Resistin: A reappraisal.

From a biological point of view, aging can be considered a progressive inability of an organism to react to stress, maintain homeostasis, and survive unfavourable changes during post-maturational life. The expression of several adipokines changes during aging and for some changes, a role in the onset of chronic disease and frailty has been proposed. Among adipokines, resistin was shown in recent studies to play a key role in aging. Resistin is a small secreted protein that regulates glucose metabolism in mammalians. High resistin levels induce insulin resistance and exert proinflammatory effects. Consistently, resistin has been shown to play a pivotal role in various metabolic, inflammatory, and autoimmune diseases. Herein, the role of resistin as a molecular link between aging and age-related conditions was reviewed and the clinical implications of this knowledge discussed.

Validation of the Photography Method for Nutritional Intake Assessment in Hospitalized Elderly Subjects.

OBJECTIVE: The aim of the present study was to validate the photographic indirect method as an accurate and specific tool to assess nutritional intake in a cohort of elderly hospitalized patients. DESIGN: this is a prospective observational study. SETTING: hospital (geriatric acute ward and transitional care of IRCCSS AUO San Martino Hospital, Genoa, Italy). PARTICIPANTS: 255 consecutive elderly hospitalized patients. MEASUREMENTS: assessment of malnutrition by: Mini nutritional assessment (MNA) and abbreviated Comprehensive geriatric assessment (CIRS; Barthel index, SPMSE). The direct method (Gold standard): food dish weight (before lunch) and residual (after lunch) food dish weight and estimation of the percentage of eaten food and of residual food for each dish. The percentages of food intake and residual food were calculated according to the following formula: intake %= initial weight of the dishes- residual food weight)/ initial weight dish x100. The unit of variable was the percentage. The indirect photographic method with extrapolation of the lunch food intake by photographic method confronting initial meal and residual meal (25% quartile food dish estimation). RESULTS: The results showed a significant correlation between the direct method (weighing residual food) and the indirect photographic method(n=255; r=0.9735; p<0.001) as well as a significant positive correlation between the indirect photographic method and the food caloric estimation calculated by the direct method (n=255; r= 0.6489, p<0.001). Intraclass coefficient (ICC), showed a highly significant degree of agreement between the gold standard and the indirect photographic method (ICC: 0.69; p<0.0001). Additionally, the results showed a good inter rater agreement of the indirect photographic method (kappa-statistic measure of interrater agreement: (Z=13.04; p<0.001); agreement 70.29% e Kappa=0.5965) and a good specificity of the indirect method as it was independent on the single food item. CONCLUSIONS: The study originally provided the validation of the indirect photographic method for the assessment of nutritional intake in a vast cohort of hospitalized elderly subjects. The present results moved a step forward in the appropriate assessment of nutrition intake in frail elderly, providing an easy to use tool that may be incorporate in routine clinical practice for early and targeted therapeutic interventions.

Pathophysiological role of neutrophils in acute myocardial infarction.

The pathogenesis of acute myocardial infarction is known to be mediated by systemic, intraplaque and myocardial inflammatory processes. Among different immune cell subsets, compelling evidence now indicates a pivotal role for neutrophils in acute coronary syndromes. Neutrophils infiltrate coronary plaques and the infarcted myocardium and mediate tissue damage by releasing matrix-degrading enzymes and reactive oxygen species. In addition, neutrophils are also involved in post-infarction adverse cardiac remodelling and neointima formation after angioplasty. The promising results obtained in preclinical modelswith pharmacological approaches interfering with neutrophil recruitment or function have confirmed the pathophysiological relevance of these immune cells in acute coronary syndromes and prompted further studies of these therapeutic interventions. This narrative review will provide an update on the role of neutrophils in acute myocardial infarction and on the pharmacological means that were devised to prevent neutrophil-mediated tissue damage and to reduce post-ischaemic outcomes.

Evidence on the pathogenic role of auto-antibodies in acute cardiovascular diseases.

Atherothrombosis is the major determinant of acute ischaemic cardiovascular events, such as myocardial infarction and stroke. Inflammatory processes have been linked to all phases of atherogenesis In particular, the identification of autoimmunity mediators in the complex microenvironment of chronic inflammation has become the focus of attention in both early and advanced atherogenic processes. Auto-antibodies against self-molecules or new epitopes generated by oxidative processes infiltrate atherosclerotic plaques and were shown to modulate the activity of immune cells by binding various types of receptors. However, despite mounting evidence for a pathophysiological role of autoantibodies in atherothrombosis, the clinical relevance for circulating autoantibodies in cardiovascular outcomes is still debated. This review aims at illustrating the mechanisms by which different types of autoantibodies might either promote or repress atherothrombosis and to discuss the clinical studies assessing the role of auto-antibodies as prognostic biomarkers of plaque vulnerability.

