Renal graft survival has improved over the past years, mainly owing to better immunosuppression. Vascular thrombosis, though rare, therefore accounts for up to one third of early graft loss. We assess current literature on transplantation, identify thrombosis risk factors, and discuss means of avoiding thrombotic events and saving thrombosed grafts. The incidence of arterial thrombosis was reported to 0.2–7.5% and venous thrombosis 0.1–8.2%, with the highest incidence among children and infants, and the lowest in living donor reports. The most significant risk factors for developing thrombosis were donor-age below 6 or above 60 years, or recipient-age below 5-6 years, per- or postoperative hemodynamic instability, peritoneal dialysis, diabetic nephropathy, a history of thrombosis, deceased donor, or >24 hours cold ischemia. Multiple arteries were not a risk factor, and a right kidney graft was most often reported not to be. Given the thrombosed kidney graft is diagnosed in time, salvage is possible by urgent reoperation and thrombectomy. Despite meticulous attentions to reduce thrombotic risk factors, thrombosis cannot be entirely prevented and means to an early detection of this complication is desirable in order to save the kidneys through prompt reoperation. Microdialysis may be a new tool for this. In 1905, the first paper on kidney transplantation was published [Fifty years after the pioneering experiments, the first successful human renal transplantation was carried out in Boston by Dr. Joseph Murray in 1954 . The donIn 2012, kidney transplantation remains the best treatment of end-stage renal disease (ESRD) resulting in improved health and quality of life  as well The surgical technique has only seen minor changes since the first renal transplantations were accomplished. Usually the kidney is placed extraperitoneally in the iliac fossa with end-to-side anastomoses to the external iliac artery and vein. In small children and infants, however, this approach is often associated with an increased risk of thrombosis . To avoiVascular thrombosis is a devastating complication because it most often leads to renal graft loss. Overall, the incidence of arterial thrombosis is reported to be 0.2–7.5% and venous thrombosis to be 0.1–8.2% as seen from Tables In one of the studies with the lowest number of thrombosis , only liUnfortunately, only few of the study data available are prospectively collected , 31, 34,Historically, renal grafts with multiple arteries have been associated with an increased risk of thrombosis ; howeverSurplus renal arteries are found in 10–20% of the population. In order to succeed in overcoming the technical challenges, it requires an experienced surgeon with an in-depth knowledge of possible variations in renal arterial anatomy and of microsurgical techniques for vascular reconstruction . The artThere is no general consensus about whether a right renal graft has increased thrombosis risk compared to the left. A few centers, with both high and low thrombosis incidence, find no association , 17, 32, The renal graft is often positioned in the contralateral iliac fossa, but also the same side as the original position can be used as there is no association between complications and the side of graft placement , 15, 17.On average, around 35% of all renal transplantations are carried out with a kidney from a living donor with large variation from country to country. When using a living donor, the anatomy is well known preoperatively and the procedure is scheduled. This often leads to better results and fewer thromboses , which iA female donor doubles the risk of thrombosis in the case-control study from the Australian and New Zealand Dialysis and Transplant Registry , probablAlthough few studies find no association between donor age and thrombosis , 12, 17,Warm ischemic time (WIT) has not been found to be a risk factor for thrombosis development , 17, 21.As previously mentioned, the placement of the renal graft in small children may influence the thrombotic risk. In a retrospective German study, children aged 6 years or younger had kidney grafts transplanted either retroperitoneally to the iliac vessels in the fossa iliaca or extraperitoneally with end-to-side anastomoses to the distal aorta and caval vein . The proIntra- and postoperative hemodynamic instability is rarely mentioned in transplantation outcome reports, maybe because the data are not always accessible. Nevertheless, this may be an important contributor to thrombotic risk. In a retrospective report from The Netherlands, it was found to be an independent risk factor . It was Due to transplantation being a relatively infrequent operation in many centers, some reports only have few patients of extreme ages and a difference was not always found , 17, 25.Also elderly recipients are more prone to graft thrombosis . NeverthIn the preceding paragraph, it was concluded that donor gender was of less importance. The same is the case for recipient gender, with only one large case-control study finding a 46% higher likelihood of thrombosis in women , while sThe underlying disease that causes ESRD in the patient also plays a role in the transplantation outcome. Of significance is diabetic nephropathy , 13, 15 The progression of the renal disease and possible comorbidities determine when and how the patient should be treated. There are three renal replacement options when reaching ESRD: transplantation, hemodialysis (HD), and peritoneal dialysis (PD). The choice, order, and duration of treatment can have a substantial impact on transplantation outcome. Preemptive transplantation, that is, when the new kidney is transplanted before initiation of dialysis, while the native kidneys still have a high urine production, is preferred to avoid dialysis and associated especially cardiovascular risks, but it has been found to be associated with increased thrombosis risk compared to transplantation after initiation of HD , 13. FurSeveral studies with large patient cohorts find substantial more thrombotic graft loss among patients who have been on PD prior to transplantation compared to HD , 13, 55,Inherited or acquired forms of thrombophilia increase the risk of thrombosis in general and hence also in a newly received renal graft and anticoagulation may be a sound consideration. Screening for thrombophilia has shown to decrease the number of thrombotic events , 56. ThiSome studies are too underpowered to detect an effect of anticoagulation among unselected recipients. For instance, heparin is used prophylactic in case of vague risk factors , but it Delayed graft function is strongly correlated to thrombosis and a vascular cause for decreased graft function should always be considered , 14, 21.Diagnosis of vascular thrombosis can be time consuming, from request to implementation, and may thus be delayed. Consequently, kidney grafts developing thrombosis are most often lost, and the patient is returned to dialysis treatment. However, with early diagnosis and intervention, it is possible to save the kidney , 58–60. Venous thrombosis classically presents itself with a swollen tender graft and hematuria, while arterial thrombosis can be more silent with a sudden drop in urine output. Intrarenal thrombosis may also be involved in severe rejections. Symptoms are often sparse, and the risk of delayed graft function is high. This can be masked by a prior low urine production or native kidney function in preemptive patients. Usually thrombotic events occur within the first 10 postoperative days, in 93% of the cases within day seven . ArteriaWhen vascular thrombosis is suspected clinically or in a routine screening, the diagnosis is confirmed by color Doppler flow examinations, which may also be used within the first hours after transplantation if the kidney is not functioning at once after revascularization. This technique detects vascular complications in general, including renal artery stenosis, with an overall sensitivity and specificity of 88% and 85%, respectively . The accDespite meticulous attentions to the risk factors discussed, thrombosis is not completely preventable and means to a fast early detection of this complication are desirable in order to implement the above-mentioned opportunities for salvage. Some steps have been taken in this direction, and thermodiffusion has been suggested as a possible new dynamic diagnostic technique. A small thermodiffusion electrode placed in the tissue continuously measures the absolute microcirculation in mL/100 g tissue per minute. It has been applied to ischemic porcine kidneys  and liveMicrodialysis is another minimally invasive technique for dynamic monitoring of the local metabolism, which is being increasingly used experimentally as well as in the clinic. A small catheter is placed in the tissue of interest and by simple diffusion through a semipermeable membrane; the concentration of local metabolites can be estimated. The reconstructive surgeons have used it for more than ten years for free flap monitoring, for example, when the esophagus is reconstructed by a free jejunal flap after carcinoma resection. Critical ischemia locally within the flap is detected by microdialysis; accordingly, the flap can be reoperated and revascularized in time . The twoArterial and venous renal graft thromboses are rare, but nevertheless devastating complications that usually lead to graft loss and accounts for a considerable fraction of early graft losses. Especially in the situation with delayed graft function, there is a need for continuous monitoring to facilitate salvage and avoid explorative surgery when no thrombosis is present.The most significant risk factors for thrombosis are donor-age below 6 or above 60 years or recipient-age below 5–6 years, per- or postoperatively hemodynamic instability, PD, diabetic nephropathy, a history of thrombosis, deceased donor, or more than 24 hours CIT. Multiple arteries and right kidney are not as severe risk factors as earlier thought, but should, nevertheless, be handled very carefully by the surgeon. A lot of the risk factors are not adjustable, and the kidneys are to be used as there is universal organ shortage and a need for kidneys even from marginal donors. When risk factors are present, the number of thrombotic events may be reduced, for example, by aggressive fluid management of pediatric patients or anticoagulation of thrombophilic patients. Furthermore, there is a need for early detection as thrombosis is not completely preventable. Microdialysis may be suitable for this.


