ROLE OF CCL2 AND ITS RECEPTORS CCR2 AND D6 IN THE ACTIVATION AND POLARIZATION OF TUMOR-ASSOCIATED MACROPHAGES

Chemokines are well known to play a major role in tumor progression and metastasis. In particular CCL2 is over-expressed in several human cancers and their higher levels correlate with poor prognosis and shorter outcomes. Here we reported two different studies in which CCL2 receptors, the canonical CCR2 and the atypical D6 (or ACKR2) were examined for their involvement in tumor progression. In particular D6 was investigated for its expression and its ability to shape CCL2 gradient in Kaposi\u2019s sarcoma, whereas CCR2 has been analyzed as potential modeler of TAM polarization. D6 is an atypical chemokine receptor acting as a decoy and scavenger for inflammatory CC chemokines expressed in lymphatic endothelial cells. Here, we report that D6 is also expressed by Kaposi\u2019s sarcoma (KS) which is a tumor ontogenetically related to lymphatic endothelium, yet its role in tumor progression was hitherto unknown. D6 expression was evaluated by immunohistochemistry in a cohort of KS patients and its role in cancer progression was investigated in an in vivo KS model. Both in human tumors and in the experimental model, D6 expression levels were inversely correlated with tumor aggressiveness, and directly correlated with increased chemokine-driven infiltration of macrophages and their acquisition of a pro-angiogenic phenotype. Inhibition of monocyte recruitment reduced growth of D6-incompetent tumors, while adoptive transfer of wt but not CCR2-/- macrophages increased the growth rate of D6-competent neoplasms. In the KS model, which presents the B-Raf V600E activating mutation, inhibition of B-Raf or downstream ERK pathway induced D6 expression, and in progressing human KS tumors activation of the K-Ras/B-Raf/ERK pathway correlate with reduced levels of D6 expression. These results indicate that activation of the K-Ras/B-Raf/ERK pathway during KS progression down-regulates D6 expression, which unleashes chemokine-mediated macrophage recruitment and their acquisition of an M2-like phenotype supporting angiogenesis and tumor growth. Thereafter, we wanted to deeper investigate how CCR2 support TAM M2 polarization firstly by using an in vitro system. Wt and CCR2-/- macrophages were polarized with M1 and M2 stimuli and analyzed for gene expression and cytokines production. While no difference was found in M2 polarized macrophages, CCR2-/- M1 or LPS activated macrophages showed higher expression of inflammatory genes and reduced production of the anti-inflammatory cytokine IL-10 and of the pro-angiogenic cytokine VEGF when compared to wt macrophages. The impaired IL-10 production was also confirmed by treating human monocytes with the CCR2 antagonist RS-504393. After LPS stimulation, CCR2-/- macrophages showed reduced activation of NF-kB and p38 MAPK when compared to wt macrophages indicating a cross talk between CCR2 and TLR4 signaling pathways. The contribution of CCR2 to cancer growth was evaluated with a transplantable lung cancer model that grew slower when co-injected with CCR2-/- macrophages, presenting a marked M1 phenotype of infiltrating TAM and a higher number of both CD4+ and CD8+ T cells, correlated with a decreased number of splenic T regulatory cells when compared to wt macrophages holding-tumors. Taken together these data indicate that CCR2 expression by macrophages not only induce their recruitment to tumor site but also affect their polarization and anti-tumor potential

Variabilità spazio-temporale della microalga <i>Chrysophaeum taylorii</i> Lewis &amp; Bryan lungo le coste nord-orientali della Sardegna = Spatio-temporal variabilty of the microalga <i>Chrysophaeum taylorii</i> Lewis &amp; Bryan along North-Eastern Sardinian coasts

In the summer months of 2010 the cell density of the alien benthic microalga Chrysophaeum taylorii Lewis &amp; Bryan (Pelagophyceae) was assessed on hard benthic substrates in sixteen study sites along the north-eastern coast of Sardinia in order to obtain useful information on spatial and temporal variations of the species in the area during summer

