Everything you always wanted to know about SYNEMIN and never dared to ask

Synemin, a type IV intermediate filament (IF) protein, forms a bridge between IFs and cellular membrane, by interacting with Desmin, Vinculin, Talin and Dystrophin. An A-kinase anchoring protein, it also provides temporal and spatial targeting of protein kinase A. This protein likely functions to integrate mechanical stress and maintain structural integrity and tissue homeostasis in skeletal muscle and other cells.

In our recent article, entitled "Synemin acts as a regulator of signalling molecules during skeletal muscle hypertrophy" (Li et al. J Cell Sci, 2014) we provide novel evidence on its role by phenotyping the synemin null mice. All the additional hints and informations on synemin role in development and postnatal life in PubMed.

Inflammation in Muscle Repair, Aging, and Myopathies

I have recently had the honor to co-edit a special issue of BioMed Research International dedicated to the role of inflammation in various acute and chronic conditions of muscle loss and disease. Our group in Rome participated to the special issue with a paper (from which the figure is extracted) confirming in vivo a role for the neurohypophyseal hormone vasopressin in muscle repair and homeostasis. The paper by Alessandra Costa et al., entitled "Local Overexpression of V1a-Vasopressin Receptor Enhances Regeneration in Tumor Necrosis Factor-Induced Muscle Atrophy" can be found by fillowing the link.

Figure legend. V1aR overexpression counteracts TNF-dependent protein degradation by stimulating the Akt pathway. (a) Western blots of phosphorylated Akt and native and phosphorylated FoxO3a expression demonstrate that in muscle overexpressing TNF, phospho-Akt and phospho-FoxO3a are downregulated, while the native Foxo3a is increased. In V1aR overexpressing muscles, the expression levels of phospho-FoxO3a and phospho-Akt is increased compared with TNF alone, while the native Foxo3a is reduced. (b–d) Densitometric analysis of three independent experiments of phospho-Akt, phospho-FoxO3a, and native FoxO3a expression levels. (e) Real-time PCR analysis revealed that the strong upregulation of atrogin-1 expression observed in the sample overexpressing TNF alone is downregulated in V1aR+TNF-transfected muscles. ; by Student’s -test.

Here is the Table of content of the special issue, a nice mix of original research and review articles. Inflammation in Muscle Repair, Aging, and Myopathies Guest Editors: Marina Bouché, Pura Muñoz-Cánoves, Fabio Rossi, and Dario Coletti Inflammation in Muscle Repair, Aging, and Myopathies, Marina Bouché, Pura Muñoz-Cánoves, Fabio Rossi, and Dario Coletti Volume 2014 (2014), Article ID 821950, 3 pages Stem Cell Transplantation for Muscular Dystrophy: The Challenge of Immune Response, Sara Martina Maffioletti, Maddalena Noviello, Karen English, and Francesco Saverio Tedesco Volume 2014 (2014), Article ID 964010, 12 pages From Innate to Adaptive Immune Response in Muscular Dystrophies and Skeletal Muscle Regeneration: The Role of Lymphocytes, Luca Madaro and Marina Bouché Volume 2014 (2014), Article ID 438675, 12 pages Cardioprotective Effects of Osteopontin-1 during Development of Murine Ischemic Cardiomyopathy, Georg D. Duerr, Bettina Mesenholl, Jan C. Heinemann, Martin Zoerlein, Peter Huebener, Prisca Schneider, Andreas Feisst, Alexander Ghanem, Klaus Tiemann, Daniela Dewald, Armin Welz, and Oliver Dewald Volume 2014 (2014), Article ID 124063, 15 pages IL-6 Impairs Myogenic Differentiation by Downmodulation of p90RSK/eEF2 and mTOR/p70S6K Axes, without Affecting AKT Activity, Michele Pelosi, Manuela De Rossi, Laura Barberi, and Antonio Musarò Volume 2014 (2014), Article ID 206026, 12 pages Local Overexpression of V1a-Vasopressin Receptor Enhances Regeneration in Tumor Necrosis Factor-Induced Muscle Atrophy, Alessandra Costa, Angelica Toschi, Ivana Murfuni, Laura Pelosi, Gigliola Sica, Sergio Adamo, and Bianca Maria Scicchitano Volume 2014 (2014), Article ID 235426, 14 pages Influence of Immune Responses in Gene/Stem Cell Therapies for Muscular Dystrophies, Andrea Farini, Clementina Sitzia, Silvia Erratico, Mirella Meregalli, and Yvan Torrente Volume 2014 (2014), Article ID 818107, 16 pages Vitamin D Receptor Agonists: Suitable Candidates as Novel Therapeutic Options in Autoimmune Inflammatory Myopathy, Clara Crescioli Volume 2014 (2014), Article ID 949730, 10 pages 7-Tesla Magnetic Resonance Imaging Precisely and Noninvasively Reflects Inflammation and Remodeling of the Skeletal Muscle in a Mouse Model of Antisynthetase Syndrome, Clara Sciorati, Antonio Esposito, Lara Campana, Tamara Canu, Antonella Monno, Anna Palmisano, Francesco De Cobelli, Alessandro Del Maschio, Dana P. Ascheman, Angelo A. Manfredi, and Patrizia Rovere-Querini Volume 2014 (2014), Article ID 879703, 8 pages Understanding the Process of Fibrosis in Duchenne Muscular Dystrophy, Yacine Kharraz, Joana Guerra, Patrizia Pessina, Antonio L. Serrano, and Pura Muñoz-Cánoves Volume 2014 (2014), Article ID 965631, 11 pages Inflammation Based Regulation of Cancer Cachexia, Jill K. Onesti and Denis C. Guttridge Volume 2014 (2014), Article ID 168407, 7 pages Macrophage Plasticity in Skeletal Muscle Repair, Elena Rigamonti, Paola Zordan, Clara Sciorati, Patrizia Rovere-Querini, and Silvia Brunelli Volume 2014 (2014), Article ID 560629, 9 pages

Our editorial "Inflammation in Muscle Repair, Aging, and Myopathies" briefly summarizes the main focus of this special issue, i.e. bringing together studies that used different experimental approaches in vivo or in vitro to dissect the dynamic changes taking place in specific immune cell populations, their cross talk with other cell types within the muscle milieu, and their contribution to normal versus pathological muscle repair. While the number of scientific publications on the topic of skeletal muscle inflammation has steadily grown over the last two decades, the notion of inflammation as a common feature in muscle degeneration occurring in aging and myopathies and its association with altered muscle has to our knowledge never previously been addressed and discussed in dedicated journal issues before.