The objective of the present study was to evaluate whether placental mesenchymal stromal cells (PDMSCs) derived from normal and preeclamptic (PE) chorionic villous tissue presented differences in their cytokines expression profiles. Moreover, we investigated the effects of conditioned media from normal and PE-PDMSCs on the expression of pro-inflammatory Macrophage migration Inhibitory Factor (MIF), Vascular Endothelial Growth Factor (VEGF), soluble FMS-like tyrosine kinase-1 (sFlt-1) and free β-human Chorionic Gonadotropin (βhCG) by normal term villous explants. This information will help to understand whether anomalies in PE-PDMSCs could cause or contribute to the anomalies typical of preeclampsia.Chorionic villous PDMSCs were isolated from severe preeclamptic (n = 12) and physiological control term (n = 12) placentae. Control and PE-PDMSCs’s cytokines expression profiles were determined by Cytokine Array. Control and PE-PDMSCs were plated for 72 h and conditioned media (CM) was collected. Physiological villous explants (n = 48) were treated with control or PE-PDMSCs CM for 72 h and processed for mRNA and protein isolation. MIF, VEGF and sFlt-1 mRNA and protein expression were analyzed by Real Time PCR and Western Blot respectively. Free βhCG was assessed by immunofluorescent.Cytokine array showed increased release of pro-inflammatory cytokines by PE relative to control PDMSCs. Physiological explants treated with PE-PDMSCs CM showed significantly increased MIF and sFlt-1 expression relative to untreated and control PDMSCs CM explants. Interestingly, both control and PE-PDMSCs media induced VEGF mRNA increase while only normal PDMSCs media promoted VEGF protein accumulation. PE-PDMSCs CM explants released significantly increased amounts of free βhCG relative to normal PDMSCs CM ones.Herein, we reported elevated production of pro-inflammatory cytokines by PE-PDMSCs. Importantly, PE PDMSCs induced a PE-like phenotype in physiological villous explants. Our data clearly depict chorionic mesenchymal stromal cells as central players in placental physiopathology, thus opening to new intriguing perspectives for the treatment of human placental-related disorders as preeclampsia. Preeclampsia (PE) is a severe placenta-related syndrome exclusive of human pregnancy that represents the main cause of feto-maternal mortality and morbidity worldwide The trophoblast is generally considered the main placental site of the morphological and molecular alterations responsible for the onset of preeclampsia. Nevertheless, the human placenta is a heterogeneous organ composed of several cellular populations each contributing to the organ physiology. Indeed, beside the trophoblast, placental villi are composed of endothelial cells and progenitors, myofibroblasts and mesenchymal stromal cells forming a complex network that concertedly interacts to maintain placental functionality.Recently, the human placenta has been identified as source of a particular type of mesenchymal stromal cells (MSCs) that retain stem cell-like properties. Placental MSCs (PDMSCs) have been successfully isolated from the placental basal plate, the amnion and the decidua. They are characterized by different degrees of plasticity and possess unique immunologic and immune-regulatory properties MSCs plasticity is of great interest for regenerative medicine and several groups are exploring the possibilities offered by the placenta as a new therapeutic tool. On the other hand, the investigation of placental MSCs immune-modulatory, pro-angiogenic and anti-inflammatory properties could open new perspectives into the understanding of placenta-related disorders due to maternal immune maladaptation and aberrations of trophoblast development as preeclampsia. The secretion of cytokines and chemoattractant by the first trimester placenta with consequent recruitment of lymphocytes, natural killer cells and macrophages is necessary to maintain proper pregnancy physiology Our hypothesis is that anomalies in PDMSCs anti-inflammatory and pro-angiogenic properties might cause or contribute to the defective placental development and/or immune maladaptation typical of preeclampsia”. In the present study, we characterized cytokine expression profiles of PDMSCs isolated from the basal plate of normal and severe PE placentae. Moreover, we investigated the effect of conditioned media from normal and PE PDMSCs on the expression of Macrophage migration Inhibitory Factor (MIF), Vascular Endothelial Growth Factor (VEGF), soluble FMS-like tyrosine kinase-1 (sFlt-1) and β-human Chorionic Gonadotropin (βhCG) in normal placental villous explants. MIF is a pro-inflammatory cytokine key player in PE inflammatory response This study was conducted according to the principles expressed in the Declaration of Helsinki. The study was approved by the Institutional Review Board of O.I.R.M. S.Anna Hospital and “Ordine Mauriziano di Torino”  . All patients provided written informed consent for the collection of samples and subsequent analysis.th weeks of gestation in previously normotensive women The study groups included singleton pregnancies complicated by severe preeclampsia (n = 12) and physiological control term pregnancies (n = 12). The diagnosis of PE was made according to the following criteria: presence of pregnancy-induced hypertension  and proteinuria (≥300 mg/24 h) after the 202.PDMSCs were isolated by using a protocol modified from Brooke and colleagues 5′-CGT GAA GCTG GAG AAG GAG AAG CTG-3′) RV(5′-CAA GGG CCG CAG CTT ACA CAT GTT C-3′); Nanog FW(5′-AAT ACC TCA GCC TCC AGC AGA TG-3′) RV(5′-CTG CGT CAC ACC ATT GCT ATT CT-3′).After passages three to five, the cells were evaluated for specific mesenchymal stromal cells antigens by flow cytometry. The following markers were investigated: HLA-DR, CD105, CD166, CD90, CD34, CD73, CD133, CD20, CD326, CD31, CD45, CD14. All the antibodies were purchased from Myltenyi Biotec . Moreover, Oct4 and Nanog mRNA expression levels were assessed in both control and PE PDMSCs by semi-quantitative RT PCR in order to verify cellular stemness. Primers sequences were the following: Oct4 FW. At passage five, physiological (3 different cell lines) and PE (3 different cell lines) PDMSCs were seeded in 96-well plates at a density of 6000 cells per well. Each cell lines was plated in six different wells. At time 0, 48 h and 120 h of culture, cells were incubated with 5 mg/ml of MTT for 4 h. Next, medium was removed and formazan salts were dissolved with 100 µl of dimethylsulfoxide . 570 nm absorbance at each time point was determined using a microplate spectrophotomether in order to establish PDMSCs cell proliferation curve.5 cells/well in DMEM LG supplemented with 10% FBS. Each cell line was seeded in six different wells. After 48 h of culture, the cells were fixed in 1X Fixation Buffer for 6 minutes and were stained with SA-b-gal–staining solution for 4 h. The senescent SA-b-gal–positive cells exhibited a blue nuclear color. Number of positive cells number was determined by phase-contrast microscopy.Senescence-associated β-galactosidase  nuclear staining was performed using a SA-b-gal staining kit  following manufacturer instructions. At passage five, normal (two different cell lines) and PE (2 different cell lines) PDMSCs were plated in 6 well plates at a density of 2×105 cells/ml . After 72 h of culture, conditioned media was collected, centrifuged, filtered to remove cellular debries and immediately stored at −80°C, while cells were processed for mRNA and protein isolation.At passage five, PDMSCs were plated in 6 well plates at a density of 2.0×10To investigate differences in cytokines production between normal and preeclamptic PDMSCs, the RayBio Human Cytokine Antibody Array  was used. This specific array was chosen because it allows contemporary detection and quantification of eighty different human cytokines, chemokines and inflammation-related growth factors. Arrays were performed on unconditioned and conditioned culture media from normal and PE PDMSCs (prepared as described above) following manufacturer instructions. Cytokines levels were quantified by densitometric analysis using ImageQuant software and normalized by setting the positive controls as 100 and negative controls as zero percent. The arrays were performed in duplicate on CMs derived from four normal term PDMSCs lines, four preeclamptic PDMSCs lines and on unconditioned media (UCM) as baseline for molecules already present in the culture media. UCM data were used to normalize Normal and PE PDMSCs CM results. Cell lines used were all derived from different control and PE patients.2 to equilibrate. Next, culture media was removed and explants were treated for 72 hours with 500 µl of conditioned media from 4 different control PDMSCs lines (n = 16 explants) and 4 different PE PDMSCs lines (n = 16 explants). Explants treated by unconditioned culture media were used as controls (n = 16 explants). Finally, culture media were collected while control and treated explants were processed for mRNA and protein isolation. Each experimental condition was performed in duplicate.Placental biopsies from physiological term placentae were processed within 2 hours from delivery. Fetal membranes and decidua were mechanically removed and placental tissues were washed in phosphate-buffered saline (PBS) solution to remove excess of blood. Small portions of placental chorionic villi  were excised and placed in a 24-well culture dish. Explants were cultured in Ham’s F12 media  and incubated overnight at 37°C and 5% COTotal RNA was extracted from placental MSCs and chorionic villous explants using TRIZOL reagent according to manufacturer instructions  and treated with DNAse I to remove genomic DNA contamination. Two µg of total RNA was reverse transcribed using random hexamers approach . The resulting templates were quantified by Real-time PCR . TaqMan primers and probes for ribosomal 18S, MIF, VEGF, HCGB, IL-8, IL-6 and TNF-α were purchased from Applied Biosystems as TaqMan® Gene Expression Assays. sFlt-1 primers and probe were designed as previously described by Nevo and colleagues Total proteins were isolated from PDMSCs and chorionic villous explants using 1X Radio Immuno-precipitation Assay (RIPA) buffer. Fifty µg of total protein from PDMSCs and villous explants were processed by SDS-page electrophoresis on 4–12% polyacrylamide pre-cast gradient gels . Next, proteins were transferred on Polyvinylidene fluoride (PVDF) membranes and probed at room temperature with primary antibodies using the SnapID system  following manufacturer instructions. Primary antibodies were: mouse monoclonal anti-human MIF , rabbit polyclonal anti-human VEGF , rabbit polyclonal anti-human soluble VEGF Receptor , rabbit polyclonal anti-human Ki67 , goat polyclonal anti-human Cytokeratin 7 , mouse monoclonal anti-human CD45  and mouse monoclonal anti-human CD166 . Biotinylated secondary antibodies were goat anti-mouse for MIF, CD45 and CD166  and donkey anti-rabbit for VEGF, sFlt-1 and Ki67  and donkey anti-goat for Cytokeratin 7 . Blots were visualized using the QDot reagent kit  following manufacturer instructions.Free β hCG released by physiological villous explants treated with normal or PE PDMSCs conditioned media was measured using the B·R·A·H·M·S Free βhCG KRYPTOR immunofluorescent assay  following manufacturer instructions.Concentrations of TNF-α and VEGF proteins in normal and PE-PDMSCs CM and in protein lysates of CM-treated villous explants were evaluated by commercially available ELISA kits (Peprotech), according to the manufacturer’s recommendations. Values are given as the mean concentration ± SEM of fourteen (PDMSCs CM) and twelve (CM-treated villous explants) independent experiments, all repeated in duplicate.All data are represented as mean ± SE. For comparison of data between multiple groups we used one-way analysis of variance (ANOVA) with posthoc Dunnett’s test. For comparison between 2 groups we used paired and unpaired Student’s t-test as appropriate. Statistical test were carried out using Graph Pad Prism 5 statistical software and significance was accepted at P<0.05.Clinical data of the study population are summarized in PDMSCs were isolated from both physiological term and preeclamptic placentae and, starting from passages 3 to 5, they were characterized for the expression of typical MSCs markers by flow cytometry. All PDMSCs lines investigated were positive for CD105, CD166, CD90 and CD73, while they were negative for HLAII, CD34, CD133, CD20, CD326, CD31, CD45 and CD14 , thus shPDMSCs cell proliferation at 48 h and 120 h was evaluated by MTT assay. We reported a significant cell proliferation increase in Normal PDMSCs at both 48 h  and 120 h , whereas PE PDMSCs showed a slower and not significant proliferation rate at both 48 h and 12 h . ImportaMesenchymal stem cells are a promising tool for regenerative medicine because of their unique immunomodulatory, pro-vasculogenic and anti-inflammatory properties. Since preeclampsia is characterized by an exacerbated feto-maternal inflammatory response accompanied by generalized endothelial damage and aberrant placental vascular development, we investigated whether PDMSCs derived from normal and PE placentae showed differences in cytokines production profiles. These data would help us to understand whether or not PDMSCs could contribute to physiological placenta development and/or to the aberrant placentation and inflammatory response typical of PE. To reach our goal, we performed a cytokine array able to detect 80 different cytokines  present In addition, Interferon-inducible protein-10 (IP-10), Leukemia inhibitory factor (LIF), Neurotrophin-3 (NT-3), Neutrophil-activating Protein-2 (NAP-2), Macrophage-migration Inhibitory Factor (MIF), Placental Growth Factor (PlGF) and Pulmonary and Activation-regulated Chemokine (PARC) were markedly increased (range of increase: 8–19%) in preeclamptic PDMSCs conditioned media .Furthermore, we performed Real Time PCR analysis on normal and PE-PDMSCs in order to confirm increased expression of IL-8, IL-6, MIF, TNF-α and VEGF as detected by Cytokine Array analysis . We seleSince we determined that preeclamptic PDMSCs released augmented concentrations of pro-inflammatory cytokines and chemotactic factors, we next investigated whether they were able to negatively influence inflammation and/or vasculogenesis on the placental tissue. To reach our goal, we treated term physiological chorionic villous explants with conditioned media from normal and PE PDMSCs. As markers of inflammation and vasculogenesis, we investigated MIF and VEGF expression. We found significantly increased MIF mRNA expression levels  in PE PDMSCs CM explants relative to both untreated controls and explants treated with normal PDMSCs conditioned media . ResultsVillous explants are representative of the whole placental tissue and composed of trophoblast, mesenchymal and endothelial cells. Thus, we investigated the contribution of these cell populations to the above mentioned results by determining the expression of Cyt7 (trophoblast marker), CD45  and CD166 (MSCs marker) in our experimental model. Cyt7 was strongly expressed without differences in untreated, normal and PE PDMSCs CM explants  brought us to the hypothesis that MSCs themselves could contribute to the pathophysiology of placentation in virtue of their immune-regulatory and anti-inflammatory properties. MSCs exert an immunosuppressive effect on T-cells Overall, we found higher expression of inflammatory molecules in media conditioned by PE relative to normal PDMSCs. In particular, preeclamptic PDMSCs released considerably elevated levels of IL-8, TGF-β2, TIMP-2, Osteoprotegerin, MCP-1, Osteopontin and, to a lesser extent, IL-6, TIMP-1, ENA-78, VEGF, Angiogenin, IGFBP-4, BDNF, LIF, IP10, PDGF-BB, MCP-3, TARC, TNF-α, Leptin and PlGF.Hwang and colleagues recently characterized cytokine production profiles of MSCs isolated from amnion and decidua of healthy versus preeclamptic placentae in vitroIn line with our results, Szarka and colleagues Other molecules abundantly over-expressed in PE-PDMSCs media are TGF-β2, TIMP-2, Osteoprotegerin and Osteopontin. Transforming Growth Factor-βs have been indicated as main regulators of trophoblast invasion TIMPs are matrix metalloproteinase inhibitors that suppress proliferation of endothelial cells, thus exerting an anti-angiogenic activity. TIMP-2 has a key role during pregnancy in maintaining the homeostasis of trophoblast invasivity. Thus, TIMP-2 expression is low during the first stages of pregnancy, to increase with advancing gestation in vivoOsteoprotegerin (OPG) inhibit bone resorption, regulates vascular integrity PE-PDMSCs expressed also higher levels of pro-angiogenic growth factors VEGF, Angiogenin and PlGF. Besides being pivotal for placentation, these angiogenic molecules can be strong endothelial cell activators. VEGF role in PE pregnancy has been controversially discussed, as its levels in PE maternal serum have been reported as increased Leptin, LIF, PDGF-BB and BDNF are as well augmented in PE-PDMSCs relative to control PDMSCs media. During pregnancy, these cytokines are involved in critical processes as placental invasion Taken together, our data clearly indicate that preeclamptic MSCs resident in the chorionic placental villi over-produce a repertoire of cytokines, chemokines and growth factors that could directly contribute to the anomalies of trophoblast development and placental angiogenesis and to the exacerbated inflammatory response typical of PE and FGR. These molecules, crucial for proper pregnancy outcome under normal conditions, when abnormally expressed are detrimental for placenta physiology. In line with our results, Waterman and colleagues described two different immune-modulating profiles in human MSCs isolated from bone marrow. They found pro-inflammatory Toll-like Receptor 4 (TLR4) primed MSCs, that expressed high levels of IL-8, IL-6 and TGF-β2 molecules, and immunosuppressive TLR3-primed MSCs, characterized by marked expression of IL-4. IL-10, RANTES and IP-10 Therefore, we next investigated whether PE-PDMSCs could alter the physiological placental equilibrium. We treated normal chorionic villous explants with media conditioned by normal or PE PDMSCs and investigated the expression of MIF, VEGF, sFlt-1, Ki67 and βhCG molecules. MIF is a central regulator of inflammation and immune response in vivo condition typical of preeclampsia, where the VEGF placental mRNA increase induced by the hypoxic and pro-inflammatory environment is antagonized by the over-expression of anti-angiogenic sFlt-1.As discussed above, VEGF orchestrates angiogenesis and it is vital for early embryonic development Moreover, preeclamptic placentae are characterized by immature hyper-proliferative trophoblast phenotype. Thus, we investigated Ki67 expression in villous explants treated by normal or PE PDMSCs conditioned media. We reported significant Ki67increase in PE-PDMSCs CM explants, indicating the induction of an hypertrophic PE-like status by preeclamptic PDMSCs.As a further confirmation of the PE-like environment provoked by PE-PDMSCs, we reported augmented release of free βhCG by normal villous explants treated with preeclamptic PDMSCs media. hCG is normally secreted by trophoblast cells to maintain maternal vascular supply during pregnancy and its increased circulating levels have been associated with a dramatically higher risk of developing preeclampsia We are currently investigating whether normal placental MSCs are able to revert the pathological inflammatory conditions typical of PE placentae.In conclusion, our data clearly depict chorionic placental mesenchymal stromal cells as central players in placental physiopathology, thus opening to new intriguing perspectives for the treatment of human placental-related disorders as preeclampsia and fetal growth restriction.Figure S1Placenta-derived Mesenchymal Stromal Cells Characterization. A) Representative phenotype of human chorionic PDMSCs at passage 5 as assessed by flow cytometry. All cells were positive for CD166, CD105, CD90, CD73 and negative for HLA II, CD34, CD133, CD20, CD326, CD31, CD45 and CD14, thus displaying proper mesenchymal profile and no contamination from epithelial, hematopoietic, immune or endothelial cells. B) Representative Oct4 and Nanog PCR analysis in control and PE PDMSCs. All cell lines expressed both gene markers of stemness.(TIF)Click here for additional data file.Figure S2Gene expression levels of key differentially expressed molecules detected by Cytokine Array in normal (C) and preeclamptic (PE) PDMSCs. MIF (A), VEGF (B), TNF-α (C), IL-8 (D) and IL-6 (E) mRNA expression levels in normal and PE-PDMSCs as assessed by Real Time PCR. Statistical significance (*) has been considered as p<0.05.(TIF)Click here for additional data file.Figure S3TNF-α and VEGF protein expression in Normal and PE-PDMSCs Conditioned Media and in physiological placental villous explants treated by normal or PE PDMSCs CM as detected by ELISA Assay. (A) TNF-α (left panel) and VEGF (right panel) protein levels in media conditioned by Normal [N-cm] or preeclamptic [PE-cm] PDMSCs. (B) TNF-α (left panel) and VEGF (right panel) protein levels in untreated control explants [C] and explants treated by normal [N-cm] and preeclamptic [PE-cm] PDMSCs conditioned medium. Results are expressed as means ± SE. Statistical significance (*) has been considered as p<0.05.(TIF)Click here for additional data file.Table S1Cytokines Expression in Normal vs Preeclamptic PDMSCs Conditioned Media.(DOC)Click here for additional data file.


