5^th World Congress on Cell & Stem Cell Research March 23-25, 2015 Chicago, Illinois, USA

Guastali completed his undergraduate and master's degree at the Universidade Estadual Paulista Júlio de Mesquita Filho, currently is a doctoral student at the same university, working with embryonic stem cells and induced pluripotency stem cells in bovines and equines, and production of bovine embryos in vitro, and is revising of the Cell Biology International since 2013.

Introduction: An alternative to creating banks of pluripotent stem cells involves reprogramming the somatic cell nucleus. This induced reprogramming results from the action of different molecules, including transcription factors and other proteins. Recently, several groups have reported the induction of pluripotency in human fibroblasts through the transduction with viral vectors expressing OCT4, C-MYC, KLF4 and SOX2 genes. The cells called induced pluripotent stem cells (iPS) have a morphology characteristic of embryonic stem cells, express pluripotent markers and are capable to differentiate in tissues from the three germ layers in vitro and in vivo. Thus, the generation of iPS cells from the reprograming of different cell types by specific factors, could represent an alternative to obtaining a bank of stem cells with pluripotent characteristics. Objective: Equine fibroblasts and umbilical cord cells (EUCC) were genetically modified using lentiviral vectors containing STEMCCA excitable polycistronic human cDNA of OCT4, SOX2, c-MYC and KLF4 (EF1a-hSTEMCCA) in order to transform unipotent and multipotent cells in pluripotent cells. Materials and methods: EUCC and fibroblasts were plated at a concentration of 5x103 cells per well of 9.6 cm2 and cultured in DMEM high glucose with 20% FBS, 1% penicillin/streptomycin and 1.2% amphotericin. When cultures reached approximately 60% confluence they were transducted in a medium containing 50 mL of viral concentrate plus 8 ηg/ml polybrene (hexadimethrine bromide, Sigma). The medium was renewed after 14 hours of incubation. Five days post-transduction, the cells were transferred into MEFs. During reprogramming period cell were cultured in a specific medium for iPS. Between days 6 and 15 post-transduction we evaluate the time to the beginning of the appearance of colonies, colony morphology and number of colonies (greater than 1 mm diameter) formed per well. Results: In fibroblasts´ cultures the colonies began to form on day 7 and in EUCC´ cultures on day 6. Both cultures developed thin colonies,with well-defined edges, formed by small cells of uniform size and hexagonal appearance, with evident nucleus containing one or two nucleoli. In each well 18 colonies were formed, on average, independently of the cell type. The cultures remained viable and colonies multiply in the course of 15 days. Conclusion: Lentiviral vectors were efficient to deliver the transcription factors to cells, formed colonies showed morphological characteristics similar to embryonic stem cells between 6 and 7 days post-transduction. Future experiments will be conducted for immuno genotypic characterization of these cells.

Graduated in Veterinary Medicine at Federal University of Viçosa (UFV) - Brazil, with honors. In 2008 completed his Masters in Clinical and Surgery of Large Animals by UFV and later Specialization in Physical Therapy and Veterinary Rehabilitation by the Faculty of Jaguariuna (2011). In 2012 became Doctor in Clinical and Surgery of Large Animals at the Faculty of Veterinary Medicine and Animal Science (FMVZ) UNESP Botucatu, Brazil. He is currently a postdoctoral researcher at FMVZ-UNESP with realization of research activities related to cultivation and stem cell biology, cryopreservation and proteomic analysis.

The aim of the present work was to evaluate the colony-forming unit-fibroblast (CFU-F) efficiency of mesenchymal stem cells from intervascular matrix of equine umbilical cord (MSC-UCMI) after cryopreservation with different mediums (M). For this, MSC-UCMI (n=5) previously characterized were cryopreserved using Mr frosty (Nalgene®) with the following protocols: M1: DMEM high glucose, 20% FBS, 10% DMSO; M2 (free from FBS): DMEM high glucose, 10% PVA, 10% DMSO; M3: 90% FBS, 10% DMSO; M4: 90% conditioned medium (DMEM high glucose + 20% FBS), 10% DMSO. After three months storage in liquid nitrogen, samples from M1, M2, M3, and M4 were thawed and seeded in low density (210 cells/ cm2) for the assay of CFU-F in triplicate. The calculation of CFU-F efficiency was carried out using the formula: CFU-F efficiency= (counted CFU-F /cells originally seeded) x 100. The results obtained for the different protocols (M1, M2, M3 and M4) were analyzed using ANOVA followed by post-test Student-Newman-Keuls Method taking P<0.05 as significant. The results revealed good CFU-F efficiency for the cryopreserved samples with M1 (9.9±1.4%), M3 (10.1±1.0%) and M4 (10.9±0.9%) without difference among these protocols (P>0, 05). On the other hand, MSC-UCMI cryopreserved with M2 (free from FBS) did not adhere and had no ability to form fibroblast colonies because of the low cell viability observed. It can be concluded that MSC-UCMI cryopreserved with medium containing FBS (M1, M3, M4) have adequate capacity of in vitro self renewal and can be cultured after thawing.