Restoration versus reconstruction: how cell anatomy and extra‐cellular matrix affect tissue regeneration

Sadly, in Italy we have a significant experience of repair following heart quakes. Two main strategies are possible and have been alternatively chosen along the decades, as shown below. Similarly, tissues respond to damage by either restoring their structure or reconstructing it ex novo, depending on the extend of the lesions as well as on the tissue type.

Here is the lecture given to the 2013 Congress of the Italian Society of Anatomy and Hsitology, entitled "Restoration versus reconstruction: how cell anatomy and extra‐cellular matrix affect tissue regeneration". Abstract published in Italian Journal of Anatomy and Embryology http://dx.doi.org/10.13128/IJAE-13961 Tissue regeneration replaces damaged cells and is involved in tissue remodeling. In order to investigate the existence of a leitmotif of tissue regeneration, we compared the cellular aspects of regeneration of epidermis, neurons and skeletal muscle, characterized by different types of histological and functional organization. Following damage, all the tissues here analyzed go through three phases: inflammation, regeneration and maturation. Another common feature is the occurrence of cellular de-differentiation and/or differentiation events, including gene transcription, which are typical of embryonic development. Nonetheless, various strategies are used by different tissues to replace their lost parts. The epidermis regenerates ex novo, whereas neurons restore their missing parts; muscle fibers use a mixed strategy, based on the regrowth of missing parts through reconstruction by means of newborn fibers. The choice of either strategy is influenced by the physical, chemical and structural features of the cells as well as by the extracellular matrix typical of a given tissue, which points to the existence of differential, evolutionary-based mechanisms for specific tissue regeneration. Taking skeletal muscle as a model, we present evidence about the importance of the fiber niche to direct and favour tissue regeneration, a phenomenon of particular relevance for highly hierarchized tissues such as striated muscles. Niche properties are accounted for by cell-cell contacts, cell-matrix interaction and paracrine effects in skeletal muscle. The ordered sequence of steps that characterizes the regeneration processes, shared by several tissues, suggests it may be possible to model this extremely important phenomenon to improve guided in situ regeneration interventions.

ARTCILES: Coletti et al. Regenerative Medicine Research 2103

Figure legend. Examples of focal injuries. (LEFT) Hematoxilin-and eosin-stained murine skeletal muscle, longitudinally sectioned to show the gaps in three adjacent fibers. The injury likely occurred following an intense exercise session (wheel running). Upon leakage of the broken sarcolemma, factors such as Wnt are released before a fast repair process known as patch repair occurs. In turn, Wnt factors trigger the activation of satellite cells and other resident interstitial cells with myogenic potential, which proliferate, migrate and fuse into small myotubes that ultimately fuse with the damaged fibers. (RIGHT) Toluidine blue-stained semithin section of a murine carotid showing damage, likely due to smooth muscle cell-restricted inactivation of the serum response factor gene. A rupture of the endothelial layer, as well as of the elastin matrix, with exposure of underlying cells is visible; release of intracellular factors (von Willebrand Factor) and exposure of undisclosed antigens (collagen) are essential for the subsequent phases of clot formation, remodeling and repair of the wall defect. Bar = 25 micron.

Figure legend. Macrophages infiltrate necrotic muscle fibers. Serial section of murine skeletal muscle in an area of necrosis; corresponding fibers are highlighted by matching color arrows. Evans Blue Dye (EBD) highlights muscle fibers whose plasma membrane is leaking owing to damage; hematoxylin and eosin staining (H&E), showing cellular infiltration in EBD + fibers; histochemistry for esterase staining highlights esterase-enriched macrophages; immunofluorescence analysis for activated macrophages expressing F4/80 (green) confirms the invasion of the muscle fibers by macropohages: laminin (red) and nuclei (blue) are also shown. Coletti et al. Regenerative Medicine Research 2013 1:4 doi:10.1186/2050-490X-1-4

In this review article we present and compare the cellular aspects of regeneration in skin, nerve and muscle, three organs characterized by differences not only in anatomical and functional organization, but also in the number and location of stem cell niches and populations, which ultimately result in varying regenerative potential. By discussing the common traits and the specific features of regeneration in three model tissues, we propose general models of regeneration and highlight various strategies adopted to cope with damage and repair in mammals.The strategies used by a wide range of tissues to replace their lost parts vary, probably as result of evolutionary-based mechanisms for specific tissue regeneration. While the epidermis regenerates ex novo, neurons restore their missing parts; muscle fibers instead use a mixed strategy, based on the reconstruction of missing parts and on the generation of new fibers. These differential strategies are represented by the two terms used in the title to refer to different forms of regeneration: restoration, the attempt to re-establish the status quo ante, and reconstruction, a more radical response, characterized by ex novo cell colonization and tissue formation. The choice of either strategy is deeply influenced by the anatomy and the distribution/features of stem cell niches typical of a given organ. In addition, the energetic costs for either regenerative strategy are also likely to play an important role. The abstraction of divergences and analogies between different types of tissue regeneration might pave the way for the mathematical modeling of this process, thereby making a major contribution to both pathology and regenerative medicine. When talking of wound healing, a distinction is made by some authors between regeneration and repair. Regeneration is used to refer to the complete replacement of damaged tissue with new tissue not associated with scar tissue, while repair is used to refer to the re-establishment of tissue continuity [1]. Regeneration can be attained by two means: a) restoration, defined as “putting together what is broken”; b) reconstruction, defined as “replacing and rebuilding what is torn down” (according to the Merriam-Webster Dictionary). To grant homeostasis, most tissues undergo continuous or cyclic processes of regeneration. Which of the afore-mentioned strategies tissues adopt depends on the histological features discussed in the article entitled "Restoration versus reconstruction: cellular mechanisms of skin, nerve and muscle regeneration compared".