Inequality in health and ophthalmic servicesUnfortunately, the medical and welfare facilities are not homogenously distributed to all parts of the world. In many countries, malnutrition could be considered as the major problem while in many other countries, the side effects of obesity are the causes of mortality and morbidity. Within the sum total of all countries there are huge inequalities as well. The same is true in ophthalmic diseases. In many parts of the world, patients have access to the best surgical corneal operations and receive the benefits of the latest methods of minimal invasive surgeries by modern technologists. In many cases, vision improves a few minutes after surgeries, the occuloplastic operations make figures appear younger and more attractive and expensive glasses and high quality intraocular lenses make life more comfortable. However, in other parts of the world many people are deprived of minimal levels of healthcare and ophthalmic services, including the correction of refractive status with simple glasses.Socioeconomic development and improved living standards in many societies have resulted in greater control of the major communicable causes of blindness, such as trachoma and ocular infectious diseases which has caused significant shifts in the pattern of visual impairment presentation. It has been shown while it is essential to continue to invest efforts in eliminating communicable eye diseases as the cause of visual impairments and blindness, the future challenge lies in the provision of eye care for the effects of non-communicable eye diseases .The relationship between socioeconomic status and various diseases have been well presented in the literature. An inverse association between socioeconomic status and rate of blindness has been reported as well. This is mostly due to diseases that are related with deprivation, such as trachoma and vitamin A deficiency, or through insufficient access to healthcare services, such as cataract removal. It has been established that health care inequalities can be attributed to social factors in many conditions . Many caIdeas to assist poor peopleInnovations and discoveries such as vaccination, antibiotics, MRI, and phacoemulsification continuously change the face of healthcare. But what’s next discovery especially in the field of ophthalmology and visual sciences? Not long ago, a contest was arranged by British Medical Journal to appreciate outstanding ideas and innovations. The winner of the contest was a designer of low priced glasses that could improve patient’s vision to some extent in low socioeconomic countries. Inventor of the world’s first adjustable glasses has confidence in bringing corrective spectacles to those who need it around £1 without the prerequisite for either ophthalmologists or optometrists. He believes this instrument will have major impacts on quality of life worldwide  Ophthalmology Journal which focuses especially on new ideas and important hypotheses in the glaucoma and optic nerve diseases. In addition to publishing articles in general ophthalmology and visual sciences, the journal tries to prove feasibility of publishing ideas in subspecialty fields too. When the journal was first being established, some of our colleagues argued that there would be only a few articles submitted to the journal because of its highly specialized focus. With all due respect to our colleagues, we have found that our journal has performed very successfully. Based on the number of manuscripts that have been submitted, it has been demonstrated that there are many capable, enthusiastic and effective researchers in the field of ophthalmology who are happy to have their ideas published in a journal specializing in innovation and ideas.One of the major areas we are looking to publish papers is in the interdisciplinary studies. As an example, we will include studies that share scope in various experimental sciences with mathematics, computer science, chemistry and visual sciences. Our journal intends to create a forum in which distribution and discussions of different types of research are studied. The importance of reform in science and research is essential. Experience has shown that policy makers who are open to new ideas and active listeners of the disenfranchised are more successful. They create the discussions and ideas that lead to growth and evolution. Actually, the world is on the threshold of abundant reforms. Scientists should not allow political or economic powers to dominate their respective worlds. The world must be run by scientists’ expertise in a logical and scientific manner. Therefore, our goal is to create an unrestricted environment for the free and open exchange of information. The editors of this journal believe that any idea, no matter how simple, if it has been written in logical and scientific way, could contribute to the progress of science around the world. This journal publishes research articles with innovations, and we welcome manuscripts that address the needs of underprivileged Individuals.We hereby extend our sincere thanks to the authors, editorial board members, and the staff of the journal. Without their help the journal could not be published continuously. Our journal not only encourages your comments and feedback; it will publish them if they are written scientifically. And as a conclusion, our major goal is to encourage the development of ideas that is much needed for attaining future evolutionary goals especially to help the underprivileged dispossessed individuals. We all have wishes and ideas to make the world better. We believe there is enormous influence in the sharing of those ideas by distributing in scientific publications. The Future we need is a universal dialogue through a positive vision.The authors report no conflicts of interest in this work.


Acinetobacter are gram negative, catalase positive, oxidase negative, nonmotile, non fermenting cocobacilli. The emergence of Acinetobacter infection is uncommon in organ systems that have a high fluid content. The nosocomial infections including, respiratory tract, CSF, peritoneal fluid, urinary tract, and endotracheal aspirtes (ET) are the place where they have the capabilities to accumulate and cause in-hospital and in-community infections. The rate of emergence of Acinetobacter spp with multi drug resistance property is increasing in different geographical regions. Due to the properties of MDR the recent treatments of such infections has become difficult. In the present study evaluation of emergence of MDR nonfermentative Acinetobacter spp was done from the niche of organ system of patients with high fluid content.Acinetobacter spp were isolated from different clinical samples according to standard procedures and Gilardi schemes. The antibiotyping of Acinetobacter was done by disk diffusion method as per CLSI standards.The nonfermentative Acinetobacter spp were isolated from different clinical samples. The ratio of resistance against different antibiotics among Female:Male was found to be 1:1.38. All Acinetobacter spp were multi-resistant and showed different multi drug resistance pattern. Acinetobacter spp from samples showed varied resistance including, 60% resistant strain from wound swabs, 50% from Pus, 28-30% from ET aspirates and blood.31.12% (62/193) of nonfermentative Acinetobacter spp posing significant problem worldwide and increasingly responsible for numerous infections. Our study shows the emergence of high rate of MDR of Acinetobacter spp against 18 antibiotics belonging to different groups.