Carolina Nogueira de Moraes Completed the graduation in 2009 by the Northern State University of Paraná, Bandeirantes, PR. Participated of the Medical Residency in Animal Reproduction, at São Paulo State University, Botucatu, SP, from 2011 to 2012. In addition, completed the Master Degree in Animal Biotechnology in 2014, and at the same institution and department currently is Doctoral student.

The aim of this study was to evaluate the effects of cryopreservation of stromal cells (SC) from bovine endometrial tissue in two moments of luteal phase, using two mediums. For this, in the slaughter house samples of endometrial tissue of animals in phase II (n=3) and III (n=3) were collected and processed at the laboratory. SC were isolated and cultured with DMEM high glucose: F12 (1:1), 10% fetal bovine serum (FBS), 1% penicillin / streptomycin, and 1.2% amphotericin B at 37.5 ºC in a humidified atmosphere, containing 5% CO2 in air. In the third passage SC were cryopreserved using Mr Frosty (-1ºC/minute until -80 ºC, 24 hours, Nalgene®) with the following medium: Medium 1 (M1): 90% SFB + DMSO, penicilin/streptomycin (10 μL/mL) and anphotericin B (3 μg/mL); Medium 2: 90% conditioned medium (culture medium) + 10% DMSO. After one month storage in liquid nitrogen, samples were thawed and analyzed on flow cytometry using annexin V APC (AN) and propidium iodide (PI). Data were analyzed using t-test or Mann-Whitney Rank Sum Test according to the normality, taking P<0.05 as significant. The viability (PI- AN-) after cryopreservation didn´t differ (P=0.52) between M1 (74.88%) and M2 (70.02%). The rate of necrosis was considered low in both groups with lower (P=0.015) values at M2 (1.75% versus 3.22). The rates of late-apoptosis + necrosis (PI+ AN+) and initial apoptosis (PI+ AN-) didn’t differ (P>0.05) between protocols. It can be concluded that the two mediums are suitable for cryopreservation, especially M2 which is commonly discarded and has a lower proportion of SFB.

Sclera is a supportive framework of outer coat on the eyeball, with quiescence sclera cell residents between fibrous layers. Previous studies indicated sclera mesenchymal stem cell express the stem cell surface markers of Sca1, CD90.2, CD44, CD105, and CD73 but negative for the hematopoietic markers. Mouse sclera stem cells highly express the cell surface marker of CD44 (98%). CD44 is a cell-surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. Since mice sclera highly express CD44 positive stem cell, we hypothesis CD44 positive stem cell in sclera is one of the factors to correlate with axial growth or form deprivation myopia (FDM). The axial growth or form deprivation myopia (FDM) mouse model is well established at 2008. This model was used to induce myopia of C57BL/6 mice at right eye and we took left eye for the internal control. Twenty-three days old C57BL/6 mice were covered at the right eyes until twenty-eight days to induce FDM and the left eyes were taken for the internal control in 22 mice. The number and function of stem cell between right and left sclera were analyzed between right and left sclera to study the correlation of stem cell and myopia. Our data indicated that the increased axial length due to FDM induction is negative correlated with 1) the number of stem cell in eyes are significantly decrease in FDM eyes; 2) the ALP acitivty of stem cell in FDM eye are decrased and 3) the differentiation abilities of stem cell toward osteogenic and chondrogenic differentiation are also decreased.

Ching-Hua Yeh has completed his PhD at the age of 40 years from Kaohsiung Medical University and postdoctoral studies from University of Virgina. She is the postdoctor of Department of Genome Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. She has published more than 9 papers in reputed journals and will publish the other two papers on JBJS and the Journal of Fertility & Sterility.