Synthetic lethality-based therapeutics: perspectives for applications in colorectal cancer.

Over the past two decades, progresses in colorectal cancer treatment have significantly improved patient survival and quality of life. However, unresectable metastatic colorectal cancer remains virtually incurable, making the search for new effective therapeutics mandatory. An important limitation to the development of new agents has been the difficulty to exploit mutated tumor suppressors or "undruggable" oncogenes as a target. Recently, evidence that mutations in tumor suppressors, such as BRCA1/2, make cancer cells highly susceptible to inhibitors of a compensatory DNA repair pathway [poly-(ADP-ribose) polymerase 1 (PARP1)] has broadened the range of possible therapeutic targets by extending it to gene products that are in a "synthetic lethal" relationship with oncogenes and tumor suppressors. Inhibition of such targets blocks specific buffer-mechanisms that are required for survival in the presence of defined oncogenic mutations, but not in their absence. As a consequence, selective elimination of mutation-bearing cells results. This approach has led to identify compounds that are highly active in the presence of different types of mutated tumor suppressors and oncogenes, including DNA repair genes, RAS, and Myc. In addition, ongoing studies promise to identify new mechanisms which, when pharmacologically interfered with, will selectively eradicate mutated cancer cells. Here, we revise and discuss these new aspects of cancer biology and highlight their potential applications in colorectal cancer treatment.

Patient-tailored treatments with anti-EGFR monoclonal antibodies in advanced colorectal cancer: KRAS and beyond.

Personalized medicine emphasizes the practice of considering individual patient characteristics as opposed to that centered on standards derived from epidemiological studies which, by definition, do not take into account the variability of individuals within a given population. When applied to oncology, personalized medicine is an even more complex concept because it extends the variability beyond the individual patient to the individual tumor. Indeed, the great genotypic and phenotypic variability (both in primary and metastatic sites of cancer) the development of targeted therapies, and the growing availability of biological assays complicate the scenario of personalized medicine in the oncological field. In this paper we review the results of anti-epidermal growth factor receptor (EGFR) monoclonal antibody (mAb) therapy in metastatic colorectal cancer (mCRC) in the context of tumor biology, delineating the future prospects of patient-tailored medicine in this area. In particular, we deal with EGFR inhibition by Cetuximab, a chimeric mouse human IgG1 mAb, and panitumumab, a fully human IgG2 mAb. We discuss the clinical impact of anti-EGFR mAbs on wild-type (WT) KRAS mCRC, also taking into account the feasibility of novel multi-marker approaches to treatment decision-making, aimed at increasing the predictive power of pre-therapy biomarkers. Experimental topics and fields of ongoing research, such as targeting microRNAs (miRNAs) with novel anticancer drugs and epigenetics in CRC are also addressed.

Current standards and future strategies in immunochemotherapy of non-Hodgkin's lymphoma.

The therapeutic options for B-cell non-Hodgkin's lymphoma (NHL) have dramatically expanded with the advent of immune-based treatments. The monoclonal antibody anti- CD20 rituximab represents the best example of these advances and has quickly become incorporated into the therapeutic armamentarium for this hematological disease. In addition, other antibodies are eventually becoming part of treatment approaches to NHL. Furthermore, the role of therapeutic vaccines continues to be an important ongoing research question. Despite this success, several questions on how to optimize the use of monoclonal antibodies in NHL remain open since the best administration schedules, as well as the optimal duration of immunotherapy still have to be determined. Finally the development of resistance to treatment remains the main limit of this innovative approach, necessitating the development of strategies to circumvent resistance itself. This review will summarize the state of the art of antibody-based immunotherapy of NHL and discuss prospective approaches to improve the benefit of these treatments in patients.

Dynamic simulations of pathways downstream of ERBB-family, including mutations and treatments: concordance with experimental results.