We have recently developed a subcutaneous anterior pelvic fixation technique (INFIX). This internal fixator permits patients to sit, roll over in bed and lie on their sides without the cumbersome external appliances or their complications. The purpose of this study was to evaluate the biomechanical stability of this novel supraacetabular pedicle screw internal fixation construct (INFIX) and compare it to standard internal fixation and external fixation techniques in a single stance pelvic fracture model.Nine synthetic pelves with a simulated anterior posterior compression type III injury were placed into three groups . Displacement, total axial stiffness, and the stiffness at the pubic symphysis and SI joint were calculated. Displacement and stiffness were compared by ANOVA with a Bonferroni adjustment for multiple comparisonsThe mean displacement at the pubic symphysis was 20, 9 and 0.8 mm for external fixation, INFIX and internal fixation, respectively. Plate fixation was significantly stiffer than the INFIX and external Fixator (P = 0.01) at the symphysis pubis. The INFIX device was significantly stiffer than external fixation (P = 0.017) at the symphysis pubis. There was no significant difference in SI joint displacement between any of the groups.Anterior plate fixation is stiffer than both the INFIX and external fixation in single stance pelvic fracture model. The INFIX was stiffer than external fixation for both overall axial stiffness, and stiffness at the pubic symphysis. Combined with the presumed benefit of minimizing the complications associated with external fixation, the INFIX may be a more preferable option for temporary anterior pelvic fixation in situations where external fixation may have otherwise been used. External fixation for unstable pelvic fractures is effective in resuscitation and reduces pelvic volume, minimizes motion between disrupted fracture surfaces or joints, allows tamponade of ongoing venous bleeding and is a fairly quick procedure-5. Side Nine synthetic pelves  with a simulated anterior posterior compression (APC) type III , by 7.0 mm × 80 mm titanium polyaxial pedicle screws and 6.0 mm stiff titanium rod  Figureb,c2. Internal fixation (n = 3), by 3.5 mm 4-hole pubic symphysis plate 3. External fixation (n = 3), by 2 supra-acetabular 5.0 mm Schanz pins, with an 11 mm carbon fiber rod A single orthopaedic surgeon prepared all the specimens. A custom device was attached to the sacrum and connected to the load cell of a uni-axial servohydraulic testing device  through a ball and socket joint which was allowed to articulate freely Figure. A unipoP = 0.017). There was also a significant difference between internal fixation and external fixation (P = 0.01). There was no significant difference in SI joint displacement between any of the groups. In terms of stiffness (N/m), internal fixation was significantly (P < 0.05) stiffer than either construct at the pubic symphysis. There was no significant difference between the stiffness of the INFIX and the external fixation device. Stiffness of the INFIX and external fixation constructs were then compared as a percentage of internal fixation stiffness  was significantly stiffer than the INFIX or the external fixator  with this single leg stance pelvic model at the symphysis pubis (p < 0.05). The INFIX construct allowed less than half the displacement at the symphysis pubis 9 vs 20 mm) when compared to the external fixator p < 017). When we compared stiffness there was a trend for the INFIX to be stiffer at the symphysis pubis verses the external fixator but this was not significant. The external fixator had slightly less displacement at the SI joint verses the INFIX (not significant). There was no significant difference in stiffness between any of the threee constructs at the SI joint. This is consistent with previous studies that have indicated that there is wide variability between fixation devices at this point,11. Many vs 20 mm. When weThe data show that both the INFIX and the external fixator are significantly weaker than internal fixation at the pubic symphysis. The INFIX was stiffer than external fixation for both overall axial stiffness, and stiffness at the pubic symphysis, the area of most clinical importance for this type of fixation. Combined with the presumed benefit of minimizing the complications associated with external fixation, the INFIX may be a more preferable option for temporary anterior pelvic fixation in situations where external fixation may have otherwise been used. Future improvements to the design may help to increase the stability of the construct.INFIX: Internal fixator; APC: Anterior posterior compression; ANOVA: Analysis of variance; SI Joint: Sacroiliac joint.Jonathan M Vigdorchik, Amanda O Esquivel, Xin Jin, King H Yang, Ndidi A Onwudiwe and Rahul Vaidya contributed equally to this work.Dr. R Vaidya Corresponding author is developing an FDA approved product with Stryker Corporation which has not been manufactured yet or produced. That is not the device in this study as these are just common pedicle screws and rods that are in any spine set. No other author has any competing interests.Role of each author - JV – First author, Participated in developing the study design, implanted the specimens, worked on writing the paper, and performed testing - AE - Second author participated in developing the study design, worked on writing the paper, testing and result evaluation - XJ: Helped with study design, testing, result evaluation and statistics - KY: Helped with study design, testing, result evaluation and statistics - NO: Participated in writing the manuscript, formatting, statistics checks and submitting of the manuscript RV: Corresponding Author, Participated in. All authors read and approved the final manuscript.1Detroit Receiving Hospital, 4201 St Antoine St, Detroit, MI 48201, USA.2Providence Hospital and Medical Centers, 16001 West 9 Mile Road, Southfield, MI, USA. 3Wayne State University, 540 E Canfield St, Detroit, MI, USA.


Thrombosis and hemostasis related disease have a heavy burden in cardiovascular disease and it is important to have a journal where research into this can be accessed by all. Thrombosis Journal is taking shape and we would like to invite all members of the community to contribute to the further development of the journal to best suit the community’s needs.At the start of our Editorship we wanted to look back and reflect upon the journal’s successes to date, but it is also important to look forward! The future of Thrombosis Journal was clearly stated by the founding Editor, Dr. Raul Altman, a highly respected and internationally known investigator in the area of thrombosis and haemostasis. On the journal’s website [Thrombosis Journal, as an open access publication, makes it possible for anyone in the world to read high-quality research in this important area”.The mission statement of  website  he says:Journal of Thrombosis and Haemostasis), or devote significant space to thrombosis and haemostasis , the “thrombosis” scope is so wide that there is ample need for additional forums. In short, our mission statement remains unchanged, we aim to support the community through the dissemination of the freely accessible original research articles published in the journal. We consider open access a critical element in the dissemination of knowledge and research data throughout the world. In the field of thrombosis (and hemostasis) many developments are relevant, not only for the developed world, but also for those in developing nations where access to subscription journals may be limited, mainly because of financial constraints. As Thrombosis Journal charges its authors to publish articles (discretionary waivers are considered on a case by case basis for authors unable to pay), the strength of the journal is that it can be accessed by anyone, anywhere, without charge. This makes it a highly competitive journal in terms of readership, throughout the world. This is also illustrated by the “Highly accessed” papers indicated on the Thrombosis Journal website and indeed, some of these papers are also well cited, including a review authored with some of my colleagues [et al.[Thrombosis Journal publications are immediately included in PubMed and PubMed Central.In a nutshell, this statement summarizes two important features. Firstly, open access is being increasingly recognised as a universal human right, in this particular case the right of clinicians and scientists to unrestricted access to new scientific data emerging from our field of interest. Secondly, the fact that thrombosis and hemostasis related diseases are the most important burden of cardiovascular disease and death worldwide ,3! Thromlleagues  that hass [et al. 53 timesThrombosis Journal as an open access journal that matters!We acknowledge that potential authors can experience pressure from their employers, grant providers, etc., to publish in journals with a high impact factor. While we recongnize that we are still small in terms of such quality indicators, we will be working closely with the journal’s publisher, BioMed Central, to ensure we are attracting more publications of sufficient interest with aim of achieving an impact factor for the journal. Dr Yukio Ozaki and I were put forward by Dr. Altman as his successors, offering him the chance to take a small step back from his very busy working life. We are taking over with the strong belief that we will be able to further strengthen For this reason we urge you to consider us when submitting your next original work in the entire area of thrombosis and hemostasis research, case reports of interest, and review articles. Since we are an online journal there are no limitations on the numbers of figures or other illustrations or colors, which creates more flexibility. We also encourage initiatives for special issues or series of articles on any topic that fits the journal. Moreover, we would like to encourage a global scope, meaning that we encourage colleagues from anywhere to consider our journal for their publications. At the same time opening up does not mean becoming less critical of the contents of submissions. As before, we aim to accept papers that may not have high interest levels but are scientifically sound and contribute to the field of research.Thrombosis Journal could also contribute to education and therefore we welcome the introduction of a “In the clinic” series of reviews providing practical answers to situations encountered in thrombosis and hemostasis clinics. We invite you to submit any manuscripts that would fall under this category to the Thrombosis Journal.In aiming for a global, open access journal for the entire community dealing with thrombosis and hemostasis related disorders , we take the baton from Raul Altman. In addition to his research, Dr Altman is also known for his continuous efforts to provide high quality education in thrombosis and hemostasis throughout South America. We feel, as a final remark, that


We report the reuse of a liver graft after brain death of the first recipient. The liver donor was an 8-year-old male who died as a result of head injury. The graft was implanted first to a 4-year-old girl for fulminant hepatic failure. Unfortunately she developed progressive coma and brain death on fifth day of transplantation. The graft functions were normal, and reuse of the liver graft was planned. After informed consent, the graft was transplanted to a 31-year-old female recipient who has hepatocellular carcinoma with an underlying cryptogenic liver cirrhosis. The patient was discharged to home on 9th day after an uneventful postoperative period. However, she was readmitted to hospital with an acute abdominal pain 30 days after the operation. Hepatic artery thrombosis was diagnosed, and the attempt to open the artery by interventional radiology was unsuccessful. She died of sepsis and multiorgan failure on 37th posttransplant day. The shortage of donors in relation to patients on the waiting list requires the consideration of any possible organ donor in order to meet the current demand. Brain death in fulminant hepatic failure is a possible but rare complication after successful orthotopic liver transplantation –4. PreviA 4-year-old girl was admitted to our hospital with fulminant hepatic failure due to hepatitis A virus. She developed progressive coma without any findings of brain death in neurologic examination. Orthotopic liver transplantation was uneventfully performed by using the normal functioning liver of an 8-year-old male cadaveric donor. Cold ischemic time of the graft was 540 minutes. Graft functions were normal after the transplantation, but the patient's neurologic conditions worsened. She was diagnosed as having brain death five days after the transplantation. Reuse of the liver graft was planned. After informed consent, the graft was transplanted to a 31-year-old female patient with hepatocellular carcinoma with an underlying cryptogenic liver cirrhosis. The recipient was 50 kg in weight. Second cold ischemic time was 85 minutes. The patient was discharged to home on postoperative 9th day after an uneventful postoperative period. She was readmitted to hospital with an acute abdominal pain 30 days after the operation. Hepatic artery was occluded on computed tomography scans. The attempt to open the thrombosed hepatic artery by interventional radiology was unsuccessful. She expired due to sepsis and multiorgan failure on 37th posttransplant day.There are several ways to enlarge the organ pool for liver transplantation. Living donor liver transplantation, splitting the liver of a deceased donor into two parts, and using grafts from marginal donors are the most popular ways to increase the organ pool. Less used ways are domino transplantation and reutilizing the previously transplanted liver. Brain death in fulminant hepatic failure is a rare complication after successful orthotopic liver transplantation , 2. In t To our knowledge, this is the youngest reused liver graft in the literature. Our donor was 4-year old. This may be the reason of late hepatic arterial occlusion. Previously, Rentsch et al. reported a 16-year-old girl (63 kg and 163 cm) as a teenage donor whose liver was successfully used twice [


