) and significantly reduced visual analog scale pain scores 3 and 6 months after treatment (P < 0.05) compared with the local anesthetic group. All primary and secondary measures were significantly improved versus baseline, but other than the above were not different from each other at 3 or 6 months. Adverse effects were minimal.

Conclusions

This comparative study suggests a beneficial effect and safety of intra-articular HA in the management of hip OA.

Trial registration number

ISRCTN39397064.     

The viscoelastic standard nonlinear solid model: predicting the response of the lumbar intervertebral disk to low-frequency vibrations

Due to the mathematical complexity of current musculoskeletal spine models, there is a need for computationally efficient models of the intervertebral disk (IVD). The aim of this study is to develop a mathematical model that will adequately describe the motion of the IVD under axial cyclic loading as well as maintain computational efficiency for use in future musculoskeletal spine models. Several studies have successfully modeled the creep characteristics of the IVD using the three-parameter viscoelastic standard linear solid (SLS) model. However, when the SLS model is subjected to cyclic loading, it underestimates the load relaxation, the cyclic modulus, and the hysteresis of the human lumbar IVD. A viscoelastic standard nonlinear solid (SNS) model was used to predict the response of the human lumbar IVD subjected to low-frequency vibration. Nonlinear behavior of the SNS model was simulated by a strain-dependent elastic modulus on the SLS model. Parameters of the SNS model were estimated from experimental load deformation and stress-relaxation curves obtained from the literature. The SNS model was able to predict the cyclic modulus of the IVD at frequencies of 0.01 Hz, 0.1 Hz, and 1 Hz. Furthermore, the SNS model was able to quantitatively predict the load relaxation at a frequency of 0.01 Hz. However, model performance was unsatisfactory when predicting load relaxation and hysteresis at higher frequencies (0.1 Hz and 1 Hz). The SLS model of the lumbar IVD may require strain-dependent elastic and viscous behavior to represent the dynamic response to compressive strain.     

Obestatin regulates cardiovascular function and promotes cardioprotection through the nitric oxide pathway

Patients with ischaemic heart disease or chronic heart failure show altered levels of obestatin, suggesting a role for this peptide in human heart function. We have previously demonstrated that GH secretagogues and the ghrelin gene‐derived peptides, including obestatin, exert cardiovascular effects by modulating cardiac inotropism and vascular tone, and reducing cell death and contractile dysfunction in hearts subjected to ischaemia/reperfusion (I/R), through the Akt/nitric oxide (NO) pathway. However, the mechanisms underlying the cardiac actions of obestatin remain largely unknown. Thus, we suggested that obestatin‐induced activation of PI3K/Akt/NO and PKG signalling is implicated in protection of the myocardium when challenged by adrenergic, endothelinergic or I/R stress. We show that obestatin exerts an inhibitory tone on the performance of rat papillary muscle in both basal conditions and under β‐adrenergic overstimulation, through endothelial‐dependent NO/cGMP/PKG signalling. This pathway was also involved in the vasodilator effect of the peptide, used both alone and under stress induced by endothelin‐1. Moreover, when infused during early reperfusion, obestatin reduced infarct size in isolated I/R rat hearts, through an NO/PKG pathway, comprising ROS/PKC signalling, and converging on mitochondrial ATP‐sensitive potassium [mitoK(ATP)] channels. Overall, our results suggest that obestatin regulates cardiovascular function in stress conditions and induces cardioprotection by mechanisms dependent on activation of an NO/soluble guanylate cyclase (sGC)/PKG pathway. In fact, obestatin counteracts exaggerated β‐adrenergic and endothelin‐1 activity, relevant factors in heart failure, suggesting multiple positive effects of the peptide, including the lowering of cardiac afterload, thus representing a potential candidate in pharmacological post‐conditioning.