Adverse conditions during pregnancy may lead to irreversible changes in the fetus by the fetal programming mechanism. Changes can be induced by maternal protein malnutrition (MPM), creating an imbalance of androgen levels thus directly influencing the morphophysiology of dependent structures, such as the prostate. Our project aim is to investigate the effects of maternal gestational protein malnutrition on the development of the rat ventral prostate (VP) by focusing on the rate of basal epithelial cell proliferation and androgen receptor expression. Briefly, 20 Sprague Dawley dams were distributed into two groups: control (NP; fed a normal diet containing 17% protein) or a restricted protein diet (RP, fed a diet containing 6% protein) during gestation. After birth, all animals were maintained on the standard diet. Euthanasia was performed at postnatal day 35 (PND35) and the ventral prostate of these animals was dissected, weighed and processed for IHC (Ki67, p63 and AR) and Western blot analysis (PCNA, p63 and AR). RP treated animals showed lower body weights compared to NP control treated animals at PND35. A reduction of testosterone plasma levels as well as the absolute and relative values of VP weights from the RP treated animals compared to the NP control treated animals was observed. Normal glandular architecture was observed in both groups. There was no change on AR protein expression in the VP tissues. However, the proliferation rate and p63+ cells were higher in the RP group indicating that the low androgenic stimulation induced by MPM promoted a delay in the VP development. As a consequence, the proliferation/differentiation cell dynamic was impaired, leading to an increase in the number of basal cells and damaging their differentiation into secretory luminal cells.

The activation process of T and B-lymphocytes involves the increase in cellular proliferation and upregulation of telomerase activity, comparable to those observed in stem cells and transformed cell lines. However, aging decline of total number of T and B-lymphocytes (immunosenescence) due decrease in the telomerase activity that is responsible to telomeric DNA region restoration. As immunosenescence is a process that presents individuals variation, the influence of genetic alteration of oxidative stress metabolism on telomerase gene expression of activated lymphocytes, need to be elucidated. Human beings present a superoxide dismutase manganese dependent (SOD2) single nucleotide polymorphism (SNP) (rs4880 in 16 codon (Ala16Val-SOD2) generating three genotypes. Both homozygous genotypes create imbalance in O-2– H2O2 levels due change in SOD2 efficiency (VV = low efficiency; AA= high efficiency) and has been associated with chronic diseases and differential response to drug and toxicants exposition. Therefore, we evaluated the effect of SOD2-SNP cell expansion, viability and telomerase expression of peripheral blood mononuclear cells (PBMCs) from subjects with different Ala16Val-SOD2 genotypes (n=12). PBMCs were cultured in RPMI 1340 in controlled conditions (CO2 5%-37oC) and the viability/cells proliferation was evaluated by MTT assay and flow cytometry (cell cycle and apoptosis induction) after 1, 3, 7 and 14 days. The telomerase modulation was analyzed by qRT-PCR and telomere shortening by ELISA immunoassay. Whereas, after 3 three days exposition AA-PBMCs presented high proliferative rate than other genotypes, from this period also occur a fast loss of proliferation and apoptosis processes. After 15 days, only VV genotype presented upregulation of telomerase gene. The present study corroborate the hypothesis that proliferation-imunosenescence is broadly regulated by O-2– H2O2 mitochondrial levels

Recently, increasing studies proved the relationship between oxidative stress and osteoporosis. And some studies have been done to detect the possible beneficial effects of Alpha-lipoic acid (ALA), a potent antioxidant, on osteoporosis in vivo and in vitro. However, the detailed mechanism(s) underlying the bone-protective action of ALA are still poorly understood. The present study aims to examine the mechanisms by which ALA produces bone-protective effects in vitro and in vivo on base of its antioxidant effects, thus the effects of ALA on H2O2-treated MC3T3-E1 pre-osteoblasts and ovariectomized osteoporosis rat model were investigated by using bone biomechanical testing, micro computed tomography, western blotting and qRT-PCR analysis. The results showed that ALA promoted osteoblastic formation, inhibited osteoblastic apoptosis, increased OPG/RANKL ratio and enhanced bone formation in vitro and prevented bone loss in vivo. And the effects of ALA were via modulating Nox4/ROS/NF-κB and Wnt/Lrp5/β-catenin signaling pathways. The current study indicated that ALA might be beneficial for the prevention and therapy of osteoporosis clinically at the end of advanced studies.

