Lorcaserin, a 5-HT2C agonist, decreases nicotine self-administration in female rats

Lorcaserin, a selective 5-hydroxytryptamine(2C) (5-HT(2C)) agonist, has been shown to facilitate weight loss in obese populations. It was assessed for its efficacy in reducing nicotine self-administration in young adult female Sprague-Dawley rats. The effect of short-term doses (subcutaneous) on nicotine self-administration (0.03 mg/kg per infusion) with a fixed ratio 1 schedule was assessed in 3-h sessions. Short-term lorcaserin doses (0.3125-20 mg/kg) were administered in a counterbalanced order. Significant reduction of nicotine self-administration was achieved with all of the short-term doses in this range. Tests of lorcaserin on locomotor activity detected prominent sedative effects at doses greater than 1.25 mg/kg with more modest transient effects seen at 0.625 to 1.25 mg/kg. Long-term effects of lorcaserin on locomotor activity were tested with repeated injections with 0.625 mg/kg lorcaserin 10 times over 2 weeks. This low lorcaserin dose did not cause an overall change in locomotor activity relative to that of saline-injected controls. Long-term lorcaserin (0.625 mg/kg) significantly reduced nicotine self-administration over a 2-week period of repeated injections. Long-term lorcaserin at this same dose had no significant effects on food self-administration over the same 2-week period of repeated injections. These studies support development of the 5-HT(2C) agonist lorcaserin to aid tobacco smoking cessation.     

Recombinant rubistatin (r-Rub), an MVD disintegrin, inhibits cell migration and proliferation, and is a strong apoptotic inducer of the human melanoma cell line SK-Mel-28

Disintegrins are low molecular weight peptides isolated from viper venom. These peptides bind to integrin receptors using a conserved binding motif sequence containing an RGD or similar motif. As a consequence, disintegrins can inhibit platelet aggregation and inhibit cell migration, proliferation, and initiate apoptosis in cancer cell lines. Rubistatin is a MVD disintegrin cloned from a Crotalus ruber ruber venom gland. The biological activity of MVD disintegrins is poorly understood. Recombinant rubistatin (r-Rub) was cloned into a pET32b plasmid and expressed in reductase-deficient Escherichia coli. Expression was induced with IPTG and the resulting fusion peptide was affinity purified, followed by thrombin cleavage, and removal of vector coded sequences. r-Rub peptide inhibited ADP-induced platelet aggregation by 54% ± 6.38 in whole blood. We assessed the ability of r-Rub to initiate apoptosis in three human cancer cell lines. Cultures of SK-Mel-28, HeLA, and T24 cells were grown for 24 h with 2.5 μM r-Rub followed by Hoechst staining. Chromatin fragmentation was observed in treated SK-Mel-28, but not in T24 or HeLA cells. A TUNEL assay revealed that 51.55% ± 5.28 of SK-Mel-28 cells were apoptotic after 18 h of treatment with 3.5 μM of r-Rub. Cell migration and proliferation assays were performed in order to further characterize the biological effects of r-Rub on SK-Mel-28 cells. At 3 μM, r-Rub inhibited cell migration by 44.4% ± 0.5, while at 3.5 μM it was able to inhibit cell proliferation by 83% ± 6.0.     

Experimental and computational analysis of soft tissue stiffness in forearm using a manual indentation device

A hand held stiffness meter can be used to measure indentation stiffness of human soft tissues, sensitively altered, e.g., by pathological tissue swelling. Under indentation load, the relative contribution of each soft tissue component (i.e., skin, adipose tissue and muscle) to the biomechanical response is not known. In the present study, we evaluated the biomechanical role of different soft tissues in relaxed, physically stressed and oedemic human forearm. Soft tissue stiffness of the forearms of nine healthy human subjects was measured under four different test protocols: (1) forearm at rest, (2) forearm under isometric flexor loading, (3) forearm under isometric extensor loading, and (4) forearm under venous occlusion. In (2) and (3) the loading forces were monitored using a dynamometer, and in (4) the soft tissue swelling was induced by venous occlusion using a pressure cuff. At the site of indentation, thickness of different tissue layers (skin, adipose tissue and muscle) was measured using B-mode ultrasound imaging. Layered, hyperelastic finite element (FE) model of the indentation measurement was created and the model response was matched with that of the stiffness meter to determine the elastic modulus for each tissue in the model. Optimized values of the elastic modulus for skin and adipose tissue at rest were 210 kPa and 1.9 kPa, respectively. Further, significance of the variations in stiffness of different tissues on the indentation response was tested. Experimentally, indentation stiffness of the forearm increased during isometric extensor and flexor loads as well as under venous occlusion by 53, 91 and 15%, respectively. The FE model could reproduce the experimental responses primarily by the increased modulus of skin; 112% (446 kPa), 210% (651 kPa) and 21% (254 kPa) under flexor and extensor loading as well as during venous occlusion, respectively. The indentation response was 9–16 times more sensitive to changes in the mechanical properties of skin than those of adipose tissue and muscle. In conclusion, the present stiffness meter may be used to quantify in vivo mechanical properties of soft tissues in the forearm, sensitively modulated by soft tissue swelling and muscle loading.     