ARTICLES: He et al. Journal of Clinical investigation 2013

Cachexia is a debilitating condition characterized by extreme skeletal muscle wasting that contributes significantly to morbidity and mortality. Efforts to elucidate the underlying mechanisms of muscle loss have predominantly focused on events intrinsic to the myofiber. In contrast, less regard has been given to potential contributory factors outside the fiber within the muscle microenvironment. In tumor-bearing mice and patients with pancreatic cancer, we found that cachexia was associated with a type of muscle damage resulting in activation of both satellite and nonsatellite muscle progenitor cells. In our paper we present extensive data demonstrating that skeletal muscle wasting in cachexia arises from the failure of myogenic progenitor cells to differentiate and fuse into skeletal muscle myofibers. The failure of myogenic progenitor cells to differentiate is due to elevated, NF-kappaB dependent expression of Pax7 leading to expansion of muscle progenitors that are prevented from differentiation and fusion to repair damaged fibers. Our findings challenge the assumption that cachexia arises from events occurring inside the muscle fibers (i.e. essentially proteasome-mediated protein degradation) leading to muscle atrophy. On the contrary, the microenvironment surrounding muscle fiber and including muscle stem cell niche is important for muscle homeostasis and can be targeted to counteract muscle wasting. The link to full text article is provided here. In addition, extensive supplemental data are available here. We are currently investigating the role of Serum Response Factor (SRF) and mechanotransduction in this stem cell mediated response. A posted position to work on this topic is currently available (see related post). Figure modified from: He et al. J Clin Invest doi:10.1172/JCI68523

ARTICLES: Galmiche et al. Circulation research 2103

Common carotid intima-media thickness and distensibility are markers of structural and functional vessel wall properties, with important consequences on cardiovascular risk. We investigated whether SRF and its target genes regulate intrinsic smooth muscle tone and, in turn, arterial stiffness, and we found that inactivating SRF in vascular smooth muscle cells (VSMC) results in profound alterations of the extra-cellular matrix composition and arterial wall properties. The link to the full text article is here.

POSTDOC POSITION IN SKELETAL MUSCLE PHYSIO-PATHOLOGY AVAILABLE IMMEDIATELY

A new postdoc position is available in the lab to work on mechanotransduction and SRF role in exercise-mediated responses in cancer cachexia. The position at the University of Paris 6 Pierre et Marie Curie is for 1 year, renewable. Full detail on the position (immediately available) are visible in the attached flyer. Please send a cover letter, CV and names and email address of two references to dario.coletti@upmc.fr

METHOD: NADH transferase staining

NADH transferase activity on muscle cryosections by histochemistry. Whilst NADH transferase is not a mithocondrial marker sensu strict , it helps visualizing the mitochondria and can help distinguishing between glycolytic (pale), oxidative (dark) and intermediate muscle fibers. Attached here is our protocol.

METHOD: INNOVATIVE RAPID PROTOCOL TO QUANTIFY NUCLEAR STAINING

I linked here our revolutionary methodology to automatically quantify several feature of immunostained cells by using the freeware software Image J. The method, to be published soon, is simply based on the novel technology called QR code and on the fact that anyone now has a smartphone. Please, feel free to use our method; your are kindly requested to acknowledge its use with the following statement " The method was originally developed at UPMC Paris 6 by D. Coletti, sponsored by April Fool's Day, AFD grant # 01011971" and possibly a reference to the publication (to be posted asap).

METHODS: Anesthesia for rodents

With the aim to shift to novel pharacological treatments, and keep those only for specific purposes, here are the guidelines for the use of avertin and pentobarbital for mouse anesthesia.

OUR TWIN BLOG: Everything You Ever Wanted To Know About SRF But Never Dared Ask

We have a dedicated blog to Serum Response Factor (SRF) and intermediated filaments in the physiopathology of muscle tissues. To access all the publications on the subject, please follow the link to the SRF blog. UPDATE: a sinthetic review on SRF (entitled "Serum response Factor in muscle tissues: from development to aging") has been recently published in European Journal of Translational Myology, open access journal publishing original data papers and reviews on muscle.

METHODS: Phosphate Buffered Solutions in our lab

With or without Ca and Mg, here is the method to make Phosphate Buffered solutions in stock and working concentrations.

METHODS: 4 color IF for extra-cellular matrix and myosin isoforms

Changes in myosin isoform expression accompanying muscle atrophy during cancer cachexia have been shown (Diffee 2002). For this reason we are very much interested in measuring the cross-sectional area as well as the percentage of muscle fiber populations differing in their physiological properties (e.g. the type of myosin isoform). In the attached immunofluorescence method we highlight the interstitial extra-cellular matrix (collagen 3 + laminin, withe) and three different form of myosin (the fast MHCIIb, green; the slow MHCI,blue; and the intermediate MHCIIa, red). Fibers purely expressing MCH type IIX are black (negative staining). EXTENSOR DIGITORUM LONGUS, top

SOLEUS, bottom

METHODS: Visualisation of myosin isoforms by elecrophoresis and silver stain

Different muscle express different myosin isoforms, for instance the Soleus is enriched in the slow myosin (type 1) while the faster Extensor Digitorum Lungos express the fast myiosins 2A and 2X but not myosin 1. Different isoforms of myosin have different molecular weights and can be separated by electrophoresis and then visualized by silver stain. Attache here is the protocol that Eleonora has learned from excellent tutors who master this technique. If interested in myosin isoform analysis, be patient and follow the link.

Figure legend. Myosin isoforms from the Extensor Digitorum Longus (EDL, first two lines) and from the Soleus (SO, lines 3 and 4) murine muscles. The isoforms 2A, 2X have the same apparent weight, the isoform B is specific pf the EDL while the isoform 1 characterizes the SO.

METHODS: Murine muscle dissection from the hinlimb

Here is the link to our illustrated,step by step method for dissecting several skeletal muscles from the hindlimb of a mouse.

EXPERIMENTAL MODELS: BALB/c substrains & running behavior

All BALB/c mice are not equal. In spite of being an inbred strain, there are several SUBstrains that diverged decades ago.In the attached notes, I summarized the names and origins of the the three main BLAB/c substrain, i.e. the Balb/c AnNCrl, the Balb/c J and the Balb/c ByJ mice. They are sold by Charles River, Jackson Laboratories and other vendors, often depending on geographical localization (CRL breeds and sells different substrain in different countries). Concerning the propensity to voluntary running (on a wheel) I could not find any information on the CRL Balb/c AnNCrl substrain,and our recently published results are probably among the first to be released on this substrain. On the contrary,a bibliographic search pinpointed strinking differences between sex, and between different BALB/c substrains, for what concerns running activity (Lightfoot et al. J Appl Physiol. 2010 September; 109(3): 623–634). The infos are summarized here. The source of the data on rod and wheel running activity come from a wonderful database, available on the Jackson website (http://phenome.jax.org/), reporting all major phenotypes of many different mouse strains. Additional data are in the post "EXPERIMENTAL MODELS: wheel running"

Grip stenght test

Here is provided the link to our standard method for measuring the force of a mouse. This method is based on the measure of the grip force by a dynamometer, while the mouse is being pulled by its tail.