Therapeutic Role of Bone Marrow-Derived Mesenchymal Stem Cells in Cyclophosphamide-Induced Cardiotoxicity in Adult Male Albino Rat: Morphological and Immunohistochemical Study

The aim of the present study was to evaluate the therapeutic role of BM-MSCs in CP-induced cardiotoxicity. Adult male albino rats weighing 200-230g were allocated into four groups: Group I received no manipulation; group II received a single dose of CP (200 mg/kg bw), i.p. on day 1 and sacrificed on day 10; group III had the same treatment as group II but sacrificed on day 40; group IV received the same treatment as group II followed by a single MSC injection (1x106) on day 10 and sacrificed on day 40. CPK serum level increased 10 days after CP administration then decreased to almost normal following MSCs treatment but not in the recovery group. Histological changes following CP administration such as distortion of the myocardial architecture, widening of the spaces, ballooning, rarefaction and vacuolation of the sarcoplasm, nuclear changes, inflammatory exudate and cellular infiltration as well as congested blood capillaries were encountered. Collagen fibers significantly increased following CP and remained high in the recovery group, but markedly decreased following MSCs treatment. Apoptotic bodies confirmed by immunohistochemical reaction of bax (pro-apoptotic) and cardiac tissue level of bax gene showed marked increase in CP-treated and recovery groups, but a decrease in MSCs-treated. On the other hand, bcl-2 reaction (anti-apoptotic) and GPx gene were markedly decreased in CP-treated and recovery groups but increase in MSCs–treated group. It could be concluded that MSCs treatment could ameliorate the histological and biochemical adverse effects in CP-induced cardiotoxicity.

Carnosic acid (CA), found in rosemary, has been reported to have antioxidant and anti-adipogenic properties. Here, we investigate the molecular mechanism by which CA inhibits hydrogen peroxide (H2O2)-induced oxidative injury in HepG2 cells. Cells were pretreated with 2.5–10 µM of CA for 2 h and then exposed to 3 mM H2O2 for an additional 4 h. CA dose dependently increased cell viability and decreased lactate dehydrogenase (LDH) activities. Pretreatment with CA completely attenuated the inhibited expression of manganese superoxide dismutase (MnSOD) and the B-cell lymphoma-extra large (Bcl-xL) caused by H2O2 exposure, whereas reversed the reactive oxygen species (ROS) accumulation and the increase in cleaved caspase-3. Importantly, sirtuin1 (SIRT1), a NAD+-dependent deacetylase, was significantly increased by CA. Considering the above results, we hypothesized that SIRT1 may play important roles in the protective effects of CA in the H2O2 injury. As expected, SIRT1 suppression by a specific inhibitor of SIRT1, Ex527, significantly aggravated H2O2-induced increased level of cleaved caspase-3, but deeply reduced the decreased expression of MnSOD and Bcl-xL. Collectively, the present study indicated that CA can alleviate H2O2-induced hepatocyte oxidative and apoptotic damage through SIRT1 pathway.

Idiberto José Zotarelli Filho has received his PhD from UNESP University in the field of Regenerative Medicine and Tissue Engineering. Currently, he is working as Researcher in the Institute of Cardiovascular Diseases and UNESP University. He has successfully completed his Administrative Responsibilities as Researcher. He has received several awards and honors, such as the International Symposium of extracellular matrix - Buzios / Brazil, International Congress of Hematology - Albert Einstein Hospital - Sao Paulo / Brazil and the International Congress of cell therapy stem cells in São Paulo / Brazil. He has published more than 100 peer-reviewed in journals and is an author of 2 books. Also actively participated in the training program in research - Research Coaching Program - Durham - Duke University - USA under the auspices of the Brazilian Cardiology Society - SBC and is a member of the Research Ethics Committee - Institute of Cardiovascular Diseases (IMC).