The pathways downstream of ErbB-family proteins are very important in BC, especially when considering treatment with onco-protein inhibitors. We studied and implemented dynamic simulations of four downstream pathways and described the fragment of the signaling network we evaluated as a Molecular Interaction Map. Our simulations, enacted using Ordinary Differential Equations, involved 242 modified species and complexes, 279 reversible reactions and 111 catalytic reactions. Mutations within a single pathway tended to be mutually exclusive; only inhibitors acting at, or downstream (not upstream), of a given mutation were active. A double alteration along two distinct pathways required the inhibition of both pathways. We started an analysis of sensitivity/robustness of our network, and we systematically introduced several individual fluctuations of total concentrations of independent molecular species. Only very few cases showed significant sensitivity. We transduced the ErbB2 over-expressing BC line, BT474, with the HRAS (V12) mutant, then treated it with ErbB-family and phosphorylated MEK (MEKPP) inhibitors, Lapatinib and U0126, respectively. Experimental and simulation results were highly concordant, showing statistical significance for both pathways and for two respective endpoints, i.e. phosphorylated active forms of ERK and Akt, p one tailed = .0072 and = .0022, respectively. Working with a complex 39 basic species signaling network region, this technology facilitates both comprehension and effective, efficient and accurate modeling and data interpretation. Dynamic network simulations we performed proved to be both practical and valuable for a posteriori comprehension of biological networks and signaling, thereby greatly facilitating handling, and thus complete exploitation, of biological data.

Peripheral blood vs. bone marrow for molecular monitoring of BCR-ABL1 levels in chronic myelogenous leukemia, a retrospective analysis in allogeneic bone marrow recipients.

Molecular monitoring of the BCR-ABL1 transcript in chronic myelogenous leukemia (CML) using quantitative real-time PCR (RQ-PCR) can be performed using either bone marrow (BM) or peripheral blood (PB). However, a recent report by Stock et al. [International Journal of Oncology 28 (2006) 1099] questioned the reliability of PB samples for BCR-ABL1 detection as performed by RQ-PCR. We report a study on 114 CML patients who received allogeneic stem cell transplantation (ASCT), and who were monitored by RQ-PCR using paired samples of BM and PB: the total number of determinations was 428, with a median follow-up after transplant of 8 years. BCR-ABL1 transcript was undetectable or <0.1%, in 106 (49.57%) and 62 (29%) paired determinations, respectively. BCR-ABL1 was >0.1% in 36 (16.8%) paired determinations and was discordant in 10 (4.7%). Agreement between PB and BM results was quantified by the kappa test (k = 0.85; 95% CI 0.76-0.94). This study shows that BCR-ABL1 RQ-PCR monitoring of CML patients after ASCT with PB is concordant with BM in 95.3% of cases, and thus may be used to monitor the disease. This may be relevant when discussing both quality of life issues and the need for post-transplant monitoring with the patient.

Molecular diagnosis and monitoring of chronic myelogenous leukemia: BCR-Abl and more.

The current treatment of chronic myelogenous leukemia (CML) is one of the most successful examples of molecularly targeted therapy in cancer. The identification of the fusion oncogene BCR-ABL allowed the discovery of small molecule inhibitors of its tyrosine kinase activity which, in turn, have literally revolutionized the treatment of this disease. However, large part of a successful clinical management of CML relies on appropriate diagnosis, molecular monitoring and identification of mutations potentially leading to drug resistance. These issues are discussed here together with an overview on how patients treated with tyrosine kinase inhibitors should be monitored.

Neurotransmitter evaluation in the hippocampus of rats after intracerebral injection of TsTX scorpion toxin

TsTX is an á-type sodium channel toxin that stimulates the discharge of neurotransmitters from neurons. In the present study we investigated which neurotransmitters are released in the hippocampus after TsTX injection and if they are responsible for electrographic or histopathological effects. Microdialysis revealed that the toxin increased glutamate extracellular levels in the hippocampus; however, levels of gamma-aminobutyric acid (GABA), glycine, 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) were not significantly altered. Neurodegeneration in pyramidal cells of hippocampus and electroencephalographic alterations caused by the toxin were blocked by pretreatment with riluzole, a glutamate release inhibitor. The present results suggest a specific activity of TsTX in the hippocampus which affects only glutamate release.

Embriotoxic effects of maternal exposure to Tityus serrulatus scorpion venom

Tityus serrulatus is the most venomous scorpion in Brazil; however, it is not known whether its venom causes any harm to the offspring whose mothers have received it. This study investigates whether the venom of T. serrulatus may lead to deleterious effects in the offspring, when once administered to pregnant rats at a dose that causes moderate envenomation (3mg/kg). The venom effects were studied on the 5th and on the 10th gestation day (GD5 and GD10). The maternal reproductive parameters of the group that received the venom on GD5 showed no alteration. The group that received the venom on GD10 presented an increase in post-implantation losses. In this group, an increase in the liver weight was also observed and one-third of the fetuses presented incomplete ossification of skull bones. None of the groups that received the venom had any visceral malformation or delay in the fetal development of their offspring. The histopathological analysis revealed not only placentas and lungs but also hearts, livers and kidneys in perfect state. Even having caused little effect on the dams, the venom may act in a more incisive way on the offspring, whether by stress generation or by a direct action.