Beilstein Journal of Organic Chemistry (BJOC) was started in 2005 [Beilstein Journal of Organic Chemistry has been to provide a new and lasting service to enable the scientific community to publish and read high-quality scientific articles, without being hindered by a subscription pay-wall, or by budget limitations.The  in 2005  as the fBeilstein Journal of Organic Chemistry to be represented in many countries. The editors also play a significant role in communicating the advantages of the Journal to colleagues, and in the selection of new Thematic Series, which have been particularly successful for the Journal. All articles are permanently stored in public archives, such as PubMed Central, and indexed by scientific databases.The Beilstein-Institut funds the Journal completely, so there are no subscriptions, or author fees, no page or color charges. The scientific community plays a significant role in the editing of the Journal; the large editorial board, currently 29 scientists, means that the submitted manuscripts are handled by experts in the field and it allows the Beilstein Journal of Organic Chemistry from the beginning to have a good impact factor, and we are now delighted that it has steadily increased over the years to reach 2.517 in 2011 [From the very beginning of the Journal, it was clear just how important impact factors have become. It is not just an interesting metric for librarians to compare journals and articles, but it has become more widely used by funding agencies, universities and the like as a measure of academic prowess and is therefore used in decision making metrics for grant applications, tenure and promotions. The merits of this practice will not be addressed here. It was the aim of the Beilstein Journal of Organic Chemistry has seen continual growth in this area as well. We have implemented strict filtering to exclude repeated views and accesses by robots when counting the HTML full text accesses and PDF downloads . CurrenThe latest mobile devices are able to be used for accessing web-content with similar performance to that of most desktop computers. This year, we brought out the Beilstein Journals App for Android and iOS smartphones and tablets –5. WhilsBeilstein Journal of Organic Chemistry has undergone rigorous internal and external review. The Beilstein-Institut has an internal editorial office for its journals, comprising a team of highly qualified scientists. The internal editorial work involves checking for plagiarism, novelty of compounds, inconsistencies and errors, and also the copyediting and laying out of the final article. The internal editors work in close contact with the external editorial board, whose expertise in specific fields of organic chemistry ensures that the manuscripts published contain novel and contemporary relevant scientific research results.Each article published in the Plagiarism is a problem that all publishers, universities and companies have to tackle. Since May 2011 we routinely check all incoming articles for plagiarism using CrossCheck  and otheBeilstein Journal of Organic Chemistry or in the Beilstein Journal of Nanotechnology. This combination allows scientists to use a visual medium to go further in the explanation of their work than is otherwise possible on paper. This can be useful in terms of overviews and perspectives, as well as for showing hands-on, just how certain experiments work or need to be set up and carried through.A further project of the Beilstein-Institut, which accompanies the journals and has valuable synergy effects, is the scientific video project – Beilstein TV . Here scBeilstein Journal of Organic Chemistry has come of age as an Open Access journal. It has been accepted by the scientific community, it is read worldwide, and is rising in its impact as judged by the citations published. The combination of the Journal with mobile access, videos and further projects bodes well for the future. The Beilstein Journal of Organic Chemistry is now established as the leading open access journal in its field, and is well placed to not only take advantage of future scientific and technical advances in publishing or media, but is flexible enough to lead the way amongst comparable journals. We are planning to expand our editorial board into other subject areas and countries, to provide the organic chemistry community with further Thematic Series of contempory interest and to develop innovative methods of communicating organic chemistry. We are looking forward with our authors, editors, reviewers and readers to an even more successful time ahead.In 2012 we have shown that the Peter H. SeebergerPotsdam, August 2012


Purpose. The aim of this study was to investigate the influence of chemical and physical properties of two graft materials on the rate of resorption. Materials and Methods. Direct sinus graft procedure was performed on 22 patients intended for implant placement. Two types of graft materials were used (Bio-Oss and Cerabone) and after 8 months healing time the implants were inserted. Radiographic assessment was performed over the period of four years. Particle size, rate of calcium release, and size and type of crystal structure of each graft were evaluated. Results. The average particle size of Bio-Oss (1 mm) was much smaller compared to Cerabone (2.7 mm). The amount of calcium release due to dissolution of material in water was much higher for Bio-oss compared to Cerabone. X-ray image analysis revealed that Bio-Oss demonstrated significantly higher volumetric loss (33.4 ± 3.1%) of initial graft size compared to Cerabone (23.4 ± 3.6%). The greatest amount of vertical loss of graft material volume was observed after one year of surgery. Conclusion. The chemical and physical properties of bone graft material significantly influence resorption rate of bone graft materials used for sinus augmentation. Maxillary sinus augmentation and placement of dental implants is a well-established technique for functional and esthetic rehabilitation of partially or completely edentulous patients with severe maxillary atrophy. Sinus pneumatization, together with poor bone quality, is one of the most challenging circumstances in implantology, a condition that will restrict implant placement in such areas. When this situation occurs, bone grafts can be used to correct bone deficits, allowing the placement of implants of adequate length and width . The firThere are diverse choices of graft materials available for replacing lost bone through atrophy, trauma, congenital or pathological processes. These graft materials include: intra or extraoral autologous bone, heterologous grafts, alloplastic grafts, xenografts or a combination of these . In geneβ-tricalciumphosphate , an alloplast, prove that all these grafting materials are biocompatible, osseoconductive and can be used successfully in conjunction during implant rehabilitation [β-tricalcium phosphate. Histological findings reported differences in the amount of newly formed bone used with each material.Clinical and histomorphometric studies done on autografts, bovine hydroxyapatite , a xenograft and litation , 8. Howelitation  evaluate The aim of this study was to evaluate the influence of chemical and physical properties of two types of bone graft materials on the rate of resorption after placement in sinus lift procedure over a period of four years. This study was conducted in coherence with the Helsinki agreement for research on humans and the study design was approved by the Institutional Review Board and Independent Ethics Committee of the Faculty of Dental Medicine, Saint Joseph University, Beirut, Lebanon. Signed informed consent forms were obtained for all participants in the study. Two xenograft materials prepared by deproteinizing technique  or high temperature decalcified freeze-dried  were selected for this study.The particle size of each graft material was calculated using particle size analyzer , and average particle size and distribution were calculated from 5 different batches for each material.θfrom 5° to 60°). The phase composition was checked using Joint Committee on Powder Diffraction standards. Crystallite size analysis was calculated using the peak broadening of XRD reflection that is used to estimate the crystallite size  using the following formula:Xs is the crystallite size in nanometer, λ is the wavelength of X-ray beam in nanometer (λ = 0.15406 nm for standard detectors), and FWHM is the full width at half maximum for the diffraction angle (2θ = 25.9° peak was selected related to (002) Miller's plane family).Crystal structure and size of crystals were calculated using X-ray diffraction (XRD) technique. 5 gram of each material was finely ground, dried, and homogenously dispersed on the measuring table of the machine . Calcium and phosphorous detectors were calibrated in standard solution before each reading. 0.25 gram of each material was immersed in 100 mL of double purified water and the amount of calcium dissolution was measured every week for a period of six months.Patients received detailed explanations of the difficulties and complications that could take place during the surgery and all patients agreed before the surgery. All of the 22 consenting patients were examined and medically compromised and uncooperative cases were excluded from the study.Local anesthesia was administered  and a horizontal incision was made along on the crestal bone in the edentulous area and then vertical incisions were made to elevate the mucoperiosteal flap. After elevation of a full-thickness mucoperiosteal flap, access was gained to the anterior bony wall of the sinus. The lateral bony wall of the sinus was cut by using a small diamond bur. All the cortical bone was removed up to the sinus membrane. After elevation of the membrane, the sinus cavity was then packed with either of the selected materials Figures 1st measurement: right after the implantation (baseline),2nd measurement: after 8 month at time of implant placement,3rd measurement: one year after implant placement,4th measurement: four years after implant placement.The implant length, alveolar crest, the original base line of the sinus floor, and the final graft height were traced by superimposition of the panoramic images. Two fixed measurement points were evaluated using image analysis software  to the accuracy of 1 um. [Height of graft material was measured at the following intervals:of 1 um. . Implantt-test (α = 0.05).Data were analyzed using computer statistical program software  to evaluate the resorption rate of graft material with time and the differences between graft materials (means and standard deviations). Changes in graft volume at different time intervals were analyzed using Student's The average particle size of Bio-Oss (1 mm) was much smaller compared to Cerabone (2.7 mm), Figures The amount of calcium release due to dissolution of the material in water was much higher for Bio-Oss compared to Cerabone. This observation was marked in the first 6 weeks after which dissolution rate of calcium ions reaches a fixed rate for both materials, t = 7.25, P < 0.001) volumetric loss  compared to Cerabone (23.4 ± 3.6%). The greatest amount of vertical loss of graft volume was observed after one year of graft surgery , which decreased almost to 10–12% per year later on for both materials (P < 0.06), Figures Four implants failed after 6 months from insertion time due to lack of adequate initial stability, these cases were replaced with new cases. All patients demonstrated adequate healing after grafting surgery without complications. X-ray image analysis revealed that Bio-Oss demonstrated significantly higher  in sinus augmentation show around 29% new bone formation after 6 months healing time. When an osseoinductive factor like platelet rich plasma (PRP) was mixed with β-TCP, the osseous regenerating capacity was increased to 38%. Nevertheless, a resorption rate of 32–43% was reported; type and quality of crystal content of graft material is a dominant factor-controlling rate of resorption.Studies , 29 usin A very recent study  performe Jensen et al.  reportedThese results should be interpreted cautiously considering the study's reduced sample size. Further in vitro and in vivo studies should be conducted to validate the results of the present study.Within limitations of this study, the physical and chemical properties of bone graft material have significant influence on rate of resorption after sinus lift procedure intended for implant placement. Careful consideration of graft properties might enhance clinical performance.