Scaffolds of chitosan and collagen can offer a biological niche for the growth of adipose derived stem cells (ADSC). The objective of this work was to characterize the physico-chemical properties of the scaffolds and the ADSC, as well as their interactions to direct influences of the scaffolds on the behavior of ADSC. The methodology included an enzymatic treatment of fat obtained by liposuction by collagenase, ASDC immunophenotyping, cell growth kinetics, biocompatibility studies of the scaffolds analyzed by the activity of alkaline phosphatase (AP), nitric oxide (NO) determination by the Griess-Saltzman reaction, and images of both optical and scanning electron microscopy of the matrices. The extent of the crosslinking of genipin and glutaraldehyde was evaluated by ninhydrin assays, solubility tests and degradation of the matrices. The results showed that the matrices are biocompatible, exhibit physical and chemical properties needed to house cells in vivo and are strong stimulators of signaling proteins (AP) and other molecules (NO) which are important in tissue healing. Therefore, the matrices provide a biological niche for ADSC adhesion, proliferation and cells activities.

Giraud-Billoud Maximiliano has completed his both PhD in Medicine and Biology (Physiology) from National University of Cuyo, Argentina and is working now in a Postdoctoral project from FONDECyT-CONICyT, Chile. He has published 11 papers in reputed journals.

Megalin and cubilin are membrane endocytic receptors that interact for endocytosis of a vast variety of filtered plasma proteins in kidney proximal tubule cells. One characteristic of diabetic nephropathy is the increase of albumin filtration and the inhibition of megalin and cubilin mediated albumin endocytosis, leading to increased albuminuria. The objective of our work wasto study the influence of mesenchymal stem cells (MSC) therapy on tubular protein transporters in kidney of type 1 and type 2 diabetic animals. We studied by Western Blot the expression of megalin (tissue) and cubilin (tissue and urine) and the functionality of megalin by the quantification of vitamin D-binding protein (BDP, urine) in control animals, type 1 and 2 diabetic animals (C57BL6+STZ and db/db C57BKS mouse, respectively) and treated with MSC (0.5x106 cells/animal) type 1 and 2 diabetic animals. The expression of megalin and cubilin decreases in T1 diabetic animals and after 2 weeks of MSC administration the expression increases. In T2 diabetic animals, only megalin decreases significantly its expression and treatment does not increase levels. Proteinuria and BDP concentration are increased in both diabetic models, but only BDP levels decrease in T2 animals after MSC administration. Finally, levels of cubilin in urinary samples are increased in both diabetic models and only decrease significantly in T1 diabetic animals that received the cell therapy. MSC administration in both experimental diabetic models improves the action of tubular protein transporters, however its potential role as a therapy and the physiopathologic mechanisms involved are currently under study.

Ample evidence has suggested that α-synuclein is a toxic species in synucleinopathies widely distributed in brain. Overexpression of α-synuclein predominantly localize to axons in the neuron of Parkinson’s disease (PD) which affects microtubule (MT) network, trafficking and actin polymerization. Furthermore, overexpression of α-synuclein and its mutant resulted in disruption of neurite morphology, microtubule cytoskeleton and the transport of endosomes as well as autophagosomes, but the specific relation of α-synuclein and different compartments of the neuronal cytoskeletal machinery has not yet been elucidated. Therefore, MT stability would play key roles in the pathogenesis and progression of PD and would be clinically relevant in PD treatment. Mesenchymal stem cells (MSCs) secrete various cytotropic factors that have neuroprotective effects through complex mechanisms, such as modulation of neuroinflammation, enhancement of cell survival signals, increased neurogenesis, and modulation of autophagy. In the present study, we identified that eukaryotic translation elongation factor 1A (eEF1A), soluble factors derived from MSCs and MSCs could exert neuroprotective effects through modulation of microtubule binding, bundling and severing on α-synuclein time-dependently. In addition, we showed that MSCs had effect on axonal transport and cell growth, which led to a prosurvival effect on neurons. Furthermore, we presented that MSCs modulate axonal transport of signaling and fuse the outer membrane of the autophagosome with lysosome formation.These studies suggest that MSCs on the microtubule system a potential target of α-synuclein, and might have an impact on neuronal structure, function and survival.

Darwesh MK Aladin received his PhD in 2010 from the University of Hong Kong. His PhD work was on the multi scale structure and mechanics of intervertebral discs. He is currently pursuing his postdoctoral studies at the Mechanobiology Institute, Singapore, in which he is studying intercellular adhesion and the role of cadherin extracellular domains in particular. He has published 6 papers in reputed journals. He has presented in more than 30 conferences and has won 18 awards including an honorable mention award from the ASME Bioengineering Division.