Indo-Italian Forum on Biomaterials and Tissue Engineering

A new space for scientific collaboration, exchange of human resources and grant rising was born in New Delhi last week: the Indo-Italina Forum on Biomaterials and Tissue Engineering. During the APA INternational congress on Advances in Human Healthcare Systems (Healthcare India 2012) we participated to the Indo-Italina Symposium on Tissue Engineering, where several aspects of cell interaction with biomaterials were addressed. In this context the Forum was born, which we hope will become a catalyzer for further exchanges between the two countries. More details on the newborn Indo-Italian Biomaterials and Tissue Engineering Forum (i2bite) have been reported on the ENEA newsletter (article in Italian). The link to the Forum web pages (under construction) is here. More details on the event (in Italian) are here.

Candidate for ISAC councilor

I am candidate as ISAC councilor. The elections are in the next few weeks (vote end by March the 30th). Are you an ISAC memeber? Please, vote. All the instructions on how to vote will be soon linked here. Not an ISAC member? Do you want to know more about ISAC? Please, click here to know more about the the International Society for Advancement of Cytometry. Below, there are a few notes on my biography and thoughts about the society. Biography Born in Latina (Italy), I performed both my undergraduate (Biological Sciences, summa cum laude, 1995) and graduate studies (Doctoral degree in Cell Science and Morphogenesis, top mention, 2000) at the Sapienza University of Rome. Since then I accumulated a quite varied experience abroad, as a visiting scholar at the Stanford University (Stanford, CA; 1999), then as a postdoctoral fellow at the Mount Sinai School of Medicine (New York, NY; 2000-2003), as invited researcher at the Myology Group, UMR S 787 Inserm, UPMC (Paris, FR; 2007), where I ultimately returned in 2010 as a Maitre de Conferences, i.e. assistant professor, at the University Paris VI/Pierre et Marie Curie. I also held in Italy the responsibility of the Laboratory of Electron Microscopy and Calcium Imaging (Rome, IT; 2004-2010). All this was a lot of fun, since I could feed myself with great science - not to mention outstanding culinary experiences - from very different environments. For all the above, I have to acknowledge several mentors, including Laura Teodori and Sergio Adamo in Rome, Marco Conti in Stanford and David Sassoon in New York. For all the details, please view my full CV, while works in progress can be followed through my blog. As a cell biologist, I dealt with analytical cytology quite early in my career. Whilst not being exclusively specialized in flow cytometry, I exploit the incredible power and the elegant performance of flow cytometry analysis to address several questions related to my scientific interests. I am mostly interested in the control of skeletal muscle differentiation and homeostasis and, more recently, in tissue engineering applications for regenerative medicine of this tissue. I had the honor to become an ISAC Scholar in 2006 and since them I am member of this society. Interests and vision I came across ISAC through my mentor, Dr. Laura Teodori at a time when I was doing my postdoctoral training about 10 years ago. Being scientifically seduced by the powerful applications of analytical cytology I started to attend ISAC international congresses and to participate more actively to the initiatives of the Society. At the XXIII ISAC congress in Quebec in 2006, I was awarded the ISAC Scholarship. As an ISAC Scholar I was encouraged to collaborate to the educational and organization strategies of the Society. I was young, mobile and without a tenured position. I felt sympathetic with the younger members of the Society and concerned about the typical issues they have to deal with: practicing, traveling, finding the resources to do that. By co-chairing a subcommittee of the MSC (Membership Services Committee) dedicated to Students' services in 2004-'08, I helped with the divulgation of skills and resources aimed to improve student members' success rate when applying to mobility grants. For instance, at the XXIV ISAC congress in Budapest in 2008 I participated to the Scientific Professional Skills Workshop with a presentation entitled “Short term mobility grants: tips and hints.” These issues found concrete development in the ISAC web page highlighting grants opportunities for short term mobility and for young fellows (originally published on the ISAC web site, http://www.isac-net.org/content/view/693/137/). I was responsible for that page, updating it twice a year and offering tutoring and advice to ISAC student members, with the aim to help their mobility and grant rising capacity. Today, from a more mature position inside the Society I wish to contribute to consolidate ISAC strengths and to further develop its potentials, hence my interest in the candidature for an ISAC Councilor position. In this position I could possibly exploit my growing experience and creative attitude to serve our common goals. I foresee two critical issues ISAC shall deal with in the incoming years: 1) geographical and intergenerational growth and 2) interaction with novel scientific and technological research areas. 1) I am convinced that our Society should invest more than ever on youngs. As far as I know ISAC educational and scholarship programs see an unprecedented success, which highlights the interests into our society by young researchers. ISAC should be even more attractive than today for them. In order to do so, we should pursue our politic of open access for young members, and of tutoring and education initiatives. Also, ISAC could set up initiatives aimed to assist its younger members in fund rising (startups, mobility). The initiatives could range from helping members to find senior partners for big grant applications to assisting members to identify calls and apply to them (a task often performed by specific services that are present only in major departments and universities). Obviously, the current programs dedicated to tutoring and visiting initiatives for young members would be synergistic with the novel actions I propose. An international society such as ISAC should become the catalyzer for exchanges and interactions not only vertically (between generations) but also horizontally (between emerging countries, where it is largely underrepresented, and consolidated scientific environments). So I would love to see novel initiatives, aimed at networking and diffusing the analytical cytology, especially targeted to the younger members and to researchers from emerging countries. 2) While being well developed in the US, tissue engineering and regenerative medicine are novel, fast growing disciplines in several countries, including european countries such as Italy and France, or Asian giants such as India and China. This area of Medicine is attracting more and more public and private financing, given its translational nature and high technological content, which in turns stimulates a growing involvement by scientists. Given the foreseen rapid shift to clinical practice in this field (indeed a reality for certain applications) it is of pivotal importance to set up at the same time innovative approaches and safety/quality control procedures for stem cell isolation and transfer, as well as for immunological stereotyping of host-implant interactions. In this context there is an important opportunity for ISAC to become the reference for such procedures and approaches. Thus, I would like to establish initiatives aimed to boost the collaboration between ISAC and non-ISAC members for regenerative medicine applications. An other level of interaction could be within the journals associated to those societies which represent the analogues of our Cytometry; politics encouraging cross-publication (and even cross-advertising for the scientific societies) could be built and I would be happy to collaborate on this. I wish that the publication of this text can be a matter of discussion and engagement by others and myself independently from my candidature as ISAC councilor. I really think that these issues are relevant for ISAC development and I am looking forward to seeing them dealt with.