Proteasome inhibitors: antitumor effects and beyond.

Proteasome inhibitors are emerging as effective drugs for the treatment of multiple myeloma and possibly certain subtypes of non-Hodgkin's lymphoma. Bortezomib (Velcade) is the first proteasome inhibitor proven to be clinically useful and will soon be followed by a second generation of small molecule inhibitors with improved pharmacological properties. Although it is now understood that certain types of malignancies have an exquisite dependence on a functional proteasome for their survival, the underlying reason(s) remain unclear as of now. In this context, addiction to nuclear factor-kappaB (NF-kappaB)-induced survival signals, activation of the unfolded protein response as well as a reduced proteasomal activity in differentiated plasma cells have all been proposed to justify proteasome inhibitors' activity in susceptible tissues. In addition to their anticancer properties, bortezomib and related drugs modulate inflammatory and immune responses by affecting function and survival of immune cells such as lymphocytes and dendritic cells. The present review offers an overview of the biological effects that have been involved in proteasome inhibitors' antitumor activity and suggests prospective future applications for these drugs based on their recently characterized anti-inflammatory and immunomodulatory effects.

Effect of different cytokines on mammaglobin and maspin gene expression in normal leukocytes: possible relevance to the assays for the detection of micrometastatic breast cancer.

In cancer patients, the ability to detect disseminated tumour cells in peripheral blood or bone marrow could improve prognosis and consent both early detection of metastatic disease and monitoring of the efficacy of systemic therapy. These objectives remain elusive mainly due to the lack of specific genetic markers for solid tumours. The use of surrogate tissue-specific markers can reduce the specificity of the assays and give rise to a clinically unacceptable false-positive rate. Mammaglobin (MAM) and maspin are two putative breast tissue-specific markers frequently used for detection of occult tumour cells in the peripheral blood, bone marrow and lymph nodes of breast cancer patients. In this study, it was evaluated whether MAM and maspin gene expression may be induced in the normal blood and bone marrow cells exposed to a panel of cytokines, including chemotactic factors (C5a, interleukin (IL)-8), LPS, proinflammatory cytokines (TNF-alpha, IL-1beta) and growth factors (IL-3, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor). The experimental data show that all cytokines included in the panel, except for IL-8, were able to induce maspin expression; on the contrary, MAM gene was never induced. These results suggest that MAM is more specific than maspin and that the possible interference of cytokines should be taken into account in interpreting molecular assays for detection of isolated tumour cells.

Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes.

Dendritic cells (DC) are professional antigen-presenting cells playing a central role in the induction of antigen-specific cytotoxic T-lymphocytes (CTL). We analyzed the efficiency of tumor RNA transfection into DC using different sources of RNA as well as delivery strategies including electroporation, lipofection and CD71-receptor-based delivery. To evaluate the sensitivity of these approaches, we utilized in vitro transcribed enhanced green fluorescence protein (EGFP)-RNA and whole tumor RNA from EGFP-transfected renal cell carcinoma cell line N43. We demonstrate that electroporation was the most effective way yielding about 30% EGFP positive cells while less than 1% of DC expressed EGFP using the transferrin receptor transfection system. Delivery of RNA with liposomes resulted in 17.5% of EGFP positive cells depending on the RNA amount. However, when these approaches were applied to transduce DC with RNA derived from the A498 cell line for T-cell priming, tumor-specific CTL could be induced using all delivery strategies suggesting that this technology has the potential to induce cytotoxic T-cell response even when low level of antigen is delivered. Furthermore, we demonstrate that amplification of whole tumor messenger RNA (mRNA) as well as the use of total instead of purified mRNA can be utilized for stimulating tumor-specific CTL responses.

New perspectives in dendritic cell-based cancer immunotherapy.