Adherens Junctions (AJs) are molecular ensembles that occur at cell–cell junctions in epithelial and endothelial tissues. Cadherins cluster during AJ formation and recruit a multitude of additional structural and regulatory proteins. The intracellular tail of cadherin binds to catenin complexes that couples cadherins with the actin cytoskeleton. Over 170 proteins have been reported to colocalize in the AJs. Our SILAC immunoprecipitation experiment aimed at identifying more proteins interacting with E-cadherin identified AQP1 as an interacting partner. AQP1 is a member of membrane water channel proteins called the Aquaporins. Silencing of AQP1 is known to dramatically affect the actin cytoskeleton organization through Lin-7/β-catenin interaction. Since actin dynamics is known to regulate E-cadherin recruitment at AJs in a mechanosensitive manner, we hypothesized that knocking down AQP1 will inhibit the adhesion strength of AJs. Using dual pipette separation force assay, we measured the force required to separate cell doublets that are formed in 18 hrs of culture and compared between the SiRNA mediated AQP1 knock down S180 cells expressing E-cadherin and the negative SiRNA control cells. As hypothesized, the separation force of AQP1 knock down cells was significantly lower than that of the controls. Furthermore, a micropipette aspiration assay of single cells revealed that the cortical shear modulus of the AQP1 knock down cells was significantly lower than the control cells. These findings suggest that AQP1 plays a role in the adhesive strength of AJs possibly by controlling the dynamics of actin cytoskeleton.

Petrenko A Yu is a Professor in Cell Biology and Cryobiology. He is the Head of Department in the Institute for Problems of Cryobiology and Cryomedicine, Professor of Kharkov University, Professor of UNESCO Chair. He is the Author of monograph “Stem cells: Properties and perspectives of clinical use” and has over 400 publications, of which about 100 are in PubMed. He is an Editorial Board Member of 3 scientific journals, a Member of several national and international societies and recipient of various awards.

Mesenchymal stromal cells (MSCs) can differentiate into various cell types, which make them attractive for regenerative medicine. Preservation of MSCs seeded and cultivated in scaffolds at cryogenic or hypothermic temperatures can serve as ready-to-use transplantation units for tissue repair. The aim of the study was to investigate the ability of proliferation and multilineage differentiation of MSCs within alginate microspheres (AMS) and porous scaffolds before and after different approaches of low temperature preservation. As porous scaffolds for MSCs alginate-gelatin macroporous cryogel sponges and plane demineralized chitinous skeletons of marine sponge Ianthella basta were used. MSCs isolated from adult human bone marrow, dermal and adipose tissues after expansion in monolayer culture had similar phenotype and growth patterns as well as ability to induce multilineage differentiation. MSCs in AMS had spherical shape and could be successfully cryopreserved by both conventional cryopreservations with 10% of dimethyl sulfoxide and vitrification protocols. Besides, encapsulation in AMS delayed cell death during storage at ambient temperatures. The response of MSCs seeded and cultivated into porous scaffolds to conventional cryopreservation protocol depended on cell adhesion and spreading as well as the structure of scaffolds per se. Between the observed three types of scaffold, the lowest cryoresistance had MSCs growing as sheets on chitinous marine demosponge Ianthella basta. After cryopreservation in various scaffolds survived MSCs retained abilities to proliferation and differentiation into osteogenic and adipogenic lineages. The data obtained indicate that cryo-banking of MSCs cultivated into tissue engineered scaffolds is feasible for the future regenerative medicine projects.

Panagiotis Zogopoulos is a resident of Neurosurgery at the General Hospital of Nikaia-Piraeus “Agios Panteleimon”, Athens, Greece. His ongoing research is in the field of drugs and their interaction with human brain and cerebral vessels. Several of his papers have been published in reputed peer-review journals and he has presented various researches in international conferences.