Quencing autofluorescence

Method for quencing background fluorescence due to aldehydes or autofluorescence

postdoctoral fellow position available (SOLD OUT!)

We are searching a postdoctoral fellow willing to join us in the frame of the founded project UPMC EMERGENCE 2011 on cardiac wasting. The location is Paris (at the UNiversity Pierre et Maire Curie) in 2012. Please, refer to the online flyer for additional information.

EXPERIMENTAL MODELS AVAILABLE IN THE LAB (2011)

A quick overview of the experimental models currently available in the lab to study the hormonal control of muscle differentiation and homostasis

LAB METHODS: Assessing cell number with a counting chamber

A Chamber with a View...each one has a different one. Supposedly trivial, estimating the number of cells in a cell suspension is matter of never-ending debates in our labs, that's why we finally wrote instructions for either a Thoma or a Malassez chamber, two widely used options in the lab.

LAB METHODS: Cardiac Stem Cell Isolation

Claudia Serradifalco, a PhD student from our collaborators' laboratory at the University of Palermo, has established in Paris their simple and elegant method to obtain Cardiac Stem Cells from adult rat hearts (link to the original paper by Di Felice et al.) by adapting the procedure to our laboratory. Here is the modified protocol...

Blind tasting session at the lab

To welcome a new PhD student in the lab, we organized a tasting session of charcuterie. This included a celebrated home-made fois gras and several salami from different countries. The latter were married to Chardonnay-based wines, a Mersault 1er cru 2008 A. Bouteller and a Chablis VV 2010 Vaucher & fils. For the fois gras I proposed an Alsace Gewurztraminer 2006, moelleux and traditional, by Schueller. A blind testing session concerned four salami and four sausages from the following countries and regions: A) Spain, Cataluna; B) France, Auvergne; C) France, Aveyron; D) Italie, Lombardia (Varzi) and Lazio (Cassino)

Most loved: Category “saucisson (big salami)”: region Avyron (France), producer Linard; Category “saucisse (small sausage)”: region cataluna (Spain)

ARTICLES: Teodori et al. Chimica e Industria 2011

THE ADVANCEMENT OF SCIENCE: SHARING OR EXCLUDING? THE “NEW BIOTECHNOLOGY DIVIDE”: AN ALARMING PERSPECTIVE OF SCIENTIFIC DUAL USE Linked to the title above, you can find the full version of our paper on biotech divide, an emerging issue related to recent advances in information technology (IT). The role of scientists is of paramount importance in understanding and predicting the impact of their research in issues related to the threat of conflicts inherent in a polarized society. They must increase their own awareness of these issues and better inform the political community by advising and helping to assess programs of cooperation that will lead to more equitable access to benefits and reduce inequalities driven by the technology divide. This article will focus on the use, distribution, and accessibility of research outcomes in one particular area of biotechnology, i.e. technology related to health care. We have identified some biotechnological barriers, given some specific examples of positive action in the field of our expertise in bridging such a divide and highlighted the direction we believe should be followed.

CLASSES, LECTURES ETC: REGENERATIVE MEDICINE

Linked here you can find a presentation dealing with regenerative medicine (in French/ oui, en Français!) for the master students in "Molecules and therapeutic targets". The presentation consists of three parts: 1) stem cells and their therapeutic use 2) what is tissue engineering 3) strategies of the regenerative medicine: in situ regeneration, stem cell transplantation, transplantation of pre-assembled organs. A similar lesson, more focused on tissue engineering (Englligh version) is visible here. Learning about the outstanding capacity of regeneration shown by the newt will allow the full regeneration of human organs? Hopefully better than what we are currently doing.

IS THIS BLOG GOING TO BE SHOT DOWN?

What is going on in Italy? Today, the Italian Wikipedia is ON STRIKE and one has only access to this sober communicate (screen image from the Wikipedia homepage http://it.wikipedia.org/wiki/Wikipedia:Comunicato_4_ottobre_2011 )

Unfortunately, this communicate is in italian, so I will try to summarize it below. The Italian parliament is discussing a bill to strongly limit the publication of texts deriving from phone call tapping used in trials. Wikipedia cites the text of the bill and remarks that a modification of the bill proposed this morning will force the responsible of a web site to rectify within 48h a given information that is possibly considered incorrect by anyone who finds this information as negatively affecting his image. All this by default, without any third party judging the dispute. It is feared that the easiness of the censoring action will discourage everybody from saying just anything on anyone, in sharp contrast with the article 27 of the Universal Declaration of Human Rights. All this happens while the bill to help the agonizing Italian economy is postponed. If you have recently read the content of hot conversations between our Prime Minister and his friends reported worldwide by media, you may imagine the reason why such bill is more urgent than boosting the economy. Indeed, I find reasonable to foresee the risk that such a proposed bill will affect any web site posting free contributions related to whatever subject, including this blog which has hosted in the past political opinions and analyses (concerning science in most of the cases). Shall I shoot it down rather than checking every 48 h not to have received an injunction of rectification?

ARTICLES: Perniconi et al. Biomaterials 2011

In this paper in press we show that one can transplant ghosts of tissues to obtain again the corresponding tissue in mice. Our "ghosts" are acellular scaffolds derived by whole organ decellularization (in 1% SDS) of skeletal muscles (the EDL and the TA). Therefore, they represent the extracellular matrix voided of the cellular component. We show that this biomaterial has niche properties, since it is able to support neomyogenesis once transplanted to replace the matching muscle.

LAB METHODS: transplantation of an acellular scaffold to replace the corresponding muscle

We are about to publish a paper where we characterize the in vivo response to a graft composed by an acellular scaffold obtained by a previously decellularized skeletal muscle. The grafting procedure is now available as a ppt - link embedded in the title of this post. The corresponding video on how to replace a TA with the corresponding acellular scaffold(iPod version) is available through the link in parentheses. For an alternative format, try to click here (avi version). The video is supplemented as Additional materilas to the Biomaterials article.

LAB METHODS: Toluidine blue staining

There is no staining method as fast and informative (two for the price of one!) as the Toluidine blue staining. We use it while cryosectioning or while doing semithin sections to monitor sample quality and orientation. Toluidine specifically stains some cell and ECM features. Linked to the title of this post, you'll find our method for Toluidine staining, with references and additional examples. Fig. legend: Toluidine-stained skeletal muscle cryosections.

Research fundings: an update...