Anticipatory postural adjustments stabilise the whole upper-limb prior to a gentle index \ufb01nger tap

Abstract Little is known about anticipatory postural adjustments (APAs) developing when body segments of tiny mass are moved. Thus, APAs in the human upper-limb were investigated during a gentle and small index \ufb01nger tap (35 mm stroke in 50 ms). This task was ful\ufb01lled by ten subjects either with prone or supine hand. EMG was recorded from Flexor Digitorum Super\ufb01cialis (FDS), the prime mover, and from several upper-limb muscles under slight tonic contraction. Regardless of hand posture, EMG was inhibited in Flexor Carpi Radialis and facilitated in Extensor Carpi Radialis well before the FDS burst. With the prone hand, the prime mover activity was preceded by Biceps inhibition and Triceps facilitation; this effect reverted in sign with the supine hand. A postural reversal was also observed in Anterior Deltoid and Trapezius which were both inhibited with the prone hand. The effect in Trapezius was present only with the unsupported forearm. It is thus demonstrated that a gentle small \ufb01nger tap produces well-de\ufb01ned anticipatory natural synergies behaving as the most \u2018\u2018classical\u2019\u2019 APAs: (1) they are distributed to several upper-limb muscles creating a postural chain aiming to prevent the effects of the interaction torques generated by the voluntary movement; (2) they change in amplitude according to the level of postural stability and (3) they revert in sign when movement direction is reverted. These results are also corroborated by data obtained from a simple mechanical model simulating \ufb01nger tapping in a \ufb01ctive upper-limb. A possible role of APAs in controlling movements\u2019 accuracy is also discussed

Determinants of depression in 111 italian patients with systemic sclerosis

Background: A high prevalence of depressive symptoms has been described in systemic sclerosis (SSc), but no clear association with organ involvement or objective indices of disease severity has been depicted. To date, no effort has been made to determine the prevalence of depressive symptoms in Italian patients with SSc or to clarify their cause. Methods: One-hundred-eleven SSc patients were asked to fill in the Beck Depression Inventory (BDI) questionnaire, the scleroderma Health Assessment Questionnaire (sHAQ) and two additional questions assessing the patient's familiar support and the social consequences of the patient's change in physical appearnace. Results: Thirty-seven subjects (33.4%) presented mild to severe depressive symptoms (BDI ³17). On univariate analysis the diffuse cutaneous form of the disease (p=0.019), higher pulmonary systolic pressures on echocardiogram (p=0.016), lower FVC percentage of predicted values (p=0.022), higher sHAQ values (p<0.001) or higher VAS values for pain (p=0.007), lung involvement (p=0.02), Raynaud's phenomenon severity (p=0.002), ulcers severity (p=0.006) or disease severity (p<0.001), were associated with the presence of pathologic depressive symptoms. On multivariate analysis only the VAS for disease severity relevant to BDI scores (p=0.016). Social behaviour changes due to SSc-related physical involvement were reported in 14 patients (38%) with depressive symptoms (p=0,006) and were more likely to be observed in younger patients (p=0.001) with a more severe Raynauds's phenomenon (p=0.013). Conclusions: Mild to severe depressive symptoms are common in SSc patients especially in those with a worse perception of disease severity, these patients should be carefully monitored and a psychological assistance counselled whenever necessary

Esperimento di rimozione degli ammassi mucillaginosi della microalga alloctona <i>Chrysophaeum taylorii</i> Lewis &amp; Bryan = Experiment of removal of the mucilaginous aggregates by the alien microalga <i>Chrysophaeum taylorii</i> Lewis &amp; Bryan

A manipulative experiment was carried out in a small bay of Tavolara Punta Coda Cavallo Marine Protected Area in order to investigate the triggers for formation and storage of the macroscopic mucilaginous aggregates of the benthic microalga Chrisophaeum taylorii Lewis &amp; Bryan (Pelagophyceae) and to test the efficacy of mucillage removal