Corticospinal output and cortical excitation-inhibition balance in distal hand muscle representations in nonprimary motor area

Transcranial magnetic stimulation (TMS) of the superior frontal gyrus in the non-primary motor area (NPMA) can evoke motor-evoked potentials (MEPs) at 20 ms latency range in contralateral distal hand muscles similar to stimulation of M1 and indicating monosynaptic corticospinal tracts. We compared the intracortical inhibitory and excitatory balance in primary motor cortex (M1) and in NPMA by navigated single- and paired-pulse TMS (ppTMS). We also evaluated the spatial stability of muscle representations in M1 and NPMA by remapping 11 healthy subjects one year after the initial mapping. Resting motor threshold (rMT) was higher in NPMA than in M1 as were the MEP amplitudes evoked by 120% rMT stimulation intensity of the local MT. Short-interval intracortical inhibition (SICI) was significantly weaker in NPMA than in M1 at ISI of 2 ms and conditioning stimulus (CS) 80% rMT. Our findings suggest that the cortical hand representations in NPMA 1) are connected to lower motoneurons monosynaptically, 2) are less strictly organized, i.e. motoneuron population representing a discrete hand muscle is sparser and less dense than in M1 and 3) have the capacity to generate powerful, rapid muscle contraction if sufficient number of motoneurones are activated. In NPMA, local intracortical inhibitory and excitatory activity is mainly similar to that in M1. The lower SICI in NPMA at an ISI of 2 ms may reflect less strict topographic organization and readiness to reorganization of neural circuits during motor learning or after motor deficits.     

Short- and intermediate-interval cortical inhibition and facilitation assessed by navigated transcranial magnetic stimulation

One of the most widely utilized in vitro models of ischemia or oxygen glucose deprivation (OGD) is the hippocampal organotypical culture (HOTC). The HOTC is used not only for the study of the mechanisms of cell death, but also has been the cornerstone of synaptic physiology. Although the intact nature of the HOTC is one of its primary advantages, some studies require a dissociated preparation in order to distinguish cell type specific responses. Typically, primary dissociated neuronal cultures are prepared from embryonic tissue. Since the HOTC is prepared from postnatal pups, we wanted to establish a primary culture of hippocampus from postnatal pups to parallel our studies in the HOTC preparation. Mixed cultures were prepared by enzymatic dissociation of hippocampus from 7-day-old mouse pups. These cultures responded to OGD with a time course of delayed cell death that was similar to that reported in HOTC. Dual label immunocytochemical staining revealed that neurons, but not astrocytes, were dying from apoptosis following OGD. To examine this vulnerability further, we also prepared neuronal enriched cultures by treating mixed cultures with cytosine-β-d-arabinofuranoside (CBA). These neuronal cultures appear to be even more sensitive to OGD. In addition, we have established primary astrocyte-enriched cultures from the same age pups to examine the vulnerability of astrocytes to OGD. These three culture preparations are useful for comparison of the responses of the two major cell types in the same culture, and the enriched cultures will allow biochemical, electrophysiological and molecular studies of homogenous cell populations.     

Motor cortical plasticity is impaired in Unverricht–Lundborg disease

Patients with Unverricht–Lundborg disease, also referred to as progressive myoclonus epilepsy type 1, exhibit widespread motor symptoms and signs in addition to epileptic seizures, which suggest abnormal excitability of the primary motor pathways. To explore the plasticity of the sensory–motor cortex, we employed a modern neurophysiological method, the paired associative stimulation protocol, which resembles the concept of long‐term potentiation of experimental studies. Seven patients with genetically verified Unverricht–Lundborg disease and 13 healthy control subjects were enrolled in the study to characterize cortical sensory–motor plasticity. In the study protocol, peripheral electric median nerve stimulation preceded navigated transcranial magnetic stimulation targeted to the representation area of thenar musculature on the contralateral primary motor cortex. The protocol consisted of 132 transcranial magnetic stimulation trials at 0.2 Hz, preceded by peripheral sensory stimulation at 25 ms. Motor‐evoked potential amplitudes were analyzed at baseline and after the paired associative stimulation protocol at an intensity of 130% of the individual motor threshold. The patients with Unverricht–Lundborg disease exhibited an average decrease of 15% in motor‐evoked potential amplitudes 30 minutes after paired associative stimulation, whereas in the control subjects, a significant increase (101%) was observed (P < .05), as expected. The results indicate a lack of normal cortical plasticity in Unverricht–Lundborg disease, which stresses the role of abnormal motor cortical functions or sensorimotor integration as possible pathophysiological contributors to the motor symptoms. The impaired cortical plasticity may be associated with the previously reported structural and physiological abnormalities of the primary motor cortex. © 2011 Movement Disorder Society     

Effects of growth and exercise on composition, structural maturation and appearance of osteoarthritis in articular cartilage of hamsters

Articular cartilage composition and structure are maintained and remodeled by chondrocytes under the influence of loading. Exercise‐induced changes in the composition, structure, mechanical properties and tissue integrity of growing and aging hamster articular cartilage were investigated. Articular cartilage samples (n = 191) were harvested from the proximal tibiae of hamsters aged 1, 3, 6, 12 and 15 months. The hamsters were divided into runners and controls. The runners had free access to a running wheel between 1 and 3 months (runner groups 3‐, 12‐ and 15‐month‐old hamsters) or 1 and 6 months (runner group 6‐month‐old hamsters) of age. Control animals were subjected to a sedentary lifestyle. Mechanical indentation tests and depth‐wise compositional and structural analyses were performed for the cartilage samples. Furthermore, the integrity of articular cartilage was assessed using histological osteoarthritis grading. Exercise affected the collagen network organization after a 5‐month exercise period, especially in the middle and deep zones. However, no effect on the mechanical properties was detected after exercise. Before the age of 12 months, the runners showed less osteoarthritis than the controls, whereas at 15 months of age the situation was reversed. It is concluded that, in hamsters, physical exercise at a young age enhances cartilage maturation and alters the depth‐wise cartilage structure and composition. This may be considered beneficial. However, exercise at a young age demonstrated adverse effects on cartilage at a later age with a significant increase in the incidence of osteoarthritis.