Dendritic cells are professional antigen-presenting cells with the unique capacity to initiate primary immune responses. Recently, several procedures to generate large numbers of dendritic cells from circulating precursors, including peripheral blood monocytes and CD34+ stem cells, have been developed. Stimulation with antigen-loaded dendritic cells was shown to break tolerance to tumour-associated antigens and to induce antitumour cytotoxic immune responses in vivo. Hence, numerous attempts to optimise delivery of tumour antigens to dendritic cells, as well as routes and schedules of administration to cancer patients, are currently under way. The first dendritic cell clinical studies have indicated this form of vaccination as feasible and safe; furthermore, in some cases, objective clinical responses were observed, even in patients heavily pretreated with standard chemo/radiotherapy approaches. These preliminary data, although encouraging, require further extensive investigations, which should address the technical and biological problems of manipulating human dendritic cells, as well as the clinical settings which could benefit from an immunotherapeutic approach.

Reverse-transcriptase polymerase chain reaction of the maspin gene in the detection of bone marrow breast carcinoma cell contamination.

BACKGROUND: Maspin is a molecular marker used for the detection of contaminating breast carcinoma (BC) cells in peripheral blood and lymph nodes. However, its specificity has been questioned recently. The objective of this study was to verify the specificity of this marker and to determine the incidence of positive bone marrow results in patients with BC who are eligible for high-dose chemotherapy (HDT) both in early and advanced disease stages and before and after treatment. METHODS: Bone marrow specimens from 41 patients with BC as well as from 35 normal volunteers and 17 patients with hematologic tumors were examined for maspin transcript expression by a modified nested reverse transcriptase-polymerase chain reaction technique. RESULTS: Maspin transcript was found in all normal and neoplastic breast tissues and in none of the 35 normal bone marrow specimens (specificity, 100%; 95% confidence interval, 90-100%). However, the transcript was found in 40% of the bone marrow samples from patients with hematologic malignancies. Thus, this marker appears very specific for discriminating between normal controls and patients with BC, but it cannot be considered disease specific. Among patients with BC, bone marrow was positive for the maspin transcript in 32% of patients with early-stage disease and in 75% of patients with metastatic disease before chemotherapy. After treatment, in 75% of patients with early-stage disease and in 50% of patients with metastatic disease, the bone marrow results became maspin negative. CONCLUSIONS: On the basis of the current data, although it is not disease specific, maspin is a reliable marker for detecting bone marrow molecular disease in patients with BC and should be considered for prospective studies as a prognostic indicator and as an assay for monitoring residual disease.

Neurotoxic effects of three fractions isolated from Tityus serrulatus scorpion venom.

Scorpion venoms contain low molecular weight basic polypeptides, neurotoxins, that are the principal toxic agents. These toxins act on ion channels, promoting a derangement that may result in an abnormal release of neurotransmitters. In the present study we investigated some of the effects of the F, H and J fractions isolated from Tityus serrulatus scorpion venom on the central nervous system of rodents. The venom was partially purified by gel filtration chromatography. The neurotoxic effect of these fractions was studied on convulsive activity after intravenous injection, and on electrographic activity and neuronal integrity of rat hippocampus when injected directly into this brain area. The results showed that intravenous injection of the F and H fractions induced convulsions, and intrahippocampal injection caused electrographic seizures in rats and neuronal damage in specific hippocampal areas. Fraction J injected intravenously reduced the general activity of mice in the open field but induced no changes when injected into the brain. These results suggest that scorpion toxins are able to act directly on the central nervous system promoting behavioural, electrographic and histological modifications.

Convulsive effects of some isolated venom fractions of the Tityus serrulatus scorpion: behavioral, electroencephalographic, and neuropathological aspects

It has been previously shown that the crude venom of Tityus serrulatus can cause convulsions. This study was designed to investigate the neurotoxic effects of B, C, G, and K fractions isolated from this venom. Intravenous injection of these fractions in mice (0.6 - 6.0 mg/kg body weight) showed that the C fraction is a potent convulsant and G fraction decreased the threshold for tonic hand limb extension elicited by transauricular electroshock. Unilateral injection of B, C, and K fractions, but not G fraction, into the spikes and epileptic discharges that began in the hippocampus and evolved to the cortex. The following motor signs were observed: movements of facial muscles, wet dog shake, immobility, myoclonus, wild-running with clonus, and in some cases, loss of postural control. Intrahippocampal injection of B, C, and K fraction, but not G fraction, caused neuronal loss at the injection site as well as in other hippocampal areas. The effect of these fractions on epileptiform activity and on neuronal loss was dose-dependent. The severity of the epileptiform activity in the ipsilateral hippocampus correlated with the severity of the neuronal loss. The electrographic, behavioral, and histological changes induced by b, C, and K fractions were similar to those obtained with other drugs that are commonly used to induce convulsion. The convulsant effects of the crude venom may be caused by the fractions studied in this work.