Well...I was too pessimistic. The fundings for the Fiscal Year 2009 ("PRIN 2009") has been released by the Italian Ministry of University and Research , with a delay of only three years and not four years, as I was foreseeing.
That's good news, worth at least a bottle of Prosecco di Valdobbiadene Giustino B. by Ruggeri!
That is also a good chance to have a look at what the USA are doing. Linked to the title is the analysis of the current presidential plan for R&D in that country. President Obama requested $ 147,696 bilion for research in the current Fiscal Year. With this rate they will DOUBLE the fundings in 11 years. Linked to the title, please find the full text of the analysis of this plan.

Left:
Research & Develoment funding path in the USA

Source:
Federal Research end Development Funding - FY 2011
JF Sargent jr., coordinator, specialist in Science and Technology Policy
June 10, 2011

Blind tasting session at the lab

To celebrate a few recent events (the UPMC Emergence 2011 grant, the Mol Endocrinol paper) and to welcome a new student in the lab, we have tasted five Bordeaux 2006 wines, from different appellations characterized by marked nuances of their terroirs and specific grape assembly. Given that the different wineyards are only about 50 Km from each other, the differences were outstanding.

Results of the blind tasting (panel : laboratory members):

1st Château-Haut Maurac, Médoc Cru Bourgeois (60 % Cabernet sauvignon, 40 % Merlot)
2nd Château Musset Chevalier , Saint Emillon Grand cru (50 % Merlot noir / 45 % Cabernet-Franc / 5 % Cabernet-Sauvignon )
3rd Les Hauts du Tertre, Margaux (55 % Cabernet sauvignon, 20 % Merlot, 20 % Cabernet franc, 5 % Petit verdot)
4th Château Prieuré-les-Tours, Graves.

We liked the winner for its intense bouquet of red fruits and its full body, with mature tannins and a long lasting aftertaste. One more cru Borgeois showing the great quality/price ratio of this category. From the color to the marked tannins it expressed the Medoc pretty well. However, I preferred the Margaux of Les Hauts de Tertre, a second wine produced by Château du Tertre, for its elegance and its more floreal bouquet. Margaux came out in the good balance between tannins, acidity and alcoolic warmth. The superb roundness of the Libournais St Emillon and the acidity of the Graves (Alas! - in such a poor interpretation) came out as well, but nobody guessed the crus for all the wines.

ARTICLES: Toschi et al. Mol Endocrinol 2011

In this paper, entitled "SKELETAL MUSCLE REGENERATION IN MICE IS STIMULATED BY LOCAL OVEREXPRESSION OF
V1a-VASOPRESSIN RECEPTOR", we identify skeletal muscle as a physiological target of hormones of the vasopressin (AVP) family and show a novel in vivo role for vasopressin-dependent pathways. FIG LEGEND In red Myc (i.e. overexpressed V1a-R) immunolocalization in skeletal muscle fibers highlighted by laminin staining in green.

In the last 10 years, we have characterized in detail AVP signaling pathways in myogenic cells in vitro. Also, we have reported that the muscle specific, V1a, AVP-receptor is modulated during myogenic differentiation in vivo, which suggest a role in muscle development. Consistently, we have shown that AVP intramuscular injection enhances muscle regeneration, a process which recapitulates muscle development in the adult.
With the last paper by Toschi et al. we formally demonstrate the biological role of AVP on skeletal muscle homeostasis and we pinpoint some molecular mechanisms underlying this effect, including calcineurin-mediated IL-4 production in the musculature in response to AVP.

FIG LEGEND role of Calcineurin-dependent effects of V1a-R overexpression on muscle regeneration. Further links to the press which cited the article: ANSA and Corriere della Sera

Against cuts in cultural funding

Again a non scientific, still relevant post insomuch as politics affect culture, research and education.

Cuts on culture and arts.
We have recently celebrated the 150th anniversary of Italian unification. As reported by the New York Times, a very intense moment occurred when Riccardo Muti conducted the "Va pensiero" at the premiere of Verdi's “Nabucco” at the Teatro dell’Opera in Rome in March, in the presence of the Prime Minister and the Mayor of the capital.
The issue was the heavy cut plan on cultural founding performed by the current government. The event had its climax at Muti's brief statements against this plan while introducing an exceptional bis of the "Va pensiero". Linked to the title of this post there is the touching video on youtube.

Cuts and management of university funding.
University budget cuts represent the other branch of the current harmful intervention on state budget, in a country which spending on university is already very low as compared to most other countries, as reported by the BBC last year. However, it is not only a matter of budget. What is even worse is the total incertitude for the CURRENT available fundings: in 2011 we are still waiting for the results of a major funding call of the Italian Ministry for the University and Research (MIUR) which is called PRIN 2009 and was released in 2010! In 2012, if and when some groups will receive the grants to which they applied three years before, what will remain to be accomplished of the proposed research projects? Won't the latter be born already aged and out to date?
A lucid analysis on the inceritude which reigns on italian university has been published a few months ago on the web pages of lavoce.info (in Italian).

CLASSES, LECTURES ETC: Mechanisms controlling skeletal muscle homeostasis

Linked to the title there is a lesson for Master students (in English) on the mechanisms controlling skeletal muscle homeostasis.

OVERVIEW:

SKELETAL MUSCLE HOMEOSTASIS, HYPERTROPHY AND ATROPHY
The skeletal muscle tissue accounts for the majority of our body mass, nonetheless, the amount of skeletal muscle can vary significantly throughout life. There are specific mechanisms finely tuning the exact amount of muscle that we have at a given time.These are apparent in conditions far from homeostasis, i.e. when we have an excessive growth (hypertrophy) or reduction (atrophy) of muscle fibers. Throughout the presentation, I also try to state the case that not only muscle protein metabolism is important for controlling muscle homeostasis but also muscle stem cells support a "flow" of myogenic cells contributing to the maintenance of muscle fibers.

EXPERIMENTAL MODELS FOR STUDYING SKELETAL MUSCLE HOMEOSTASIS
Where I presents different approaches to study the regulation of muscle differentiation, growth and repair in vitro and in vivo.

MUSCLE ATROPHY, WASTING, CACHEXIA
Where I present different forms of muscle fiber atrophy and present in detail the features of the most severe form of muscle wasting, the syndrome of cachexia.


ENDURANCE EXERCISE & PROTEIN METABOLISM
Where I present some experimental data on exercise effects on muscle metabolism and homeostasis in physiological and pathological conditions.

MUSCLE REGENERATION IN PATHOLOGICAL CONDITIONS
Where I presents mechanisms whereby skeletal muscle regeneration is affected in cachexia, ultimately providing the molecular explanation for an important deficit in muscle regenerative capacity accounting for loss of muscle mass.