References 21, 51, 52, and 53 are the incorrect references. The corrected references are as follows:21. Narendra A, Raderschall CA, Robson SKA (2013) Homing abilities of the Australian intertidal ant Polyrhachis sokolova. Journal of Experimental Biology: 216, 3674-3681. doi:10.1242/jeb.08964951. Leggett LMW, Stavenga DG (1981) Diurnal changes in angular sensitivity of crab photoreceptors. Journal of Comparative Physiology 144: 99-109.52. Nässel DR, Waterman TH (1979) Massive diurnally modulated photoreceptor membrane turnover in crab light and dark-adaptation. Journal of Comparative Physiology 131: 205-216.53. Williams DS (1982) Ommatidial structure in relation to turnover of photoreceptor membrane in the locust. Cell and Tissue Research 225: 595-617.


Objective physical assessment of patients with lumbar spondylosis involves plain film radiographs (PFR) viewing and interpretation by the radiologists. Physiotherapists also routinely assess PFR within the scope of their practice. However, studies appraising the level of agreement of physiotherapists’ PFR interpretation with radiologists are not common in Ghana.Forty-one (41) physiotherapists took part in the cross-sectional survey. An assessment guide was developed from findings of the interpretation of three PFR of patients with lumbar spondylosis by a radiologist. The three PFR were selected from a pool of different radiographs based on clarity, common visible pathological features, coverage body segments and short post production period. Physiotherapists were required to view the same PFR after which they were assessed with the assessment guide according to the number of features identified correctly or incorrectly. The score range on the assessment form was 0–24, interpreted as follow: 0–8 points (low), 9–16 points (moderate) and 17–24 points (high) levels of agreement. Data were analyzed using one sample t-test and fisher’s exact test at α = 0.05.The mean score of interpretation for the physiotherapists was 12.7 ± 2.6 points compared to the radiologist’s interpretation of 24 points (assessment guide). The physiotherapists’ levels were found to be significantly associated with their academic qualification (p = 0.006) and sex (p = 0.001). However, their levels of agreement were not significantly associated with their age group (p = 0.098), work settings (p = 0.171), experience (p = 0.666), preferred PFR view (p = 0.088) and continuing education (p = 0.069).The physiotherapists’ skills fall short of expectation for interpreting PFR of patients with lumbar spondylosis. The levels of agreement with radiologist’s interpretation have no link with year of clinial practice, age, work settings and continuing education. Thus, routine PFR viewing techniques should be made a priority in physiotherapists’ continuing professional education. Spondylosis is a degenerative disease which can occur at any level of the spine but commonly detected at the cervical and lumbar regions . It is cPlain film radiograph (PFR) is one of the common imaging techniques used to confirm the diagnosis of lumbar spondylosis by the radiologists in most clinical settings in Ghana and it remains the mainstay in most developing African countries given its availability and cost effectiveness. Studies assessing the level of agreement of interpretation of imaging techniques among allied health professionals have been documented in literature. For instance, the outcome of meta-analysis on the degree of agreement between radiographers and radiologists to determine the radiographers’ accuracy in radiographic reports revealed accuracy level of the radiographers at 92.6% and 97.7% sensitivity and specificity respectively while nurses working at remote units also showed their respective accuracy levels at 96% for sensitivity and 87% for specificity ,6. AlthoBased on anedoctal observation from the Ghanaian health care setting, at least two orthogonal PFR views  are commonly examined in the preliminary assessment of patients with lumbar spondylosis by the physicians and orthopaedic specialists. Eventhough the interpretation of PFR remains within the purview of the radiologists, physiotherapists often perform this task either as extended scope of practice or as routine procedure during objective assessment. With the emergence of physiotherapy practice from under the cover of traditional referral system to self-referral methods of health care delivery particularly in wellness and musculoskeletal fields, Ghanaian physiotherapists are required to extend their horizon to be relevant in the ongoing global health reform . CorrectGiven the growing reliance on radiographic imaging and the evolvement of first contact practice in some allied health professionals, it is necessary to appraise the skills of the relevant available health care professionals in the interpretation of imaging techniques. This study therefore determined the level of agreement in the interpretation of PFR of lumbar spondylosis between the physiotherapists and radiologists in Ghana.Forty-one (41) clinical physiotherapists participated in this cross-sectional survey. They were recruited through sample of convenience from the Government hospitals and private physiotherapy clinics located at four of the ten Regions in Ghana. Clinical physiotherapists who were registered members of Ghana Physiotherapy Association and those who had practiced for at least one (1) year were included in the study. Retired physiotherapists and those in academics were excluded.Three separate PFR  were selected from a pool of PFR of patients undergoing physiotherapy for lumbar spondylosis at the selected hospitals and clinics. The selection was based on the clarity of the films, availability of common lumbar spondylosis pathology, coverage of peripheral body segments , and post production period not exceeding three (3) months. The selected films were shown to an experienced independent radiologist who in turn identified the pathological features and listed them as gold standard to serve as an assessment guide (Appendix 1). The listed pathological features were found to be comparable and consistent to that obtained from twenty retrospective radiologists’ reports on patients with lumbar spondylosis. A total of 24 features were identified by the radiologist from three PFR with each consisting 8 features. The assessment guide was subjected to test re-tests reliability among seven experienced radiologists whose reports were not included in the retrospective reports to scrutinize the list. A good internal consistency of 0.79 Chrombach’s alpha value was obtained through the evaluation process [The Ethics and Protocol Review Committee of the School of Allied Health Sciences, University of Ghana approved the proposal for this study. An information sheet containing the title and the intent of the study was made available to the participants and their written consents were obtained. A self-designed data capturing form was used to obtain demographic data such as age, sex and academic qualification and clinical data including number of years of clinical experience, work settings and the number of continuing professional development (CPD) programme attended in the last 5 years (Appendix 2). The three plain film radiographs (PFR) of lumbar spondylosis from which the assessment guide was developed were viewed by the physiotherapists in the presence of the researchers and in no particular order to avoid stereo-typed interpretation. They were required to list the identifiable patho-anatomical features observed in the PFR. A time commitment of 30 minutes was expected of the participants for the completion of the entire procedure. On completion of the interpretation and documentation process, the researchers collected the findings and evaluated each physiotherapist using the assessment guide.Data were analyzed using the SPSS version 19.0 software. Descriptive statistics of frequency, percentage, mean and standard deviation were used to summarize the data. One sample Kolmogorov-Smirnov test was explored to test the data for normalcy. Comparison of PFR interpretation between the physiotherapists and the radiologist was tested with one sample t test while Fisher’s exact test was used to determine the associations between levels of agreement and the physiotherapists’ demographic and clinical variables at alpha level of 0.05.Forty-one 41) physiotherapists participated in this study comprising 17 (41.5%) males and 24 (58.5%) females. The demographic and clinical profiles of the participants are presented in Table  physiothResults of the categorical levels of assessment shows that 4 (9.8%) physiotherapists scored between 0 and 8 points (Low); 35 (85.4%) scored between 9 and 16 points (Moderate); 2 (4.9%) scored between 17 and 24 points (High). The levels of agreement were significantly associated with academic qualification (0.006) and the sex (p = 0.001) of the physiotherapists. However, the levels of agreement were not significantly associated with the age group (p = 0.098), work setting (p = 0.171), year of clinical practice (p = 0.666), preferred PFR views (p = 0.088) and the number of CPD attended (p = 0.069)  thereby justifying the adoption of one sample t test analysis. The mean score of agreement (12.7 ± 2.6) for the physiotherapists was significantly lower (p = 0.001) than the gold standard point of 24 (Radiologist’s score).The main focus of this study was to assess the level of agreement of interpretation of plain radiographs (PFR) between the radiologists and physiotherapists in Ghana. The mean score of interpretation of the physiotherapists compared to that of the radiologists in this study was 12.7  which was statistically lower than the required scores. This finding falls short of expectation giving the high agreement of other health care professionals with the radiologists in the previous studies . For insIn addition, approximately 85.4% of the physiotherapists’ scores were within the moderate range (9–16 points) which implies that majority of the participants’ performances were within the average in PFR interpretation. Although PFR viewing is a common routine procedure in physiotherapy, it seems little priority has been placed on its techniques in the series of their continuing education programmes. Presumably, they might have acquired their rudimentary skills through personal efforts and interest. These speculations may be justifiable given the crops of young generation of the practicing physiotherapists in Ghana.In the same vein, the levels of agreement in the interpretation of PFR by the physiotherapists’ were significantly associated with their academic qualification and sex. Most of the physiotherapists were first degree holders implying that they were still at the minimum entry level of the profession as stipulated by the World Confederation for Physical Therapy . Post-grA shift in the focus of health care delivery from the hospital to the community, are placing increased demands on physiotherapists to embrace primary-care systems. Available evidence suggests