Distribution and density of the benthic microalga <i>Chrysophaeum taylorii</i> Lewis &amp; Bryan from Northern to central-Eastern Sardinian coasts = Distribuzione e densità della microalga bentonica <i>Chrysophaeum taylorii</i> Lewis &amp; Bryan dalle coste nord a quelle centro orientali della Sardegna

In August 2009 the distribution and density of the alien microalga Chrysophaeum taylorii Lewis &amp; Bryan (Pelagophyceae) were investigated on hard benthic substrates in seventeen sites from northern to central-eastern Sardinia, in order to estimate the distribution and abundance of this species in the area

Mechanical and focal electrical stimuli applied to the skin of the index fingertip induce both inhibition and excitation in low-threshold flexor carpi radialis motor units

It has been observed that mechanical stimulation of the skin of the index fingertip causes a weak short-latency inhibition followed by a strong long-lasting facilitation of the flexor carpi radialis (FCR) H-reflex. Based on threshold and latency, these cutaneous reflexes are thought to be routed to motoneurons by parallel pathways. As recent studies have shown predominant inhibitory potentials in slow motoneurons and predominant excitatory potentials in faster ones, the question arises as to whether or not the two cutaneous pathways converge onto the same motoneuron. The poststimulus time histogram technique was used to investigate the changes in firing frequency of low-threshold FCR motor units (MUs), induced by passive mechanical or focal electrical stimuli to the index skin. After gently tapping the finger pulp a small sharp inhibition appeared in 20 MUs. On average, inhibition started 10.2 +/- 1.6 ms from the homonymous Ia monosynaptic effect, and its central delay was estimated to be 1.2 +/- 1.6 ms. The subsequent facilitation, more consistent, had a mean latency of 13.5 +/- 1.7 ms. Inhibition and excitation were statistically significant (P < 0.05). A similar biphasic effect was observed in seven other FCR-MUs, also after focal electrical stimulation of the same skin area. Comparison with the time course of the H-reflex, representing the whole population of MUs, showed striking similarities in time course and latency to the present MU effect. It is thus suggested that cutaneous spinal pathways may have a homogeneous distribution within the FCR motoneuron pool, and that the skewed distribution of cutaneous afferents onto motoneurons should be not taken as a ruleIt has been observed that mechanical stimulation of the skin of the index fingertip causes a weak short-latency inhibition followed by a strong long-lasting facilitation of the flexor carpi radialis (FCR) H-reflex. Based on threshold and latency, these cutaneous reflexes are thought to be routed to motoneurons by parallel pathways. As recent studies have shown predominant inhibitory potentials in slow motoneurons and predominant excitatory potentials in faster ones, the question arises as to whether or not the two cutaneous pathways converge onto the same motoneuron. The poststimulus time histogram technique was used to investigate the changes in firing frequency of low-threshold FCR motor units (MUs), induced by passive mechanical or focal electrical stimuli to the index skin. After gently tapping the finger pulp a small sharp inhibition appeared in 20 MUs. On average, inhibition started 10.2 \ub1 1.6 ms from the homonymous Ia monosynaptic effect, and its central delay was estimated to be 1.2 \ub1 1.6 ms. The subsequent facilitation, more consistent, had a mean latency of 13.5 \ub1 1.7 ms. Inhibition and excitation were statistically significant (P < 0.05). A similar biphasic effect was observed in seven other FCR-MUs, also after focal electrical stimulation of the same skin area. Comparison with the time course of the H-reflex, representing the whole population of MUs, showed striking similarities in time course and latency to the present MU effect. It is thus suggested that cutaneous spinal pathways may have a homogeneous distribution within the FCR motoneuron pool, and that the skewed distribution of cutaneous afferents onto motoneurons should be not taken as a rule