SUGGESTED READINGS

Glass D. 2003 Molecular mechanisms modulating muscle mass

Moseri V. 2010 Myogenin and calss II HDACs control neurogenic muscle atrophy by inducing E3 ubiquitin ligases

Musaro` A. 2004 Stem cell mediated muscle regeneration is enhanced by local isoform of Insulin-like Growth Factor 1

Zhou X. 2010 Reversal of cancer cachexia and muscle wasting by ActRIIB antagonism leads to prolonged survival

The Real Face of Death

A creative, funny interpretation of a real TEM image representing an eucariotic cell in culture undergoing apoptosis (programmed cell death). Apoptosis major features are represented in this photomicrograph: loss of cell attachment, but maintenance of cell integrity, cell shrinkage, nuclear fragmentation and chromatin condensation.

ARTICLES: The Problem of Subjective/Objective Genitive in Matters of Heart

Following recent discoveries of cardiac wasting in cachexia of both cardiac and non-cardiac origin, it appears that the heart can be both the trigger and the target of cachexia. By analogy to muscle cachexia the use of “cardiac cachexia” may arise in a double subjective and objective sense inducing misleading interpretations. We recently got the following comment published on Cell online.

Comment on:
Cell, 20 August 2010, Volume 142, Issue 4, 531 - 543
doi:10.1016/j.cell.2010.07.011
Article
Reversal of Cancer Cachexia and Muscle Wasting by ActRIIB Antagonism Leads to Prolonged Survival
Xiaolan Zhou, Jin Lin Wang, John Lu, Yanping Song, Keith S. Kwak, Qingsheng Jiao, Robert Rosenfeld, Qing Chen, Thomas Boone, W. Scott Simonet, David L. Lacey, Alfred L. Goldberg, and H.Q. Han

by:
Dario Coletti
Barbara Perniconi, Sergio Adamo, Zhenlin Li, Denise Paulin, Mathias Mericskay
19 novembre 2010
10:21:04 HNEC
Affiliation:University Pierre et Marie Curie Paris 6, France & Sapienza University of Rome, Italy

The Problem of Subjective/Objective Genitive in Matters of Heart
 
Zhou et al. uncovered a previously unappreciated loss of heart mass in cachectic mice for which, from our point of view, they correctly used the expression “atrophy of the heart”. This study is likely to generate a new line of research, which is distinct from cardiac cachexia, i.e. the atrophy of the skeletal muscle induced by cardiac pathologies. We urge to clarify the terminology to describe these phenomenons, since we foresee the risk of misleading use of related expressions, e.g. cardiac cachexia and cardiac atrophy, sounding alike but very different de facto one from the other. In particular, “cardiac cachexia” poses a problem of ambiguity, thus its use might be abandoned.  
The old problem of the subjective/objective genitive case.  
Does amor patris (father's love) mean that the father (pater) loves his children (subjective genitive) or that the children love their father (objective genitive)? The father can be either the subject or the object of the action of loving. There is no difference in form between the subjective and the objective genitive. Only context can make a final determination. In addition, English does not typically mark nouns for a genitive case morphologically. Rather, it uses the Saxon genitive “ 's” for people or the preposition “of” like in “Molecular Biology of the Cell”. Biology of the cell can also be referred to as “Cell biology” by using the noun as adjective.  
What is cardiac cachexia, then?  
About 200 papers to date referred to cardiac cachexia as a syndrome of skeletal muscle wasting associated to a specific pathology, i.e. Chronic Heart Failure (CHF). Cardiac cachexia is not meant as cachexia of the heart, i.e. cardiac atrophy. However, the existence of a cardiac component of cardiac cachexia has also been reported (Florea et al., 2002). Several papers demonstrate the growing use of “muscle cachexia” referred to as skeletal muscle wasting associated to a chronic disease, including CHF (Pajak et al., 2008). Following the recent discoveries of cardiac wasting in cachexia of both cardiac and non-cardiac origin (Florea et al., 2002, Zhou et al., 2010), it appears that the heart can be both the trigger and the target of cachexia. We are concerned that, by analogy to muscle cachexia, the use of “cardiac cachexia” may arise in a double, subjective/objective sense.   
Different names for cardiac muscle wasting: PROs and CONs.  
The following are alternative ways to refer to the phenomenon of cardiac muscle wasting.  
1)  Atrophy of the heart. This expression is unambiguous, even though relatively long. It is very general and implies the need to clarify the context in which the atrophy arises, e.g. cancer-induced atrophy of the heart to specify that the latter is triggered by cancer.  
2)  Heart atrophy, which is formally ambiguous in terms of  subjective/objective genitive.  
3) Cardiac atrophy, which has the same risk and sounds dangerously similar to cardiac cachexia.   
4) Cachectic heart, a more holistic expression not entirely described to date.   
5) Cardiac wasting, which has a nuance toward pathology and appears as a specific, easily discernible expression. Therefore, we are in its favor.   
Highlighting the atrophy of the heart in the definition of cachexia.  
 The current  consensus definition of cachexia, “Cachexia is a complex metabolic syndrome associated with underlying illness and characterized by loss of muscle  [...]” (Evans et al., 2008), does not explicitly refers to the loss of cardiac muscle. We think opportune to refer to both skeletal and cardiac musculature when reporting about muscle wasting in cachexia.  
References  
Evans, W.J. et al. (2008). Clinical nutrition (Edinburgh, Scotland) 27, 793-799  
Florea, V.G. et al. (2002).  American heart journal 144, 45-50  
Pajak, B. et al. (2008). J Physiol Pharmacol 59 Suppl 9, 251-264  

Published online on 11/19/2010
http://www.cell.com/comments/S0092-8674(10)00780-4

LAB METHODS: isolation of skeletal muscle fibers

The procedure after Dr. Peter Zammitt's...linked here are the figures of our version of the method

LAB METHODS: the simplest mycoplasma test

Mycoplasma is a common contaminant of cell cultures throughout the wolrd. It affects experimental results and it perturbs cell behaviour. Mycoplasma is a genus of bacteria that lack a cell wall.Without a cell wall, they are unaffected by many common antibiotics such as penicillin or other beta-lactam antibiotics that target cell wall synthesis. It is very difficult to get rid of mycoplasma: the best is to screen regularly for contamination and to throw out the contaminated cells! Here is the method for the screening....in italian

RELOCATING IN PARIS - UPMC

NEWS! Effective September the 1st I am at the University Paris VI Pierre et Marie Curie. I do teaching and research as Maître de Conférences. When I left Italy for the USA I had in my luggage a bottle of Barolo Granbussia 1982 by Aldo Conterno. It was a great experience - both the wine and postdoc! This time, coming to Paris, I put in my bag a Barolo Vigneto Arborina 2005 by Elio Altare - thanks Elio! I got to teach something to these guys...