that physiotherapists’ involvements in the primary health care setting are beneficial to patients suffering from musculoskeletal problems such as arthritis and low back pain . Indeed,The findings from this study are limited by the use of very few plain film radiographs of patients suffering from lumbar spondylosis with which the participants were assessed. Sampling over sufficient numbers of patients’ x-rays would have ensured external validity of the results. These observations should be the focus of the future study.We conclude that the sampled Ghanaian physiotherapists had disproportionate level of agreement in the interpretation of PFR of patients with lumbar spondylosis compared to that of the Radiologist. The sex and academic qualification of the sampled physiotherapists had significant bearing on their level of agreement. The outcomes of this study thus underscore the urgent need to include basic training in the interpretation of plain radiographs into the continuing professional development programme of physiotherapists in Ghana.The underlisted three plain film radiographs (PFR) A, B, and C contain the common degenerative features found in patients diagnosed with lumbar spondylosis in two views :PFR A: Lumbar spine only (anterio-posterior view)PFR B: Lumbar spine + Pelvis (anterio-posterior view)PFR C: Lumbar spine only The following (8) features were the pathological features found in each of the PFR.1. Marginal osteophytes on lumbar vertebrae2. Decreased vertebral body height3. Decreased lumbar intervertebral disc space4. End plate sclerosis of articular surfaces in the lumbar region5. Scoliotic lumbar vertebrae6. Exaggerated lumbar lordosis7. Straightening of the lumbar spine 8. Vacuum phenomenonMark each correctly identified pathological feature as 1 point and wrong identification or non-identification as 0 point. The total score is 24 ranging from 0–24 in which the minimum obtainable score is zero (0) while the maximum obtainable score is 24. The scoring is interpreted as follow: 0–8 scores (low level of agreement), 9–16 scores (moderate level of agreement) and 17–24 scores (high level of agreement).Instruction: Kindly supply the following information as applicable to youSection A: Demographics1. AGE: 21–25 □ 26–30 □ 31–35 □ 36–40 □ 41–45 □ 46–50 □ 51–55 □ 55 □2. SEX: Male □ Female □3. ACADEMIC QUALIFICATIONS (kindly tick more than one if applicable)Diploma □ B. Sc. □ M. Sc. □ Ph. D □Others (specify)……………………………………………..1. Are you a registered member of the Ghana Association of Physiotherapists?Yes □ No □Section B: Clinical variables1. In which of the following work settings do you practise?Teaching Hospital □ General Hospital □ Regional Hospital □ Private □1. How many years of Clinical experience (excluding internship) do you have? ……………..2. How many Orthopaedics related Continuous Professional Development (C.P.D.) programme  have you attended in the last 5 years? ………3. Which view of the plain Radiograph would you request for when assessing a patient suspected to have Lumbar spondylosis?Antero-Posterior □ Lateral □The authors declare that they have no competing interests.AB responsible for the design of the manuscript for intellectual presentation. He also contributed to the collection, management, analysis and interpretation of data. EO, assisted in the collection, management, analysis and interpretation of data. OA, participated in the collection of data and final editing of the manuscript. DA participated in data interpretation and analysis. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2342/14/13/prepub


After the usual year of editorial silence, I come back to comment the Impact Factor (IF) attribution for the year 2011. 28 June 2012 has been an important day for every scientific journal, and the wish to share the taste of the results of an entire year of work is strong.® (Thomson Reuters 2011), and data say that The Journal of Headache and Pain has seen a growth of the 21 % compared to the previous year. The absolute value of the IF of The Journal of Headache and Pain for the year 2011 is 2.427, a result of excellence, both per se and compared to the IF of the other journals working in the same area [The IF, the more known bibliometric index, universally accepted for stating the quality of a journal, has been published on that day on the 2011 Journal Citation ReportsThe Journal of Headache and Pain to deliver a great performance.I publicly thank everyone who, in this occasion, has expressed appreciation on the obtained result. Nothing anyway would have been possible without the weighty management structure of Springer, the continuous and qualified contribution of the reviewers, the shared effort from the Editorial and the Advisory Board, creating the conditions for this extraordinary vehicle that is The distribution of the citations per year has increased of 43 %The ranking in the subject category Clinical Neurology is 81/191 (+10)The ranking in the subject category Neuroscience is 142/243 (+19)The percentage of self-citations has remained steady (21 %)An analysis of the result is due, and some simple data (2011 vs. 2010) are self-descriptive:The total number of submissions has increased by 10 %The number of downloaded papers has increased by 62 %The rejection rate has increased by 18 %Additionally, more data related to 2011 versus 2010 should be reported:More technical detailed analyses are released as Supplemental Material to this article, for those who wish to have a more in-depth information.These data must be contextualized in a global economic recession which strongly penalizes biomedical research, and produced an overt slowdown in filling the pipeline of new drugs for headache.The headache research has changing borders, and this peculiarity has always produced ideas and results of excellence. As for the headache health care services, now undeniably worthy if organized in a multi-disciplinary way, also the headache research can gain new lymph by looking to its next-door neighbours and by sharing with them its own backyard.viaticum for the management success is by now ascertained. Togetherness, needless to say, has its clashes but carries a potential for synergies much greater than when different scientists remain in their own backyards. Genetics of common headache disorders, electrical neuromodulation, toxicology, rehabilitation, etc. are the borders where our research must continue to face its challenges tomorrow. The Journal of Headache and Pain will focus on all the innovative areas which, together with the historical ones, shape the forthcoming future of the research.The notion that a multiple approach to chronic headache is a I am optimistic about the future of The Journal of Headache and Pain. The basis of the journal growth wasn’t politically determined. The fundamentals have revolved around an ecosystem of independency, efficiency, and innovation with appropriate levels of regulations.The solid relationship with the affiliated societies, European Headache Federation and Lifting The Burden will be further strengthened, thus to complete the journal, which strongly aims to contribute to the birth of strategies which model the foundations of new ideas, instead of being a simple receptacle for them.Furthermore, the choice to render the journal freely accessible as Open Access has proved to be successful , 3, and Finally, in this last year of my second 5-year-tenure as Editor-in-Chief, the journal will keep its can-do-attitude operative, accepting innovative ideas for promoting a most wide-ranging and inter-cultural headache research.Supplementary material 1 (DOC 34 kb)Below is the link to the electronic supplementary material.


Southern Med Review (SMR) into the Journal of Pharmaceutical Policy and Practice[Journal of Pharmaceutical Policy and Practice (JoPPP) will reach a greater number of readers. Open access will lead to a wider readership, higher downloads, higher citations, and a higher Impact Factor [We are happy to announce the metamorphosis of the  Practice-3. As an Practice-7, Journt Factor . ArticleSMR was published on 23rd Dec 2008 [access  as well as medicines use  [While rejoicing the start of an exciting new phase, it seems appropriate to review where we have come from and where are we heading with this new Journal. The first issue Vol 1 Issue 1) of  Issue 1 SMR was to promote pharmaceutical policy research from low and middle income countries - a job which the journal did well. SMR provided opportunity and support to emerging researchers with 30 to 40% of the contributors being first time authors [SMR was commended by the World Health Organisation (WHO) and its policy impact was evident from published studies which were instrumental in changes in the medicines pricing policies of Vietnam and Thailand [The main aim of  authors . The jou authors -12. SMR Thailand .Journal of Pharmaceutical Policy and Practice (JoPPP) [JoPPP.Now the journal is entering a new phase and is renamed the  (JoPPP) . This chSMR was an internationally focussed journal with a wide geographic and socioeconomic flavour; although most of the research centred on the developing world [JoPPP flagship. However, in line with the broadening remit we expect to receive more pharmaceutical policy related work from those in high-income countries.ng world ,2. AcadePharmaceutical was also chosen with care; to be intentionally broad and to differentiate from Pharmacy per se. The term pharmacy encompasses the physical structure of the community or hospital pharmacy and the practices within that. However, pharmacies are only one component of a larger complex health system which influences global access to, and use of, medicines. The term pharmaceutical encompasses pharmacy but is broader in that it involves wider aspects of medicines access and use including: technology, the pharmaceutical industry, other health professionals that influence use such as doctors and nurse prescribers and so forth. Pharmaceutical policy includes such areas as national pharmaceutical policy, pharmacoeconomics [The word conomics , regulatFrom a practice viewpoint, international policy has been churned out of government departments in high income countries; particularly in the commonwealth . HoweverJoPPP title warrants consideration. There are a number of contexts that encompass pharmaceutical practice and we will be particularly interested in the studies related to community pharmacy setting. This could range from the papers discussing policies and their impact on pharmaceutical practice. Also, the interest would be in medicines compliance, concordance, rationale use of medicines, medicines use among minorities and the disadvantaged groups as well those in the disaster or conflict zones.Finally, adding the word ‘practice’ to the JoPPP continues to grow and morph. We encourage our readers to keep their contributions rolling in!We believe pharmaceutical policy is encompassing and directing activities which are related to access to medicines and their use. We look forward to receiving more of these papers as