Cannabinoids are chemical compounds that have recently been added to the therapeutic armamentarium of cancer both for their palliative effects (inhibition of chemotherapy-induced nausea and vomiting, stimulation of appetite) but also as antitumor drugs. Δ9-Tetrahydrocannabinol (the primary psychoactive component of cannabis plant), cannabinol and cannabidiol are the most studied natural cannabinoids. They can induce cancer cell apoptosis and reduce neoangiogenesis as well as tumor cell invasiveness and metastasis in various cancer types such as gliomas, breast, prostate, skin, lung, thyroid, gastric, colon, hepatocellular, pancreatic, lymphoma and leukemia as has been shown in experimental studies. Furthemore, the endogenous signalling system of endo cannabinoids which share common receptors and manifest similar actions with cannabinoids is also a promising field for the treatment of cancer. They include anandamide and 2-arachidonoylglycerol, which are the most studied but also 2-arachidonoylglyceryl ether (noladin ether), N-arachidonoyldopamine and O-arachidonoylethanolamine (virodhamine). Besides their antitumorigenic action, the lack of severe adverse side effects of cannabinoids and endocannabinoids compared to the generalized toxic actions of conventional chemotherapeutics which makes them promising candidates for the future treatment of various cancer types in human and clinical trials should be planned towards that direction.

César Vinícius Gil Braz do Prado is a Certified Veterinary Acupuncturist. He has completed his Veterinary Graduation in 2010 and has specialized in Veterinary Acupuncture on 2012, working on Traditional Chinese Veterinary Medicine in small and wild animals since then. He is a PhD student of University of São Paulo and his main research area is stem cells and acupuncture.

Studies have shown a possible synergism between electro acupuncture and stem cells grafts in the treatment of acute Spinal Cord Injury (SCI). This study aims to evaluate the association among those therapiesin dogs with severe chronic SCI. 20 paraplegic dogs with thoracolumbar intervertebral disc disease were divided randomly into 4 groups: Stem Cells (ST), Electro Acupuncture (EA), Stem Cells + Electro Acupuncture (ST+EA) and Control group (C). 11 of 20 dogs have been treated already. Functional improvement was observed on Group ST (2 of 4 animals), Group EA (1 of 3 animals) and Group EA+ST (1 of 4 animals). Changes on neurological assessment were observed on: Group ST (2 points in 4 of 4 animals), Group EA (2 points in 2 animalsand 1 point in 1 animal, of 3 animals) and Group ST+EA (6 points in 2 animals, 5 points in 1 animal and 1 point in 1 animal, of 4 animals). The 9 remaining animals will still be treated, including the animals from Control Group and more tests will still be analyzed (e.g., magnetic resonance images and neurotrophic factors levels on cerebrospinal fluid and blood serum). Although, the partial results showed that all animals had some kind of neurological improvement (including recovery of urinary control and voluntary tail movement), which is more evident on animals with hind limb medullar reflexes preserved prior to the study. Non statistical inference can be done for now until all animals are tested.

Background: Chronic leg ulcer is a major health problem which may associate different diseases as vascular, neoplastic, infectious diseases and trauma as well. The patients suffer from many complications as infection, septicemia, psychology troubles and physical inability leading to many economic troubles the last line of treatment is amputation with its high rate of morbidity and mortality. Aim: The study was design for randomized controlled clinical trial in an attempt to assess the potential treatment of chronic leg ulcer through the use of the autologous bone marrow stem cells. Patients & Methods: Fifty patients with chronic leg ulcer were included from vascular surgery department Tanta University Hospital. Patients were divided randomly into two equal groups, each group included 25 patients. In group I, the patients were subjected to autologous transplantation of bone marrow derived CD34+ mononuclear cells into the depth and around the ulcer. In group II (control group), saline dressing to the leg ulcer was done. Results: In group I,14 patients were males and 11 were females, age ranged from 19 to 66 years with mean age of 48.7 years while in group II 13 patients were males and 12 patients were males with mean age of 51.2 years and age ranged from 21 to 63 years. In group I, the follow up period ranged from 6 to 24 months. Complete ulcer healing occurred in 12 patients (48%), partial ulcer healing in 10 patients (40%) and no healing in 3 patients (12%).In group II, the follow up period ranged from 8 to 24 months. Complete ulcer healing occurred in 2 patients (8%), partial ulcer healing in 3 patients (12%) and no healing in 20 patients (80%). In group I, the overall patients satisfaction and pain relief was found in 18 patients (72%) while in group II only 4 patients (16%). Conclusion: Autologous transplantation of bone marrow derived CD34+ mononuclear cells is a simple, safe and effective modality of treatment for chronic leg ulcer.