ARTICLES: link to Aulino et al. BMC Cancer 2010

In the paper linked above we perform an extensive molecular, cellular and physiological characterization of the cancer cachexia-inducing C26 colon carcinoma in mouse. This widely diffesed murin model of cancer and cancer associated cachexia was originally developed by Corbett et al. in 1975, during an effort to establish an animal colon tumor model for biological and chemotherapy studies. Colon tumors were induced and transplanted in different inbred mouse strains. Four tumors survived the first transplant, which displayed a variety of histological and malignancy features. These four tumors included the colon tumor 26, described as an undifferentiated Grade IV carcinoma.
Since then, the model spread around the world and was exploited for a pletora of studies in the absence of an organic description of its main features.This gave rise to misunderstandings and mistakes, such as describing the C26 tumor as an adenocarcinoma without showing histological images.Today, the communities of scientists exploiting the C26 model to study either cancer or cachexia are not fully aware of each other’s works (as shown by cross-reference analysis on PubMed) and this may have deleterious consequences for the progress of integrative medicine applied to a complex syndrome associated with underlying illness. We suggest that “C26” be included among the keywords whenever work is conducted on this experimental model to provide adequate visibility.

Among the NOVEL DATA that we shown in this paper, I'd like to stress the functional analysis of cachectic muscles. We found that while the drop in muscle force is a hallmark of cachexia, it is simply due to muscle atrophy. In fact, no differences exist between normalized force (i.e. specific force) of control and cachectic animals. On the other hand, murine cachectic muscles are characterized by fatigue, which is in agreement with clinical observations. Left: C26 H&E staining. The measures shown below refer to the EDL muscle of ctr and C26-bearing mice.

CLASSES, LECTURES ETC: Materials for the students of Tissue engineering School of Dentistry

Attached to the link there is a summary (in Italian) of basic concepts of Tissue engineering for the sutudents of the School of Dentistry.
IN allegato - link nel titolo - una dispensa con i concetti base di ingegneria tissutale presentati nell'ADE per gli studenti di OPD.
Linked to this text there are several articles on this topic.

A review...

LAB METHODS: 3D acellular scaffold from skeletal muscle

The method linked here is a simplified procedure to obtain an acellular scaffold suitable for cell culture of myoblasts into a 3D scaffold.

Cachexia, sarcopenia, inactivity: the three Fates (Moirae) of muscle atrophy

A common question is: what is the difference, if any, between muscle atrophy conditions which look all similar? What happens to our muscles when one lays in bed with a brokne leg, as opposed to getting cancer or happily ageing? Dr. Evans, one of the wolrd experts on muscle atrophy, simply and concisely pinpoints the differences in this review...



from:
Am J Clin Nutr. 2010 Apr;91(4):1123S-1127S. Epub 2010 Feb 17.
Skeletal muscle loss: cachexia, sarcopenia, and inactivity.
by Evans WJ.
Division of Geriatrics, Department of Medicine, Duke University Medical Center, Durham, NC 27709, USA. william.j.evans@gsk.com

LAB METHODS: CMFDA cell tracker

The protocol linked here is a method to label live cells and assess whether they are VITAL and/or STRESSED. The dye in fact accumulates inside the cells due to a chemical modification that makes it non-cell permeant. This esterase reaction requires glutathione. Therefore a shift toward low fluorescence characterizes a cell population with depleted levels of glutathione, i.e. that have been subjected to oxidative stress - GSH being the principal buffer of the redox state.

caveat for the use of the Molecular Probes anti-Mouse Alexa Fluor 350 Ab

Linked here is the comparison between two Molecular Probes secondary antibodies: anti-M AF350-conjugated vs the classical AF488-conjugated. Thanks Paola for trying...
Looking at something in blue is cool but rather blues!

Il caso e la necessità, Le hasard et la nécessité, Chance and Necessity

"Tutto ciò che siste nell'universo è il frutto del caso e della necessità"
"Tout ce qui existe dans l'univers est le fruit du hasard et de la nècessité"
"Everything in the universe is the fruit of chance and necessity"
Democritus, circa 460-370 BC
IN: Jacques Monod, Le Hasard et al nécessité: Assai sur la phyolosophie naturelle.



Léon-Alexandre Delhomme (1841-1895)
Description
Democritus che medita sull'anima, bronzo, 1868.
Démocrite méditant sur le siège de l'âme, bronze, 1868.
Democritus meditating on the seat of the soul, bronze, 1868.

CLASSES, LECTURES ETC: Regenerative Medicine & Tissue Engineering

Here is the link to the ENGLISH VERSION of an introductory lecture to tissue engineering. The principles of the latter are summarized, and various strategies for regenerative medicine are presented through discussion of the most recent outbreaking reports in the field. For the 1st y medical students and biotechnology students. Talking about students, I have to acknowledge the great work done in the lab by Barbara Perniconi and Alessandra Costa (some of their data on acellular skeletal muscle are shown). A similar, broader lesson on regenerative medicine in FRENCH has been published here.


Figure legend: confocal image of a 20 micron tick cryosection of a murine acellular skeletal muscle matrix (laminin staining, red)

A flu dealed with a secret treatment - article by Nerina Dirindin

Linked to the tile is an article, published by our great online magazine Lavoce.info, on the incredible management of the flu crisis by the Italian Minister of Health. The article is in Italian, but I wish to summarize it in English asap. Below is a little taste of the content...

About 4% of the Italian population has been vaccinated against the flu virus, which has recenlty given rise to pandemic emergency. On the other hand, the Italian government has bought from Novartis viruses for 40% of the population. This is not the real issue, though (except for singificant stocking and expiration problems). What is unbeliavable, is that the Ministry of Health has purchased the vaccines by using a procedure allowed by the Prime Minister (ordinanza n. 3275 del presidente del Consiglio del 2003) for cases of risk of terrorist attack. This procedure is secret, direct and charges almost all the burden on the government rather than on the company - the criticism comes from our Corte dei Conti (the top organ of suveillance of public expenses). For istance, the government will take care for all the expenses due to adverese effects or aother complications in the population underging vaccination. To date, it is not clear the cost of such operation.